Thuan Nguyen Huy, Tatipamula Vinay Bharadwaj, Trung Nguyen Thanh, Van Giang Nguyen
Center for Pharmaceutical Biotechnology, Duy Tan University, Da Nang 550000, Vietnam.
Faculty of Biotechnology, Vietnam National University of Agriculture, Trau Quy, Hanoi 100000, Vietnam.
J Ind Microbiol Biotechnol. 2023 Feb 17;50(1). doi: 10.1093/jimb/kuad030.
Genkwanin has various significant roles in nutrition, biomedicine, and pharmaceutical biology. Previously, this compound was chiefly produced by plant-originated extraction or chemical synthesis. However, due to increasing concern and demand for safe food and environmental issues, the biotechnological production of genkwanin and other bioactive compounds based on safe, cheap, and renewable substrates has gained much interest. This paper described recombinant Escherichia coli-based co-culture engineering that was reconstructed for the de novo production of genkwanin from d-glucose. The artificial genkwanin biosynthetic chain was divided into 2 modules in which the upstream strain contained the genes for synthesizing p-coumaric acid from d-glucose, and the downstream module contained a gene cluster that produced the precursor apigenin and the final product, genkwanin. The Box-Behnken design, a response surface methodology, was used to empirically model the production of genkwanin and optimize its productivity. As a result, the application of the designed co-culture improved the genkwanin production by 48.8 ± 1.3 mg/L or 1.7-fold compared to the monoculture. In addition, the scale-up of genkwanin bioproduction by a bioreactor resulted in 68.5 ± 1.9 mg/L at a 48 hr time point. The combination of metabolic engineering and fermentation technology was therefore a very efficient and applicable approach to enhance the production of other bioactive compounds.
芫花素在营养、生物医学和药物生物学领域具有多种重要作用。此前，这种化合物主要通过植物提取或化学合成来生产。然而，由于人们对食品安全和环境问题的关注度及需求不断增加，基于安全、廉价且可再生底物的芫花素及其他生物活性化合物的生物技术生产受到了广泛关注。本文描述了基于重组大肠杆菌的共培养工程，该工程被重建用于从d -葡萄糖从头生产芫花素。人工芫花素生物合成链被分为两个模块，其中上游菌株包含从d -葡萄糖合成对香豆酸的基因，下游模块包含一个产生前体芹菜素和最终产物芫花素的基因簇。采用响应面法中的Box - Behnken设计对芫花素的生产进行经验建模并优化其生产率。结果表明，与单培养相比，所设计的共培养应用使芫花素产量提高了48.8±1.3mg/L，即提高了1.7倍。此外，在生物反应器中扩大芫花素生物生产规模，在48小时时间点产量达到68.5±1.9mg/L。因此，代谢工程与发酵技术的结合是提高其他生物活性化合物产量的一种非常有效且适用的方法。
