C-Terminal proline-rich sequence broadens the optimal temperature and pH ranges of recombinant xylanase from Geobacillus thermodenitrificans C5 |
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| Institution: | 1. Department of Microbiology, Faculty of Biological Sciences, Quaid-I-Azam University, Islamabad, Pakistan;2. Department of Molecular Biology and Genetic, Faculty of Sciences, Karadeniz Technical University, 61080 Trabzon, Turkey;3. Department of Biology, Faculty of Sciences, Karadeniz Technical University, 61080 Trabzon, Turkey;1. Industrial Enzymes National Engineering Laboratory, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, China;2. Institute of Biological Chemistry, Academia Sinica, Taipei 11529, Taiwan;3. Key Laboratory for Feed Biotechnology of the Ministry of Agriculture, Feed Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, China;4. College of Biotechnology, Tianjin University of Science and Technology, Tianjin 300457, China;5. School of Biotechnology, Jiangnan University, Wuxi 214122, China;1. Department of Bioprocess and Polymer Engineering, Faculty of Chemical and Energy Engineering, Universiti Teknologi Malaysia, 81310, Skudai, Johor, Malaysia;2. Malaysia Genome Institute, Jalan Bangi Lama, 43000, Bangi, Selangor, Malaysia;1. Beijing Advanced Innovation Center for Food Nutrition and Human Health, Beijing Technology and Business University (BTBU), Beijing 100048, China;2. School of Food and Chemical Engineering, Beijing Technology and Business University, No.33, Fucheng Road, Beijing 100048, China;3. Beijing Engineering and Technology Research Center of Food Additives, Beijing Technology and Business University (BTBU), Beijing 100048, China;4. Department of Biology, Emory University, 1510 Clifton Road, Atlanta, GA 30322, USA;1. Key Laboratory of Recycling and Eco-Treatment of Waste Biomass of Zhejiang Province, Zhejiang University of Science and Technology, Hangzhou, 310023, PR China;2. School of Food and Biological Engineering, Jiangsu University, Zhenjiang 212013, PR China;1. Federal State Institution “Federal Research Centre “Fundamentals of Biotechnology” of the Russian Academy of Sciences”, Leninsky Pr. 33, Moscow 119071, Russia;2. Department of Chemistry, M.V. Lomonosov Moscow State University, Vorobyovy Gory 1/11, Moscow 119991, Russia;3. G.K. Skryabin Institute of Biochemistry and Physiology of Microorganisms, Russian Academy of Sciences, Pushchino 142292, Moscow region, Russia;1. Department of Chemical and Biological Engineering, South Dakota School of Mines and Technology, Rapid City, SD 57701, USA;2. Renewable Product Technology Research Unit, Agricultural Research Service, National Center for Agricultural Utilization Research, U.S. Department of Agriculture, Peoria, IL 61604, USA;3. Department of Chemical Engineering and Materials Science, DOE Great Lakes Bioenergy Research Center, Michigan State University, Lansing, MI 48823, USA |
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| Abstract: | Efficient utilization of hemicellulose entails high catalytic capacity containing xylanases. In this study, proline rich sequence was fused together with a C-terminal of xylanase gene from Geobacillus thermodenitrificans C5 and designated as GthC5ProXyl. Both GthC5Xyl and GthC5ProXyl were expressed in Escherichia coli BL21 host in order to determine effect of this modification. The C-terminal oligopeptide had noteworthy effects and instantaneously extended the optimal temperature and pH ranges and progressed the specific activity of GthC5Xyl. Compared with GthC5Xyl, GthC5ProXyl revealed improved specific activity, a higher temperature (70 °C versus 60 °C) and pH (8 versus 6) optimum, with broad ranges of temperature and pH (60–80 °C and 6.0–9.0 versus 40–60 °C and 5.0–8.0, respectively). The modified enzyme retained more than 80% activity after incubating in xylan for 3 h at 80 °C as compared to wild ?type with only 45% residual activity. Our study demonstrated that proper introduction of proline residues on C-terminal surface of xylanase family might be very effective in improvement of enzyme thermostability. Moreover, this study reveals an engineering strategy to improve the catalytic performance of enzymes. |
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| Keywords: | Xylanase Proline tagged xylanase (GthC5ProXyl) Characterization Thermostability |
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