Thermostable chitinase from Cohnella sp. A01: isolation and product optimization.
Twelve bacterial strains isolated from shrimp farming ponds were screened for their growth activity on chitin as the sole carbon source. The highly chitinolytic bacterial strain was detected by qualitative cup plate assay and tentatively identified to be Cohnella sp. A01 based on 16S rDNA sequencing and by matching the key morphological, physiological, and biochemical characteristics. The cultivation of Cohnella sp. A01 in the suitable liquid medium resulted in the production of high levels of enzyme. The colloidal chitin, peptone, and K2HPO4 represented the best carbon, nitrogen, and phosphorus sources, respectively. Enzyme production by Cohnella sp. A01 was optimized by the Taguchi method. Our results demonstrated that inoculation amount and temperature of incubation were the most significant factors influencing chitinase production. From the tested values, the best pH/temperature was obtained at pH 5 and 70°C, with Km and Vmax values of chitinase to be 5.6mg/mL and 0.87μmol/min, respectively. Ag+, Co2+, iodoacetamide, and iodoacetic acid inhibited the enzyme activity, whereas Mn2+, Cu2+, Tweens (20 and 80), Triton X-100, and EDTA increased the same. In addition, the study of the morphological alteration of chitin treated by enzyme by SEM revealed cracks and pores on the chitin surface, indicating a potential application of this enzyme in several industries.
['Aliabadi|Nasrin|N|', 'Aminzadeh|Saeed|S|', 'Karkhane|Ali Asghar|AA|', 'Haghbeen|Kamahldin|K|']
D001407:Bacillus D002244:Carbon D002688:Chitinases D004789:Enzyme Activation D004795:Enzyme Stability D006863:Hydrogen-Ion Concentration D007477:Ions D007700:Kinetics D008670:Metals D009584:Nitrogen D010758:Phosphorus D012336:RNA, Ribosomal, 16S D017422:Sequence Analysis, DNA D013696:Temperature
Thermostable Chitinase Cohnella sp. A01 Isolation Biochemical characterization