Ps6-19) was commonly utilized in the de novo biosynthesis of PLA from biomass. This study tested alternative class I PHA synthase (PhaCCs) from Chromobacterium sp. USM2 in engineered Escherichia coli for the de novo biosynthesis of PLA from glucose. The results indicated that PhaCCs had better performance in PLA production than that of class II synthase PhaC1Ps6-19. In addition, the sulA gene was engineered in PLA-producing strains for morphological engineering. The morphologically engineered strains present increased PLA production. This study also tested fused propionyl-CoA transferase and lactate dehydrogenase A (fused PctCp/LdhA) in engineered E. coli and found that fused PctCp/LdhA did not apparently improve the PLA production. After systematic engineering, the highest PLA production was achieved by E. coli MS6 (with PhaCCs and sulA), which could produce up to 955.0 mg/L of PLA in fed-batch fermentation with the cell dry weights of 2.23%, and the average molecular weight of produced PLA could reach 21,000 Da.Polylactic acid (PLA), a homopolymer of lactic acid (LA), is a bio-derived, biocompatible, and biodegradable polyester. The evolved class II PHA synthase (PhaC1 Biopolyesters have been developed in recent years as alternatives to petroleum-based synthetic polyesters . As a code novo biosynthesized from renewable biomass by engineered strains  to gain the ability to synthesize PLA and LA-containing copolymers. Although the engineered strains (with engineered class II synthase) can efficiently produce LA-containing copolymers  (PHB) (from 8 to 9 g/L) and poly (3HB-co-4HB) (from 8.2 to 9.2 g/L)  .Chromobacterium sp. USM2, which has been used in the polymerization of 3-hydroxypropionic acid   , and evaAll the information on strains and plasmids is listed in ackA gene from the genome of E. coli BL21 (DE3). The homolog arms (600 bp upstream and downstream of the ackA gene) were cloned into the suicide plasmid of pRE112. The constructed pRE112-△ackA was transformed into E. coli χ7213 to construct the donor strain of E. coli χ7213/pRE112-△ackA. Then, the conjugation occurred between the donor strain and the receptor strain of E. coli BL21 (DE3) for the deletion of the ackA gene. The strain of E. coli MS1  were constructed using the following methods. As shown in sulA gene was expressed in strains for larger cell space.Based on the chassis strain of E. coli MS1, E. coli MS2 additionally expressed the ldhA gene from E. coli BL21 . From E. coli MS2, E. coli MS3 additionally expressed the evolved PctCp gene  from Clostridium propionicum DSM 1682 and the evolved PhaC1Ps6-19 gene  from Pseudomonas sp. MBEL 6-19. From E. coli MS3, E. coli MS4 additionally expressed the sulA gene from E. coli str. K-12 substr. MG1655 (NCBI No. NC_000913.3).From E. coli MS2, E. coli MS5 additionally expressed the evolved PctCp gene from C. propionicum and the PhaCCs gene  from Chromobacterium sp. USM2. From E. coli MS5, E. coli MS6 additionally expressed the sulA gene from E. coli str. K-12.From E. coli MS1, E. coli MS7 additionally expressed the gene of the fused enzyme of PctCp/LdhA 2SO4, 3 g K2HPO4.3H2O, 1.9 g KCl, 5 g yeast extract, 1 g sodium citrate, 1 g citric acid, 1 g glycine betaine, 0.24 g MgSO4, and 1 ml of the stored solution of trace element. The stored solution of trace element consists of (per liter) 3.7 g (NH4)6Mo7O24·4H2O, 2.9 g ZnSO4 7H2O, 24.7 g H3BO3, 2.5 g CuSO4 5H2O, and 15.8 g MnCl2 4H2O. Variable antibiotics were supplemented in the cultivation medium for different strains (E. coli MS1.All strains utilized in this study are cultivated in the Luria–Bertani (LB) medium (flask-level) or M9 medium (fermenter-level). The LB medium contains (per liter) 10 g tryptone, 5 g yeast extract, and 10 g NaCl. The M9 medium consists of (per liter) 1 g  was added in the fermenter, and then the cultivation temperature was decreased to 30°C. Then, 100 ml of 50% glucose (w/v) was supplemented every 12 h. The cell growth value of OD600 was monitored using the spectrophotometer at 600 nm. The duration time of fermentation was 72 h.For fermentation of PLA, individual strains were cultivated in 10 ml LB medium at 37°C overnight in a rotary shaker at 220 rpm. Then, 5% (V/V) seed cultures were added into 100 ml LB medium (in a flask) and cultivated at 37°C overnight in a rotary shaker at 220 rpm. The secondary seed cultures were inoculated into a 5-L fermenter  containing 2 L M9 medium, and 20 g/L glucose was added as a starting carbon source. Fed-batch fermentation starts at 37°C. During the fermentation process, ammonium hydroxide solution (6M) was automatically added to maintain the pH at 7. The dissolved oxygen concentration (DOC) was maintained at 10–20% by changing the agitation speed and ventilatory capacity (VC). Ten hours after inoculation, 0.5 mM of isopropyl-The solvent extraction method  was util1H and 13C nuclear magnetic resonance (NMR) spectra using a Bruker AM-500 MHz spectrometer at 500 and 125 MHz, respectively. The sample was solved in CDCl3 with tetramethylsilane (TMS) as an internal chemical shift standard. The number-average molecular weight (Mn) and the weight-average molecular weight (Mw) of PLA were determined by gel permeation chromatography (GPC) equipped with TSKgel SuperMutipore HZ-M*2 column and GPC data processing software. The PLA sample (1 mg/ml) was eluted using tetrahydrofuran (THF) before injection (20 μl), and GPC was operated at a flow rate of 0.35 ml/min at 40°C. Polystyrene with a narrow range of polydispersity (1.03-1.05) was used for calibration.To qualitatively determine the polymer structure, the samples were analyzed by E. coli was characterized using a scanning electron microscope (SEM) on a Hitachi S-4800 instrument. The cells were harvested by centrifugation at 5000 rpm for 5 min and subsequently washed with phosphate-buffered saline (PBS) (pH 7.4) three times. Then, the washed cells were fixed with 2.5% (V/V) glutaraldehyde overnight at 4°C. The fixed cells were washed again with phosphate-buffered saline (PBS) (pH 7.4) three times (30 min each). Afterward, ethanol gradients of 30%, 50%, 70%, 80%, 90%, and 95% (V/V) solutions were used to dehydrate the fixed cells in a sequential way. The cell samples were further dehydrated with 100% ethanol three times. After that, tertiary butyl alcohol was mixed with ethanol in a ratio of 1:1 and pure tertiary butyl alcohol was used to achieve metathesis of ethanol in the cells  on a Hitachi H-7650 instrument. The cells were harvested by centrifugation at 5000 rpm for 5 min and subsequently washed with phosphate-buffered saline (PBS) (pH 7.4) three times. Then, the washed cells were fixed with 2.5% (V/V) glutaraldehyde overnight at 4°C. The fixed cells were washed again with phosphate-buffered saline (PBS) (pH 7.4) three times (30 min each), and then 1% (V/V) osmic acid was added to fix the cell for 1 h. The fixed cells were washed again with PBS (pH 7.4) three times (30 min each). Afterward, acetone gradient solutions of 30%, 50%, 70%, 80%, 90%, and 95% (V/V) were used to dehydrate the fixed cells in a sequential way. The resulting cell samples were further dehydrated with 100% acetone three times. Finally, the cell samples are embedded with Spurr resin for imaging.The PLA granules in p < 0.05 and significant if p < 0.01.The significance of differences between the mean values of testing samples was compared using Student’s t-test. Differences were considered statistically obvious if 1H and 13C assay of polymer product  could produce up to 323.4 mg/L of PLA (0.85% of CDW), obviously higher (p < 0.05) than the PLA production  by the control strain of E. coli MS3 (with evolved PhaC1Ps6-19). The results indicated that class I PHA synthase (from Chromobacterium sp. USM2) exhibited better performance in the biosynthesis of the PLA homopolymer. This study further compared the Mw of the produced PLA with the strains of MS5 and MS3 , the lower Mw/Mn value of MS5 indicated that the PLA produced with MS5 has more centralized molecular weight distribution than that of MS3.The qualitative  product  confirme strains  showed t and MS3 . AlthougCs) could catalyze the polymerization of 3-hydroxypropionic acid , and the CDW of the produced P (3HP) can reach 40%  can improve their substrate specificities toward LA monomers . Class Ieach 40% , indicatE. coli MS4  and MS3 (control strain without sulA) were harvested for SEM/TEM analysis. The SEM results revealed that E. coli MS4 has an elongated rod cell shape (E. coli MS3 (E. coli MS4 (E. coli MS3 (E. coli MS4. The results of fed-batch fermentation (E. coli MS4 significantly (p < 0.01) had higher production of PLA (CDW of 1.96%) than E. coli MS3 (CDW of 0.54%). The highest PLA production was achieved by E. coli MS6 (contains both PhaCCs and sulA), which can produce up to 955.0 mg/L of PLA in fed-batch fermentation with the CDW of 2.23%.At the time point of 36 h after inoculation, the cells of ll shape , which pcoli MS3 . Moreovecoli MS4  occupiedcoli MS3 , indicatentation  confirmeE. coli MS4 and MS6 than their control strains of E. coli MS3 and MS5  (E. coli strains (Pseudomonas (The significantly increased PLA production in  and MS5  indicateutyrate) . It has  strains . The Suludomonas , indicatCp are two key enzymes in the biosynthetic pathway of lactyl-CoA. In order to supply the increased level of lactyl-CoA for PLA biosynthesis, the fused PctCp/LdhA enzyme was engineered in E. coli MS7 (E. coli MS4 (CDW of 1.96%), the PLA production of E. coli MS7 (CDW of 2.08%) was not apparently increased (p > 0.05). In addition, E. coli MS7 (containing PctCp and LdhA fusion enzyme) had a lower dry cell weight  than E. coli MS4 , which indicated that the presence of fused enzyme exhibits growth stress toward the engineered strain of MS7.As shown in coli MS7 . CompareCp and LdhA can spatially restrain multiple catalytic domains in one fused enzyme. The results of this study indicated that fused PctCp/LdhA enzyme retained the biocatalytic functions of individual PctCp and LdhA. However, fused PctCp/LdhA did not apparently improve the general biosynthesis of PLA, which indicated that the final step (polymerization of lactyl-CoA into PLA by PHA synthase) could be the bottleneck step in the biosynthesis of the PLA homopolymer, as reported by earlier studies  is feasible to catalyze the polymerization of the PLA homopolymer from lactyl-CoA. In de novo PLA fermentation from glucose, engineered E. coli strains having PhaCCs present improved PLA production than control strains, which expressed the evolved class II PHA synthase PhaC1Ps6-19. In addition, sulA-mediated morphological engineering could enlarge the cell space and further improve PLA production of the engineered E. coli strains.The present study demonstrated that class I PHA synthase from


