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| Expression of Thermomyces lanuginosus Xylanase 1YNA and Its Disulphide Bridge Mutant in Pichia Pastoris |
| LI Si-jia1, WANG Ya-wei1, FU Zheng1, WANG Wen-jun1, Ossi Turunen2, XIONG Hai-rong 1 |
1. Engineering Research Centre of Bioresource in Southern China, College of Life Science, South-central University for Nationalities, Wuhan 430074, China;
2. Aalto University, School of Chemical Technology, Department of Biotechnology and Chemical Technology, FI-00076 Aalto, Finland |
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Abstract The wild type xylanase gene (1yna) and its disulfide bridge mutant gene were linked to vectors pPIC9, and then transformed into Pichia pastoris GS115 to construct the recombinant strains for expression the wild type xylanase (TLX) and its disulfide bridge mutant (DSB), respectively. The recombinants express TLX and DSB at high level, with electrophoresis pure protein products. In shake flask cultivation, the protein expression levels by P. pastoris were 1.75 mg/ml (TLX) and 1.82 mg/ml (DSB), respectively. The enzymatic properties showed that DSB performed the optimum temperature at 75℃, and the TLX performed the optimum temperature at 68℃. DSB showed a better thermostabllity than TLX. After 30 min inactivation at 75℃, DSB still remained 50% of the residual activity, whereas TLX only remained less than 20% of its activity.
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Received: 28 November 2012
Published: 25 March 2013
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