文献类型：期刊文献

英文题名：Optimization of culture on the overproduction of TRAIL in high-cell-density culture by recombinant Escherichia coli

作者：Luo, Qingping[1]; Shen, Ya-Ling[1]; Wei, Dong-Zhi[1]; Cao, Wei[1]

机构：[1] State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, 130 Meilong Road, Shanghai, 200237, China

年份：2006

卷号：71

期号：2

起止页码：184

外文期刊名：Applied Microbiology and Biotechnology

收录：EI(收录号：2006259945716)

语种：英文

外文关键词：Bacteriology - Cell culture - Glucose - Glycerol - pH effects - Proteins - Tumors

摘要：Different nutrient-feeding cultures were carried out in producing recombinant protein of truncated tumor necrosis factor related apoptosis-inducing ligand (TRAIL) (114-281 amino acids of TRAIL) in Escherichia coli strain C600/pBV-TRAIL. The effects of preinduction specific growth rate, postinduction carbon source (glucose and glycerol), and feeding strategies were investigated. The higher preinduction specific growth rate (μ=0.22 h -1) contributed to the increase in the TRAIL production, at which TRAIL was accumulated in bacterial cells as 7.2% of total cellular protein, corresponding to 1.99 g l-1 in contrast with 5.1% (1.29 g l -1) at preinduction specific growth rate (μ=0.1 h-1) during high-cell-density culture. Glycerol was superior to glucose as the postinduction carbon source for TRAIL production. Under similar culture conditions, the final concentration of TRAIL was produced 1.59-fold more when glycerol was used as postinduction carbon source than when glucose was used. At the same time, the results showed that it is efficient to adopt the pH-stat feeding strategy at postinduction for the overproduction of TRAIL. The TRAIL production was increased up to 4.51 g l-1, approximately 16.1% of total cellular protein. The mechanisms behind the preinduction specific growth rate effect on the expression level may be ascribed to the leakage secretion of acetate. ? Springer-Verlag 2005.