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首页> 外文期刊>Metabolic engineering >Inactivation of aldehyde dehydrogenase: a key factor for engineering 1,3-propanediol production by Klebsiella pneumoniae.
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Inactivation of aldehyde dehydrogenase: a key factor for engineering 1,3-propanediol production by Klebsiella pneumoniae.

机译：醛脱氢酶的失活：肺炎克雷伯氏菌工程化生产1,3-丙二醇的关键因素。

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摘要

Production of 1,3-propanediol (1,3-PD) from glycerol by Klebsiella pneumoniae is restrained by ethanol formation. The first step in the formation of ethanol from acetyl-CoA is catalyzed by aldehyde dehydrogenase (ALDH), an enzyme that competes with 1,3-PD oxidoreductase for the cofactor NADH. This study aimed to improve the production of 1,3-PD by engineering the ethanol formation pathway. An inactivation mutation of the aldA gene encoding ALDH in K. pneumoniae YMU2 was generated by insertion of a tetracycline resistance marker. Inactivation of ALDH resulted in a nearly abolished ethanol formation but a significantly improved 1,3-PD production. Metabolic flux analysis revealed that a pronounced redistribution of intracellular metabolic flux occurred. The final titer, the productivity of 1,3-PD and the yield of 1,3-PD relative to glycerol of the mutant strain reached 927.6 mmol L(-1), 14.05 mmol L(-1)h(-1) and 0.699 mol mol(-1), respectively, which were much higher than those of the parent strain. In addition, the specific 1,3-PD-producing capability (1,3-PD produced per gram of cells) of the mutant strain was 2-fold that of the parent strain due to a lower growth yield of the mutant. By increasing NADH availability, this study demonstrates an important metabolic engineering approach to improve the efficiency of oxidoreduction-coupled bioprocesses.
机译：肺炎克雷伯菌（Klebsiella pneumoniae）由甘油生产1,3-丙二醇（1,3-PD）受乙醇形成的限制。由乙酰辅酶A形成乙醇的第一步是由醛脱氢酶（ALDH）催化，该酶与1,3-PD氧化还原酶竞争辅酶NADH。这项研究旨在通过设计乙醇形成途径来提高1,3-PD的产量。肺炎克雷伯菌YMU2中编码ALDH的aldA基因的失活突变是通过插入四环素抗性标记而产生的。 ALDH的失活导致几乎消除了乙醇的形成，但大大提高了1,3-PD的产量。代谢通量分析显示细胞内代谢通量发生了明显的重新分布。相对于甘油，突变体菌株的最终效价，1,3-PD的生产率和1,3-PD的产量分别达到927.6 mmol L（-1），14.05 mmol L（-1）h（-1）和0.699 mol mol（-1）分别比亲本菌株高得多。另外，由于突变体的较低生长产量，突变体菌株的特异性1，3-PD产生能力（每克细胞产生1，3-PD）是亲本菌株的2倍。通过增加NADH的可用性，本研究证明了一种重要的代谢工程方法，可以提高氧化还原偶联生物过程的效率。

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