Codon-optimized expression and characterization of a pH stable fungal xylanase in Pichia pastoris |
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| Affiliation: | 1. Oil Crops Research Institute, Chinese Academy of Agricultural Sciences, Hubei Key Laboratory of Lipid Chemistry and Nutrition, Key laboratory of Biology and Genetic Improvement of Oil Crops, Ministry of Agriculture, Wuhan 430062, China;2. Functional Oil Laboratory Associated by Oil Crops Research Institute, Chinese Academy of Agricultural Sciences and Infinite (China) Co. LTD, Guangzhou 51000, China;1. School of Industrial Technology, Universiti Sains Malaysia, Malaysia;2. Department of Microbiology, Faculty of Life Sciences, University of Ilorin, Nigeria;1. Department of Biotechnology, K.S. Rangasamy College of Technology, Tiruchengode, 637 215 Namakkal District, Tamil Nadu, India;2. Department of Biotechnology, Faculty of Engineering, Karpagam Academy of Higher Education, Coimbatore 641 021, Tamil Nadu, India;3. Department of Botany, Government Arts College (Autonomous), Salem 636 007, Tamil Nadu, India;4. Department of Biological Sciences, School of Mathematics and Natural Sciences, Copperbelt University, Riverside, Jambo Drive, P O Box 21692, Kitwe, Zambia;1. School of Light Industry and Food Engineering, Guangxi University, 100 Daxue Road, Nanning, 530004, Guangxi, China;2. Sugar Industry Collaborative Innovation Center, Guangxi University, 100 Daxue Road, Nanning, 530004, Guangxi, China |
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| Abstract: | Novel xylanase (EC 3.2.1.8) is in great demand due to its industrial significance. In this study, we have developed and characterized a novel xylanase-producing yeast strain. This mature xylanase gene xyn11A consists of 870 base pairs and belongs to GH11 family. The gene sequence was optimized and synthesized, and was then cloned into yeast vector pGAPZαA under the control of the constitutive GAP promoter. SDS-PAGE analysis indicates that Xyn11A is extracellularly expressed as a glycosylated protein in P. pastoris. Xyn11A is optimally active at 70 °C and pH 7.4. This xylanase retained more than 90% of its activity after incubation at 50 °C and 60 °C for up to 1 h. Xyn11A is also stable over a wide range of pH (2.0–11.0). Most metal ions tested such as copper (Cu2+) and lead (Pb2+) have little inhibitory effects on Xyn11A. It is also resistant to pepsin and proteinase K digestion, retaining 80% and 90% of its activity after digestion at 37 °C for 1 h, respectively. Those superior properties make Xyn11A a robust xylanase with great potential for industrial use. To the best of our knowledge, this is the first report of xylanase from the fungus Corynascus thermophilus. |
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| Keywords: | Fungal xylanase Codon optimization High cell density fermentation |
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