Lee Jae Hyung, Lim Myung-Ye, Kim Mi-Jin, Heo Sun-Yeon, Seo Jin-Ho, Kim Yeon-Hee, Nam Soo-Wan
Department of Biomaterial Control (BK21 Program), Dong-Eui University, Busan 614-714, Korea.
J Microbiol Biotechnol. 2007 Dec;17(12):2076-80.
The endoxylanase (GenBank Access No. U51675) of Bacillus spp. and endoglucanase (GenBank Access No. AY466436) of Trichoderma spp. were separately inserted downstream of the yeast constitutive ADH1 promoter, resulting in three different plasmids (pAGX1, pAGX2, and pAGX3) according to the transcription direction of two genes. When the yeast transformants, S. cerevisiae SEY2102 harboring each expression plasmid, were grown on YPD medium, the total activities of the enzymes were approximately 3.01 unit/ml, 3.24 unit/ml, and 7.56 unit/ml for endoxylanase and 0.60 unit/ml, 0.54 unit/ml, and 0.39 unit/ ml for endoglucanase, in the following order: the pAGX1, pAGX2, and pAGX3. More than 70% of the endoxylanase and endoglucanase activities was found in the extracellular media.
将芽孢杆菌属的内切木聚糖酶(GenBank登录号:U51675)和木霉属的内切葡聚糖酶(GenBank登录号:AY466436)分别插入酵母组成型ADH1启动子的下游,根据两个基因的转录方向,构建了三种不同的质粒(pAGX1、pAGX2和pAGX3)。当携带每种表达质粒的酵母转化子酿酒酵母SEY2102在YPD培养基上生长时,内切木聚糖酶的总活性分别约为3.01单位/毫升、3.24单位/毫升和7.56单位/毫升,内切葡聚糖酶的总活性分别约为0.60单位/毫升、0.54单位/毫升和0.39单位/毫升,顺序如下:pAGX1、pAGX2和pAGX3。超过70%的内切木聚糖酶和内切葡聚糖酶活性存在于细胞外培养基中。