title: Construction of an engineering strain producing high yields of α-transglucosidase via Agrobacterium tumefaciens-mediated transformation of Asperillus niger.
abstract: 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.
authors.affiliation: Key Laboratory of Industrial Fermentation Microbiology, Education Ministry of China, College of Biotechnology, Tianjin University of Science and Technology.
authors.forename: Ming
authors.initials: M
authors.lastname: Li
authors.forename: Liying
authors.initials: L
authors.lastname: Zhou
authors.forename: Meng
authors.initials: M
authors.lastname: Liu
authors.forename: Yunyan
authors.initials: Y
authors.lastname: Huang
authors.forename: Xin
authors.initials: X
authors.lastname: Sun
authors.forename: Fuping
authors.initials: F
authors.lastname: Lu
mesh_terms: D016960:Agrobacterium tumefaciens / Q000166:cytology / Q000187:drug effects / Q000235:genetics* / Q000254:growth & development; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001234:Aspergillus niger / Q000235:genetics*; D018920:Coculture Techniques; D005818:Genetic Engineering / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics; D005964:Glucosyltransferases / Q000096:biosynthesis* / Q000235:genetics; D008938:Mitosis; D013172:Spores, Fungal / Q000235:genetics; D013696:Temperature; D013997:Time Factors; D014170:Transformation, Genetic
chemical_list: D000900:Anti-Bacterial Agents; D005964:Glucosyltransferases