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首页> 外文期刊>Applied Microbiology >Metabolic Engineering of Synechocystis sp. Strain PCC 6803 for Isobutanol Production
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Metabolic Engineering of Synechocystis sp. Strain PCC 6803 for Isobutanol Production

机译：代谢囊藻的代谢工程。用于异丁醇生产的PCC 6803菌株

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Global warming and decreasing fossil fuel reserves have prompted great interest in the synthesis of advanced biofuels from renewable resources. In an effort to address these concerns, we performed metabolic engineering of the cyanobacterium Synechocystis sp. strain PCC 6803 to develop a strain that can synthesize isobutanol under both autotrophic and mixotrophic conditions. With the expression of two heterologous genes from the Ehrlich pathway, the engineered strain can accumulate 90 mg/liter of isobutanol from 50 mM bicarbonate in a gas-tight shaking flask. The strain does not require any inducer (i.e., isopropyl β-d-1-thiogalactopyranoside [IPTG]) or antibiotics to maintain its isobutanol production. In the presence of glucose, isobutanol synthesis is only moderately promoted (titer = 114 mg/liter). Based on isotopomer analysis, we found that, compared to the wild-type strain, the mutant significantly reduced its glucose utilization and mainly employed autotrophic metabolism for biomass growth and isobutanol production. Since isobutanol is toxic to the cells and may also be degraded photochemically by hydroxyl radicals during the cultivation process, we employed in situ removal of the isobutanol using oleyl alcohol as a solvent trap. This resulted in a final net concentration of 298 mg/liter of isobutanol under mixotrophic culture conditions.
机译：全球变暖和化石燃料储量减少，引起了人们对利用可再生资源合成高级生物燃料的浓厚兴趣。为了解决这些问题，我们进行了蓝藻集胞藻属的代谢工程。菌株PCC 6803，以开发一种可以在自养和混养条件下合成异丁醇的菌株。通过从Ehrlich途径表达两个异源基因，工程菌株可以在气密摇瓶中从50 mM碳酸氢盐中积累90 mg / L的异丁醇。该菌株不需要任何诱导剂（即，异丙基β-d-1-硫代吡喃半乳糖苷[IPTG]）或抗生素即可维持其异丁醇生产。在葡萄糖存在下，仅适度促进异丁醇的合成（滴度= 114 mg / L）。根据同位异构体分析，我们发现，与野生型菌株相比，该突变体显着降低了其葡萄糖利用率，并且主要利用自养代谢来促进生物量的生长和异丁醇的生产。由于异丁醇对细胞有毒，并且在培养过程中还可能被羟基自由基光化学降解，因此我们使用油醇作为溶剂捕集剂，就地去除了异丁醇。在混合营养培养条件下，最终净浓度为298 mg / L异丁醇。

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