重组人源白介素-1β发酵条件的优化及其产品的纯化

Optimization of Fermentation Conditions and Purification of rhIL-1β

目的初步建立重组人源IL-1β巴斯德毕赤酵母的培养体系;进一步优化大规模培养重组人源IL-1β巴斯德毕赤酵母的条件;探讨大规模纯化重组人源IL-1β巴斯德毕赤酵母表达产物的条件;鉴定酵母表达产物及纯化样品。方法在菌体湿重达190g.L-1左右时开始甲醇诱导表达;选用了SP Sepharose XL阳离子交换层析和反相疏水柱层析进行纯化。结果在pH4.5的含0.5%蛋白胨的FM21培养基中发酵时,放罐时间在30h较为适宜;经过纯化蛋白纯度即可达95%以上。结论最终经过鉴定,表达和纯化的样品就是我们想要得到的IL-1β。

OBJECTIVE To set up the cultivation system of rhIL-1β Pichia pastoris yeast initially）optimize conditions of large-scale culture of rhIL-1β Pichia pastoris yeast further）discuss purification conditions of rhIL-1β Pichia pastoris yeast） and identify the expression and purification product of yeast. METHODS To induce expression by methanol when wet weight arrive 190g· L^-1. Purification with SP Sepharose XL cation-exchange chromatography. RESULTS Harvest time was 30h when to cultivate rhIL-1β Pichia pastoris yeast in FM21 medium with 0. 5% peptone and pH4.5. Through the purification, the purity of protein can be above 955. CONCLUSION After expressing and purifying, we identified finally,that the product was just IL- 1β.