Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, People's Republic of China.
National Engineering Laboratory for Cereal Fermentation Technology, Jiangnan University, Wuxi 214122, People's Republic of China.
J Agric Food Chem. 2021 Mar 17;69(10):3103-3113. doi: 10.1021/acs.jafc.1c00008. Epub 2021 Mar 8.
Maximizing the flux of farnesyl diphosphate (FPP) to farnesene biosynthesis is the main challenge of farnesene overproduction in . In this study, we screened α-farnesene synthase from soybean () with a higher catalytic ability. Combining the overexpression of the mevalonate (MVA) pathway with the expression of , an engineered yeast strain producing 190.5 mg/L α-farnesene was screened with poor growth. By decreasing the copies of 3-hydroxy-3-methylglutaryl-coenzyme (HMGR) overexpressed, the titer was increased to 417.8 mg/L. Then, the coexpression of and HMGR under the control of the GAL promoter and inactivation of lipid phosphate phosphatase encoded by promoted the titer to 1163.7 mg/L. The titer was further increased to 1477.2 mg/L at the shake flask level with better growth by the construction of a prototrophic strain. Finally, the highest α-farnesene production of 10.4 g/L in was obtained by fed-batch fermentation in a 5 L bioreactor.
在 中，最大限度地提高法呢基二磷酸（FPP）向法呢烯生物合成的通量是法呢烯过量生产的主要挑战。在这项研究中，我们筛选出了一种催化能力更高的α-法呢烯合酶来自大豆（）。通过过表达甲羟戊酸（MVA）途径并表达，筛选出了一株产 190.5mg/Lα-法呢烯的工程酵母菌株，但生长较差。通过降低过表达的 3-羟基-3-甲基戊二酰基辅酶 A（HMGR）的拷贝数，将产量提高到 417.8mg/L。然后，通过共表达 和 HMGR 并受 GAL 启动子控制，以及失活编码的脂质磷酸酶，将产量提高到 1163.7mg/L。通过构建原养型菌株，在摇瓶水平上进一步提高生长速度，将产量提高到 1477.2mg/L。最后，通过在 5L 生物反应器中进行分批补料发酵，获得了 10.4g/L 的最高α-法呢烯产量。
