嗜碱嗜盐芽孢杆菌内切木聚糖酶的序列分析、克隆及过表达

Sequence analysis, cloning and over-expression of an endoxylanase from the alkaliphilic Bacillus halodurans.

作者信息

Martínez M Alejandra, Delgado Osvaldo D, Baigorí Mario D, Siñeriz Faustino

机构信息

PROIMI, Planta Piloto de Procesos Industriales Microbiológicos, Av. Belgrano y Pasaje Caseros, T-400 1MVB, Tucumán, Argentina.

出版信息

Biotechnol Lett. 2005 Apr;27(8):545-50. doi: 10.1007/s10529-005-2879-2.

DOI:10.1007/s10529-005-2879-2

PMID:15973487

Abstract

The BhMIR32 xyn11A gene, encoding an extracellular endoxylanase of potential interest in bio-bleaching applications, was amplified from Bacillus halodurans MIR32 genomic DNA. The protein encoded is an endo-1,4-beta-xylanase belonging to family 11 of glycosyl hydrolases. Its nucleotide sequence was analysed and the mature peptide was subcloned into pET22b(+) expression vector. The enzyme was over-expressed in a high density Escherichia coli culture as a soluble and active protein, and purified in a single step by immobilised metal ion affinity chromatography with a specific activity of 3073 IU mg-1.

摘要

从嗜碱芽孢杆菌MIR32基因组DNA中扩增出BhMIR32 xyn11A基因，该基因编码一种在生物漂白应用中具有潜在价值的细胞外内切木聚糖酶。所编码的蛋白质是一种属于糖基水解酶家族11的内切-1,4-β-木聚糖酶。分析了其核苷酸序列，并将成熟肽亚克隆到pET22b(+)表达载体中。该酶在高密度大肠杆菌培养物中作为可溶性活性蛋白过量表达，并通过固定化金属离子亲和色谱一步纯化，比活性为3073 IU mg-1。