Methanol production by reversed methylotrophy constructed in Escherichia coli
Methanol production by reversed methylotrophy constructed in Escherichia coli
复制标题
大肠杆菌中构建的反向甲基营养型甲醇生产菌
DOI：
10.1080/09168451.2020.1715202
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发表时间：
2020-01
期刊：
影响因子：
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通讯作者：
Tomoyuki Takeya;Miyabi Yamakita;D. Hayashi;Kento Fujisawa;Y. Sakai;H. Yurimoto
中科院分区：
文献类型：
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作者：
Tomoyuki Takeya;Miyabi Yamakita;D. Hayashi;Kento Fujisawa;Y. Sakai;H. Yurimoto
摘要我们用合成生物学的方法构建了一条反向甲基营养途径，以6-磷酸果糖(F6P)为原料，通过分解葡萄糖等生物质衍生的糖来提供甲醇，甲醇是生产有用化合物的原料。以大肠杆菌为表达宿主，异源表达了甲醇营养细菌依赖NAD+的甲醇脱氢酶(MDH)和胃分枝杆菌MB19的3-己果糖-6-磷酸合成酶和6-磷酸-6-磷酸异构酶(HPS-PHI)。我们证实了这些酶可以催化甲醇氧化和甲醛固定的反向反应。共表达MDH和HPS-PHI基因的工程菌在静息细胞反应中不仅从F6P还从葡萄糖产生甲醇。我们成功地将反向甲基化作用授予了大肠杆菌，我们的结果为利用生物质衍生的糖化合物生产生物甲醇提供了概念验证。通过将MDH和HPS-PHI导入大肠杆菌细胞，构建了以葡萄糖或F6P为原料合成甲醇的反转甲基营养体系。
ABSTRACT We constructed a reversed methylotrophic pathway that produces methanol, a promising feedstock for production of useful compounds, from fructose 6-phosphate (F6P), which can be supplied by catabolism of biomass-derived sugars including glucose, by a synthetic biology approach. Using Escherichia coli as an expression host, we heterologously expressed genes encoding methanol utilization enzymes from methylotrophic bacteria, i.e. the NAD+-dependent methanol dehydrogenase (MDH) from Bacillus methanolicus S1 and an artificial fusion enzyme of 3-hexulose-6-phosphate synthase and 6-phospho-3-hexuloisomerase from Mycobacterium gastri MB19 (HPS-PHI). We confirmed that these enzymes can catalyze reverse reactions of methanol oxidation and formaldehyde fixation. The engineered E. coli strain co-expressing MDH and HPS-PHI genes produced methanol in resting cell reactions not only from F6P but also from glucose. We successfully conferred reversed methylotrophy to E. coli and our results provide a proof-of-concept for biological methanol production from biomass-derived sugar compounds. Graphical abstract Reversed methylotrophy to produce methanol from glucose or F6P was constructed by introducing MDH and HPS-PHI into E. coli cells.