Whitehead T R, Hespell R B
Northern Regional Research Center, Department of Agriculture, Peoria, Illinois 61604.
J Bacteriol. 1990 May;172(5):2408-12. doi: 10.1128/jb.172.5.2408-2412.1990.
Genes coding for three xylan-degrading activities, xylanase, xylosidase, and arabinosidase, were simultaneously cloned from the colonic anaerobic organism Bacteriodes ovatus. The genes for the three enzymes were located on a 3.8-kilobase EcoRI genomic insert and were cloned by using plasmid pUC18. All three activities were expressed in Escherichia coli JM83, and all were cell associated. Expression of the xylanase gene was independent from expression of the xylosidase and arabinosidase genes, whereas expression of the latter two genes appeared to be coordinated. Restriction endonuclease analysis of the arabinosidase and xylosidase genes and partial purification of these enzyme activities from E. coli suggested that these activities were catalyzed by a bifunctional protein or two proteins of very similar molecular weight. All three enzyme activities were regulated in B. ovatus in response to the carbon source used for growth. This is the first report of the cloning and expression of B. ovatus genes.
从结肠厌氧生物卵形拟杆菌中同时克隆出编码三种木聚糖降解活性的基因,即木聚糖酶、木糖苷酶和阿拉伯糖苷酶。这三种酶的基因位于一个3.8千碱基的EcoRI基因组插入片段上,并使用质粒pUC18进行克隆。所有这三种活性在大肠杆菌JM83中均有表达,且都与细胞相关。木聚糖酶基因的表达独立于木糖苷酶和阿拉伯糖苷酶基因的表达,而后两者基因的表达似乎是协同的。对阿拉伯糖苷酶和木糖苷酶基因进行限制性内切酶分析,并从大肠杆菌中对这些酶活性进行部分纯化,结果表明这些活性是由一种双功能蛋白或两种分子量非常相似的蛋白催化的。卵形拟杆菌中的所有这三种酶活性均根据用于生长的碳源进行调节。这是关于卵形拟杆菌基因克隆和表达的首次报道。