Miettinen-Oinonen Arja, Suominen Pirkko
Primalco Ltd. Biotec, Rajamäki, Finland.
Appl Environ Microbiol. 2002 Aug;68(8):3956-64. doi: 10.1128/AEM.68.8.3956-3964.2002.
Trichoderma reesei strains were constructed for production of elevated amounts of endoglucanase II (EGII) with or without cellobiohydrolase I (CBHI). The endoglucanase activity produced by the EGII transformants correlated with the copy number of the egl2 expression cassette. One copy of the egl2 expression cassette in which the egl2 was under the cbh1 promoter increased production of endoglucanase activity 2.3-fold, and two copies increased production about 3-fold above that of the parent strain. When the enzyme with elevated EGII content was used, an improved stonewashing effect on denim fabric was achieved. A T. reesei strain producing high amounts of EGI and -II activities without CBHI and -II was constructed by replacing the cbh2 locus with the coding region of the egl2 gene in the EGI-overproducing CBHI-negative strain. Production of endoglucanase activity by the EG-transformant strain was increased fourfold above that of the host strain. The filter paper-degrading activity of the endoglucanase-overproducing strain was lowered to below detection, presumably because of the lack of cellobiohydrolases.
构建了里氏木霉菌株,用于在有或没有纤维二糖水解酶I(CBHI)的情况下生产更多的内切葡聚糖酶II(EGII)。EGII转化体产生的内切葡聚糖酶活性与egl2表达盒的拷贝数相关。在cbh1启动子控制下的一个egl2表达盒拷贝使内切葡聚糖酶活性的产量提高了2.3倍,两个拷贝使产量比亲本菌株提高了约3倍。当使用EGII含量升高的酶时,牛仔布的石洗效果得到改善。通过在过量产生EGI的CBHI阴性菌株中用egl2基因的编码区替换cbh2基因座,构建了一种不产生CBHI和CBII但产生大量EGI和-II活性的里氏木霉菌株。EG转化体菌株产生的内切葡聚糖酶活性比宿主菌株提高了四倍。过量产生内切葡聚糖酶的菌株的滤纸降解活性降低到检测限以下,可能是因为缺乏纤维二糖水解酶。