Chen Fei, Chen Xiu-Zhen, Qin Li-Na, Tao Yong, Dong Zhi-Yang
State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, No. 1 West Beichen Road, Chaoyang District, Beijing 100101, China; University of Chinese Academy of Sciences, Beijing 100049, China.
State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, No. 1 West Beichen Road, Chaoyang District, Beijing 100101, China.
Biochem Biophys Res Commun. 2015 Apr 3;459(2):184-188. doi: 10.1016/j.bbrc.2014.12.066. Epub 2014 Dec 19.
Trichoderma reesei is thought to be a promising recombinant host for the production and secretion of complex proteins due to its ability to secrete large amounts of proteins. In this study we identified a functional N-acetyl-β-glucosaminidase (NAGase) gene Nag1 in T. reesei. Nag1, a putative gene encoding a GH 20 family NAGase in T. reesei, was cloned and homologous overexpressed in the T. reesei RutC30ΔU3 with a strong cellobiohydrolase1 gene (cbh1) promoter. Nag1 was secreted in its active form and the highest expression level was around 499.85IU/ml. Nag1 has a molecular mass of 80kDa. The optimum pH and temperature were 4.0 and 60°C, respectively.
由于里氏木霉具有分泌大量蛋白质的能力，它被认为是生产和分泌复杂蛋白质的一种很有前景的重组宿主。在本研究中，我们在里氏木霉中鉴定出一个功能性的N-乙酰-β-氨基葡萄糖苷酶（NAGase）基因Nag1。Nag1是里氏木霉中一个推定的编码GH 20家族NAGase的基因，它被克隆并在具有强纤维二糖水解酶1基因（cbh1）启动子的里氏木霉RutC30ΔU3中进行同源过表达。Nag1以其活性形式分泌，最高表达水平约为499.85IU/ml。Nag1的分子量为80kDa。最适pH和温度分别为4.0和60°C。
