| 重组毕赤酵母高密度发酵生产碱性果胶酶的策略 |
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| 引用本文： | 王芸,华兆哲,刘立明,张朝晖,堵国成,陈坚.重组毕赤酵母高密度发酵生产碱性果胶酶的策略[J].微生物学报,2008,24(4):635-639. |
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| 作者姓名： | 王芸 华兆哲 刘立明 张朝晖 堵国成 陈坚 |
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| 作者单位： | 江南大学工业生物技术教育部重点实验室, 无锡 214122;江南大学工业生物技术教育部重点实验室, 无锡 214122;江南大学工业生物技术教育部重点实验室, 无锡 214122;浙江工业大学生物与环境工程学院, 杭州 310014;江南大学工业生物技术教育部重点实验室, 无锡 214122;江南大学工业生物技术教育部重点实验室, 无锡 214122; 江南大学食品科学技术国家重点实验室, 无锡 214122 |
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| 基金项目： | 国家杰出青年基金(No. 20625619)、国家自然科学基金 (973计划) (Nos. 2007CB714306, 2007CB714303)和浙江省重点科技项目(No. 2006C21117)资助。 |
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| 摘 要： | 重组Pichia pastoris GS115表达碱性果胶酶的诱导阶段, 最佳初始菌体浓度和甲醇诱导浓度分别为122 g/L和20 g/L, 两者之间最佳比值范围是0.16~0.20 g/g (甲醇/菌体浓度). 在此基础上通过生长阶段甘油的指数流加, 以及诱导阶段基于甲醇比消耗速率和溶氧等参数进行甲醇流加的方式, 将甲醇与菌体浓度比例控制在0.171~0.195 g/g之间. 此时, 酶活达到430 u/mL, 生产强度为4.34 u/mL/h, 实现了碱性果胶酶高效生产。
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| 关 键 词： | 重组毕赤酵母 碱性果胶酶 甲醇与菌体比例 流加策略 |
High-level Production of Alkaline Polygalacturonate Lyase in Recombinant Pichia pastoris |
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| Yun Wang,Zhaozhe Hu,Liming Liu,Zhaohui Zhang,Guocheng Du and Jian Chen.High-level Production of Alkaline Polygalacturonate Lyase in Recombinant Pichia pastoris[J].Acta Microbiologica Sinica,2008,24(4):635-639. |
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| Authors: | Yun Wang Zhaozhe Hu Liming Liu Zhaohui Zhang Guocheng Du and Jian Chen |
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| Institution: | Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, China;Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, China;Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, China;College of Biological & Environmental Engineering, Zhejiang University of Technology, Hangzhou 310014, China;Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, China;Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, China; State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi 214122, China |
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| Abstract: | In order to increase the production of alkaline polygalacturonate lyase (PGL) by recombinant Pichia pastoris GS115, the effect of cell and methanol concentration on the PGL production was carefully investigated by single factor experiment. The optimum conditions were listed as follows: the cell concentration 122 g/L, the methanol concentration 20 g/L, and the ratio of methanol and cell concentration 0.16~0.20 g/g (methanol/cell). With the glycerol and methanol feeding strategies, the ratio of methanol and cell concentration could be controlled at the range of 0.171 to 0.195 g/g. And the highest PGL activity (430 u/mL) and highest PGL productivity (4.34 u/mL/h) were achieved. |
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| Keywords: | Pichia pastoris alkaline polygalacturonate lyase ratio of methanol and cell concentration feeding strategy |
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