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Identification of a β-glucosidase from the Mucor circinelloides genome by peptide pattern recognition

Paper ID Volume ID Publish Year Pages File Format Full-Text
17117 42643 2014 6 PDF Available
Title
Identification of a β-glucosidase from the Mucor circinelloides genome by peptide pattern recognition
Abstract

•PPR can simultaneously identify protein subfamily grouping and predict function.•PPR was applied to mine the Mucor circinellolides genome for new GH genes.•12 different genes encoding different GHs were found in the genome.•A functional β-glucosidase was expressed in Pichia pastoris.

Mucor circinelloides produces plant cell wall degrading enzymes that allow it to grow on complex polysaccharides. Although the genome of M. circinelloides has been sequenced, only few plant cell wall degrading enzymes are annotated in this species. We applied peptide pattern recognition, which is a non-alignment based method for sequence analysis to map conserved sequences in glycoside hydrolase families. The conserved sequences were used to identify similar genes in the M. circinelloides genome. We found 12 different novel genes encoding members of the GH3, GH5, GH9, GH16, GH38, GH47 and GH125 families in M. circinelloides. One of the two GH3-encoding genes was predicted to encode a β-glucosidase (EC 3.2.1.21). We expressed this gene in Pichia pastoris KM71H and found that the purified recombinant protein had relative high β-glucosidase activity (1.73 U/mg) at pH5 and 50 °C. The Km and Vmax with p-nitrophenyl-β-d-glucopyranoside as substrate was 0.20 mM and 2.41 U/mg, respectively. The enzyme was not inhibited by glucose and retained 84% activity at glucose concentrations up to 140 mM. Although zygomycetes are not considered to be important degraders of lignocellulosic biomass in nature, the present finding of an active β-glucosidase in M. circinelloides demonstrates that enzymes from this group of fungi have a potential for cellulose degradation.

Keywords
PPR, peptide pattern recognition; GH family, glycoside hydrolase family; pNPG, p-nitrophenyl-β-d-glucopyranoside; MUG, 4-methylumbelliferyl β-d-glucopyranosidePeptide pattern recognition; Mucor circinelloides; β-Glucosidase; Glycoside hydrolase 3
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Identification of a β-glucosidase from the Mucor circinelloides genome by peptide pattern recognition
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Publisher
Database: Elsevier - ScienceDirect
Journal: Enzyme and Microbial Technology - Volume 67, December 2014, Pages 47–52
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us