These four phenolic acids inhibited the activity of MS-related enzymes. Valerenic acid demonstrated more of an inhibitory ability for ACE  and α-glucosidase  than phenolic acids. Valerian extract inhibited key enzyme activities that were associated with obesity (lipase), hypertension (ACE), and type 2 diabetes (α-amylase and α-glucosidase), suggesting that it is a potential candidate for the development of functional supplements.Extractions of the underground parts of valerian were prepared with water and ethanol (25–95%) at 25–75 °C. Extraction yields, bioactive compounds, and the 1,1-Diphenyl-2-picrylhydrazyl (DPPH) radical scavenging ability of lyophilized extracts were determined. The inhibitory effects of the extracts, valerenic acid derivatives and phenolic acids, on metabolic syndrome (MS)-related enzymes activities were further examined. Both roots and rhizomes extracted with 95% ethanol at 75 °C had the highest levels of bioactive compounds. The antioxidant capacity and inhibition of MS-related enzymes of the roots extract were better than those of the rhizomes. The roots extract more strongly inhibited pancreatic lipase (inhibition of 50% of enzyme activity (IC Valeriana officinalis  is a flowering plant of the Caprifoliaceae family that is commonly used to relieve sleep and anxiety disorders. It is safe for short-term use, but its side effects include headache, stomach upset, etc. [et, etc. ,2. Valeret, etc. . Among tet, etc. ,5,6.Phenolic compounds inhibit free radicals to link with antioxidant activity, limiting nutrient oxidation by inhibiting oxidative chain reactions, which positively affect cardiovascular health and have anti-inflammatory, antioxidant, and anti-cancer properties . CliniciTheoretically, valerian could be utilized to treat metabolic syndrome (MS); however, to date, few studies ,12,13 haV. officinalis. Further, the inhibitory effects of the extracts on the specific MS-related enzymes activities were examined: pancreatic lipase, ACE, α-amylase, and α-glucosidase.MS is a group of syndromes that increase the risk of coronary heart disease, diabetes, stroke, and other serious health problems . At leasV. officinalis) parts, including the roots and rhizomes , and filtered through Advantec No.1 filter paper. The residue was re-extracted with 150 mL portions of the solvent, as described above. The combined filtrate was concentrated under reduced pressure at a 50 °C water bath and freeze-dried. The lyophilized extracts obtained by extracting the samples with water, and 25%, 50%, 75%, and 95% ethanol in a 25 °C water bath were labeled 25C0E, 25C25E, 25C50E, 25C75E, and 25C95E, respectively. The first set of numbers and letters indicates the bath temperature in Celsius, and the second set of numbers and letters indicates the percentage of ethanol. All treatments were randomly generated and performed in quadruplicate.Valerian powder (15 g) was added to 150 mL of water or various concentrations  of ethanol in a shaking water bath at 150 rpm for 30 min at 25–75 °C, then centrifuged at 6932× Methanol (purity > 99.9%), acetonitrile (purity > 99.9%), and phosphoric acid (86.2%) were purchased from Avantor Performance Material . Valerenic acid (purity > 99.9%) and hydroxyvalerenic acid (purity > 99.9%) were purchased from ChromaDex . Chlorogenic acid (purity > 95%), caffeic acid (purity > 97%), protocatechuic acid (purity > 97%) and gallic acid (purity > 98%), Folin–Ciocalteu’s phenol reagent (2N), DPPH (purity > 95%), DMSO (purity > 99.5%), sodium dodecyl sulfate (purity > 97%), starch (purity > 99.9%), acarbose (purity > 95%), ascorbic acid (purity > 99%), α-tocopherol (purity > 96%), BHA (purity > 98.5%), 3,5-dinitrosalicylic acid (purity > 98%), porcine pancreatic α-amylase (EC 3.2.1.1), p-nitrophenyl-α-D-glucopyranoside (purity > 98%), p-nitrophenyl laurate (purity > 98%), α-glucosidase (EC 3.2.1.20), rabbit lung ACE (EC 3.4.15.1), hippuryl-l-histidyl-l-leucine (purity > 98%), hippuric acid (purity > 98%), captopril (purity > 99.5%), Orlistat (purity > 98%), acetoxyvalerenic acid (purity > 95%), and Triton X-100 were purchased from Sigma-Aldrich . Sodium chloride (purity > 98.8%) was purchased from Shimakyu’s Pure Chemicals . Anhydrous sodium carbonate (purity > 99%), sodium dihydrogen phosphate (purity > 98%), sodium phosphate dibasic dodecahydrate (purity > 98%), and hydrochloric acid (37%) were purchased from Union Chemical Work . Formic acid (purity > 98%) was purchased from Honeywell International . Potassium sodium tartrate tetrahydrate (purity > 99.5%) was purchased from Wako Pure Chemical . Tris(hydroxymethyl) aminomethane (purity > 98%) was purchased from Amresco . Ethanol (95%) was purchased from Taiwan Tobacco & Liquor .gallic acid (μg/mL) + 0.0077, R² = 0.9993). The results were expressed as milligrams of gallic acid equivalent (GAE) per gram of lyophilized extract. The phenolic acid levels in the lyophilized extract were determined based on the method of Sarikurkcu et al. [g for 5 min. The solution was filtered with a nylon syringe filter (13 mm × 0.45 μm) before injection into an HPLC system consisting of a Hitachi 5110 pump, a Hitachi 5430 Diode array detector , and a Phenomenex Luna C18 (2) column . The mobile phase was composed of 0.1% (v/v) formic acid (solvent A) and methanol (solvent B) at a flow rate of 0.4 mL/min. Elution was carried out using a linear gradient as follows: 2% B from 0 min to 6 min, 2–25% B from 6 min to 12 min, 25–50% B from 12 min to 20 min, 50–95% B from 20 min to 28 min, 95% B from 28 min to 34 min, 95–2% B from 34 min to 35 min, and 2% B from 35 min to 45 min. The sample injection volume was 10 μL. Gallic acid and protocatechuic acid were detected at a wavelength of 280 nm. Chlorogenic, caffeic, and rosmarinic acids were detected at 330 nm.The extraction yield of all treatments is calculated as (freeze-dried extract weight/sample weight) × 100%. Total phenols levels of the extracts were analyzed following the method described by Mau et al. . The totu et al. , with soValerenic acid derivatives in the extracts were determined using a modified method by Donovan et al. . The sam50 value (mg extract/mL) is the effective concentration at which the DPPH radicals were scavenged by 50%. Ascorbic acid, BHA, α-tocopherol, acetoxyvalerenic acid, and valerenic acid were compared. Each analysis was carried out in quadruplicate.The scavenging ability of each extract on DPPH radicals was determined based on the method by Shimada et al. . Each ex50 value (mg/mL) is the inhibition concentration at which enzyme activity was inhibited by 50%. Each analysis was carried out in quadruplicate. The analytical method is briefly described below. In the lipase inhibitory activity, 25 μL of extracts (roots 0–60 mg/mL and rhizomes 0–100 mg/mL), orlistat (0–125 ng/mL), phenolic acids , valerenic acid derivatives (0–2 mg/mL) dissolved in 5% DMSO solution, were incubated with 50 μL substrate and 25 μL lipase (150 U/mL) for 30 min at 37 °C. The supernatant of the reacted solutions was read at 405 nm using an ELISA reader. Orlistat was used as a positive control. The inhibitory abilities of the extracts, phenolic acids, or valerenic acid derivatives; or orlistat, acarbose, or captopril solutions on lipase, α-amylase, α-glucosidase, and ACE were determined according to a previous study ,24. The In the ACE-inhibitory activity, a mixture of 100 μL of ACE solution (2.5 mU/mL), 100 μL of extract (0–20 mg/mL), captopril (0–3.33 ng/mL), phenolic acids , or valerenic acid derivatives (0–1 mg/mL) solution was preincubated for 30 min at 37 °C. The above mixture was added with 100 μL of 3 mM hippuryl-L-histidyl-L-leucine , and the mixture was incubated for 30 min at 37 °C. The reaction was stopped by adding 100 μL of 12% phosphoric acid. The hippuric acid was determined via HPLC. Captopril was used as a positive control. In the α-amylase inhibitory activity, a total of 200 μL of extract (0–50 mg/mL), acarbose (0–25 μg/mL), phenolic acids (0–2.5 mg/mL), or valerenic acid derivative (0–2 mg/mL) solutions; and 200 μL of 0.02 M sodium phosphate buffer (pH 6.9 with 0.006 M NaCl) containing α-amylase solution (10 U/mL) were incubated in a shaking bath (37 °C) at 100 rpm for 45 min. Then, 400 μL of a 0.5% starch solution was added to each tube and incubated in a shaking bath (37 °C) for 10 min. The reaction was stopped with 1.0 mL of dinitrosalicylic acid color reagent. The test tubes were incubated in a boiling water bath for 10 min and cooled to room temperature. The reaction mixture was then diluted after adding 3 mL of distilled water, and the absorbance was measured at 540 nm using a spectrophotometer. The readings were compared with the controls containing buffer instead of the sample extract. Acarbose was used as a positive control. In the α-glucosidase inhibitory activity, 100 μL of extract (0–30 mg/mL), acarbose (0–400 μg/mL), phenolic acids (0–6 mg/mL), or valerenic acid derivatives (0–2 mg/mL) solution, and 100 μL of α-glucosidase solution (1 U/mL) in a 0.1 M phosphate buffer (pH 6.9) was incubated at 25 °C for 10 min. Then, 50 μL of 5 mM p-nitrophenyl-α-d-glucopyranoside solution was added to the 0.1 M phosphate buffer (pH 6.9). The mixture was incubated at 25 °C for 5 min, followed by the addition of 25 μL of 0.1 M sodium carbonate solution to stop the reaction. The absorbance was measured at 405 nm using an ELISA reader. Acarbose was used as a positive control.All measurements were performed in quadruplicate. The data were subjected to analysis of variance with SAS software . When a significant difference was found among the treatment groups, Duncan’s multiple range tests were performed to determine the differences among the mean values at a level of α = 0.05.The extraction was performed using solvents  to separate the desired natural products from the raw materials . In thisThe total phenols extracted from the roots and rhizomes are detailed in 50 values (mg extract/mL) of roots extracts for scavenging DPPH free radicals were 0.352 (75C95E), 0.374 (75C75E), 0.535 (75C50E), 0.639 (75C25E), and 0.708 (75C0E)  progression is linked to free radicals . Natural (75C0E) . Thus, t40 μg/mL . In anot40 μg/mL , which w40 μg/mL ,32.50 values (µg/mL) of ascorbic acid, BHA, and α-tocopherol in scavenging DPPH radicals were 15.67, 16.48, and 16.09, respectively. Valerian extracts were less effective than ascorbic acid, BHA, and α-tocopherol at DPPH scavenging, but these standards are additives that are used in foods or that are present at milligram levels. However, valerian extract can be used as a food supplement at the gram level.Roots extracts had better antioxidant properties, which may be attributed to the fact that the levels of phenolic compound in the roots extracts were higher than those in the rhizomes extracts . Valeren50 values (mg extract/mL) for