| 克氏担孢酵母脂肪酶基因在毕赤酵母中的高效表达 |
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| 引用本文： | 王建荣,刘丹妮,李鹏,李阳源.克氏担孢酵母脂肪酶基因在毕赤酵母中的高效表达[J].食品科学,2015,36(3):104-108. |
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| 作者姓名： | 王建荣 刘丹妮 李鹏 李阳源 |
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| 作者单位： | 广东溢多利生物科技股份有限公司，广东 珠海 519060 |
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| 基金项目： | 国家高技术研究发展计划(863计划)项目(2014AA093514) |
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| 摘 要： | 以克氏担孢酵母V3脂肪酶基因为研究对象，构建毕赤酵母稳定的高效表达系统。采用同源克隆法从克氏担孢酵母V3中获得一个脂肪酶基因，并将其构建到表达载体pPICZαA，转入毕赤酵母X33中，成功实现了克氏担孢酵母V3脂肪酶基因在毕赤酵母X33中的表达。重组工程菌摇瓶培养120 h后，酶活力可达18 U/mL。重组酶的最适反应温度为60 ℃，最适pH值为5.0。1 mmol/L的Mg2+、K+和Na+以及质量浓度为1 g/100 mL的曲通-100、吐温-80、吐温-20对重组KLIP具有激活作用。重组酶的最适底物为三硬脂酸甘油酯（C18）。
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| 关 键 词： | 克氏担孢酵母 脂肪酶 毕赤酵母 |
Overexpression of the Lipase Gene from Kurtzmanomyces in Pichia pastoris |
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| WANG Jianrong;LIU Danni;LI Peng;LI Yangyuan.Overexpression of the Lipase Gene from Kurtzmanomyces in Pichia pastoris[J].Food Science,2015,36(3):104-108. |
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| Authors: | WANG Jianrong;LIU Danni;LI Peng;LI Yangyuan |
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| Affiliation: | Guangdong VTR Bio-tech Co. Ltd., Zhuhai 519060, China |
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| Abstract: | A stable and highly effective expression system was constructed for the expression of the lipase gene from
Kurtzmanomyces sp. V3. The lipase gene was isolated by homologous cloning, inserted into the expression vector pPICZαA
and transformed into Pichia pastoris X33. The results showed that the gene was expressed successfully in Pichia pastoris X33.
The optimal temperature and pH of the recombinant lipase were 60 ℃ and 5.0, respectively. Its activity was stimulated by
1 mmol/L Mg2+, K+ and Na+, and 1 g/100 mL Triton-100, Tween-80 and Tween-20. The recombinant lipase showed high
activity toward tristearin (C18). |
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| Keywords: | Kurtzmanomyces lipase Pichia pastoris |
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