对大肠杆菌进行基因改造以用于乙醇生产。
Re-engineering Escherichia coli for ethanol production.
作者信息
Yomano L P, York S W, Zhou S, Shanmugam K T, Ingram L O
机构信息
Department Microbiology and Cell Science, University of Florida, Gainesville, FL 32611, USA.
出版信息
Biotechnol Lett. 2008 Dec;30(12):2097-103. doi: 10.1007/s10529-008-9821-3. Epub 2008 Sep 5.
A lactate producing derivative of Escherichia coli KO11, strain SZ110, was re-engineered for ethanol production by deleting genes encoding all fermentative routes for NADH and randomly inserting a promoterless mini-Tn5 cassette (transpososome) containing the complete Zymomonas mobilis ethanol pathway (pdc, adhA, and adhB) into the chromosome. By selecting for fermentative growth in mineral salts medium containing xylose, a highly productive strain was isolated in which the ethanol cassette had been integrated behind the rrlE promoter, designated strain LY160(KO11, Deltafrd::celY(Ec) DeltaadhE DeltaldhA, DeltaackA lacA::casAB(Ko) rrlE::(pdc( Zm)-adhA(Zm)-adhB(Zm)-FRT-rrlE)pflB(+)). This strain fermented 9% (w/v) xylose to 4% (w/v) ethanol in 48 h in mineral salts medium, nearly equal to the performance of KO11 with Luria broth.
大肠杆菌KO11的产乳酸衍生物菌株SZ110，通过删除编码所有NADH发酵途径的基因，并将含有完整运动发酵单胞菌乙醇途径（pdc、adhA和adhB）的无启动子mini-Tn5盒（转座体）随机插入染色体，进行了乙醇生产的基因工程改造。通过在含木糖的矿物盐培养基中选择发酵生长，分离出了一个高产菌株，其中乙醇盒已整合到rrlE启动子之后，命名为菌株LY160（KO11，Deltafrd::celY(Ec) DeltaadhE DeltaldhA，DeltaackA lacA::casAB(Ko) rrlE::(pdc(Zm)-adhA(Zm)-adhB(Zm)-FRT-rrlE)pflB(+)）。该菌株在矿物盐培养基中48小时内将9%（w/v）的木糖发酵为4%（w/v）的乙醇，几乎与KO11在Luria肉汤中的表现相当。
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