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| 灵芝锰过氧化物酶毕赤酵母工程菌培养条件的优化 |
| 徐慧1,郭梦圆1,靳通1,高艳华1,路慧丽2,李伟3,周选围1,2* |
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(1. 上海交通大学农业与生物学院植物生物技术研究中心, 上海 200240;
2. 上海交通大学药学院细胞工程及抗体药物教育部工程研究中心, 上海 200240;
3. 上海交通大学上海系统生物医学研究中心, 上海 200240
*通信作者) |
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| 摘要: |
| 目的 研究毕赤酵母生产重组灵芝锰过氧化物酶（recombinant Ganoderma lucidum manganese peroxidase，rGlMnP）的最优培养条件。方法 在单因素实验基础上，采用中心组合设计和响应曲面法对rGlMnP工程酵母产rGlMnP的发酵条件进行优化，探讨初始pH、温度、甲醇浓度和血红素浓度对重组毕赤酵母生产的rGlMnP的酶活力的影响，分析上述4个因子的交互作用及其最佳水平范围，并验证模型的有效性。结果 初始pH、温度、甲醇浓度和血红素浓度对毕赤酵母生产rGlMnP的酶活力有显著影响；毕赤酵母生产rGlMnP最优的培养条件为：初始pH 5.5、诱导温度30℃、甲醇浓度1.5%以及血红素浓度1.0 mmol/L。结论 本研究为利用毕赤酵母工程菌大规模生产rGlMnP奠定了基础。 |
| 关键词: 灵芝 锰过氧化物酶 毕赤酵母 优化 响应曲面法 |
| DOI：10.16781/j.0258-879x.2016.08.0955 |
| 投稿时间：2016-01-31修订日期：2016-03-19 |
| 基金项目:国家自然科学基金（30771500）. |
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| Optimization of fermentation conditions of recombinant Pichia pastoris for the production of Ganoderma lucidum manganese peroxidase |
| XU Hui1,GUO Meng-yuan1,JIN Tong1,GAO Yan-hua1,LU Hui-li2,LI Wei3,ZHOU Xuan-wei1,2* |
(1. Plant Biotechnology Research Center, Shanghai Jiaotong University School of Agriculture and Biology, Shanghai 200240, China;
2. Engineering Research Center of Cell and Therapeutic Antibody of Ministry of Education, Shanghai Jiaotong University School of Pharmacy, Shanghai 200240, China;
3. Shanghai Center for System Biomedicine, Shanghai Jiaotong University, Shanghai 200240, China
*Corresponding author) |
| Abstract: |
| Objective To search for the optimal fermentation conditions for producing recombinant Ganoderma lucidum manganese peroxidase (rGlMnP) by Pichia pastoris. Methods Based on the single factorial experiment, an optimum fermentation condition for rGlMnP production by recombinant Pichia pastoris was obtained through a central composite design (CCD) and response surface methodology (RSM) in the present work. The effects of initial pH, temperature, concentrations of methanol and heme on the enzyme activity of rGlMnP produced by Pichia pastoris were explored. The interactive effects and the optimal range levels of the 4 factors were studied and the validity of the model was also verified. Results The results showed that initial pH, temperature, the concentrations of methanol and heme had significant effects on the enzymic activity. The optimum conditions for enzymic activity of rGlMnP produced by Pichia pastoris were as follows: initial pH 5.5,temperature 30℃, concentration of methanol 1.5% and concentration of heme 1.0 mmol/L. Conclusion This study has laid a foundation for producing rGlMnP by Pichia pastoris in industrial scale. |
| Key words: Ganoderma lucidum manganese peroxidase Pichia pastoris optimization response surface methodology |
