华南农业大学学报2011，Vol.32Issue(1)：103-107,5.
重组毕赤酵母高密度发酵表达猪表皮生长因子及其纯化鉴定
Expression, Purification and Bioactivity Analysis of Porcine Epidermal Growth Factor in Recombinant Pichia pastoris by High-Density Fermentation
摘要
Abstract
The recombinant Pichia pastoris strain infected with pPIC9-pEGF/GS115 for expression of strain porcine epidermal growth factor was high cell density cultured in a 5.5 L fermentor. Following prefermentation and reaching D600nm density to 150, the yeast cells were induced by addition of methanol,which elevated cell density D600nm to 315. As the result, the medium concentration of secreted pEGF reached 300 mg/L. The pEGF fermentation product was purified to 90％ by ultrafiltration, His-tag affinity column chromatography and dialysis. In vitro biological activity test results showed that the expressed pEGF fusion protein could enhance significantly the proliferation of BALB/c 3T3 cells. The above results demonstrated successful preparation of biologically active pEGF by the P. pastoris eukaryotic expression system.关键词
猪表皮生长因子/毕赤酵母/高密度发酵/纯化/生物活性Key words
porcine epidermal growth factor/ Pichia pastoris/ high density culture/ purification/ bioactivity分类
农业科技引用本文复制引用
焦晓军,黄显会,刘帅帅,谭清甜..重组毕赤酵母高密度发酵表达猪表皮生长因子及其纯化鉴定[J].华南农业大学学报,2011,32(1):103-107,5.基金项目
国家自然科学基金(30771627) （30771627）
