Systematic Engineering of Genistein Biosynthetic Pathway through Genetic Regulators and Combinatorial Enzyme Screening

• Published in: Journal of agricultural and food chemistry Vol. 72; no. 11; pp. 5842 - 5848
• Main Authors: Hwang, Yunhee, Hwang, Hyun Gyu, Lee, Ji Yeon, Jung, Gyoo Yeol
• Format: Journal Article
• Language: English
• Published: United States American Chemical Society 20.03.2024
• Subjects: Biotechnology and Biological Transformations; enzyme screening; metabolic engineering; genistein; transcriptional repressor
• ISSN: 0021-8561; 1520-5118
• DOI: 10.1021/acs.jafc.3c09687

Microbial production of genistein, an isoflavonoid primarily found in soybeans, is gaining prominence in the food industry due to its significant nutritional and health benefits. However, challenges arise in redesigning strains due to intricate regulatory nodes between cell growth and genistein production and in systematically exploring core enzymes involving genistein biosynthesis. To address this, this study devised a strategy that simultaneously and precisely rewires flux at both acetyl-CoA and malonyl-CoA nodes toward genistein synthesis. In particular, naringenin, the primary precursor of genistein, was accumulated 2.6 times more than the unoptimized strain through transcriptional repressor-based genetic regulators. Building upon this, a combination of isoflavone synthase and cytochrome P450 reductase with the remarkable conversion of naringenin to genistein was screened from enzyme homologue libraries. The integrated metabolic engineering strategy yields the highest reported production (98 mg/L of genistein) to date, providing a framework for the biosynthesis of diverse flavonoids, including genistein.