Purification and characterization of cycloisomaltotetraose-forming glucanotransferases from Agreia sp. D1110 and Microbacterium trichothecenolyticum D2006.
Glucanotransferases that can synthesize cyclo-{→6)-α-d-Glcp-(1→6)-α-d-Glcp-(1→6)-α-d-Glcp-(1→6)-α-d-Glcp-(1→} (CI4) from dextran were purified to homogeneity from the culture supernatant of Agreia sp. D1110 and Microbacterium trichothecenolyticum D2006. The molecular mass of both enzymes was estimated to be 86 kDa by SDS-PAGE. The glucanotransferase, named CI4-forming enzyme, from Agreia sp. exhibited the highest activity at pH 6.0 and 40 °C. The enzyme was stable on the pH range of 4.6-9.9 and up to 40 °C. On the other hand, the enzyme from M. trichothecenolyticum exhibited the highest activity at pH 5.7 and 40 °C. The enzyme was stable on the pH range of 5.0-6.9 and up to 35 °C. Both enzymes catalyzed 4 reactions, namely, intramolecular α-1,6-transglycosylation (cyclization), intermolecular α-1,6-transglycosylation, hydrolysis of CI4, and coupling reaction. Furthermore, the CI4-forming enzyme produced CI4 from α-1,6-linked glucan synthesized from starch by 6-α-glucosyltransferase. These findings will enable the production of CI4 from starch.
['Fujita|Akihiro|A|', 'Kawashima|Akira|A|', 'Mitsukawa|Yuuki|Y|', 'Kitagawa|Noriaki|N|', 'Watanabe|Hikaru|H|', 'Mori|Tetsuya|T|', 'Nishimoto|Tomoyuki|T|', 'Aga|Hajime|H|', 'Ushio|Shimpei|S|']
D039903:Actinobacteria D003470:Culture Media D003500:Cyclization D004591:Electrophoresis, Polyacrylamide Gel D006004:Glycogen Debranching Enzyme System D006031:Glycosylation D006358:Hot Temperature D006863:Hydrogen-Ion Concentration D000083822:Microbacterium D008970:Molecular Weight D009844:Oligosaccharides
glucanotransferase cycloisomaltooligosaccharide cycloisomaltotetraose transglycosylation α-1,6 linkage