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首页> 外文期刊>African Journal of Biotechnology >Engineering thermostable xylanase enzyme mutant from Bacillus halodurans
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Engineering thermostable xylanase enzyme mutant from Bacillus halodurans

机译：嗜盐芽孢杆菌的工程热稳定木聚糖酶突变体

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Endo-1,4-beta-xylanase is the main enzyme in xylan-backbone hydrolysis and has received attractable research interest due to its significant application in various industrial processes such as food, feed, waste treatment, fuel and chemical production, paper and pulp industries; but these applications require thermostable xylanase enzymes. Error-prone polymerase chain reaction (PCR) and site-directed mutagenesis were used to engineer new thermostable mutants of?Bacillus halodurans. The results showed that the rate of mutagenesis in the error-prone PCR was at least 1%, resulting in more than 1000 mutated colonies. About one third of mutants lost the enzymes’ activities and no sign of thermal stability improvement was observed in other mutants; less than 2% of mutants were active at temperature higher than 75°C and they lost their activities quickly at temperature higher than 80°C. In site-directed mutagenesis at position 274,?threonine?(T) amino acid changed to proline (P) but the extent of thermostability improvement in newly engineered mutants were not sufficient. Various screening, clustering, decision tree and generalized rule induction models used to search for patterns of thermostability and the frequency of glutamine was the most important feature in many bioinformatics models. The importance of these methods has been discussed here.
机译：Endo-1,4-β-木聚糖酶是木聚糖主链水解中的主要酶，由于其在食品，饲料，废物处理，燃料和化工生产，造纸和纸浆等各种工业过程中的重要应用而受到了广泛的研究兴趣。行业;但是这些应用需要热稳定的木聚糖酶。易错的聚合酶链反应（PCR）和定点诱变被用来设计新的嗜盐芽孢杆菌的热稳定突变体。结果表明，易错PCR的诱变率至少为1％，导致超过1000个突变菌落。约有三分之一的突变体丧失了酶的活性，其他突变体均未观察到热稳定性改善的迹象。不到2％的突变体在高于75°C的温度下具有活性，并且在高于80°C的温度下迅速丧失活性。在位点274的定点诱变中，苏氨酸α（T）氨基酸变为脯氨酸（P），但是在新改造的突变体中热稳定性提高的程度不足。用于搜索热稳定性和谷氨酰胺频率的各种筛选，聚类，决策树和广义规则归纳模型是许多生物信息学模型中最重要的特征。这些方法的重要性已在此处讨论。

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