Department of Biochemistry, Kangwon National University, Chunchon 200-701, Republic of Korea.
J Microbiol. 2010 Oct;48(5):706-8. doi: 10.1007/s12275-010-0272-x. Epub 2010 Nov 3.
Coprinellus congregatus generates several chitinases during its entire life cycle: at the growing hyphal stage and at the mushroom autolysis stage. We have isolated a chitinase gene (chi1) from the mushroom tissue at the autolysing stage, and constructed a chitinase expression vector to get large amount of enzyme protein. Chitinase 1 (chi1) cDNA was heterologously expressed in Saccharomyces cerevisiae by gal1 promoter. The transformants showed no specific change in growth characteristics under normal growth conditions. However the expression of the gene by the gal1 promoter in the yeast transformants resulted in complete growth inhibition, while laccase expression by the gal1 promoter showed normal growth. The chitinase activities from the transformants were also more than 3 times higher than that of the recipient strain, and the chitinase expression by the real time-PCR also showed increased expression of the chi1 in the yeast transformant. Expression of a chitinase which was produced at the mushroom autolysing stage of C. congregatus resulted in yeast growth inhibition.
在生长菌丝阶段和蘑菇自溶阶段。我们已经从自溶阶段的蘑菇组织中分离出一种几丁质酶基因（chi1），并构建了几丁质酶表达载体以获得大量酶蛋白。几丁质酶 1（chi1）cDNA 通过 gal1 启动子在酿酒酵母中异源表达。在正常生长条件下，转化体的生长特性没有特定变化。然而，gal1 启动子在酵母转化体中表达该基因导致完全生长抑制，而漆酶表达则表现出正常生长。转化体的几丁质酶活性也比受体菌株高出 3 倍以上，实时 PCR 也显示出酵母转化体中 chi1 的表达增加。丛生侧耳蘑菇自溶阶段产生的几丁质酶的表达导致酵母生长抑制。
