5ayb: Difference between revisions

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. This article is protected by copyright. All rights reserved.

Crystal structure and identification of a key amino acid for glucose tolerance, substrate specificity and transglycosylation activity of metagenomic beta-glucosidase Td2F2.,Matsuzawa T, Jo T, Uchiyama T, Manninen JA, Arakawa T, Miyazaki K, Fushinobu S, Yaoi K FEBS J. 2016 Apr 19. doi: 10.1111/febs.13743. PMID:27092463<ref>PMID:27092463</ref>

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[[Category: Large Structures]]

[[Category: Metagenomes]]

[[Category: Arakawa T]]

[[Category: Fushinobu S]]

[[Category: Jo T]]

[[Category: Manninen JA]]

[[Category: Matsuzawa T]]

[[Category: Uchiyama T]]

[[Category: Yaoi K]]