pancreatic lipase were 50.75 (0E), 41.84 (25E), 38.28 (50E), 30.67 (75E), and 17.59 (95E) (Pancreatic lipase is a key enzyme that regulates lipid absorption . Pancrea59 (95E) . The inh50 was achieved  increased, a more potent inhibition of pancreatic lipase was observed A. The ICectively . Thus, g50 value (mg extract/mL) of the inhibition of ACE activity by the roots extracts  of the inhibition of roots extracts on α-amylase was 75C95E (12.53), 75C75E (20.65), 75C50E (28.23), 75C25E (30.41), and 75C0E (32.32). The R75C95E demonstrated the most strongly inhibitory ability of α-amylase. Compounds that inhibit α-amylase activity in methanol extracts from the upper part of V. officinalis may result from phenolic compounds [Citrus aurantium (L) peel strongly inhibit α-amylase activity; at 1.0 mg extract/mL, α-amylase activity is inhibited by up to 76% [Type 2 diabetes treatment can decrease postprandial hyperglycemia by inhibiting carbohydrate hydrolases in the gastrointestinal tract . Compareompounds . Other sp to 76% .50 value was 5.40 μg/mL  for inhibiting α-amylase  is higher than that in rhizomes . The total phenols (33.16 mg GAE/g lyophilized extract) and total contents of four phenolic acids (3.08 mg/g lyophilized extract) in roots R75C95E were significantly higher than those in rhizomes 75C95E  of valerenic, acetoxyvalerenic, gallic, protocatechuic, chlorogenic, and caffeic acid on α-glucosidase inhibition was 0.617, 1.827, 2.164, 3.721, 5.524, and 1.289, respectively  when the added doses were increased to 3.75 and 4.87 mg/mL, respectively (50% inhibition). Additionally, at doses of 12.53 mg/mL (R75C95E) and 21.99 mg/mL (Rh75C95E), these extracts demonstrate an equivalent level of inhibition against α-amylase, compared to that of acarbose applied at a dose of 5.404 μg/mL (50% inhibition). For α-glucosidase, the estimated IC50 values for R75C95E and Rh75C95E were 15.40 and 17.10 mg/mL, respectively, while the acarbose dose was 95.47 μg/mL. Therefore, theoretically, a combination of 49.27 mg R75C95E or 84.88 mg Rh75C95E may result in a 50% inhibition of lipase, ACE, α-amylase, and α-glucosidase activities. To increase the inhibition of pancreatic lipase, ACE, α-amylase, and α-glucosidase to 70%, the dosage of 75C95E in roots and rhizome extracts should be 104.45 and 141.52 mg extract/mL (Based on the ICtract/mL . The 75CWith the rapid development of food processing technologies, researchers have been increasingly interested in developing natural products as dietary supplements to meet consumers’ health needs. Valerian extracts prepared with 95% ethanol as the extraction solvent and in a 75 °C water bath produced the highest bioactive component levels. The advantage of the specific extraction method is that ethanol is non-toxic, easy to mix with water, and low-cost; however, disadvantages exist since ethanol is flammable. There are other possible extraction methods, such as ultrasonic-assisted extraction, which is low-cost and high-yield; however, special equipment is required. The total phenols content of the roots extract was higher than that of the rhizomes. The phenolic acids in the roots and rhizomes mainly consisted of protocatechuic acid, but the total valeric acid content of the rhizomes was higher than that of the roots. The inhibition of R75C95E on pancreatic lipase, α-amylase, α-glucosidase, and ACE was higher than that of Rh75C95E. It is important to pay attention to these specific extracts as they have the highest inhibitory effects on MS-related enzymes activities. In conclusion, valerian roots and rhizomes extracts can be considered as a source for functional food development. Additional in vivo studies are required to investigate valerian extract, for preventing and managing MS.


Escherichia coli EC1390 from a UTI patient by using whole-genome sequencing (WGS) in combination with phenotypic assays.Urinary tract infection (UTI) is one of the most common outpatient bacterial infections. In this study, we isolated and characterized an extensively-drug resistant (XDR) NDM-5-producing E. coli recipient C600. Phenotypic assays, including growth curve, biofilm formation, iron acquisition ability, and cell adhesion, were performed to characterize the function of EC1390 plasmids.Antimicrobial susceptibility to 23 drugs was determined by disk diffusion method. The genome sequence of EC1390 was determined by Nanopore MinION MK1C platform. Conjugation assays were performed to test the transferability of EC1390 plasmids to E. coli. A de novo genome assembly was generated using Nanopore 73,050 reads with an N50 value of 20,936 bp and an N90 value of 7,624 bp. WGS analysis showed that EC1390 belonged to the O101-H10 serotype and phylogenetic group A E. coli. Moreover, EC1390 contained 2 conjugative plasmids with a replicon IncFIA  and IncFII , respectively. No significant difference was observed in the bacterial growth rate in LB broth and iron acquisition ability between C600, C600 containing pEC1390-1, C600 containing pEC1390-2, and C600 containing pEC1390-1 and pEC1390-2. However, the bacterial growth rate in nutrition-limited M9 broth was increased in C600 containing pEC1390-2, and the cell adhesion ability was increased in C600 containing both pEC1390-1 and pEC1390-2. Moreover, these plasmids modulated the biofilm formation under different conditions.Our results showed that EC1390 was only susceptible to tigecycline and colistin, and thus was classified as XDR E. coli EC1390 and identified two plasmids contributing to the antimicrobial resistance, growth of bacteria in a nutrition-limited medium, biofilm formation, and cell adhesion.In summary, we characterized the genome of XDR-The online version contains supplementary material available at 10.1186/s12866-022-02562-6. Escherichia coli, Klebsiella pneumoniae, Proteus mirabilis, Enterococcus faecalis, and Staphylococcus saprophyticus, are common pathogens isolated from UTI patients [E. coli (UPEC) is the dominant infectious pathogen in both uncomplicated and complicated UTIs [Urinary tract infection (UTI) is one of the most common outpatient bacterial infection, with a lifetime incidence of 50–60% in adult women. Therefore, UTI causes a substantial financial burden on society . Escheripatients . Among tted UTIs . The greted UTIs . Carriagted UTIs , 5.E. coli-causing UTI symptoms in the last decade [E. coli isolated from patients with UTIs showed relatively low resistance to carbapenems , therefore, carbapenems were considered the drug of last resort for the treatment of UTIs caused by extended-spectrum β-lactamases-producing E. coli [Enterobacteriaceae are strongly associated with carbapenemase production, overexpression of efflux pumps, and porin loss [Enterobacteriaceae and is a matter of major clinical concern [E. coli strain EC1390 isolated from a UTI patient.Cephalosporins, fosfomycin, trimethoprim, fluoroquinolones, and amoxicillin in combination with β-lactamase inhibitors, are considered effective antibiotics to reduce the duration of t decade . Howevert decade . E. coli E. coli . The mecrin loss . Moreove concern . Current concern . TherefoE. coli isolates were recovered from patients with UTIs at National Cheng Kung University Hospital (NCKUH) with the approval of the NCKUH Research Ethics Committee . Informed consent was waived because of the retrospective nature of the study, the isolates were remnants from patient samples, and the analysis used anonymous clinical data. A total of 844 non-duplicate E. coli isolates were identified in the clinical laboratory by colony morphology, Gram stain, biochemical tests, and the Vitek system  according to the manufacturer's recommendations [E. coli isolates were stored at − 80 °C in lysogeny broth (LB) containing 20% glycerol (v/v) until tested.ndations . E. coliE. coli ATCC 25922 was used as a quality control strain. The interpretation of resistance to these antimicrobial agents was determined according to the recommendations of the CLSI [The antimicrobial susceptibility was determined by disk diffusion assay against 23 antibiotics ((amikacin (30 μg), amoxicillin (30 μg), ampicillin (10 μg), ampicillin/sulbactam, cefazolin (30 μg), cefepime (30 μg), cefixime (5 μg), cefmetazole (30 μg), cefoperazone/sulbactam (10/10 μg), cefoxitin (30 μg), ceftazidime (30 μg), ceftriaxone (30 μg), cefuroxime (30 μg), ciprofloxacin (5 μg), ertapenem (10 μg), gentamicin (10 μg), imipenem (10 μg), meropenem (10 μg), levofloxacin (5 μg), piperacillin/tazobactam (100/10 μg), sulfamethoxazole/trimethoprim (23.75/1.25 μg), tetracycline (30 μg), and tigecycline (15 μg))  following the Clinical and Laboratory Standards Institute (CLSI) guidelines . The Vitthe CLSI .https://www.bioinformatics.babraham.ac.uk/projects/fastqc/). Raw signals were translated into a DNA sequence by using ONT Gussy basecalling program (version 4.2.3). EC1390 genome was constructed with Flye de novo assembler (version 2.9), and the options for backward compatibility (plasmids) and uneven coverage mode (meta) were used in this analysis [The genomic DNA (gDNA) of EC1390 was extracted using a Presto™ gDNA Bacteria Advanced Kit  from a 5 mL broth culture grown in LB, followed by the protocol for Gram-negative bacteria. One μg gDNA was used to construct the sequencing library by using a Ligation Sequencing Kit . KAPA HyperPure beads purchased from Oxford Nanopore techonologies were used to purify the gDNA fragments. The Oxford Nanopore MinION MK1C was used to determine the whole genome sequence of EC1390. A total of 300 ng gDNA was loaded onto R9.4.1 flow cell. The quality of reads generated was assessed using Fast QC v0.11.5 . CGView Server was used to generate graphical maps of circular genomes and plasmids . Rapid Annotations using Subsystems Technology  was used to predict the subsystem of genes in EC1390 genome . The multilocus sequence typing (MLST), ResFinder, and PlasmidFinder databases (http://www.genomicepidemiology.org/) were used to find the sequence type (ST), serotype, antibiotic resistance genes, and the plasmid types present in EC1390. Insertion sequences (ISs) and CRISPR-Cas were characterized using the IS Finder database (https://www-is.biotoul.fr/)  and CRISPR Finder database (https://crispr.i2bc.paris-saclay.fr/Server/), respectively. CRISPRTarget was used to identify the target of spacers .Genome annotation was performed by NCBI Prokaryotic Genome Annotation Pipeline (PGAP) , respectively, were selected to perform CulstalW alignment  to construct Maximum Likelihood trees inferred with the Jukes-Cantor model by using Molecular Evolutionary Genetics Analysis (MEGA) software version 11.0. Bootstrap confidence values were obtained applying 300 replications.E. coli C600 as described previously [Salmonella OU7526 and E. coli EC974/EC1515 contained 2 (50 and 90 kbp) and 3  plasmids, respectively, were used as plasmid size controls [blaNDM primers was used to detect the presence of blaNDM in the TCGs [The liquid mating-out assay was used to determine the conjugation ability of plasmids pEC1390-1 and pEC1390-2, to rifampin-resistant eviously . TCGs weeviously . Salmonecontrols . PCR witthe TCGs .E. coli ATCC 25922 strain. The plates were incubated at 35 °C for 16 to 20 h and the mCIM and eCIM results were interpreted as previously described [K. pneumoniae ATCC BAA-1706 (carbapenemase-negative), K. pneumoniae ATCC BAA-1705 (blaKPC-positive), and K. pneumoniae ATCC BAA-2146 (blaNDM-positive) were used as internal controls for mCIM and eCIM tests.mCIM and eCIM were performed on EC1390 and TCGs to detect the presence of carbapenemase . A 1-μL escribed . K. pneu600 nm) in the microplates over the 24 h period. Each strain was analyzed in triplicate wells on at least three separate occasions to give nine data sets for analysis.The rate of growth was determined over 24 h at 37 °C using a BioTek Synergy HTX multimode reader . 