School of Energy and Chemical Engineering, Ulsan National Institute of Science and Technology, Ulsan 44919, Republic of Korea.
Department of Chemical and Biochemical Engineering, North Carolina State University, Raleigh, NC 27606, USA.
Bioresour Technol. 2023 May;376:128911. doi: 10.1016/j.biortech.2023.128911. Epub 2023 Mar 17.
The production of 1,3-butanediol (1,3-BDO) from glucose was investigated using Escherichia coli as the host organism. A pathway was engineered by overexpressing genes phaA (acetyl-CoA acetyltransferase), phaB (acetoacetyl-CoA reductase), bld (CoA-acylating aldehyde dehydrogenase), and yqhD (alcohol dehydrogenase). The expression levels of these genes were optimized to improve 1,3-BDO production and pathways that compete with 1,3-BDO synthesis were disrupted. Culture conditions were also optimized, including the C: N ratio, aeration, induction time, temperature, and supplementation of amino acids, resulting in a strain that could produce 1,3-BDO at 257 mM in 36 h, with a yield of 0.51 mol/mol in a fed-batch bioreactor experiment. To the best of our knowledge, this is the highest titer of 1,3-BDO production ever reported using biological methods, and our findings provide a promising strategy for the development of microbial cell factories for the sustainable synthesis of other acetyl-CoA-derived chemicals.
利用大肠杆菌作为宿主生物，从葡萄糖生产 1,3-丁二醇（1,3-BDO）。通过过表达phaA（乙酰辅酶 A 乙酰转移酶）、phaB（乙酰乙酰辅酶 A 还原酶）、bld（CoA 酰化醛脱氢酶）和 yqhD（醇脱氢酶）基因来构建途径。优化这些基因的表达水平以提高 1,3-BDO 的产量，并破坏与 1,3-BDO 合成竞争的途径。还优化了培养条件，包括 C:N 比、通气、诱导时间、温度和氨基酸的补充，最终在分批补料生物反应器实验中，该菌株能够在 36 小时内产生 257mM 的 1,3-BDO，产率为 0.51mol/mol。据我们所知，这是使用生物方法生产 1,3-BDO 的最高浓度，我们的研究结果为开发可持续合成其他乙酰辅酶 A 衍生化学品的微生物细胞工厂提供了有前途的策略。
