产碱性纤维素酶芽孢杆菌ZA-05的分离及功能基因克隆

Isolation and Functional Gene Cloning of Alkaline Glucanase Producing Bacillus Strain ZA-05

从造纸污泥中分离到一株芽孢杆菌ZA-05,该菌可产生内切β-1,4-葡聚糖酶酶活性,且在碱性条件下酶活较高,经生理生化和分子生物学鉴定,该菌株为淀粉液化芽孢杆菌(Bacillus amyloliquefaciens);根据GenBank登录的内切-β-1,4-葡聚糖酶基因(DQ782954.1,M28332.1,AY859492.1)的同源性序列,利用PCR方法克隆到该菌株的功能基因,并对其进行测序。测序结果显示其全长为1 500 bp,推测其含499个氨基酸。

An alkaline glucanase producing strain ZA-05 which had the enzyme activity of endo-β-1,4-glucanase was isolated from paper mill sludge by Congo-red staining method.It was identified as Bacillus amyloliquefaciens based on its morphological characters and 16S rDNA sequence.According to the homologous sequence of endo-β-1,4-glucanase gene（GenBank No.DQ782954.1,M28332.1,AY859492.1）,an alkali-tolerant glucanase gene was amplified by PCR and then sequenced.The full length of this alkali-tolerant glucanase gene was 1 500 bp and presumably encoded 499 amino acids.