Department of Forest Resources, Gyeongnam National University of Science and Technology, Jinju 52725, Republic of Korea.
J Microbiol Biotechnol. 2019 May 28;30(5):770-776. doi: 10.4014/jmb.1911.11009.
Genistein is a type of isoflavonoid found predominantly in leguminous plants. Genistein has diverse biological activities, such as anthelmintic and antioxidant effects, as well as inhibitory effects on the growth of several cancers. In addition, genistein is well known as a phytoestrogen. In this study, we attempted to biologically synthesize genistein from either -coumaric acid or naringenin using as a biotransformation host. Four genes, , , , and , were used for genistein production. To functionally express RcIFS and OsCPR, two members of the cytochrome P450 family, in , the membrane-binding anchor domain of each gene was removed, and and were translationally fused to generate an hybrid. Os4CL and PeCHS, or the RcIFS-OsCPR hybrid, were then transformed into BL21(DE3). Using these strains, we optimized our culture system at a laboratory scale in terms of the cell density, concentrations of substrate and isopropyl-β-D-thiogalactoside, temperature, and culture medium. Under the optimized culture conditions, genistein was produced at up to 35 mg/l and 18.6 mg/l using naringenin and -coumaric acid, respectively.
染料木黄酮是一种主要存在于豆科植物中的异黄酮。染料木黄酮具有多种生物活性，如驱虫和抗氧化作用，以及对几种癌症生长的抑制作用。此外，染料木黄酮是众所周知的植物雌激素。在这项研究中，我们试图使用作为生物转化宿主，从对香豆酸或柚皮苷生物合成染料木黄酮。为了生产染料木黄酮，使用了四个基因、、、和。为了在中功能性表达细胞色素 P450 家族的两个成员 RcIFS 和 OsCPR，去除了每个基因的膜结合锚定结构域，并将和翻译融合以产生杂种。然后将 Os4CL 和 PeCHS 或 RcIFS-OsCPR 杂种转化到 BL21(DE3)中。使用这些菌株，我们在实验室规模上根据细胞密度、底物和异丙基-β-D-硫代半乳糖苷的浓度、温度和培养基优化了我们的培养系统。在优化的培养条件下，分别使用柚皮苷和对香豆酸可生产高达 35 mg/L 和 18.6 mg/L 的染料木黄酮。