200 μL E. coli was assayed using a 96-well flat-bottom polystyrene microtiter plate, as described previously [Biofilm formation by eviously , with soE. coli, incubated at 37˚C for 16 h, and monitored for the formation of an orange halo. CAS remains blue when complexed with iron, but it turns orange when iron is chelated by other iron chelators. Siderophore activity was expressed as the halo diameter.Chrome azurol S (CAS) agar plates were prepared as described previously . The modE. coli strains, centrifuged at 600 g for 5 min to synchronize bacteria-host cell contact, and incubated at 37 °C for 2 h. The cells were washed three times with phosphate buffered saline (PBS). The cells were lysed by incubation with saponin at 37 °C for 15 min and plated on LB plates. The resulting colonies were counted to determine the adherent bacteria [The bladder epithelial cell line 5637 was a kind gift of Prof. Ching-Hao Teng . 5637 cells were seeded into 12-well plates in RPMI-1640 media (Thermo Fisher Scientific) with 10% fetal bovine serum (FBS)  and 1% penicillin–streptomycin (HyClone) and grown to a monolayer. Before 2 h of infection, the culture medium was replaced with fresh medium without 1% penicillin–streptomycin. The cells were infected with a MOI of 10 of the bacteria .t test was performed to evaluate the statistical differences in phenotypic assay experiments. A value of p < 0.05 was considered statistically significant.All analyses were performed in GraphPad Prism 8.0 software . Student’s E. coli strain, EC1390, was resistant to three carbapenems, including imipenem, ertapenem, and meropenem. Moreover, EC1390 showed resistance to ampicillin, amoxicillin, amikacin, ceftazidime, ciprofloxacin, cefmetazole, ceftriaxone, cefuroxime, cefazolin, cefepime, cefoxitin, gentamicin, levofloxacin, ampicillin/sulbactam, sulfamethoxazole/trimethoprim, tetracycline, and piperacillin/tazobactam. EC1390 was only susceptible to tigecycline and colistin, therefore, EC1390 was classified as extensively-drug resistant (XDR) E. coli.Among 844 UTI isolates, our antimicrobial susceptibility tests showed that 11 (1.3%), 221 (26.2%), 640 (75.8%), 265 (31.4%), 244 (28.9%), 30 (3.6%), 7 (0.8%), 9 (1.1%), 4 (0.5%), 259 (30.7%), 249 (29.5%), 99 (11.7%), 249 (29.5%), 199 (23.6%), 121 (14.3%), 154 (18.2%), 359 (42.5%), 286 (33.9%), 462 (54.7%), and 408 (48.3%) isolates were non-susceptible to amikacin, gentamicin, ampicillin, amoxicillin, ampicillin/sulbactam, piperacillin/tazobactam, imipenem, ertapenem, meropenem, cefazolin, cefuroxime, cefmetazole, ceftriaxone, ceftazidime, cefepime, cefoxitin, ciprofloxacin, levofloxacin, tetracycline, and sulfamethoxazole/trimethoprim, respectively. All 844 isolates were susceptible to tigecycline and colistin. Moreover, only one EC1390 was isolated from a 50-year-old woman admitting to NCKUH on September 4, 2020, for acute hypoxemic respiratory failure with acute respiratory distress syndrome. The patient also had chronic lymphocytic leukemia. The patient was hospitalized at NCKUH from September 4 to December 3. The timeline of the first 4 weeks of history of drug use and pathogen isolation of this patient was described in Fig. N50 value of 20,936 bp and an N90 value of 7,624 bp (mean coverage was 316). Reads were assembled and returned three contigs with the head segment was almost identical to the tail segment, indicating the circular nature of the contigs. The closed EC1390 genome comprises a 4,779,543-bp chromosome, a 156,286-bp plasmid named pEC1390-1, and a 71,840-bp plasmid named pEC1390-2  confirmed that EC1390 belonged to a novel sequence type, and the results were further verified by sanger’s sequencing.Serotype was determined by the genotype of To characterize CRISPR-Cas systems in EC1390, the presence of true CRISPRs in the EC1390 genome was assessed by the CRISPRCasFinder tool. To discriminate spurious CRISPR-like elements from the true CRISPRs, only CRISPRs classified with evidence levels 4 were considered for further analyses. Based on the selection criteria, one CRISPR type IE loci/arrays containing 10 spacers were detected in EC1390 -Ib-cr, blaCTX-M-15, blaOXA-1, blaNDM-5, catB3, catA1, sul1, dfrA17, tetA, and mph(A), and therefore conferred resistance to streptomycin, fluoroquinolone, aminoglycosides, β-lactams, chloramphenicol, sulfamethoxazole, trimethoprim aminoglycosides, tetracycline, trimethoprim, and azithromycin  and IncFII pEC1390-2) were found in EC1390 , E. coli strain NDM4 plasmid unnamed1 , NDM6 plasmid unnamed1 , NDM5 plasmid unnamed3 , and pIP1206 . However, the E. coli strains NDM3, NDM4, NDM5, and NDM-6 belonged to the same clone. pIP1206 was identified in France. The phylogenetic tree indicated that pEC1390-1 showed the highest similarity to plasmid unnamed1 in E. coli strain NDM4, which was identified in E. coli containing blaNDM-5 in Austria, followed by NDM3 plasmid unnamed1 and pDA33133-157  , pHN3A11 , pYHCC , plasmid 31 , and pKP_160-4 . Interestingly, pNCYU-29–69-2 was identified in a E. coli recipient C600  (C600 contained pEC1390-1), and EC1390-TCG(S + L) (C600 contained pEC1390-1 and pEC1390-2)  Fig. S. The res2) Fig. S. MoreoveE. coli strains were measured in triplicate for each of M9 broth (nutrition-limited) and LB broth , compared to C600, EC1390-TCG(S), and EC1390-TCG(S + L)  Fig. A. Our re L) Fig. B. The re L) Fig. C. All 3  L) Fig. C. Moreov L) Fig. D.Fig. 4CE. coli isolated from UTI patient. EC1390 was isolated from the urine sample on day 14 of patient’s hospitalization. Cefepime, doxycycline, and sulfamethoxazole/trimethoprim, were used for the treatment of bacterial infections before EC1390 was isolated  was isolated from the patient’s rectal swab, however, we did not further determine the clonality of CREC isolated from urine and rectal swab.In this study, we combined Nanopore WGS with phenotypic assays to characterize an NDM-5-producing The inconsistency of annotation results from NCBI PGAP and RAST server suggests the assemblies generated only from MinION data contained many indels and substitutions that affected the following analyses Table . In addiE. coli. The phylogenetic groups A, B1, and D had fewer virulence determinants than B2 strains, and O101-H10 serotype phylogenetic group A E. coli was relatively rare reported [E. coli bloodstream isolates in a Spanish hospital, 96 (4.0%) were extended-spectrum β-lactamase producers. Moreover, only 2 out of 96 ESBL-producer belonged to the O101-H10 serotype  [E. coli were rarely determined. Currently, XDR-E. coli ST7624 belonged to O − :H34 serotype harboring blaCTX−M−14b was identified from milk samples [E. coli that can cause human diseases should be monitored.Our results showed that EC1390 belonged to the O101-H10 serotype, phylogenetic group A, and untypable sequence type reported , 28. Mam samples . The chaE. coli was reported but only a few UPEC isolates harboring blaNDM-5 were identified [E. coli carrying blaNDM-5 in Taiwan [E. coli from 2012 to 2015, and only one ST117 E. coli contained blaNDM-5 [E. coli clinical isolates collected at National Taiwan University Hospital were carbapenem-non-susceptible. Moreover, among the 78 carbapenem-nonsusceptible E. coli isolates, 23 (29.5%) were carbapenemase-producing E. coli, and 5 NDM-5-producers were identified ) [blaNDM-5-positive plasmids worldwide. Therefore, it is worth monitoring the spread of blaNDM-5-positive plasmids in Taiwan.Currently, international circulation of NDM-5-producing entified –32. To on Taiwan , 34. Cha(n = 2)) . HoweverblaNDM-5 is less investigated. Although our results showed that the plasmid pEC1390-2 promoted the growth of bacteria in nutrition-limited M9 broth  and 3  plasmids, respectively, were used as plasmid size controls. GeneRuler 1 kb DNA ladder was used as a size marker. The experiment was conducted in duplicate.Fig. S2. (A). Subsystem distribution of E. coli EC1390 chromosome based on the RAST annotation server. Out of 10,738 coding sequences predicted by RAST server, the subsystem coverage is 64% which contributes to a total of 599 subsystems. (B). Subsystem distribution of E. coli pEC1390-1 plasmid based on RAST annotation server. Out of 395 coding sequences predicted by RAST server, the subsystem coverage is 33% which contributes to a total of 13 subsystems. The green bar of the subsystem coverage indicates the percentage of the proteins included in the subsystems while the blue bar refers to the percentage of the proteins that are not included in the subsystems. Fig. S3. Verification of EC1390 transconjugants. (A). Random amplified polymorphic DNA (RAPD) patterns of E. coli recipient C600, EC974, EC1515, and TCGs. EC974 was used as a conjugation positive control. (B). PCR analysis to detect the blaNDM-5 gene. (C). Plasmid profiles of E. coli recipient and transconjugants. E. coli C600 was used as a negative control. Salmonella OU7526, E. coli EC974, and EC1515, contained 2 (50 and 90 kbp), 3 , and 3  plasmids, respectively, were used as plasmid size controls. The experiment was conducted in duplicate. NC, negative control; TGC, transconjugant; Marker, 100-bp DNA ladder. Fig. S4. mCIM and eCIM tests of EC1390 and its derived transconjugants. The inhibition zone diameter (mm) is shown in parentheses. According to the CLSI guidelines, K. pneumoniae ATCC BAA-1706 (carbapenemase negative), K. pneumoniae ATCC BAA-1705 (blaKPC positive), and K. pneumoniae ATCC BAA-2146 (blaNDM positive) were used as internal controls for mCIM and eCIM tests. The mCIM and eCIM tests were replicated by two independent investigators to ensure reproducibility. Table S1. EC1390 CRISPR type IE spacer sequences and their plasmid and/or phage targets. Table S2. Antimicrobial susceptibility of E. coli recipient and transconjugants.


