Improved Expression and Characterization of a Multidomain Xylanase from Thermoanaerobacterium aotearoense SCUT27 in Bacillus subtills
Improved Expression and Characterization of a Multidomain Xylanase from Thermoanaerobacterium aotearoense SCUT27 in Bacillus subtills
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DOI：
10.1021/acs.jafc.5b01259
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发表时间：
2015-07-22
影响因子：
6.1
通讯作者：
Li, Shuang
中科院分区：
文献类型：
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作者：
Huang, Xiongliang;Li, Zhe;Li, Shuang
从Thermoanerobacter aotearoense SCOT27中克隆并鉴定了木聚糖酶基因，该基因被证明由信号肽、一个糖苷水解酶家族10结构域、四个碳水化合物结合模块和三个表面层同源结构域组成。将表达宿主从大肠杆菌改为枯草芽孢杆菌导致截短的XynA Delta SLH的比活性增加4.1倍。枯草芽孢杆菌中五种不同版本的分泌信号表明它优选通过 Sec 依赖性途径传递。纯化的 XynA Delta SLH 对山毛榉木聚糖显示出 37.8 U/mg 的高活性。 XynA Delta SLH 在 80 摄氏度、pH 6.5 时具有最佳活性。薄层色谱结果表明，木二糖和推测的甲基葡萄糖醛酸木三糖(MeGlcAXyl(3))是重组木聚糖酶催化山毛榉木聚糖释放的主要产物。所有结果表明，XynA Delta SLH 是从纤维素材料生产工业用低聚木糖的合适候选者。
A xylanase gene was cloned and characterized from, Thermoanerobacterium aotearoense SCOT27, which was attested to consist of a signal peptide, one glycoside hydrolase family 10 domain, four carbohydrate binding modules, and three surface layer homology domains. The change of expression host from Escherichia coli to Bacillus subtilis resulted hi a 4.1, fold increase of specific activity for the truncated XynA Delta SLH. Five different versions of secretion signals in B. subtilis indicated that it was preferably routed via a Sec dependent pathway. Purified XynA Delta SLH showed a high activity of 37.8 U/mg on beechwood xylan. XynA Delta SLH was optimally active at 80 degrees C, pH 6.5. Thin layer chromatography results showed that) xylobiose and the presumed methylglucuronoxylotriose (MeGlcAXyl(3)) were the main products liberated-from beechwood xylan catalyzed by the recombinant xylanase. All of the results suggest that XynA Delta SLH is a suitable candidate for generating xylooligosaccharides from cellulosic material's for industrial uses.