Producing malonate in Saccharomyces cerevisiae via theβ‑alanine pathway  

作　　者：Shiyun Li Wenxuan Fu Ruifang Su Yunying Zhao Yu Deng 

机构地区：[1]National Engineering Research Center of Cereal Fermentation and Food Biomanufacturing,Jiangnan University,1800 Lihu Road,Wuxi 214122,Jiangsu,China [2]Jiangsu Provincial Research Center for Bioactive Product Processing Technology,Jiangnan University,1800 Lihu Road,Wuxi 214122,Jiangsu,China

出　　处：《Systems Microbiology and Biomanufacturing》2023年第2期328-338,共11页系统微生物学与生物制造（英文）

基　　金：supported by the National Key R&D Program of China(2019YFA0905502);the National Natural Science Foundation of China(21877053);Tianjin Synthetic Biotechnology Innovation Capacity Improvement Project(TSBICIP-KJGG-015);the Open Foundation of Jiangsu Key Laboratory of Industrial Biotechnology(KLIB-KF201807).

摘　　要：Malonate is a high-value chemical that can be used to produce value-added compounds.Due to the toxic by-products and low product yield for malonate production through hydrolysis of cyanoacetic acid,microbial production methods have attracted significant attention.Previously,theβ-alanine pathway has been engineered in Escherichia coli for malonate production.In this study,theβ-alanine pathway was constructed in Saccharomyces cerevisiae by introducing the heterologous genes of BcBAPAT and TcPAND to convert l-aspartate to malonic semialdehyde,combining with co-expression genes of AAT2 and UGA2 to improve precursor supply and malonate producing.Through delta sequence-based integration of the two heterologous genes,the engineered strain produced with 7.21 mg/L malonate was screened.Further,replaced the succinic semialdehyde dehydrogenase gene UGA2 with yneI from E.coli which was utilized to produce malonate in previous study,increased the malonate titer to 7.96 mg/L in flask culture.Following optimization,fermentation of the final engineered strain in shake flasks yielded a maximum malonate titer of 12.83 mg/L,and this was increased to 91.53 mg/L during fed-batch fermentation in a 5 L bioreactor which increased by two-fold compared with that of the engineered strain overexpressing UGA2.

关 键 词：MALONATE Saccharomyces cerevisiae Β-ALANINE Metabolic engineering Delta-sequence integration 

分 类 号：Q815[生物学—生物工程]