Journal
FEBS JOURNAL
Volume 283, Issue 12, Pages 2340-2353Publisher
WILEY-BLACKWELL
DOI: 10.1111/febs.13743
Keywords
beta-glucosidase; glycoside hydrolase; metagenomics; substrate specificity; X-ray crystallography
Categories
Funding
- JSPS [26850067]
- Grants-in-Aid for Scientific Research [15K07383, 15H02443, 26850067, 26660083] Funding Source: KAKEN
Ask authors/readers for more resources
beta-Glucosidase Td2F2 isolated from a compost metagenome has high glucose tolerance and transglycosylation activity. In this study, we determined the high-resolution crystal structure of Td2F2. It has a unique structure at the -1 subsite that is important for substrate specificity but not for glucose tolerance. To elucidate the mechanism(s) of glucose tolerance, we isolated a glucose-sensitive Td2F2 mutant using random mutagenesis. In this mutant, Asn223 residue located between subsites +1 and +2 was mutated. The Asn223 mutation resulted in reduced glucose tolerance and transglycosylation activity, and drastically changed substrate specificity. These results indicate that the structure between subsites +1 and +2 is critical for the glucose tolerance and substrate specificity of Td2F2. Our findings shed light on the glucose tolerance and transglycosylation activity mechanisms of glycoside hydrolase family 1 beta-glucosidases.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available