Abstract
Bacterial chitinases recruited multiple accessory domains for the conversion of recalcitrant polysaccharides to simple soluble sugars/amino sugars. Here, we report detailed properties of a multi-domain GH18 chitinase from Enterobacter cloacae subsp. cloacae (EcChi2) that preferred β-chitin as substrate. EcChi2 exhibited transglycosylation (TG) activity on oligomeric substrates from DP4-DP6. The high amount of DP2 is indicative of exo mode activity of EcChi2. We generated EcChi2 variants (truncated and fusion chimeras) and elucidated the role of catalytic and accessory domains. The catalytic efficiency of truncated GH18 and fusion chimera of GH18+ChBD1-ChBD2 decreased to 22 and 17-fold, respectively, than EcChi2, and lost the hydrolytic activity on polymeric substrates, except colloidal chitin. On the other hand, the catalytic activity of truncated PKD1-GH18-PKD2 on polymeric and oligomeric substrates was similar to EcChi2, suggesting that PKD domains are essential for increasing the rate of hydrolysis. Moreover, the truncated ChBD1-ChBD2 and fusion PKD1 + PKD2 participated in chitin-binding.
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
