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首页> 外文期刊>Biotechnology Progress >Metabolic Engineering of the Anaerobic Central Metabolic Pathway in Escherichia coli for the Simultaneous Anaerobic Production of Isoamyl Acetate and Succinic Acid
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Metabolic Engineering of the Anaerobic Central Metabolic Pathway in Escherichia coli for the Simultaneous Anaerobic Production of Isoamyl Acetate and Succinic Acid

机译：大肠杆菌厌氧中心乙酸代谢途径同时代谢的乙酸异戊酯和琥珀酸的代谢工程

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An in vivo method of producing isoamyl acetate and succinate simultaneously has been developed in Escherichia coli to maximize yields of both high value compounds as well as maintain the proper redox balance between NADH and NAD~+. Previous attempts at producing the ester isoamyl acetate anaerobically did not produce the compound in high concentrations because of competing pathways and the need for NAD~+ regeneration. The objective of this study is to produce succinate as an example of a reduced coproduct to balance the ratio of NADHINAD~+ as a way of maximizing isoamyl acetate production. Because the volatility of the two compounds differs greatly, the two could be easily separated in an industrial setting. An 1dhA, adhE double mutant strain (SBS110MG) served as the control strain to test the effect of an additional ackA-pta mutation as found in SBS990MG. Both strains overex-pressed the two heterologous genes pyruvate carboxylase and alcohol acetyltransferase (for ester production). The triple mutant SBS990MG was found to produce higher levels of both isoamyl acetate and succinate. At the optimal condition of 25°C, the culture produced 9.4 mM isoamyl acetate and 45.5 mM succinate. SBS990MG produced 36% more ester and over 700% more succinate than SBS110MG. In addition, this study demonstrated that a significantly higher isoamyl acetate concentration can be attained by simultaneously balancing the carbon and cofactor flow; the isoamyl acetate concentration of 9.4 mM is more than seven times higher than an earlier report of about 1.2 mM.
机译：已经在大肠杆菌中开发了一种同时生产乙酸异戊酯和琥珀酸酯的体内方法，以最大化两种高价值化合物的收率，并维持NADH和NAD〜+之间的适当氧化还原平衡。由于竞争途径和对NAD +再生的需要，先前生产厌氧乙酸异戊酯的尝试并未以高浓度生产该化合物。这项研究的目的是生产琥珀酸酯作为减少副产物的实例，以平衡NADHINAD〜+的比例，从而最大限度地提高乙酸异戊酯的产量。由于两种化合物的挥发性差异很大，因此在工业环境中很容易将二者分离。以1dhA，adhE双突变菌株（SBS110MG）作为对照菌株，以测试SBS990MG中发现的其他ackA-pta突变的作用。两种菌株都过表达两个异源基因丙酮酸羧化酶和醇乙酰基转移酶（用于酯生产）。发现三突变体SBS990MG产生更高水平的乙酸异戊酯和琥珀酸酯。在25°C的最佳条件下，培养物产生9.4 mM乙酸异戊酯和45.5 mM琥珀酸酯。与SBS110MG相比，SBS990MG产生的酯多36％，琥珀酸酯的产生多700％。此外，这项研究表明，通过同时平衡碳和辅助因子的流量，可以获得明显更高的乙酸异戊酯浓度。 9.4 mM的乙酸异戊酯浓度比早先报道的约1.2 mM高出七倍以上。

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