Wang Shengli, Zhan Chuanling, Nie Shengxin, Tian Daoguang, Lu Juane, Wen Mingzhang, Qiao Jianjun, Zhu Hongji, Caiyin Qinggele
Department of Pharmaceutical Engineering, School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072, People's Republic of China.
Key Laboratory of Systems Bioengineering (Ministry of Education), Tianjin University, Tianjin 300072, People's Republic of China.
J Agric Food Chem. 2023 Aug 23;71(33):12452-12461. doi: 10.1021/acs.jafc.3c03677. Epub 2023 Aug 13.
α-Farnesene, a type of acyclic sesquiterpene, is an important raw material in agriculture, aircraft fuel, and the chemical industry. In this study, we constructed an efficient α-farnesene-producing yeast cell factory by combining enzyme and metabolic engineering strategies. First, we screened different plants for α-farnesene synthase (AFS) with the best activity and found that AFS from (CsAFS) exhibited the most efficient α-farnesene production in 4741. Second, the metabolic flux of the mevalonate pathway was increased to improve the supply of the precursor farnesyl pyrophosphate. Third, inducing site-directed mutagenesis in CsAFS, the CsAFS variant was obtained, which considerably increased α-farnesene production. Fourth, the N-terminal serine-lysine-isoleucine-lysine (SKIK) tag was introduced to construct the SKIK∼CsAFS variant, which further increased α-farnesene production to 2.8 g/L in shake-flask cultures. Finally, the α-farnesene titer of 28.3 g/L in was obtained by fed-batch fermentation in a 5 L bioreactor.
α-法尼烯是一种无环倍半萜烯，是农业、航空燃料和化学工业中的重要原料。在本研究中，我们通过结合酶工程和代谢工程策略构建了一个高效生产α-法尼烯的酵母细胞工厂。首先，我们筛选了不同植物中具有最佳活性的α-法尼烯合酶（AFS），发现来自[具体植物名称]（CsAFS）的AFS在[具体酵母菌株名称]4741中表现出最有效的α-法尼烯生产能力。其次，增加甲羟戊酸途径的代谢通量以改善前体法尼基焦磷酸的供应。第三，在CsAFS中诱导定点诱变，获得了CsAFS变体，其显著提高了α-法尼烯的产量。第四，引入N端丝氨酸-赖氨酸-异亮氨酸-赖氨酸（SKIK）标签构建SKIK∼CsAFS变体，在摇瓶培养中进一步将α-法尼烯产量提高到2.8 g/L。最后，通过在5 L生物反应器中进行补料分批发酵，在[具体发酵条件]下获得了28.3 g/L的α-法尼烯滴度。
