毕赤酵母高密度发酵产Ⅲ型类人胶原蛋白及其胃粘膜修复功能

发布时间：2019-07-22 16:39

【摘要】：为提高重组毕赤酵母Pichia pastoris发酵重组人Ⅲ型胶原蛋白产量,采用响应面对其生长阶段的BMGY培养基(Buffered minimal glycerol-complex medium)组成进行优化。通过Placket-Burman试验,得出酵母提取物、蛋白胨、甘油对胶原蛋白产量有显著影响,通过Box-Behnken中心组合实验以及Design-Expert分析软件建立了以胶原蛋白产量为响应值的响应方程,得到最适浓度分别为酵母提取物1.13%,蛋白胨1.61%,甘油0.86%。经实验验证,在最优BMGY培养基条件下培养12 h后,所得到毕赤酵母干重为4.41 g/L,生长阶段的菌重量增加了26%。在22 L发酵罐中通过高密度发酵,产量达到4.71 g/L。该重组胶原蛋白对大鼠乙酸灼伤胃粘膜有明显修复作用,为生物医用材料的应用制备奠定了基础。
[Abstract]:In order to increase the yield of recombinant human type III collagen fermented by recombinant Pichia pastoris, the composition of BMGY medium (Buffered minimal glycerol-complex medium in response to the growth stage was optimized. Through Placket-Burman test, it was concluded that yeast extract, peptone and glycerol had significant effects on collagen production. The response equation with collagen yield as response value was established by Box-Behnken central combination experiment and Design-Expert analysis software. The optimum concentrations were 1.13%, 1.61% and 0.86%, respectively. The results showed that the dry weight of Pichia pastoris was 4.41 g 路L ~ (- 1) and the weight of Pichia pastoris increased by 26% after 12 h culture in the optimal BMGY medium. Through high density fermentation in 22 L fermentor, the yield reached 4.71 g 路L ~ (- 1) 路L ~ (- 1). The recombinant collagen can repair the gastric mucosa of rats with acetic acid burn, which lays a foundation for the preparation of biomedical materials.
【作者单位】：西北大学化工学院陕西省可降解生物医用材料重点实验室;西北大学化工学院陕西省生物材料与发酵工程技术研究中心;
【基金】：陕西省教育厅重点实验室科研计划项目(No.16JS104) 博士后科学基金面上项目(No.2015M582698) 西北大学科研启动金(No.338050025)资助~~
【分类号】：TQ931;R318.08

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