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Cloning, expression, characterization, and high cell-density production of recombinant endo-1,4-β-xylanase from Aspergillus niger in Pichia pastoris

Paper ID Volume ID Publish Year Pages File Format Full-Text
17963 42708 2007 7 PDF Available
Title
Cloning, expression, characterization, and high cell-density production of recombinant endo-1,4-β-xylanase from Aspergillus niger in Pichia pastoris
Abstract

A recombinant gene XylB (564 bp) encoding endo-1,4-β-xylanase, obtained from Aspergillus niger BCC14405, was successfully cloned and secreted as a 21 kDa in Pichia pastoris under the control of AOX1 promoter. The activity of the recombinant xylanase was highest at 55 °C which was 5 °C higher than native xylanase. In addition, the recombinant xylanase was active over the range of pH 3.6–6.5 with maximal activity at pH 5 (8007 U/mg). When compared to a commercial enzyme, in vitro digestibility of the recombinant enzyme was 1.8- and 2.4-folds higher digesting rates of rice bran and soybean meal fibers, respectively.Two-liter production of xylanase was performed with BSM medium which increased cell concentration up to 84.5 gdry-weight/L via the 80% μmax exponential feed strategy. This process provided maximum xylanase production (3676 U/mL) with highest specific activity (7352 U/mgprotein) and volumetric productivity (22,832 U/L/h) at 3.0% (v/v) methanol induction. By far, this was the highest xylanase expression in P. pastoris host system being reported. Thus, this BCC14405 recombinant xylanase could be produced and used effectively as a feed additive for animals.

Keywords
Xylanase; Aspergillus niger; Pichia pastoris; In vitro digestibility; Productivity; Fed-batch
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Cloning, expression, characterization, and high cell-density production of recombinant endo-1,4-β-xylanase from Aspergillus niger in Pichia pastoris
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Publisher
Database: Elsevier - ScienceDirect
Journal: Enzyme and Microbial Technology - Volume 41, Issues 1–2, 2 July 2007, Pages 19–25
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