Extending Carbon Chain Length of 1-Butanol Pathway for 1-Hexanol Synthesis from Glucose by Engineered Escherichia coli
Extending Carbon Chain Length of 1-Butanol Pathway for 1-Hexanol Synthesis from Glucose by Engineered Escherichia coli
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DOI：
10.1021/ja203814d
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发表时间：
2011-08-03
影响因子：
15
通讯作者：
Liao, James C.
中科院分区：
文献类型：
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作者：
Dekishima, Yasumasa;Lan, Ethan I.;Liao, James C.
通过延长外源酶催化辅酶A（CoA）依赖的1-丁醇合成反应序列，对大肠杆菌菌株进行工程改造，以从葡萄糖合成1-己醇。C4-酰基-CoA中间体首先通过来自各种生物体的乙酰-CoA乙酰转移酶（AtoB）、3-羟基丁酰-CoA脱氢酶（Hbd）、巴豆酸酶（Crt）和反式-烯酰-CoA还原酶（Ter）合成。合成的丁酰-CoA通过β-酮硫解酶（BktB）、Hbd、Crt和Ter进一步延伸为己酰-CoA。最后，通过醛/醇脱氢酶（AdhE 2）还原己酰-CoA以产生1-己醇。C6中间体的酶活性通过使用HPLC和GC的测定来确认。1-己醇在厌氧条件下分泌到发酵培养基中。此外，共表达来自博伊丁假丝酵母的甲酸脱氢酶（Fdh）增加了1-己醇滴度。这种通过延长1-丁醇途径生产1-己醇的证明提供了使用相同策略生产其他中链长醇的可能性。
An Escherichia coli strain was engineered to synthesize 1-hexanol from glucose by extending the coenzyme A (CoA)-dependent 1-butanol synthesis reaction sequence catalyzed by exogenous enzymes. The C4-acyl-CoA intermediates were first synthesized via acetyl-CoA acetyltransferase (AtoB), 3-hydroxybutyryl-CoA dehydrogenase (Hbd), crotonase (Crt), and trans-enoyl-CoA reductase (Ter) from various organisms. The butyryl-CoA synthesized was further extended to hexanoyl-CoA via beta-ketothiolase (BktB), Hbd, Crt, and Ter. Finally, hexanoyl-CoA was reduced to yield 1-hexanol by aldehyde/alcohol dehydrogenase (AdhE2). Enzyme activities for the C6 intermediates were confirmed by assays using HPLC and GC. 1-Hexanol was secreted to the fermentation medium under anaerobic conditions. Furthermore, co-expressing formate dehydrogenase (Fdh) from Candida boidinii increased the 1-hexanol titer. This demonstration of 1-hexanol production by extending the 1-butanol pathway provides the possibility to produce other medium chain length alcohols using the same strategy.