产酸克雷伯氏菌(K.oxytoca) α-乙酰乳酸脱羧酶基因(BudA)的克隆及生物信息学分析

Cloning and bioinformatical analysis of α-acetolactate decarboxylase gene( BudA) in Klebsiella oxytoca

以Klebsiella oxytoca(K.oxytoca)HD79为出发菌株,根据α-乙酰乳酸脱羧酶基因序列相似的特点,利用Primer5.0设计一对引物,以K.oxytoca HD79为模板,PCR扩增,获得K.oxytoca HD79基因片段Bud A,此片段经测序显示长度为780 bp,无碱基缺失。利用生物信息学软件对该序列进行了同源性分析、氨基酸组成分析、疏水性分析、磷酸化位点预测、亚细胞定位分析及二、三级结构预测。结果表明,该片段为K.oxytoca HD79的α-乙酰乳酸脱羧酶基因序列。对该酶进行的生物信息学特异性分析为其构建产2,3-丁二醇的工程菌株提供一定的理论参考。

BudA gene from Klebsiella oxytoca （K. oxytoca） HD79 was cloned with PCR and the genome of K. oxytoca HD79 was acted as the template. The primers of BudA sequence were designed according to the sequence similarity of the sequence by using Primer 5.0 software. The fragment was sequenced with the length of 780 bp and without base deletion. Furthermore, the homology, amino acid composition, hy- drophobicity, phosphorylation site calculation, sub cellar localization, and secondary and tertiary struc- ture prediction of the sequence were analyzed by using the bioinformatics software. The results showed that the fragment was α-acetolactate decarboxylase gene from K. oxytoca HD79. Thus, bioinformatics spe- cific analysis of this enzyme provided a referential value and laid a solid foundation of construction of 2, 3 -BD high-producing engineering strains.