Li Ming, Zhou Liying, Liu Meng, Huang Yunyan, Sun Xin, Lu Fuping
Key Laboratory of Industrial Fermentation Microbiology, Education Ministry of China, College of Biotechnology, Tianjin University of Science and Technology.
Biosci Biotechnol Biochem. 2013;77(9):1860-6. doi: 10.1271/bbb.130281. Epub 2013 Sep 7.
In this study, Agrobacterium tumefaciens-mediated transformation (ATMT) was used in breeding industrial strains for the purpose of improving α-transglucosidase production. Firstly, an efficient ATMT system for Asperillus niger was established by optimization of several influencing factors, in which transformation efficiency was improved up to 14-fold compared with the initial conditions. Furthermore, binary vector pBI-Glu containing an α-transglucosidase expression cassette was constructed and transferred into Agrobacterium tumefaciens LBA4404 in order to infect A. niger. By the efficient ATMT method, the gene for α-transglucosidase, driven by strong promoter PglaA (the glucoamylase gene promoter), had a high expression level in A. niger A-8 (25.02 U/mL). The optimized ATMT system was found to be effective and suitable for A. niger, and should be a useful tool for studying the function of A. niger genes and for industrial breeding of this strain.
在本研究中，为提高α-转葡萄糖苷酶的产量，采用根癌农杆菌介导的转化（ATMT）方法培育工业菌株。首先，通过优化多个影响因素，建立了一种高效的黑曲霉ATMT系统，与初始条件相比，转化效率提高了14倍。此外，构建了包含α-转葡萄糖苷酶表达盒的二元载体pBI-Glu，并将其转入根癌农杆菌LBA4404中以感染黑曲霉。通过高效的ATMT方法，由强启动子PglaA（糖化酶基因启动子）驱动的α-转葡萄糖苷酶基因在黑曲霉A-8中具有高表达水平（25.02 U/mL）。结果表明，优化后的ATMT系统对黑曲霉有效且适用，应成为研究黑曲霉基因功能和该菌株工业育种的有用工具。
