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Cloning and characterization of a new β-mannosidase from Streptomyces sp. S27

Paper ID Volume ID Publish Year Pages File Format Full-Text
17485 42672 2011 7 PDF Available
Title
Cloning and characterization of a new β-mannosidase from Streptomyces sp. S27
Abstract

A new β-mannosidase gene, designated as man2S27, was cloned from Streptomyces sp. S27 using the colony PCR method and expressed in Escherichia coli BL21 (DE3). The full-length gene consists of 2499 bp and encodes 832 amino acids with a calculated molecular mass of 92.6 kDa. The amino acid sequence shares highest identity of 62.6% with the mannosidase Man2A from Cellulomonas fimi which belongs to the glycoside hydrolase family 2. Purified recombinant Man2S27 showed optimal activity at pH 7.0 and 50 °C. The specific activity, Km, and kcat values for p-nitrophenyl-β-d-mannopyranoside (p-NP-β-MP) were 35.3 U mg–1, 0.23 mM, and 305 s–1, respectively. Low transglycosylation activity was observed when Man2S27 was incubated with p-NP-β-MP (glycosyl donor) and methyl-α-d-mannopyranoside (p-NP-α-MP) (acceptor) at 50 °C and pH 7.0, and a small amount of methylmannobioside was synthesized. Using locust bean gum as the substrate, more reducing sugars were liberated by the synergistic action of Man2S27 and β-mannanase (Man5S27), and the synergy degree in sequential reactions with Man5S27 firstly and Man2S27 secondly was higher than that in the simultaneous reactions.

Keywords
Streptomyces sp. S27; β-Mannosidase; Transglycosylation; Synergistic action
First Page Preview
Cloning and characterization of a new β-mannosidase from Streptomyces sp. S27
Publisher
Database: Elsevier - ScienceDirect
Journal: Enzyme and Microbial Technology - Volume 49, Issue 3, 10 August 2011, Pages 277–283
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering