摘要
以报道的黑曲霉耐酸性α-淀粉酶基因(ANasamy)为基础设计两条引物,通过PCR等分子生物学手段克隆了一段全长为1975 bp的ANasamy基因,构建了关于ANasamy的表达质粒pPTH-ANasamy,并在黑曲霉菌株中获得表达。分离、纯化重组耐酸性α-淀粉酶并对该酶的基本酶学性质进行研究,该酶蛋白质分子量约为70 kDa,在65℃及pH 4.5条件下催化活力最高,具有较高的热稳定性及低pH耐受性。Cu^(2+)和Fe^(3+)对该淀粉酶活力有较强的抑制作用。
An acid-stable a-amylase gene (ANasamy) of Aspergillus niger was cloned, sequenced and expressed by a series of molecular method. An expressing plasmid pPTH-ANasamy carrying the ANasamy was constructed and expressed in A. niger. The molecular weight of the purified α-amylase was about 70 kDa. Enzyme characterization revealed that the a-amylase had an optimal temperature of 65℃ and an optimal pH of 4.5, and had prominent temperature stability and low pH stability. The purified ANasamy was strongly inhibited by Cu^2+ and Fe^3+ .
出处
《工业微生物》
CAS
CSCD
2010年第4期63-68,共6页
Industrial Microbiology
基金
国家高技术研究发展计划(863项目
2006AA020204)
关键词
黑曲霉
耐酸性廿淀粉酶
表达载体
酶学性质
AspergiUus niger
acid-stable a-amylase
expression vector
enzyme characterization