Promotion of Monacolin K production by Agrobacterium tumefaciens-mediated transformation in Monascus albidus 9901.
The binary vector pCAMBIA3300-gpdA-hph-trpC with hygromycin B phosphotransferase (hph) was constructed and transformed into Monascus albidus 9901 by Agrobacterium tumefaciens-mediated transformation, with gene hph as the selective marker. In order to improve the efficiency of A. tumefaciens-mediated transformation in M. albidus 9901, we optimized various factors including concentration of M. albidus 9901 spores, cell density of A. tumefaciens, co-cultivation time, temperature, and acetosyringone concentration. Most transformants of M. albidus 9901 could grow stably on media containing 50 μg ml⁻¹ hygromycin B up to five generations. The presence of hph was identified by PCR. Two transformants H1 and H2 which produced more Monacolin K than M. albidus 9901 were screened, and the concentration of Monacolin K in the fermented millet by H1 and H2 increased by 42.15% and 40.34% respectively compared with that produced by M. albidus 9901.
Wang
Lu
L
The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, Jiangsu Province, China. wang_lu1234@163.com
Wang
Wu
W
Xu
Ganrong
G
D000098:Acetophenones / Q000378:metabolism; D016960:Agrobacterium tumefaciens / Q000235:genetics*; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D008148:Lovastatin / Q000096:biosynthesis*; D032741:Monascus / Q000235:genetics / Q000378:metabolism*; D030781:Organisms, Genetically Modified / Q000235:genetics / Q000378:metabolism; D013696:Temperature; D013997:Time Factors; D014170:Transformation, Genetic
D000098:Acetophenones; C051667:acetosyringone; D008148:Lovastatin