ggpS基因过表达对集胞藻PCC 6803甘油葡萄糖苷和甘油合成的影响

Effects of ggpS over-expression on glycosylglycerol and glycerol biosynthesis of Synechocystis sp. PCC 6803

摘要:

为了研究甘油葡萄糖苷磷酸合成酶 (GgpS) 在集胞藻PCC 803甘油葡萄糖苷和甘油合成中的作用，本研究在前期获得高产甘油葡萄糖苷藻株的基础上分别过量表达来自于集胞藻PCC 6803自身和聚球藻PCC 7002的甘油葡萄糖苷磷酸合成酶基因ggpS，并测定了在不同浓度NaCl胁迫时突变藻株的甘油葡萄糖苷和甘油积累量。结果发现获得的突变株甘油葡萄糖苷合成没有提高，但是甘油合成显著增强。此外，当培养基NaCl浓度从600 mmol/L提高到900 mmol/L时，集胞藻PCC 6803自身ggpS过表达藻株的甘油合成进一步提高75%。这些结果显示了GgpS在将碳代谢流导入集胞藻甘油合成途径中的作用。研究成果也为进一步通过基因工程改造提高集胞藻甘油葡萄糖苷和甘油合成效率奠定了基础。

关键词:甘油葡萄糖苷，甘油，集胞藻PCC6803，ggpS基因，基因工程

Abstract:

To study the roles of glucosylglycerol phosphate synthase (Ggps) in glucosylglycerol (GG) and glycerol biosynthesis, we over-expressed Ggps from either Synechocystis sp. PCC 6803 or Synechococcus sp. PCC 7002 in a Synechocystis strain with a high GG titer, and determined the GG and glycerol accumulation in the resultant mutants grown under different NaCl-stress conditions. Ion chromatography results revealed that GG yield was not improved, but glycerol production was significantly enhanced by over-expression of Ggps from Synechocystis sp. PCC 6803 (6803ggpS). In addition, increasing the NaCl concentration of medium from 600 to 900 mmol/L led to a further 75% increase of glycerol accumulation in the mutant strain with 6803ggpS over-expression. These findings show the role of ggpS in driving the carbon flux to the glycerol biosynthesis pathway, and will be helpful for further improvement of GG and glycerol production in Synechocystis.

Key words:glucosylglycerol, glycerol, Synechocystis sp. PCC 6803, ggpS, genetic engineering