Engineering a high-performance,metagenomic-derived novel xylanase with improved soluble protein yield and thermostability |
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| Affiliation: | 1. Beijing Advanced Innovation Center for Food Nutrition and Human Health, Beijing Technology and Business University, Beijing 100048, China;2. School of Food and Chemical Engineering, Beijing Technology and Business University, No.11, Fucheng Road, Beijing 100048, China;3. Beijing Engineering and Technology Research Center of Food Additives, Beijing Technology and Business University, Beijing 100048, China;4. School of Foreign Language, Beijing Technology and Business University, No.33, Fucheng Road, Beijing 100048, China;1. Department of Systems Biology, Agricultural Biotechnology Research Institute of Iran (ABRII), Agricultural Research Education and Extension Organization (AREO), Karaj, Iran;2. Institute of Biochemistry and Biophysics (IBB), University of Tehran, Tehran, Iran |
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| Abstract: | The novel termite gut metagenomic-derived GH11 xylanase gene xyl7 was expressed in Escherichia coli BL21, and the purified XYL7 enzyme exhibited high specific activity (6340 U/mg) and broad pH active range of 5.5–10.0. Directed evolution was employed to enhance the thermostability of XYL7; two mutants (XYL7-TC and XYL7-TS) showed a 250-fold increase in half-life at 55 °C, with a 10 °C increase in optimal temperature compared to that of wild-type XYL7. A truncated enzyme (XYL7-Tr3) acquired by protein engineering showed similar catalytic properties as the wild-type, with a tenfold increase in soluble protein yield by the mutant. The reducing sugar produced by XYL7-TC was about fourfold greater than that produced by their parents when incubated with xylan at 60 °C for 4 h. The engineered novel xylanase exhibited superior enzymatic performance and showed promise as an excellent candidate for industrial application due to its high specific activity, stability and soluble protein yield. |
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| Keywords: | Directed molecular evolution GH11 xylanase Metagenomic Protein engineering Thermostability |
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