Yang Fan, Gong Yanfen, Liu Gang, Zhao Shengming, Wang Juan
Shenzhen Key Laboratory of Microbial Genetic Engineering, College of Life Sciences, Shenzhen University, Shenzhen, Guangdong 518060, P.R. China.
Shenzhen Key Laboratory of Marine Bioresources and Ecology, Shenzhen, Guangdong 518060, P.R. China.
J Microbiol Biotechnol. 2015 Jul;25(7):1101-7. doi: 10.4014/jmb.1501.01049.
The role of CRE1 in a thermophilic fungus, Myceliophthora thermophila ATCC42464, was studied using RNA interference. In the cre1-silenced strain C88, the filter paper hydrolyzing activity and β-1,4-endoglucanase activity were 3.76-, and 1.31-fold higher, respectively, than those in the parental strain when the strains were cultured in inducing medium for 6 days. The activities of β-1,4-exoglucanase and cellobiase were 2.64-, and 5.59-fold higher, respectively, than those in the parental strain when the strains were cultured for 5 days. Quantitative reverse-transcription polymerase chain reaction showed that the gene expression of egl3, cbh1, and cbh2 was significantly increased in transformant C88 compared with the wild-type strain. Therefore, our findings suggest the feasibility of improving cellulase production by modifying the regulator expression, and an attractive approach to increasing the total cellulase productivity in thermophilic fungi.
利用RNA干扰技术研究了嗜热真菌嗜热毁丝霉ATCC42464中CRE1的作用。在cre1基因沉默菌株C88中,当菌株在诱导培养基中培养6天时,滤纸水解活性和β-1,4-内切葡聚糖酶活性分别比亲本菌株高3.76倍和1.31倍。当菌株培养5天时,β-1,4-外切葡聚糖酶和纤维二糖酶的活性分别比亲本菌株高2.64倍和5.59倍。定量逆转录聚合酶链反应表明,与野生型菌株相比,转化体C88中egl3、cbh1和cbh2的基因表达显著增加。因此,我们的研究结果表明通过修饰调节因子表达来提高纤维素酶产量是可行的,这是提高嗜热真菌纤维素酶总生产力的一种有吸引力的方法。