Engineering of acetate recycling and citrate synthase to improve aerobic succinate production in Corynebacterium glutamicum.
Engineering of acetate recycling and citrate synthase to improve aerobic succinate production in Corynebacterium glutamicum.
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乙酸回收和柠檬酸合成酶工程提高谷氨酸棒杆菌好氧琥珀酸产量
DOI：
10.1371/journal.pone.0060659
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Chen T
中科院分区：
文献类型：
--
作者：
Zhu N;Xia H;Wang Z;Zhao X;Chen T
缺乏琥珀酸脱氢酶复合物的谷氨酸棒杆菌能够在有氧条件下产生琥珀酸，乙酸是主要副产物。为提高琥珀酸产量所做的努力包括缺失乙酸形成途径以及过表达回补途径，但乙酸的形成无法完全消除。为解决这一问题，我们在产琥珀酸的谷氨酸棒杆菌中构建了一个回收浪费碳的途径。来自枯草芽孢杆菌的乙酰辅酶A合成酶首次被异源引入谷氨酸棒杆菌。工程菌株ZX1（pEacsA）不分泌乙酸，琥珀酸产量为0.50摩尔/（摩尔葡萄糖）⁻¹。此外，为了使更多的碳流向琥珀酸生物合成，过表达由gltA编码的天然柠檬酸合酶，得到菌株ZX1（pEacsAgltA），与菌株ZX1（pEacsA）相比，该菌株的琥珀酸产量提高了22%，丙酮酸产量降低了62%。在补料分批培养中，菌株ZX1（pEacsAgltA）产生241毫摩尔琥珀酸，平均体积生产率为3.55毫摩尔/小时⁻¹，平均产量为0.63摩尔/（摩尔葡萄糖）⁻¹，使其成为大规模有氧生产琥珀酸的一个有前景的平台。
Corynebacterium glutamicum lacking the succinate dehydrogenase complex can produce succinate aerobically with acetate representing the major byproduct. Efforts to increase succinate production involved deletion of acetate formation pathways and overexpression of anaplerotic pathways, but acetate formation could not be completely eliminated. To address this issue, we constructed a pathway for recycling wasted carbon in succinate-producing C. glutamicum. The acetyl-CoA synthetase from Bacillus subtilis was heterologously introduced into C. glutamicum for the first time. The engineered strain ZX1 (pEacsA) did not secrete acetate and produced succinate with a yield of 0.50 mol (mol glucose)−1. Moreover, in order to drive more carbon towards succinate biosynthesis, the native citrate synthase encoded by gltA was overexpressed, leading to strain ZX1 (pEacsAgltA), which showed a 22% increase in succinate yield and a 62% decrease in pyruvate yield compared to strain ZX1 (pEacsA). In fed-batch cultivations, strain ZX1 (pEacsAgltA) produced 241 mM succinate with an average volumetric productivity of 3.55 mM h−1 and an average yield of 0.63 mol (mol glucose) −1, making it a promising platform for the aerobic production of succinate at large scale.
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影响因子：
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