科微学术
生物工程学报
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。
- Weina Li
Weina Li
1 Shaanxi Key Laboratory of Degradable Biomedical Materials, School of Chemical Engineering, Northwest University, Xi’an 710069, Shaanxi, China; 2 Shaanxi R&D Center of Biomaterials and Fermentation Engineering, College of Chemical Engineering, Northwest University, Xian 710069, Shaanxi, China
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在本站中查找 - Zifang Shang
Zifang Shang
1 Shaanxi Key Laboratory of Degradable Biomedical Materials, School of Chemical Engineering, Northwest University, Xi’an 710069, Shaanxi, China; 2 Shaanxi R&D Center of Biomaterials and Fermentation Engineering, College of Chemical Engineering, Northwest University, Xian 710069, Shaanxi, China
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在本站中查找 - Zhiguang Duan
Zhiguang Duan
1 Shaanxi Key Laboratory of Degradable Biomedical Materials, School of Chemical Engineering, Northwest University, Xi’an 710069, Shaanxi, China; 2 Shaanxi R&D Center of Biomaterials and Fermentation Engineering, College of Chemical Engineering, Northwest University, Xian 710069, Shaanxi, China
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在本站中查找 - Linbo Li
Linbo Li
1 Shaanxi Key Laboratory of Degradable Biomedical Materials, School of Chemical Engineering, Northwest University, Xi’an 710069, Shaanxi, China; 2 Shaanxi R&D Center of Biomaterials and Fermentation Engineering, College of Chemical Engineering, Northwest University, Xian 710069, Shaanxi, China
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在本站中查找 - Jing He
Jing He
1 Shaanxi Key Laboratory of Degradable Biomedical Materials, School of Chemical Engineering, Northwest University, Xi’an 710069, Shaanxi, China; 2 Shaanxi R&D Center of Biomaterials and Fermentation Engineering, College of Chemical Engineering, Northwest University, Xian 710069, Shaanxi, China
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在本站中查找 - Daidi Fan
Daidi Fan
1 Shaanxi Key Laboratory of Degradable Biomedical Materials, School of Chemical Engineering, Northwest University, Xi’an 710069, Shaanxi, China; 2 Shaanxi R&D Center of Biomaterials and Fermentation Engineering, College of Chemical Engineering, Northwest University, Xian 710069, Shaanxi, China
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Educational Commission of Shaanxi Province of China (No. 16JS104), China Postdoctoral Science Foundation (No. 2015M582698), Scientific Research Foundation in Northwest University (No. 338050025).
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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。
To improve collagen production by recombinant Pichia pastoris, we applied Placket-Burman and Box-Behnken design to optimize the fermentation medium. Through Placket-Burman design, three variables in the medium (concentration of yeast extract, peptone and glycerol) were selected for having significant effect on cell dry weight. Through Box-Behnken design regression coefficients analysis, a secondary degree polynomial equation was established, and the optimum levels of the three variables were yeast extract 1.13%, peptone 1.61% and glycerol 0.86%. During the growth period, an average cell dry weight of 4.41 g/L was obtained after 12 h fermentation, increased by 26%. Through high density fermentation, the production of recombinant human collagen (Ⅲ) was up to 4.71 g/L in 22 L fermentor. The recombinant human collagen (Ⅲ) exhibited good results to repair acetic acid induced gastric ulcer in rats.
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李伟娜,尚子方,段志广,李林波,贺婧,范代娣. 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[J]. 生物工程学报, 2017, 33(4): 672-682
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- 收稿日期:2016-10-14
- 在线发布日期: 2017-03-31
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