Matsubara Mitsuru, Urano Nobuyuki, Yamada Shohei, Narutaki Ai, Fujii Misaki, Kataoka Michihiko
Division of Applied Life Sciences, Graduate School of Life and Environmental Sciences, Osaka Prefecture University, 1-1 Gakuen-cho, Naka-ku, Sakai, Osaka 599-8531, Japan.
Division of Applied Life Sciences, Graduate School of Life and Environmental Sciences, Osaka Prefecture University, 1-1 Gakuen-cho, Naka-ku, Sakai, Osaka 599-8531, Japan.
J Biosci Bioeng. 2016 Oct;122(4):421-6. doi: 10.1016/j.jbiosc.2016.03.011. Epub 2016 Apr 9.
Fermentative production of 1-propanol, which is one of the promising precursors of polypropylene production, from d-glucose, l-rhamnose and glycerol using metabolically engineered Escherichia coli was examined. To confer the ability to produce 1-propanol from 1,2-propanediol (1,2-PD) in recombinant E. coli, a part of the pdu regulon including the diol dehydratase and the propanol dehydrogenase genes together with the adenosylcobalamin (AdoCbl) regeneration enzyme genes of Klebsiella pneumoniae was cloned, and an expression vector for these genes (pRSF_pduCDEGHOQS) was constructed. Recombinant E. coli harboring pRSF_pduCDEGHOQS with 1,2-PD synthetic pathway (pKK_mde) genes, which was constructed in our previous report (Urano et al., Appl. Microbiol. Biotechnol., 99, 2001-2008, 2015), produced 16.1 mM of 1-propanol from d-glucose with a molar yield of 0.36 mol/mol after 72 h cultivation. 29.9 mM of 1-propanol was formed from l-rhamnose with a molar yield of 0.81 mol/mol using E. coli carrying only pRSF_pduCDEGHOQS. In addition, 1-propanol production from glycerol was achieved by addition of the ATP-dependent dihydroxyacetone kinase gene to E. coli harboring pKK_mde and pRSF_pduCDEGOQS. In all cases, 1-propanol production was achieved by adding only a small amount of AdoCbl.
研究了使用代谢工程改造的大肠杆菌，从d-葡萄糖、l-鼠李糖和甘油中发酵生产1-丙醇的过程，1-丙醇是聚丙烯生产中一种很有前景的前体物质。为了使重组大肠杆菌具备从1,2-丙二醇（1,2-PD）生产1-丙醇的能力，克隆了肺炎克雷伯菌的部分pdu操纵子，包括二醇脱水酶和丙醇脱氢酶基因以及腺苷钴胺素（AdoCbl）再生酶基因，并构建了这些基因的表达载体（pRSF_pduCDEGHOQS）。携带pRSF_pduCDEGHOQS和1,2-PD合成途径（pKK_mde）基因的重组大肠杆菌，在我们之前的报告（Urano等人，《应用微生物学与生物技术》，99，2001 - 2008，2015）中构建，在培养72小时后，从d-葡萄糖中产生了16.1 mM的1-丙醇，摩尔产率为0.36 mol/mol。仅携带pRSF_pduCDEGHOQS的大肠杆菌从l-鼠李糖中形成了29.9 mM的1-丙醇，摩尔产率为0.81 mol/mol。此外，通过向携带pKK_mde和pRSF_pduCDEGOQS的大肠杆菌中添加ATP依赖性二羟基丙酮激酶基因，实现了从甘油生产1-丙醇。在所有情况下，仅添加少量的AdoCbl就能实现1-丙醇的生产。
