3WH5

Crystal structure of GH1 beta-glucosidase Td2F2

Summary for 3WH5

• Entry DOI: 10.2210/pdb3wh5/pdb
• Related: 3WH6 3WH7 3WH8
• Descriptor: beta-glucosidase, 2-[N-CYCLOHEXYLAMINO]ETHANE SULFONIC ACID, GLYCEROL, ... (5 entities in total)
• Functional Keywords: tim barrel, hydrolase
• Biological source: metagenomes
• Total number of polymer chains: 1
• Total formula weight: 51169.23

Authors

Jo, T.,Fushinobu, S.,Uchiyama, T.,Yaoi, K. (deposition date: 2013-08-21, release date: 2014-09-03, Last modification date: 2023-11-08)

Primary citation

Matsuzawa, T.,Jo, T.,Uchiyama, T.,Manninen, J.A.,Arakawa, T.,Miyazaki, K.,Fushinobu, S.,Yaoi, K.
Crystal structure and identification of a key amino acid for glucose tolerance, substrate specificity, and transglycosylation activity of metagenomic beta-glucosidase Td2F2.
Febs J., 283:2340-2353, 2016

PubMed Abstract: β-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 β-glucosidases.

PubMed: 27092463
DOI: 10.1111/febs.13743

Experimental method

X-RAY DIFFRACTION (1.6 Å)