The hexagonal structures proved to consist of alternating ···B-CO3··· and ···A-F··· layers in planes ab, interconnected along axis c by infinite chains ···F-B-F···, where cations formed polyhedra AOnF3 and BOmF2. The calculations included the band energy structure, the total and partial density of electron states, the energy and band widths of the upper ns- and np-states of alkali and alkaline-earth metals, as well as nd-zinc and nd-cadmium. For hydrostatic compression, we calculated the parameters of the Birch–Murnaghan equation of state and the linear compressibility moduli along the crystal axes and bond lines. We also defined the elastic constants of single crystals to obtain the Voigt–Reuss–Hill approximations for the elastic moduli of polycrystalline materials. The study also revealed the relationship between the elastic properties and the nature of the chemical bond. Hybrid functional B3LYP made it possible to calculate the modes of normal long-wavelength oscillations, which provided the spectra of infrared absorption and Raman scattering. Intramolecular modes ν1 and ν4 with one or two maxima were found to be intense, and their relative positions depended on the lengths of nonequivalent C–O bonds.The present work applied the methods of density functional theory and the van der Waals interaction PBE + D3(BJ) on the basis of localized orbitals of the CRYSTAL17 package. It featured the effect of interactions between structural elements of fluorocarbonates ABCO These new NLO materials demonstrated better mechanical and chemical stability. Zou et al. also measured second-harmonic generation (SHG) coefficients: they were by 1.11 , 3.33 , and 3.61 (KCaCO3F) times greater than those of the potassium dihydrogen phosphate (KDP) crystal KH2PO4. In their other study ∞. Yang et al. 3F and Rb3F  and KCaCO3F. They crystallize into a hexagonal crystal system with space group P-6m2 (No 187) with one formula unit (Z = 1) per unit cell. Isostructural ACdCO3F  and KZnCO3F also crystallize into a hexagonal lattice with symmetry group P-6c2 (No 188) and Z = 2 [3F  and CsCaCO3F with symmetry group P-62m (No 189) contain three formula units [Structural studies are important for creating new compounds with excellent optical characteristics. Reference  providestured in , while [tured in  focus on-6m2 No 17 with on62m No 18 contain 3F  with and without the van der Waals (vdW) correction to the standard density functional theory (DFT) methods. These crystals proved to be insulators with a wide bandgap and they were all relatively harder than KDP. The same study also introduced their linear optical characteristics, e.g., dielectric function, refractive indices, birefringence, and absorption spectra. In [3F  as entropy, Debye temperature, heat capacity, enthalpy, thermal expansion, and thermal conductivity. Another publication, [3F in two structural phases. KCdCO3F appeared to be a direct-gap insulator with fairly flat bands below the Fermi level. However, we found no study that featured the effect of pressure on the structural properties of fluorocarbonates.Fluorocarbonate crystals are hard to synthesize and grow. As a result, all their physical properties but not optical remain experimentally understudied. Therefore, computer simulation methods usually serve this purpose. In , Rao et ctra. In , DFT calication, , feature3F  and some of their main physical properties.Therefore, previous studies established that fluorocarbonates have good prospects for searching for new optical materials in the ultraviolet range. They are part of a wide class of NLOs, some of which science is already familiar with and some have yet to be created. A smart search for such materials requires predictive models that can describe the macroscopic properties of a substance based on its microscopic parameters. Computer methods are a convenient tool for modeling the nature of the interaction between atoms and molecules. They can establish the effect of chemical bonding on the physical properties of crystals. If scientists know the mechanisms of chemical bonding, they can design crystal systems with desired properties. As a result, the present research objective was to analyze the features of chemical bonding in crystals of fluorocarbonates ABCOWe used the density functional theory (DFT) methods in the CRYSTAL17 software to study the structure and electronic properties of metal carbonates . The sofDFT is an exact theory, provided that the exact exchange–correlation function is available . Since tk-points in the irreducible part of the Brillouin zone. The accuracy of the self-consistency procedure was ≥10−9 a.u. (1 a.u.= 27.21 eV).The reciprocal space was discretized using the Monkhorst–Pack grid  with 216DFT-D3 = DFTE + Edisp, where EDFT is the usual self-consistent Kohn–Sham energy obtained by the functional and Edisp is the empirical dispersion correction. In the present study, we used the well-known Grimm scheme [A correct description of non-covalent interactions relies on long-range correlation effects, which DFT methods lack. To take into account the missing dispersion interaction energy, one has to increase the total energy calculated for a given density approximation : EDFT-D3m scheme  and chosm scheme .η in the Voigt notation: V is the lattice cell volume, α, β = 1–6 . The second derivative is calculated numerically, while the first one is from the analytical gradient of the total energy.The elastic constant tensor was calculated using the algorithm described in ,33 as thE(V): x = 1. E0, V0, K0, and K1 are the four EoS parameters, where V0 is the volume at the energy minimum, E0 defines the depth of the curve E(V), and K0 and K1 define its shape. Knowing these parameters, we could easily calculate the pressure as To study the effect of pressure on the structure and electron properties, we used the equations of state (EoS) in the Birch–Murnaghan parametrization . The park = 0, the center of the first Brillouin zone) were obtained by diagonalizing the mass-weighted matrix of the second derivatives of energy with respect to atomic displacements u [a and b that weigh aM and bM shift in the lattice cell (index 0) along Cartesian directions i and j from the equilibrium state, respectively.The harmonic frequencies of phonons at point Г ; group II—KZnCO3F, KCdCO3F, RbCdCO3F (P-6c2); group III—RbMgCO3F, RbCaCO3F, CsCaCO3F (P-62m). Each group clearly demonstrates the roles of its alkali metal, alkaline-earth metal, or the one that contained a completely full nd10 shell in addition to the full ns2 shell.Based on their crystal structure, all the fluorocarbonates in this research were divided into three groups: group I—KCaCO3F are layered as follows: strontium (Sr), carbon (C), and oxygen (O) are located in plane z = c/2, whereas potassium (K) and fluorine (F) atoms are in plane z = 0. Here and below, coordinate z is chosen along axes c, x, and y in plane ab. Atomic position coordinates are as follows: K, Sr, C, F, and O in lattice vectors a = b, c. Thus, the independent parameters of the lattice were determined by optimizing its geometry and involved three quantities: a, c, and xO. Atoms of crystal KSrCO3F obtained by subtracting the density of non-interacting atoms from the crystal density. It illustrates the redistribution of the electronic charge during the chemical bond formation. Positive Δρ areas are red, while the areas of negative values are blue. Density values here and below are given in atomic units.e|, where e is the electron charge. The ion charge was calculated according to the Mulliken scheme, which splits it between atoms, which is not entirely true. The charge flows out from the inner regions of the carbon and oxygen atoms. Then, it flows into the region between them and behind the oxygen nuclei symmetrically to bond line C–O (1.308 Å). As a result, carbon acquires a positive charge of +0.39 |e|, while oxygen acquires a negative charge of −0.70 |e|. The overlap population of electron clouds on bond line C-O (PC-O) is 0.539 e, which indicates that they develop a strong covalent chemical bond and a stable carbonate ion. The excess charge behind the oxygen nuclei in plane ab, z = c/2 produces a stable SrCO3 structure. All carbonate groups line up in parallel, which gives the maximal contribution to the macroscopic SHG effect [Each Sr atom is surrounded by six oxygen atoms separated by 2.622 Å. All distances are given for functional PBE + D3, and ab, z = 0  in the primitive cell of KSrCO3F. It has strontium and fluorine atoms, as well as closely spaced oxygen and potassium atoms, so that the layers of Sr, C, and O alternate with those of K and F. The fluorine atoms are located above and below the strontium atoms at 2.354 Å. The overlap population for Sr-F is 0.003 e, which is even less than Sr-O at 0.008 e with a larger distance. Thus, the resulting ionic chemical bond provides infinite ···F-Sr-F··· chains. Each potassium atom is surrounded by six oxygen atoms at 2.909 Å. As maxima Δρ are located behind the oxygen nuclei, the overlap population on line K-O is 0.018 e. Thus, the nearest environment of negatively charged ions is polyhedra KO6F3 for potassium and SrO6F2 for strontium. According to [NOF is nine, the radius of the potassium ion RA equals 1.55 Å, and for a strontium ion with NOF = 8, RB equals 1.26 Å. As a result, the average radius of cation RC = (RA + RB)/2 is 1.405 Å. This value allowed us to establish all ordinary patterns in this research.rding to , if the 3F, the replacement triggers no fundamental change in the parameters of the crystal structure. Constants a and c increase, as do all interatomic distances, but the quantitative crystallographic environment remains the same. The cationic charge of strontium changes insignificantly and reaches +1.81 |e|. For the rubidium ion, the charge is +0.84 |e| larger than for potassium, and its cationic radius is 1.63 Å. In KCaCO3F, the calcium ion radius is 1.12 Å smaller than that of strontium. Therefore, lattice constants a and c and the interatomic distances are also smaller than in other crystals. The charge of the calcium ion is much less than that of strontium and equals +1.45 |e|, while the charge of potassium reaches +0.76 |e|. Due to the smaller calcium radius, the overlap populations of Ca-O and Ca-F increase by 0.049 and 0.036 e, respectively. Unlike KSrCO3F, the charge of the Ca-CO3 layer is negative and equals −0.024 |e|.When the alkali metal cation Rb replaces K in isostructural RbSrCORC in the series of KCaCO3F, KSrCO3F, and RbSrCO3F increases as 1.335, 1.405, and 1.445 Å, respectively. They have a stable linear dependency on the geometric parameters of the lattice. Lattice constant a increases as a(Å) = 2.815 + 1.713·RC, and so do the interatomic distances: RA-O(Å) = 0.405 + 1.778·RC, RB-O(Å) = 1.371 + 0.888·RC. The change in the distances triggers the change in the parameters of the