文献类型：期刊文献

英文题名：Biosynthesis of Limonene Using Acetate as the Carbon Source in Metabolically Engineered Escherichia coli

作者：Zhao, Weiying[1];Shang, Yanzhe[2];Pan, Meiyu[1];Fei, Peng[1];Wang, Yuying[1];Luo, Yuanchan[1];Wu, Hui[1,2]

机构：[1]East China Univ Sci & Technol, Shanghai Collaborat Innovat Ctr Biomfg Technol, Sch Biotechnol, Key Lab Biobased Mat Engn China Natl Light Ind Cou, 130 Meilong Rd, Shanghai 200237, Peoples R China;[2]Dalian Univ Technol, Sch Bioengn, MOE Key Lab Biointelligent Mfg, Dalian 116024, Peoples R China

年份：2026

卷号：74

期号：11

起止页码：9526

外文期刊名：JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY

收录：;EI(收录号：20261320355853);WOS:【SCI-EXPANDED(收录号:WOS:001712098800001)】；

基金：This work was supported by the National Key R&D Program of China (2022YFC2105400), the Science and Technology Commission of Shanghai Municipality (24HC2820800), the Fundamental Research Funds for the Central Universities (DUT24RC(3)029), the Project Program of Key Laboratory of Industrial Fermentation Microbiology, Ministry of Education, and Tianjin Key Laboratory of Industrial Microbiology, China (2024KF01).

语种：英文

外文关键词：limonene; acetic acid; metabolic engineering; Escherichiacoli

摘要：Limonene is a high-value monoterpenoid used in food, pharmaceutical, and fragrance applications. To overcome limitations of conventional production, this study developed a biosynthesis platform using acetate as the sole carbon source. Acetate is an ideal nonfood substrate that can be sustainably derived from industrial waste streams, syngas, and CO2 reduction. Initially, the heterologous synthetic pathway was constructed to yield 0.13 mg/L limonene from acetate. By introducing the mutated EfMVAS A110G to enhance catalytic efficiency, along with promoter engineering and substitution of the synthesis substrate, the production titer was enhanced to 49.24 mg/L. Subsequent host optimization through NADPH regeneration and pathway knockout, combined with fermentation optimization, increased the titer to 167.45 mg/L. Ultimately, whole-cell biocatalysis achieved 450.72 mg/L, establishing the highest reported acetate-based limonene titer. Overall, this study establishes a framework for biosynthesis of high-value terpenoids from nonfood feedstocks and demonstrates acetate as a promising carbon source for advanced bioproduction systems.