| 采用混合策略提高葡萄糖氧化酶在毕赤酵母中的表达水平 |
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| 引用本文： | 牟庆璇,胡美荣,陈飞,江贤章,陶勇,黄建忠. 采用混合策略提高葡萄糖氧化酶在毕赤酵母中的表达水平[J]. 生物工程学报, 2016, 32(7): 986-990 |
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| 作者姓名： | 牟庆璇 胡美荣 陈飞 江贤章 陶勇 黄建忠 |
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| 作者单位： | 1 福建师范大学 生命科学学院，福建 福州 350108,2 中国科学院微生物研究所，北京 100101,1 福建师范大学 生命科学学院，福建 福州 350108,1 福建师范大学 生命科学学院，福建 福州 350108,2 中国科学院微生物研究所，北京 100101,1 福建师范大学 生命科学学院，福建 福州 350108 |
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| 基金项目： | 福建省产业化关键技术项目 (No. 闽财指[2010]358号) 资助。 |
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| 摘 要： | 为了提高葡萄糖氧化酶 (GOD) 在毕赤酵母中的表达水平，提出了甲醇/山梨醇混合碳源诱导和共表达分子伴侣二硫键异构酶 (PDI) 和透明颤菌血红蛋白 (VHb) 两种策略。利用对照菌株X33/pPIC9k–GOD 在5 L发酵罐放大培养时，采用甲醇/山梨醇混合碳源诱导，GOD最终酶活为456 U/mL，比只采用甲醇作为单一碳源诱导时GOD最终酶活提高了20%。利用整合伴侣蛋白菌株X33/pPIC9k-GOD/pPICZ-PDI-VHb在5 L发酵罐进行高密度发酵，采用甲醇/山梨醇混合碳源诱导，GOD最终酶活达到716 U/mL，蛋白浓度为7.4 g/L。研究结果对提高外源蛋白在毕赤酵母中的表达有重要参考价值。
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| 关 键 词： | 毕赤酵母，葡萄糖氧化酶，二硫键异构酶，透明颤菌血红蛋白酶，山梨醇 |
| 收稿时间： | 2015-10-23 |
High-level production of glucose oxidase by recombinant pichia pastoris using a combined strategy |
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| Qingxuan Mu,Meirong Hu,Fei Chen,Xianzhang Jiang,Yong Tao and Jianzhong Huang. High-level production of glucose oxidase by recombinant pichia pastoris using a combined strategy[J]. Chinese journal of biotechnology, 2016, 32(7): 986-990 |
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| Authors: | Qingxuan Mu Meirong Hu Fei Chen Xianzhang Jiang Yong Tao Jianzhong Huang |
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| Affiliation: | 1 College of Life Sciences, Fujian Normal University, Fuzhou 350108, Fujian, China,2 Institute of Microbiology, Chinese Academy of Scienses, Beijing 100101, China,1 College of Life Sciences, Fujian Normal University, Fuzhou 350108, Fujian, China,1 College of Life Sciences, Fujian Normal University, Fuzhou 350108, Fujian, China,2 Institute of Microbiology, Chinese Academy of Scienses, Beijing 100101, China and 1 College of Life Sciences, Fujian Normal University, Fuzhou 350108, Fujian, China |
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| Abstract: | To enhance the production of glucose oxidase by recombinant Pichia pastoris, two strategies were developed, which were namely co-feeding of methanol and sorbitol and co-expressing of the protein disulfide isomerase (PDI) and Vitreoscialla hemoglobin (VHb). The volumetric activity reached 456 U/mL by using the strain X33/pPIC9k-GOD, in 5 liter fermentator, with the co-feeding of methanol and sorbitol, it was 0.2 fold higher than that only feeding by methanol. The improved strain was obtained by co-expressing PDI-VHb with GOD. While fermented in a 5 liter fermentator by feeding methanol and sorbitol, the activity of the improved strain reached 716 U/mL with a yield of 7 400 mg/L total soluble protein concentration. These results indicated that heterologous protein expression level can be enhanced by optimizing fermentation condition and co-expression molecular chaperon in Pichia pastoris. |
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| Keywords: | Pichia pastoris glucose oxidase protein disulfide isomerase Vitreoscialla hemoglobin sorbitol |
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