chemical bond. As RC increases, the charges of CO3 anions  and fluorine anions  also increase. The overlap population of bond C–O also goes up , which means that the covalent chemical bond in the carbonate group also increases.The average radius of cation d10 and 4d10 electron shells, which are energetically close to the upper full 4s2 shell. Therefore, d-electrons participate in the hybridization of electron states. As a result, fluorocabonates with zinc and cadmium have different properties.Zinc and cadmium atoms have completely full 33F contains two formula units. Unlike group I, lattice constant c is twice as large, and its alternating layers are more numerous. The coordinates of nonequivalent atoms in units a and c are as follows: K, Cd, C, F, and O, i.e., a total of four independent quantities . Potassium atom charge falls down to +0.75 |e|, while that of fluorine rises as high as −0.68 |e|. The crystalline environment of the zinc atom differs from that of cadmium in KCdCO3F. It is surrounded by three oxygen atoms at 2.002 Å, two fluorine atoms, and three carbons, while three more oxygen atoms come last at 3.028 Å. This pattern occurs because the carbonate groups change their direction axis relative to the cations , which provides a significant overlap of electron shells with oxygen at 0.089 e and fluorine at 0.073 e, as well as a high share of the covalent component in the chemical bond.When rubidium replaces potassium, the resulting RbCdCO3F, KCdCO3F, and RbCdCO3F. For constant c(Å) = 1.918 + 5.234·RC and distances RC-O(Å) = 1.274 + 0.022·RC (0.99), the increasing bond length increases the population of overlap PC-O, which is 0.483, 0.493, and 0.506 e in this series.Group II demonstrates the linear dependency of the crystal parameters on the average cation radius, which increases as 1.225, 1325, and 1.365 Å in the series of KZnCO3F, atoms occupy the following positions: Rb, Ca, carbon C1 , C2, C1, C2, F, O1, and O2  = 6.90 + 1.671·RC and RA-O(Å) = 1.880 + 0.828·RC demonstrate a linear dependence.Like other groups, the average cation radius in the series of RbMgCO3F [3F, and CsCaCO3F [VR: a, c, V, RA-O, RA-F, RB-O, RB-F, and RC-O (mean for group III) is 0.42% for PBE + D3 functional, 1.53% for PBE, and 1.26% for B3LYP for nine fluorocarbonates. PBE + D3 gives a minimal deviation of 0.35% for lattice constant a, 0.3% for constant c, 0.58% for the shortest distances between cations A and oxygen, 0.36% for cations B and oxygen, and 0.79% for hybrid functional B3LYP for C-O. PBE + D3 decreases constant a in KSrCO3F (0.6%), RbSrCO3F (0.25%), KZnCO3F (0.3%), and KCdCO3F (0.6%). It decreases constant c in KZnCO3F (0.4%), RbCdCO3F (0.2%), RbMgCO3F (0.2 %), and RbCaCO3F (0.04%). Functional PBE in all fluorocarbonates increases both constant a and constant c. Functional B3LYP leads to a similar result, except for KZnCO3F and RbCdCO3F, where constant c is lower by 0.1%. Thus, van der Waals interactions provide an agreement between the calculated and experimental parameters of the crystal structure in fluorocarbonates ABCO3F.We compared the experimental crystallographic data for fluorocarbonates of group I , group I3F , RbCaCO3CsCaCO3F . The meaVR calculated by different functionals. For KCaCO3F, it is VRPBE = −0.052 + 1.025·VRPBE + D3. For volume V, the value calculated by functional PBE + D3 is 100.794 Å3, while the value obtained by the interpolation formula for PBE is 103.279 Å3, its precise value being 103.281 Å3. For functional B3LYP, the formula is VRB3LYP = −0.047 + 1.022·VRPBE + D3. On the contrary, gradient functional PBESOL and hybrid functional PBESOL0 lead to decreased lattice constants and actual interatomic distances: VRPBESOL = 0.038 + 0.982·VRPBE + D3, VRPBESOL0 = 0.057 + 0.972·VRPBE + D. Their mean-square deviations are 0.7 and 1.0%, which exceeds those for PBE + D3 by 0.45%.All fluorocarbonates demonstrate a simple linear relationship between the structural parameters V/Z(Å3) = −59.75 + 121.16·RC (0.99), the average distance from cations A and B to the nearest oxygen (RA–O + RB-O)/2(Å) = 0.253 + 1.738·RC (0.91), and the average distance between cations A and B and fluorine (RA–F + RB-F)/2(Å) = 1.13 + 1.1·RC (0.98) and RC-O(Å) = 1.264 + 0.03·RC (0.86). These formulae can predict the corresponding values for other fluorocarbonates. 3F (Z = 3), RbZnCO3F (Z = 2), RbPbCO3F, and CsPbCO3F (Z = 1), which were beyond the scope of this work. All fluorocarbonates demonstrated a stable linear dependence. The lattice cell volume per formula unit depends on the average cation radius E(k). The relevant points of the Brillouin zone were Г, M, L, A, K, H. 3F, KZnCO3F, and RbCaCO3F.The energy distribution of electrons in a crystal has a band structure that reflects the dependence of the electron energy on wave vector E and practically flat, depending on k. This pattern is typical of crystals with an ionic chemical bond. The conduction bands have a greater dispersion than the valence bands. The energy distance between the top of the valence band and the bottom of the conduction band determines an important parameter we defined as the band gap gE. The zones are divided into direct and indirect. In direct zones, the electron transition occurs from an occupied to an unoccupied state, both belonging to the same wave vector k. In indirect zones, they belong to different ones.The last full state is taken as the energy reference point. Therefore, the occupied  states have negative energies, and the unoccupied (conduction bands) have positive ones. Here and below, we give PBE + D3 calculations by default. The valence bands are narrow in energy pxy-oxygen. The bottom unoccupied bands are also separated from the subsequent ones by a band gap of 0.08, 0.40, and 1.67 eV. They are formed of about 60% zp- carbon and of about 39% oxygen. Only the following conduction bands are metallic in nature for 50–70% of s-potassium, rubidium, calcium, and strontium. The share of CO3 is 15–30%.In the fluorocarbonate group I, the top of the valence band and the bottom of the conduction band occur at point A. The band gaps in this series are 5.18, 5.12, and 5.32 eV. The uppermost valence bands are 0.78, 0.50, and 0.02 eV wide. Each is separated from the previous band by a band gap of 0.6–0.7 eV. This band is formed of 98% 3F and RbCdCO3F, the top of the valence band and the bottom of the conduction band are at point Г. In this series, its width is 5.18 eV  and 5.2 in  and 3F, the top of the valence band and the bottom of the conduction band are at point A. The band gap is 5.07 eV. A band gap of 0.75 eV separates the uppermost two valence bands from the previous ones and their width is 0.5 eV. They are formed of 98% of the xyp-states of O1 oxygen and have an anti-bonding character. The lower three unoccupied bands are also separated from the subsequent ones by a band gap of 0.4 eV. The lowest one is 87% formed of zp- states of carbon and oxygen from C2O3, and the other two are 92% formed of C1O3. Only the next conduction band has a metallic character at 29% s-rubidium and 37% magnesium.In RbMgCO3F and CsCaCO3F fundamentally differs from that of the rubidium–magnesium fluorocarbonate. The top of the valence band is at point Г, while the bottom of the conduction band is at point A and its indirect band gap is 4.97 and 5.16 eV. The top three valence bands are 1.1 and 1.0 eV wide. They are separated by a band gap of 0.35 eV from the preceding ones. The top one of these bands is 95% formed of pxy-oxygen from C2O3, whereas the other two are 92% formed of C1O3. The bottom unoccupied three bands are separated from the subsequent bands by band gaps 0.44 and 0.11 eV. The two bottom ones are formed of zp- carbon (68%) and oxygen from C1O3, while the third one is formed of C2O3 (88%). The following conduction bands are metallic: s- rubidium—39%, cesium—73%, and calcium—26.8%, respectively.The band structure of RbCaCON(E) reflect the hybridization degree.The total and partial density distribution can characterize the nature of the electronic states . The ene3F and KSrCO3F, the K3s- and K3p-states of potassium are located in the region of −27.8, −28.4 and −11.9, −12.2 eV, respectively. Their respective widths are about 0.01 and 0.16 eV. Therefore, they participate very little in the chemical bond formation, hence the insignificant overlap population of their electron shells with other atoms. In KZnCO3F and KCdCO3F, the energy positions of bands Ks3  and Kp3  shift towards higher values. The width of the former remains insignificant, while that of the latter one rises to 0.28 and 0.17 eV, respectively. The energies of 4s- and 4p-states of rubidium shift towards lower values as the cation B radius increases: from −25.6 and −9.3 eV with widths of 0.05 and 0.71 eV in RbMgCO3F and −26.1 and −9.8 eV with widths of 0.02 and 0.36 eV in RbCaCO3F to −28.4 and −12.3 eV with widths of 0.00 and 0.15 eV in RbSrCO3F. The energy positions of Css5 and Csp5 are −20.2 and −6.8 eV, and their widths are the most significant so far, namely 0.37 and 1.42 eV. This means that as the cation A radius increases at a fixed B, its states shift towards higher energies and the bandwidths increase.The states of alkali metals hardly affect the formation of the upper valence region between −5 and 0 eV. In KCaCOs2-, Mgp2-, Cas3-, and Srs4-lie in the energy region of the core states and are not marked in p3- maintains its energy position as −19.6 eV, while its width increases from 1.33 eV in KCaCO3F to 1.57 eV in RbCaCO3F. These energies coincide with the region of states Cs2-, Os2-, and Fs2-. Therefore, their considerable width indicates mutual hybridization. The energies of Srs4- states in KSrCO3F are at −15.3 eV with a width of 0.74 eV, while in RbSrCO3F they are at −15.6 eV with a width of 0.62 eV, which is higher in energy than the bands of hybrid anion s-states.Mgd states range from −5.5 to −4.0 eV with a maximal N(E) at −4.3 eV. They overlap with the region of Cp2-Op2 hybrid states from below and Fp2-Op2 states from above. Zinc states have a high degree of hybridization not only in this energy region but even in the upper valence states. For KCdCO3F, the band of cadmium 4d states ranges from –7.2 to –5.4 eV with maxima at −6.2 and −5.2 eV. The total share of cadmium for this energy range is 70%, oxygen—20%, and fluorine—5%. It is this bundle of energy bands that is responsible for the overlap population of Cd-F electron shells at 0.046 e and Cd-O at 0.057 e, which is approximately 70% of their total population. In RbCdCO3F, cadmium d-bands are located in the same energy region with a maximum of 6.3 eV. Anion–cation hybridization p-d is responsible for the overlap population of the Cd-O bond as 0.057 e and Cd-F as 0.037 e. The bottom hybrid zones with maxima at −6.7 and −7.5 eV provides 0.03 e of Cd-O overlap population. However, the upper bands with maxima at −5.1 eV are anti-bonding for Cd-O and bonding