| 产胆固醇酯酶毕赤酵母发酵条件优化及酶学性质研究 |
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| 引用本文： | 权浩严,王鹏,位正鹏,杜春影,沈照鹏,张京良,乔乐克.产胆固醇酯酶毕赤酵母发酵条件优化及酶学性质研究[J].食品工业科技,2021,42(17):94-99. |
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| 作者姓名： | 权浩严 王鹏 位正鹏 杜春影 沈照鹏 张京良 乔乐克 |
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| 作者单位： | 1.中国海洋大学食品科学与工程学院，山东青岛 2660032.青岛市市立医院，山东青岛 2660113.海洋功能食品国家地方联合工程实验室，山东荣成 2643094.青岛海洋生物医药研究院，山东青岛 266071 |
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| 基金项目： | 海洋经济创新发展示范城市产业链协同创新类项目（威海）牡蛎活性肽系列产品开发及产业链构建（2019WHZ008）；舟山市长江生态环境保护修复驻点跟踪研究项目（SZGXZS2020068） |
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| 摘 要： | 为实现胆固醇酯酶的高效发酵，以成功表达的毕赤酵母工程菌株P.pastoris X-33为研究对象，通过单因素和正交试验对其发酵条件进行优化。结果表明:最适培养基组成为:酵母浸粉含量1.0%、蛋白胨浓度1.5%、甘油含量1.0%、KH2PO4 1.18%、K2HPO4 0.3%、生物素4×10-5%、YNB 1.34%；在初始pH7.0，30 ℃发酵96 h条件下，酶活力可达11.21 U/mL，较于优化前提高了3.65倍。酶学性质结果显示，该酶最适pH和最适反应温度分别为8.0和50 ℃，Zn2+对酶有较强的抑制性。该胆固醇酯酶酶学性质较为稳定，可为今后的规模化应用提供技术基础。
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| 关 键 词： | 毕赤酵母 胆固醇酯酶 发酵条件 正交优化 酶学性质 |
| 收稿时间： | 2020-11-11 |
Optimization of Fermentation Conditions and Enzymatic Characteristics of the Cholesterol Esterase from Pichia pastoris |
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| QUAN Haoyan,WANG Peng,WEI Zhengpeng,DU Chunying,SHEN Zhaopeng,ZHANG Jingliang,QIAO Leke.Optimization of Fermentation Conditions and Enzymatic Characteristics of the Cholesterol Esterase from Pichia pastoris[J].Science and Technology of Food Industry,2021,42(17):94-99. |
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| Authors: | QUAN Haoyan WANG Peng WEI Zhengpeng DU Chunying SHEN Zhaopeng ZHANG Jingliang QIAO Leke |
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| Affiliation: | 1.College of Food Science and Engineering, Ocean University of China, Qingdao 266003, China2.Qingdao Municipal Hospital, Qingdao 266011, China3.National and Local United Engineering Lab of Marine Functional Food, Rongcheng 264309, China4.Marine Biomedical Research Institute of Qingdao, Qingdao 266071, China |
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| Abstract: | In order to achieve the high-efficiency fermentation of cholesterol esterase, Pichia pastoris was selected as the research object, and its fermentation conditions were optimized by single factor and orthogonal experiments. The results showed that the optimum medium was yeast extract 1.0%, peptone 1.5%, glycerin 1.0%, KH2PO4 1.18%, K2HPO4 0.3%, biotin 4×10-5%, YNB 1.34%. Under the condition of initial pH7.0, fermentation temperature 30 ℃, fermentation time 96 h, the enzyme activity could reach 11.21 U/mL. And it was 3.65 times higher than before. The results of enzymatic properties showed that the optimum pH and reaction temperature of the enzyme were 8.0 and 50 ℃, respectively. Zn2+ had a strong inhibition on the enzyme. The cholesterol esterase had stable enzymatic properties and could provide technical basis for large-scale production in the future. |
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