Wattanachaisaereekul Songsak, Lantz Anna Eliasson, Nielsen Michael Lynge, Nielsen Jens
Center for Microbial Biotechnology, Technical University of Denmark, Department of Systems Biology, Denmark.
Metab Eng. 2008 Sep;10(5):246-54. doi: 10.1016/j.ymben.2008.04.005. Epub 2008 May 10.
The heterologous production of fungal polyketides was investigated using 6-methylsalicylic acid synthase (6-MSAS) as a model polyketide synthase and Saccharomyces cerevisiae as a host. In order to improve the production of 6-MSA by enhancing the supply of precursors, the promoter of the gene (ACC1) encoding acetyl-CoA carboxylase, which catalyzes the conversion of acetyl-CoA to malonyl-CoA, was replaced with a strong, constitutive promoter (TEF1p) in a strain harboring two plasmids carrying the genes encoding 6-MSAS from Penicillium patulum and PPTase from Aspergillus nidulans, respectively. The strain was characterized in batch cultivations with a glucose minimal media (20 g/L), and a 60% increase in 6-MSA titer was observed compared to a strain having the native promoter in front of ACC1. The production of 6-MSA was scaled up by the cultivation in minimal media containing 50 g/L of glucose, and hereby a final titer of 554+/-26 mg/L of 6-MSA was obtained.
以6-甲基水杨酸合酶(6-MSAS)作为模型聚酮合酶,酿酒酵母作为宿主,对真菌聚酮化合物的异源生产进行了研究。为了通过增加前体供应来提高6-MSA的产量,在携带分别编码来自展青霉的6-MSAS和来自构巢曲霉的PPTase基因的两个质粒的菌株中,将编码乙酰辅酶A羧化酶(催化乙酰辅酶A转化为丙二酰辅酶A)的基因(ACC1)的启动子替换为强组成型启动子(TEF1p)。该菌株在含20 g/L葡萄糖的基本培养基的分批培养中进行表征,与ACC1前具有天然启动子的菌株相比,观察到6-MSA滴度提高了60%。通过在含50 g/L葡萄糖的基本培养基中培养来扩大6-MSA的生产,从而获得了最终滴度为554±26 mg/L的6-MSA。