Graduate School of Science, Technology and Innovation, Kobe University, 1-1 Rokkodai, Nada, Kobe 657-8501, Japan.
Engineering Biology Research Center, Kobe University, 1-1 Rokkodai, Nada, Kobe 657-8501, Japan.
ACS Synth Biol. 2022 Jun 17;11(6):2098-2107. doi: 10.1021/acssynbio.2c00047. Epub 2022 May 16.
Bioactive plant-based secondary metabolites such as stilbenoids, flavonoids, and benzylisoquinoline alkaloids (BIAs) are produced from l-tyrosine (l-Tyr) and have a wide variety of commercial applications. Therefore, building a microorganism with high l-Tyr productivity (l-Tyr chassis) is of immense value for large-scale production of various aromatic compounds. The aim of this study was to develop an l-Tyr chassis in the nonconventional yeast () to produce various aromatic secondary metabolites (resveratrol, naringenin, norcoclaurine, and reticuline). Overexpression of feedback-inhibition insensitive variants of 3-deoxy-d-arabino-heptulosonate-7-phosphate synthase () and chorismate mutase () enhanced l-Tyr titer from glycerol in . These engineered strains increased the titer of resveratrol, naringenin, and norcoclaurine by 258, 244, and 3400%, respectively, after expressing the corresponding heterologous pathways. The titer of resveratrol and naringenin further increased by 305 and 249%, resulting in yields of 1825 and 1067 mg/L, respectively, in fed-batch fermentation, which is the highest titer from glycerol reported to date. Furthermore, the resveratrol-producing strain accumulated intermediates in the shikimate pathway. l-Tyr-derived aromatic compounds were produced using crude glycerol byproducts from biodiesel fuel (BDF) production. Constructing an l-Tyr chassis is a promising strategy to increase the titer of various aromatic secondary metabolites and is an attractive host for high-yield production of l-Tyr-derived aromatic compounds from glycerol.
生物活性植物源性次生代谢产物，如芪类、类黄酮和苯并异喹啉生物碱（BIAs），是由 l-酪氨酸（l-Tyr）产生的，具有广泛的商业应用。因此，构建一个具有高 l-Tyr 生产能力（l-Tyr 底盘）的微生物对于各种芳香族化合物的大规模生产具有巨大的价值。本研究旨在开发非传统酵母（）中的 l-Tyr 底盘，以生产各种芳香族次生代谢产物（白藜芦醇、柚皮素、去甲可乐定和瑞替丁）。3-脱氧-d-阿拉伯庚酮糖-7-磷酸合酶（）和分支酸变位酶（）的反馈抑制不敏感变体的过表达提高了甘油中 l-Tyr 的产量。这些工程菌株在表达相应的异源途径后，分别将白藜芦醇、柚皮素和去甲可乐定的产量提高了 258%、244%和 3400%。在分批补料发酵中，白藜芦醇和柚皮素的产量进一步提高了 305%和 249%，分别达到 1825 和 1067mg/L，这是迄今为止从甘油报告的最高产量。此外，白藜芦醇生产菌株在莽草酸途径中积累了中间体。利用生物柴油（BDF）生产中的粗甘油副产物生产 l-Tyr 衍生的芳香族化合物。构建 l-Tyr 底盘是提高各种芳香族次生代谢产物产量的有前途的策略，而 是从甘油生产 l-Tyr 衍生的芳香族化合物的高产量生产的有吸引力的宿主。
