大豆异黄酮代谢途径在大肠杆菌中的构建及表达

Construction and Expression of the Soybean Isoflavonoid Biosynthetic Pathway in Escherichia coli

摘要:

自然界异黄酮合成途径主要存在于豆科植物中。以微生物为宿主研究异黄酮代谢，则需要将整个相关代谢途径的多酶体系组装到工程菌种，从而进行表达及代谢研究，这就需要用到多基因的转化和共表达技术。综合应用了多基因单载体和多基因多载体方法，将大豆异黄酮代谢途径中的五个关键酶基因导入到大肠杆菌中，对异黄酮代谢途径在大肠杆菌中的构建和表达进行了研究和探索，获得了含有五个外源基因的重组大肠杆菌；重组菌经IPTG诱导，以L-酪氨酸为底物进行发酵，发酵产物经过HPLC测定，结果表明和空白对照相比有新的代谢产物生成，初步断定为异黄酮类化合物。

Abstract:

The natural isoflavones biosynthetic pathway is only limited in legumes plant. To study the isoflavone in bacteria by metabolic engineering requires transformation of multi-gene of the whole pathway into the host strain to resembling the expression and metabolism of the genes. The multi-gene transformation and expression strategy become necessary because of this. This article talks about the multi-gene transformation strategy using one or many vectors, taking the five genes of isoflavonoid biosynthetic pathway to E. coli. The recombinant bacteria carry five genes with two vectors, the whole Isoflavonoid biosynthetic pathway was constructed into E. coli. Fermented with L-tyrosine as substrate and IPTG as an inducer, the recombinant bacteria can produce a new isoflavone related metabolite showed on HPLC analysis profile.