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Inducible and constitutive expression of a novel thermostable alkaline β-mannanase from alkaliphilic Bacillus sp. N16-5 in Pichia pastoris and characterization of the recombinant enzyme

Paper ID Volume ID Publish Year Pages File Format Full-Text
17990 42709 2008 6 PDF Available
Title
Inducible and constitutive expression of a novel thermostable alkaline β-mannanase from alkaliphilic Bacillus sp. N16-5 in Pichia pastoris and characterization of the recombinant enzyme
Abstract

A novel thermostable alkaline β-mannanase from alkaliphilic Bacillus sp. N16-5 was expressed successfully in Pichia pastoris GS115. The combined usage of inducible and constitutive promoters (AOX1 and GAP) enhanced the expression of β-mannanase. Among the parameters investigated in shaking flask cultures, the pH value of medium had significant influence on the production of β-mannanase by recombinant P. pastoris. β-Mannanase produced at pH 7.0 was 6.7 times of that at pH value of 6.0. The highest β-mannanase activity of 32.2 IU/ml in culture supernatant was achieved at 120 h of cultivation in BMGY medium (pH 7.0). The recombinant β-mannanase was purified and characterized. The purified β-mannanase produced by P. pastoris has optimum pH of 10.0 and optimum temperature of 70 °C, which are very close to those of the native enzyme from alkaliphilic Bacillus sp. N16-5. However, much higher thermal stability and pH stability were observed in recombinant β-mannanase. These properties make the recombinant β-mannanase more useful in the detergent industries, the pulp and paper processing and other industrial processes.

Keywords
Pichia pastori; Alkaline β-mannanase; Alkaliphilic Bacillus sp.; Secretory expression; Characterization
First Page Preview
Inducible and constitutive expression of a novel thermostable alkaline β-mannanase from alkaliphilic Bacillus sp. N16-5 in Pichia pastoris and characterization of the recombinant enzyme
Publisher
Database: Elsevier - ScienceDirect
Journal: Enzyme and Microbial Technology - Volume 43, Issue 1, 7 July 2008, Pages 13–18
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering