透明颤菌血红蛋白基因在产TRAIL蛋白大肠杆菌中的克隆表达

Cloning and Expression of Vitreoscilla Hemoglobin in Escherichia coli for TRAIL Production

摘要: 为解决大肠杆菌高密度发酵生产TRA IL过程中的供氧矛盾,提高菌体的发酵密度,在大肠杆菌中引入透明颤菌血红蛋白基因(vgb)。聚合酶链反应(PCR)检验和CO差光谱法检验表明:vgb基因能够在C 600(pBV 220-TRA IL)中表达并受溶氧调控,重组菌CTV具有良好的遗传稳定性。3.7 L发酵罐实验表明:在同样的发酵条件下,含vgb基因的重组菌CTV的最高菌体密度为对照菌的1.64倍,达到50.6 g/L,乙酸积累为对照菌的55.6%,TRA IL蛋白产量提高了70%,达到2.8 g/L。

Abstract: Vitreoscilla hemoglobin gene(vgb) was cloned and expressed in Escherichia coli C600(pBV220-TRAIL) for improving the oxygen uptake rate(OUR) and cell density.PCR reactions and carbon monoxide different spectra analysis showed that vgb was successfully expressed in C600(pBV220-TRAIL),and this expression system was regulated by dissolved oxygen(DO).Recombinant engineering strain CTV maintained genetic stability well.Under the same culture conditions,the highest cell density of CTV(50.6 g/L) and the yield of TRAIL(2.8 g/L)was 1.64-fold and 1.7-fold higher than that of control in 3.7 L bioreactor,respectively,whereas the acetic acid concentration was decreased by 45%.