Graduate School of Biotechnology and Bioengineering, Yuan Ze University, 135 Yuan-Tung Rd, Chung-Li, 320 Taiwan.
Appl Biochem Biotechnol. 2012 Feb;166(3):796-804. doi: 10.1007/s12010-011-9469-7. Epub 2011 Dec 28.
Cupriavidus necator is well known for its ability to accumulate polyhydroxybutyrate (PHB). When supplemented with propionic acid (or sodium propionate) in the growth medium, the bacterium is also able to synthesize polyhydroxybutyrate-co-hydroxyvalerate (PHBV). In order to increase the fraction of 3-hydroxyvalerate (3HV) in PHBV, we cloned the propionate permease gene prpP from C. necator and the propionyl-CoA synthase gene prpE from Cupriavidus taiwanensis and transformed into an Escherichia coli containing phaCAB operon of C. necator. The effects on PHBV accumulation in cells co-expressed with phaCAB and prpE or prpP in the media contained mixed carbon sources (glucose and sodium propionate) were evaluated. The HV fraction in PHBV increased when prpE or prpP was overexpressed in the cells. Concentrations of yeast extracts could also affect the fraction of HV. In addition, when glucose was replaced by sodium pyruvate, sodium succinate, or sodium gluconate, only PHB were detected in the recombinant strains.
贪铜菌以其积累聚羟基丁酸酯(PHB)的能力而闻名。当在生长培养基中补充丙酸(或丙酸钠)时,该细菌还能够合成聚羟基丁酸酯-共-羟基戊酸酯(PHBV)。为了增加 PHBV 中 3-羟基戊酸酯(3HV)的比例,我们从贪铜菌中克隆了丙酸转运蛋白基因 prpP 和 Cupriavidus taiwanensis 中的丙酰辅酶 A 合酶基因 prpE,并将其转化到含有贪铜菌的 phaCAB 操纵子的大肠杆菌中。在含有混合碳源(葡萄糖和丙酸钠)的培养基中共同表达 phaCAB 和 prpE 或 prpP 的细胞中,评估了对 PHBV 积累的影响。当细胞中过表达 prpE 或 prpP 时,PHBV 中的 HV 比例增加。酵母提取物的浓度也会影响 HV 的比例。此外,当用丙酮酸、琥珀酸钠或葡萄糖酸钠代替葡萄糖时,重组菌株中仅检测到 PHB。