for C-O with 0.115 e of overlap population.Zinc 3N(E) of RbMgCO3F reflects the anion states since magnesium has almost no effect on its formation. The lowest bands with maxima at −22.1 and −21.4 eV are 0.74 eV wide. They are formed by hybridized s-states of carbon and oxygen, which are responsible for the C1-O1 overlap population of 0.106 e and C2-O2 of 0.101 e. Three bands at −20.8 eV are 0.12 eV wide. This group is formed by 95% of the fluorine s-states. The bands with N(E) maxima at −18.9 and −18.3 eV are 0.69 eV wide. They are formed by two-thirds by C1O3 s-states and by one-third by C2O3. They provide an overlap population of 0.178 e for C1-O1 and 0.16e for C2-O2. The energies of the p-states of oxygen and carbon  range from −8.3 to −5.4 eV with N(E) maxima at −8.0, −6.6, −6.2, and −5.5 eV. They provide an overlap of 0.071 e for Mg-O1, 0.014 e for Mg-O2, 0.131 e for C1-O1, and 0.178 e for C-O2. The bands in the energy range from –4.2 to –3.5 eV are formed of the pz-states of fluorine (73%) and magnesium (6%). They contribute 0.037 e to overlap population Mg-F. These crystalline orbitals provide π-conjugation of the layers in the crystal by developing ···F-Mg-F···chains.Structure E(V) was approximated by the third-order Birch–Murnaghan analytical equation of state. The distances–pressure dependence R(P) are described using compressibility modulus RK = R0/(dR/dP), where the derivative is calculated from a quadratic dependence. E(V), P(V), a(P)/a0, and c(P)/c0. The equilibrium values of the crystal parameters  = −59.27 + 120.83·RC (0.99) following the increase in the average cation radius. The volume modulus decreases, which provides a reliable correlation for the entire K0(GPa) = 168.37 − 78.24·RC (0.81). Fluorocarbonates have similar values of moduli K0. They are lower than those of calcite or dolomite and approach 65.24 GPa for aragonite [3F and KZnCO3F with the smallest grade B cations have the maximal K0 values . KCdCO3F and RbMgCO3F have the maximal SGH coefficients. Their maximal volume modulus increase rate with K1 pressure is maximal. Their linear correlation coefficient SGH = −16.8 + 4.11·K1 is 0.70. The ion exceeds D for the dispersive energy per one formula unit Edisp/Z, which is also at its maximum in these carbonates: −22.96 and −23.04 kJ/mol.For each group of fluorocarbonates, the cell volume per formula unit increases as ragonite . RbMgCO3aK along axis a and cK along axis c decrease as the average cation radius increases. However, group II is an exception: this rule does not work for module cK in RbCdCO3F. As for groups I and III aK > cK, this ratio increases in the former following the growing average cation radius and decreases in the latter. In group II, aK < cK and their ratio also decrease following the growing cation radius. In KZnCO3F, both directions along axes a and c also change with pressure.For each group of crystals, compressibility moduli KA-O for the distance between metal A and oxygen O decreases in each group with an increase in the average cation radius, while the distances RA-O grow. The linear dependency between them is stable, e.g., in group I: KA-O(GPa) = 425.8–97.92·RA-O. This pattern also exists for RB-O distances in groups I and III. However, group II has a much higher compressibility modulus KB-O than other fluorocarbonates with alkaline-earth metals. As mentioned above, Zn-O and Cd-O have high overlap populations. The fact that the region between the nuclei contains electrons prevents the distance from decreasing. This tendency is especially obvious in bond C-O, where compressibility modulus varies from 1000 to 1500 GPa, i.e., it is practically incompressible. In KC-O value between two non-equivalent CO3 groups to illustrate the comparison. The compressibility modulus KC1-O1 for C1O3 in RbMgCO3F is 922 GPa. For KC2-O2, it is as high as 2042 GPa, which is almost 2.5 times bigger. On the contrary, in RbCaCO3F and CsCaCO3F, compressibility modulus KC1-O1 is 1753 (1572) GPa for C1O3 group and ≤871 (830) GPa for KC2-O2. This fact can be explained by the different values of distances C–O, which have already manifested themselves in the band structure.Compressibility modulus RA-F occur in plane ab  ≈ 0.577·a and RR-F(Å) ≈ 0.577·c. Linear moduli have a similar relationship, which can be used to calculate them: KA-F(GPa) = −44.89 + 1.22·Ka (0.97), KB-F(GPa) = −9.95 + 1.05·Kc.The shortest distances between alkali metal atom A and fluorine atom e ab see , and thog axis c . Under pElastic properties determine how a certain material deforms or changes shape under applied forces. In microscopic theory, the elastic behavior of a crystal is related to the second derivative of the free energy with respect to reversible physical deformation. Thus, crystal elasticity is a very sensitive tool that can detect interactions and forces that occur between atoms and determine the structure and stability of crystals.ijC  for hexagonal crystals contains five independent constants and C66 = (C11-C12)/2 . As C12 > C13, it means that if the same normal stress acts in direction x, the crystals in direction z shrink more than in direction y. Small C44 values indicate that shift deformation occurs more easily if stresses are applied to horizontal planes ab. Since C66 > C44, a shift in plane ab causes less response than in the perpendicular direction. C44 value can serve as an indicator to determine the cracking of a hexagonal crystal along axis c.Elastic constants determine the properties of mechanical stability, which look as follows in hexagonal crystals  C44 > 0,C11 decreases as the average cation radius increases. Group I has the following dependence: C11(GPa) = 306.0 − 137.8·RC. This downward trend in C11 indicates that the bond between In fluorocarbonates, C11 and C33 always satisfy the following condition: C11,PBE + D3 > C11,B3LYP > C11,PBE, which means that the van der Waals interaction enhances the bond between BCO3 molecules and between layers. The shift constant, on the contrary, always satisfies C44,B3LYP > C44,PBE > C44,PBE + D3. Elastic constants ijC calculated by different functionals have a linear dependency. For example, for KSrCO3F, it looks like CPBE(GPa) = 1.38 + 0.90·CPBE + D3, CB3LYP(GPa) = 1.01 + 0.94·CPBE + D3; for KCdCO3F, it is CPBE(GPa) = 1.20 + 0.91·CPBE + D3, CB3LYP(GPa) = 0.97 + 0.92·CPBE + D3; and for RbMgCO3F, it is CPBE(GPa) = 1.23 + 0.92·CPBE + D3, CB3LYP(GPa) = 1.13 + 0.97·CPBE + D3.No experimental data for elastic constants are available; therefore, we have to compare their values obtained with different functionals. Constants KV, GV) [KR, GR) [KH = (KV + KR)/2, GH = (GV + GR)/2. These values [EH = 9KHGH/(3KH + GH) and the Poisson ratio µ = (3KH − 2GH)/(2(3KH + GH))  any uniform deformation in a polycrystalline aggregate is equated to the value of external deformation; (2) uniform stress is equated to external stress. These two assumptions of uniform local strain and uniform local stress are known as the Voigt   and Reus[KR, GR)  approxim[KR, GR) . The Hil[KR, GR) : volume e values  make it H + GH)) .KV/KR < 1 and GV/GR < 1. For all fluorocarbonates, these ratios are ≥1, but by a very small amount. The KV/KR ratio is minimal for KZnCO3F (1.000) and KCaCO3F (1.0003); it is maximal for RbMgCO3F (1.034) and RbCdCO3F (1.0205). Other crystals have intermediate values. These ratios are included in the so-called universal anisotropy index [UA = 5·GV/GR + KV/KR-6, which takes into account elastic anisotropy and is equally valid for all types of crystals (symmetries). The orthorhombic phase of carbon has the highest anisotropy, where UA is 397.3. Trigonal carbon has 284.0 [3F (1.6) and KSrCO3F has UA ≥ 1 (1.18). It is at its lowest in CsCaCO3F (0.27) and RbCdCO3F (0.41).A crystal cannot physically exist outside py index  AU = 5·Gas 284.0 . Only RbKH), the direction a(b) has a minimal value of βmin, and axis c has a maximal value of βmax (3F (1.75) and RbMgCO3F (1.71), while the minimal value belongs to KZnCO3F (1.00). The minimal value of shift modulus Gmin in all fluorocarbonates is in the direction of axis c and the maximal value is in various combinations of a(b)c. The maximal anisotropy coefficient for the shift modulus belongs to RbCaCO3F (3.33): it is ≥ 2 everywhere except RbCdCO3F (1.82) and CsCaCO3F (1.67). The maximal Young modulus Emax is along axis c. Its maximal anisotropy coefficient belongs to RbCaCO3F (2.58) and RbSrCO3F (2.45), whereas the lowest value is in CsCaCO3F (1.44). The Poisson ratio also has anisotropy but it is impossible to indicate any single selected direction for all materials.Anisotropy is an important property of any polycrystalline material. The anisotropy coefficient for each modulus can be calculated through its maximal and minimal values for each direction . For lin of βmax . The maxKH, GH, and EH decrease as the average cation radius increases. For each modulus, its value is maximal in group III and minimal in group I. Fluorocarbonates with a small grade B cation radius have the maximal KH. The modulus has a reliable linear dependence on average cation radius KH(GPa) = 171.1–79.9·RC (0.81).For each group of crystals, elastic moduli G describes plastic deformation resistance and volume modulus K reflects fracture resistance. Therefore, the brittle or plastic behavior of solids can be predicted based on the simple empirical relationship that occurs between them. The critical value that separates them is 1.75. If K/G > 1.75, the material is plastic; otherwise, it is brittle. Shift modulus G/K = 0.6) is the bottom limit, while 0.5 (G/K = 0) is the top limit. At μ ≤ 0.25 (G/K > 0.6), the interatomic forces are non-central, which indicates that the bond has a covalent component. In fluorocarbonates, μ ≥ 0.28 indicates central forces and ionic bonding. The Poisson’s ratio and ratio (G/K) = 1.238–2.588 µ have a reliable linear dependency: group II fluorocarbonates with Zn and Cd and µ > 0.34 have chemical bonding with a greater covalent character than other fluorocarbonates. The same crystals also have greater plasticity, since for them K/G > 2.8.The Poisson ratio provides information about the characteristics of the bond strengths. For central forces in solids , value μ = 0.25 (vp) and transverse (vs) waves can be theoretically defined as [Pv = ((KH + 4/3GH)/ρ)/21, Sv = (GH/ρ)/21. The highest velocities of the longitudinal and transverse waves belong to KCaCO3F and RbMgCO3F. In each group, they decrease following the increase in the average cation radius. For example, it is vs(km/s) = 11.74 − 6.22·RC, Pv(km/s) = 22.10 − 11.77·RC in group I.The resulting set of elastic moduli can be used for semi-empirical estimates of some other physical properties of polycrystalline materials. For instance, acoustic velocities in a solid can be obtained from the volume modulus, shift modulus, and density ρ. In an anisotropic material, wave energy propagates in two