Enzyme and Metabolic Engineering Strategies for Biosynthesis of α-Farnesene in Saccharomyces cerevisiae

• DOI: 10.1021/acs.jafc.3c03677
• Publication Date: 2023-08-14T06:33:31Z
• Authors: Shengli Wang; Chuanling Zhan; Shengxin Nie; Daoguang Tian; Juane Lu; Mingzhang Wen; Jianjun Qiao; Hongji Zhu; Qinggele Caiyin
• Keywords: Metabolic Engineering; Metabolic pathway

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 efficient α-farnesene-producing yeast cell factory by combining enzyme metabolic engineering strategies. First, screened different plants for α-farnesene synthase (AFS) with best activity found that AFS from Camellia sinensis (CsAFS) exhibited most production Saccharomyces cerevisiae 4741. Second, flux mevalonate pathway was increased to improve supply precursor farnesyl pyrophosphate. Third, inducing site-directed mutagenesis CsAFS, CsAFSW281C variant obtained, which considerably production. Fourth, N-terminal serine-lysine-isoleucine-lysine (SKIK) tag introduced construct SKIK∼CsAFSW281C variant, further 2.8 g/L shake-flask cultures. Finally, titer 28.3 S. obtained fed-batch fermentation 5 L bioreactor.