Publication: 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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Issued Date
2007-07-02
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01410229
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2-s2.0-34248385655
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Mahidol University
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SCOPUS
Bibliographic Citation
Enzyme and Microbial Technology. Vol.41, No.1-2 (2007), 19-25
Suggested Citation
Vasimon Ruanglek, Rutchadaporn Sriprang, Nakul Ratanaphan, Pacawadee Tirawongsaroj, Duriya Chantasigh, Sutipa Tanapongpipat, Kusol Pootanakit, Lily Eurwilaichitr Cloning, expression, characterization, and high cell-density production of recombinant endo-1,4-β-xylanase from Aspergillus niger in Pichia pastoris. Enzyme and Microbial Technology. Vol.41, No.1-2 (2007), 19-25. doi:10.1016/j.enzmictec.2006.11.019 Retrieved from: https://repository.li.mahidol.ac.th/handle/123456789/24165
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Title
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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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% μmaxexponential 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. © 2006 Elsevier Inc. All rights reserved.