modes, i.e., longitudinal and transverse. As a rule, the longitudinal mode is faster: particle oscillations move parallel to wave energy propagation. The transverse mode is slower: particle oscillations are perpendicular to wave energy propagation. Acoustic velocities of longitudinal (fined as : vP = ((3h) has four main IR-active vibrational modes [ν1 (1100 cm−1), out-of-plane bending ν2 (800 cm−1), singular asymmetric stretching ν3 (1400 cm−1), and singular plane deformation mode ν4 (700 cm−1). The same types of vibrations are active in RS.The spectra of infrared light absorption (IRS) and Raman light scattering (RS) of fluorocarbonates are divided into two regions: (1) lattice vibrations that involve metal atoms, fluorine, translational, and rotational displacements of whole carbonate groups; (2) intramolecular vibrations of atoms of carbonate groups. Free al modes : symmetrA1u and A2g, which are inactive for the indicated spectra. P-6m2, only one mode where polarization vectors transform according to the irreducible representation xx, αyy, and αzz tensor components are nonzero. This vibration belongs to the intramolecular v1 type. As the number of carbonate groups increases, the total number of these vibrations for group II with symmetry P-6c2 and group III with symmetry P-62m is 2 and 6, respectively. Doubly degenerate modes of symmetry E||z and in RS with nonzero components αxx, αxy, and αyy. Group I fluorocarbonates have only five such modes: two internal modes v4 and v3 and three lattice modes. Crystals of symmetry P-62m and Z = 3 have only fourteen: six internal modes, three v4 modes, three v3 modes, and eight lattice modes.For group I fluorocarbonates of symmetry v3 is taken as 100%. Its intensity decreases in the series as 2207, 2161, and 1964 km/mol with a decrease in the average Born charge of ZC cation. For all the carbonates in this study, the maximal intensity averaged over number Z of modes v3 equals I(km/mol) = −2337 + 2727·ZC (0.91). The dynamic charge of alkali cations ZA correlates with their electronic charge ZA(|e|) = 1.85 − 0.83·QA (0.93), and this correlation is the same for ZB(|e|) = 2.83 − 0.445·QB (0.96). The intensity of line v1 of the symmetry 3F, modes v4 of v2 of symmetry v1 with wave numbers that decrease in series I as 1110, 1096, and 1096 cm−1, as well as by modes v4 (~35% intensity) and modes v3 , which makes it possible to identify fluorocarbonates.In IRS, KCaCO−1 are formed by translational vibrations of cations A with CO3 anions and cations B with fluorine anions in opposite directions of plane xy. Alkali metal atoms provide the maximal contribution of 40–50% to the displacement vector amplitudes, while fluorine atoms are responsible for 50–40%. Modes with wave numbers 164, 138, and 148 cm−1 appear when A and F atoms (50%) shift in one direction, while B and CO3 (40%) atoms shift in the other direction. These are translational vibrations of two atomic planes relative to each other. The most intense modes at 210, 208, and 200 cm−1 are formed by translational shifts of cations A and B (46–10%) and anions CO3 (50–90%) and F in opposite directions. Symmetry vibrations z. Modes with low intensity and wave numbers 306, 313, and 245 cm−1 are formed by vibrations of atoms A, CO3 (65%) and B, F. Modes of high intensity at 471, 456, and 395 cm−1 are formed by vibrations of atoms B (27–10%) and fluorine (69–90%). In RS, librational oscillations of CO3 correspond to symmetry modes Not only wave number and intensity can characterize lattice modes. In the case of carbonate ions, the squared oscillation amplitude (percentage) of an atom or group of atoms is another option. In IRS group I, lattice modes of symmetry 3F in IRS . Modes v4 with wave numbers ~720 cm−1 (730–680 cm−1) have a low intensity that starts at ~1%. Modes v2 of symmetry −1 (853 cm−1) and 847 cm−1 (843 cm−1) also have low intensity. In RS of cadmium fluorocarbonates, symmetry −1 demonstrate the most intense vibration, while modes v4 and v3 have an intensity of ~7–10%.For intramolecular vibrations of KCdCOF in IRS , mode v3432 cm−1 . In RbCd3F, symmetry modes −1 and 117 cm−1 prove to be the most intense ones (13%). In the first vibration, cadmium (3%) and CO3 (94%) atoms are displaced in opposite directions of plane xy. In the second vibration, Cd and CO3 atoms shift in the same direction, while potassium (40%) and fluorine (56%) atoms shift in the other direction (in-plane shifts). Symmetry mode −1 appears as potassium and fluorine atoms (~90%) shift in one direction z, while Cd and CO3 (~10%) shift in another direction. Symmetry mode −1 results from translational vibrations of cadmium atoms (15%) and UCO3 anions (80%).For IRS of lattice vibrations in KCdCO−1, as well as symmetry modes xy (~33%) and CO3 atoms that rotate in the direction of axis z (~65%).As for group II fluorocarbonates, the most intense (~15–20%) lattice vibrations in RS are symmetry modes 3F (v3 with wave number 1539 cm−1 is the most intense one (5852 km/mol). It is followed by a mode with 13% intensity and a wave number of 1591 cm−1. The mode with 0.5% intensity at 1624 cm−1 comes last  [−1 prove that molecules interact. The contribution of the polarization vectors to mode I is 82% of C1O3 and 17% of C2O3. For mode II, it is 67% and 32%, respectively. For mode III, it is 50% of each. Three modes v4 of symmetry −1 (680 cm−1). Their intensity is ≤0.5%. The C2O3 group is responsible for 99% of the last mode. Two modes of type v2 with intensities of ~2% have wave numbers of 842 and 846 cm−1 (890 cm−1). The third mode of this type with wave number 843 cm−1 belongs to symmetry −1 (100%) and 1107 cm−1 (52%) prove to be the most intense ones. They develop by 99.6% due to vibrations of oxygen atoms O1 and by 99.9% due to vibrations of oxygen atoms O2 along the C–O bond line.In IRS of RbMgCO3F , mode v33F has the second v1 maximum located to the left of the main one. However, in RbCaCO3F and CsCaCO3F it is on the right. The fact is that in fluorocarbonate with magnesium, bond length RC1-O1 is shorter than RC2-O2, whereas, in fluorocarbonate calcium, RC1-O1 is longer than RC2-O2. These structural features also lead to other changes in the spectrum. The positions of the wave numbers and mode v3 intensities swap places. As a result, the central maximum changes at 1536 cm−1 in CsCaCO3F. The position of the maxima in band v4 also changes, while maintaining its width.The other two group III fluorocarbonates show changes in RS. RbMgCO3F, the IRS demonstrates an intense (37%) symmetry mode −1. It depends on 60% of magnesium atom vibration, 23% on C1O3, and 16% on C2O3. Symmetry mode −1  and O2 (30%).In lattice vibrations of RbMgCOEZP(kJ/mol) = 83.89 − 22.34·RC. The wave numbers of intramolecular vibrations also depend on the average radius of the cation. For mode v4, they increase in group I while decreasing  in groups II and III. As a result, their linear dependence has a correlation coefficient of at least 0.75 of v4(cm−1) = 883 − 123·RC. Out-of-plane vibration mode v2 (cm−1) = 690 − 123·RC behaves approximately the same way. For the fully symmetric mode that dominates RS, it is v1(cm−1) = 1315 − 151·RC. These formulas can predict the IRS and RS of fluorocarbonates.The energy of zero-point vibrations 3F in [3F, functional PBE calculations of wave numbers for twenty active vibrations are connected by a linear dependency with the functional B3LYP as vPBE(cm−1) = −2.47 + 0.97·vB3LYP. For PBE + D3, it is vPBE + D3(cm−1) = 4.74 + 0.97·vB3LYP. For B3LYP, some frequency values are higher and some are lower than for PBE + D3, but they are always greater than PBE. For example, the lattice mode Mg-F of symmetry −1, and for PBE + D3 its value is 537 cm−1. Therefore, intermolecular interaction agrees with the experimental value for this lattice mode. The situation is different for intramolecular modes, e.g., for KCaCO3F, it is as follows: vPBE(cm1) = −18.2 + 0.98·vB3LYP, vPBE + D3(cm−1) = −22.9 + 0.99·vB3LYP. Thus, the functionals PBE and PBE + D3 provide lower wave numbers than B3LYP. The ratio of zero-point energy obtained by functional B3LYP is 1.001 ÷ 1.036 times greater than by functional PBE and 1.00 ÷ 1.027 times greater than the functional PBE + D3.Tran et al. measured wave numbers of some intramolecular modes for RbMgCO3F in . Their m3··· and ···A-F··· in planes ab, interconnected along axis c with infinite chains ···F-B-F···. As a result, cations form polyhedra AOnF3 and BOmF2. The average radius of cations RC = (RA + RB)/2 depends on coordination environment n + 3 and m + 2. The radius can establish linear dependences of structural and chemical bonding parameters. The resulting formulas can be used to predict the structural properties of other fluorocarbonates.Hexagonal fluorocarbonates ABCO3F  form structures of alternating layers ···B-COK0(GPa) = 168.37 − 78.24·RC. The compressibility of metal–oxygen bonding for cations B was lower than that for alkali cations A, while bonds C-O were practically incompressible. For bonds A-F and B-F, we established the dependence on the linear compressibility moduli along axes a and c as KA-F(GPa) = −44.89 + 1.22·Ka and KB-F(GPa) = −9.95 + 1.05·Kc, one of which can be used to calculate the other.In this research, we studied the effect of pressure on the structure of fluorocarbonates. For volume compression moduli, the series dependence of the cation on the radius was KH) was minimal for direction a(b) and maximal along axis c. Shift modulus GH (Young modulus EH) was minimal  along axis c. For each of the three groups of crystals with one, two, or three formula units Z, elastic moduli KH, GH, and EH decreased as the average cation radius increased. We established a linear relationship between the Poisson ratio and ratio GH/KH: crystals with a larger proportion of covalent bonds were also more plastic.The elastic properties of monocrystalline and polycrystalline fluorocarbonates were anisotropic. Linear compressibility (1/−1 in the degenerate asymmetric extension modes of carbonate ion ν3 and the Raman scattering spectra in the most intense region of 1080 ÷ 1180 cm−1 in symmetric extension modes ν1 for fluorocarbonates with two crystallographically inequivalent carbonate groups had two maxima while the rest had one. The intensity of band v3, as well as the presence of one or two maxima v1, can be used to identify fluorocarbonates by their vibrational spectra.The infrared absorption spectra for the most intense region with wave numbers 1490 ÷ 1620 cmThe physicochemical change patterns for fluorocarbonates ABCO3F were established based on the cations. These patterns can serve as a basis for a programmed search for new NLO crystals with different cations. In addition, the new approach can be applied to the systematic analysis of physical characteristics of other halogen-carbonate families with different halogen/oxygen ratios in an anion sublattice.