亚磷酸脱氢酶表达对大肠杆菌琥珀酸发酵的影响Effect of Phosphite Dehydrogenase Expression on Succinic Acid Fermentation in Escherichia coli
刘利,陈炜琪,邹诗瑶,王金华,高娃,王永泽
摘要(Abstract)：
琥珀酸在食品中有广泛的用途，但琥珀酸发酵往往受制于还原力不平衡的问题。相比于其他常用于平衡还原力的酶，亚磷酸脱氢酶催化产物为磷酸盐，对后续琥珀酸的分离纯化影响较小。文章克隆了来源于施氏假单胞菌的亚磷酸脱氢酶(编码基因为ptxD),比较ptxD在载体和基因组上表达对琥珀酸发酵的影响，同时研究了亚磷酸盐添加量对发酵的影响。结果表明，在ptxD表达条件下，亚磷酸盐在5～15 mmol/L范围内添加并未表现出对细胞生长的抑制性，且在10 mmol/L时展现出较高的糖耗速率和琥珀酸得率。基因组上表达ptxD的工程菌WS105(WS100,iclR::pJ23119-ptxd)能积累79.38 g/L琥珀酸，较载体上表达ptxD的工程菌WS104(WS100/puc19-pJ23119-ptxd)提升了4.87%,较出发菌株WS100提升了13.81%。WS105副产物乙酸含量显著下降，浓度为2.76 g/L,较WS100、WS104分别降低了42.38%、29.41%,表明亚磷酸脱氢酶的表达能有效促进琥珀酸产量的提高。
关键词(KeyWords)： 琥珀酸;亚磷酸脱氢酶;还原力平衡
基金项目(Foundation): 国家自然科学基金青年项目(31501677);; 企业合作项目(2021433)
作者(Author): 刘利,陈炜琪,邹诗瑶,王金华,高娃,王永泽
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