5U2O

Crystal structure of Zn-binding triple mutant of GH family 9 endoglucanase J30

Summary for 5U2O

• Entry DOI: 10.2210/pdb5u2o/pdb
• Related: 5U0H
• Descriptor: J30 CCH, CITRATE ANION, GLYCEROL, ... (5 entities in total)
• Functional Keywords: glycoside hydrolase, gh9, ig-like domain, cbm30, hydrolase
• Biological source: Thermobacillus composti KWC4
• Total number of polymer chains: 1
• Total formula weight: 64609.95

Authors

Ellinghaus, T.L.,Pereira, J.H.,McAndrew, R.P.,Welner, D.H.,Adams, P.D. (deposition date: 2016-11-30, release date: 2018-05-30, Last modification date: 2024-04-03)

Primary citation

Ellinghaus, T.L.,Pereira, J.H.,McAndrew, R.P.,Welner, D.H.,DeGiovanni, A.M.,Guenther, J.M.,Tran, H.M.,Feldman, T.,Simmons, B.A.,Sale, K.L.,Adams, P.D.
Engineering glycoside hydrolase stability by the introduction of zinc binding.
Acta Crystallogr D Struct Biol, 74:702-710, 2018

PubMed Abstract: The development of robust enzymes, in particular cellulases, is a key step in the success of biological routes to `second-generation' biofuels. The typical sources of the enzymes used to degrade biomass include mesophilic and thermophilic organisms. The endoglucanase J30 from glycoside hydrolase family 9 was originally identified through metagenomic analyses of compost-derived bacterial consortia. These studies, which were tailored to favor growth on targeted feedstocks, have already been shown to identify cellulases with considerable thermal tolerance. The amino-acid sequence of J30 shows comparably low identity to those of previously analyzed enzymes. As an enzyme that combines a well measurable activity with a relatively low optimal temperature (50°C) and a modest thermal tolerance, it offers the potential for structural optimization aimed at increased stability. Here, the crystal structure of wild-type J30 is presented along with that of a designed triple-mutant variant with improved characteristics for industrial applications. Through the introduction of a structural Zn site, the thermal tolerance was increased by more than 10°C and was paralleled by an increase in the catalytic optimum temperature by more than 5°C.

PubMed: 29968680
DOI: 10.1107/S2059798318006678

Experimental method

X-RAY DIFFRACTION (1.46 Å)