Cloning and expression of GH11 xylanase gene from Aspergillus fumigatus MKU1 in Pichia pastoris(ENZYMOLOGY, PROTEIN ENGINEERING, AND ENZYME TECHNOLOGY)

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A xylanase gene, xynf11a of Aspergillus fumigatus MKU1 was cloned and expressed in Pichia pastoris X33. Two exons of the xynf11a gene were amplified separately and fused by overlap extension PCR. The fused product was cloned in yeast expression vector pPICZB and expressed in P. pastoris under the control of the AOX1 promoter. P. pastoris transformants expressing recombinant xylanases were selected on xylan agar plate and their ability to produce the xylanase was evaluated in flask cultures. P. pastoris X33 (pZBxynf11aFP) efficiently secreted the recombinant xylanase into the medium and produced the high level of xylanase activity (14U/ml) after 96h of growth. The recombinant xylanase produced by P. pastoris showed maximum activity at pH6.0 and temperature 60℃. The recombinant xylanase did not exhibit any cellulase activity and hence it could be potentially used for pretreatment of paper pulp before bleaching.
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