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  • Glutamic acid 160 is the acid-base catalyst of beta-xylosidase from Bacillus stearothermophilus T-6: a family 39 glycoside hydrolase.

Glutamic acid 160 is the acid-base catalyst of beta-xylosidase from Bacillus stearothermophilus T-6: a family 39 glycoside hydrolase.

FEBS letters (2001-04-27)
T Bravman, A Mechaly, S Shulami, V Belakhov, T Baasov, G Shoham, Y Shoham
ABSTRACT

A beta-xylosidase from Bacillus stearothermophilus T-6 was cloned, overexpressed in Escherichia coli and purified to homogeneity. Based on sequence alignment, the enzyme belongs to family 39 glycoside hydrolases, which itself forms part of the wider GH-A clan. The conserved Glu160 was proposed as the acid-base catalyst. An E160A mutant was constructed and subjected to steady state and pre-steady state kinetic analysis together with azide rescue and pH activity profiles. The observed results support the assignment of Glu160 as the acid-base catalytic residue.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
4-Nitrophenyl α-D-xylopyranoside, α-xylosidase substrate
Sigma-Aldrich
4-Nitrophenyl β-D-xylopyranoside, ≥98%