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用于光合生产琥珀酸的蓝细菌代谢工程。

Metabolic engineering of cyanobacteria for the photosynthetic production of succinate.

作者信息

Lan Ethan I, Wei Crystal T

机构信息

Department of Biological Science and Technology, National Chiao Tung University, Hsinchu 30010, Taiwan.

Department of Biological Science and Technology, National Chiao Tung University, Hsinchu 30010, Taiwan.

出版信息

Metab Eng. 2016 Nov;38:483-493. doi: 10.1016/j.ymben.2016.10.014. Epub 2016 Oct 28.

Abstract

Succinate is an important commodity chemical currently used in the food, pharmaceutical, and polymer industries. It can also be chemically converted into other major industrial chemicals such as 1,4-butanediol, butadiene, and tetrahydrofuran. Here we metabolically engineered a model cyanobacterium Synechococcus elongatus PCC 7942 to photosynthetically produce succinate. We expressed the genes encoding for α-ketoglutarate decarboxylase and succinate semialdehyde dehydrogenase in S. elongatus PCC 7942, resulting in a strain capable of producing 120mg/L of succinate. However, this recombinant strain exhibited severe growth retardation upon induction of the genes encoding for the succinate producing pathway, potentially due to the depletion of α-ketoglutarate. To replenish α-ketoglutarate, we expressed the genes encoding for phosphoenolpyruvate carboxylase and citrate synthase from Corynebacterium glutamicum into the succinate producing strain. The resulting strain successfully restored the growth phenotype and produced succinate with a titer of 430mg/L in 8 days. These results demonstrated the possibility of photoautotrophic succinate production.

摘要

琥珀酸是一种重要的商品化学品,目前应用于食品、制药和聚合物行业。它还可以通过化学方法转化为其他主要工业化学品,如1,4-丁二醇、丁二烯和四氢呋喃。在此,我们对模式蓝藻聚球藻PCC 7942进行代谢工程改造,使其通过光合作用生产琥珀酸。我们在聚球藻PCC 7942中表达了编码α-酮戊二酸脱羧酶和琥珀酸半醛脱氢酶的基因,得到了一株能够产生120mg/L琥珀酸的菌株。然而,在诱导琥珀酸生产途径的基因表达后,该重组菌株表现出严重的生长迟缓,这可能是由于α-酮戊二酸的消耗所致。为了补充α-酮戊二酸,我们将谷氨酸棒杆菌编码磷酸烯醇式丙酮酸羧化酶和柠檬酸合酶的基因导入琥珀酸生产菌株。所得菌株成功恢复了生长表型,并在8天内产生了430mg/L的琥珀酸。这些结果证明了光合自养生产琥珀酸的可能性。

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