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首页> 外文期刊>International Journal of Biological Macromolecules: Structure, Function and Interactions >Efficient xylan-to-sugar biotransformation using an engineered xylanase In hyperthermic environment
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Efficient xylan-to-sugar biotransformation using an engineered xylanase In hyperthermic environment

机译：高温环境中使用工程木聚糖酶高效的木聚糖生物转化

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Hyperthermophilic xylanases play a critical role in bioconversion from xylan to sugar in the process of bio-mass utilization. The discoveiy of new or improvement of existing xylanases based on directed evolution is expected to be an effective approach to meet the increasing demand of thermostable xylanases. In this work, a xylanase B gene (CTN_0623) from Thermotoga neapolitana (Tire) was cloned and xylanase B (herein named TnexlnB) was solubly expressed in E. coli with a high amount using a beat shock vector pHsb. TnexlnB showed the highest endo-beta-1,4-xylan hydrolase activity at 75 degrees C and pH 6.0. Additionally, 1 mM Mg2+, Ba2+ and Ca2+ improved the activity of TnexlnB by 31%, 37%, and 53%, respectively. The optimal temperature reached 85 degrees C by site-directed mutation at the last three helices of TnexlnB. K-m and V-max towards birchwood xylan were determined for both wide type and the best mutant, as follow: 1.09 mg/mL, 191.76 U/mg and 0.29 mg/mL, 376.42 U/mg, respectively. Further characterization highlighted good thermal stability (>80% of enzymatic activity after 1 h at 90 degrees C) for the best mutant, which made this enzyme suitable for biomass degradation at high temperature. (C) 2020 Elsevier B.V. All rights reserved.
机译：高嗜热的木聚糖酶在生物质量利用过程中，在含木聚糖中的生物转化中发挥着致力作用。基于定向演化的现有木聚糖酶的新的或改善的动态或改善是一种有效的方法，以满足恒温木聚糖酶的不断增加的需求。在这项工作中，克隆了来自Thermotoga Neakitana（轮胎）的木聚糖酶B基因（CTN_0623），并且使用拍摄休克载体PHSB，用大量在大肠杆菌中用大量表达木聚糖酶B（本文命名为TNEXLNB）。 TNEXLNB在75℃和pH6.0中显示出最高的腹腹剂-1,4-木聚糖水解酶活性。另外，1mM Mg2 +，Ba2 +和Ca2 +改善TNEXLNB的活性分别为31％，37％和53％。在TNEXLNB的最后三个螺旋中，最佳温度达到85摄氏度达到85℃。对于宽型和最佳突变体测定K-M和V-MAX朝向Birchwood木聚糖，如下：1.09mg / ml，191.76u / mg和0.29mg / ml，376.42 U / mg。对于最佳突变体，进一步表征突出了良好的热稳定性（在90℃下在90℃下酶活性的80％），这使得该酶适用于高温下的生物质降解。 （c）2020 Elsevier B.v.保留所有权利。

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