人可溶性TRAIL蛋白表达及纯化条件优化

摘要

Objective To optimize the production and purification conditions of human soluble TNF -related apop-tosis inducing ligand ( TRAIL) , thereby increasing the productivity and activity of TRAIL .Methods ①6 different me-dium, pH, the induced concentration and time of isopropyl β-D-1-thiogalactopyranoside ( IPTG) were investigated and the optimum fermentation conditions were selected .②Better human soluble TRAIL protein was obtained through the combination of nickel ion affinity chromatography and ultrafiltration .Results ①The optimum fermentation conditions for human soluble TRAIL was YPD medium at pH 7.4.The optimum concentration of IPTG used was 0.2 mmol/L, the best time was 6 hours.②Through nickel ion affinity chromatography and ultrafiltration combination , higher effects and more purity protein could be obtained .Conclusion The fermentation conditions for human soluble TRAIL are successfully op-timized, which lays foundation for future extraction of large amount of TRAIL protein .%目的：优化人可溶性TRAIL蛋白生产菌发酵及纯化条件，从而提高TRAIL的产量及活性。方法①探索6种不同培养液、培养液pH值、异丙基硫代半乳糖苷（ IPTG）诱导浓度及时间，选择最佳发酵条件；②联合镍离子亲和层析及超滤法纯化人可溶性TRAIL蛋白。结果①人可溶性TRAIL的最佳发酵条件为pH7．4的YPD培养液，所用IPTG的最佳浓度为0．2 mmol/L，最佳诱导时间为6 h；②经镍离子亲和层析和超滤联合作用可得纯度极高的蛋白。结论成功完成了人可溶性TRAIL发酵及纯化条件的优化，为今后大量提取TRAIL蛋白奠定了基础。