| 枯草芽孢杆菌(Bacillus subtilis)碱性脂肪酶基因在毕赤酵母中的表达 |
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| 引用本文： | 戴苏,赵小兰,林影,韩双艳.枯草芽孢杆菌(Bacillus subtilis)碱性脂肪酶基因在毕赤酵母中的表达[J].生物技术通报,2012,0(3):135-140. |
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| 作者姓名： | 戴苏 赵小兰 林影 韩双艳 |
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| 作者单位： | 华南理工大学生物科学与工程学院,广州,510006 |
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| 摘 要： | 将来自枯草芽孢杆菌的碱性脂肪酶基因经密码子优化,全基因合成后克隆到pPICZαA载体,构建了pPICZαA-bsl分泌型重组质粒,该重组质粒经限制性内切酶PmeI线性化后使用LiCl法转化到毕赤酵母X-33,经过筛选获得分泌表达碱性脂肪酶的重组毕赤酵母X-33/pPICZαA-bsl。摇瓶发酵液上清酶活最高可达4.78 U/mL,初步研究了该脂肪酶的酶学性质,其最适作用温度为40-60℃,最适pH9.0,且具有高度耐碱的特性。该重组脂肪酶对旧新闻纸具备较明显的脱墨能力。
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| 关 键 词： | 枯草芽孢杆菌 碱性脂肪酶 密码子优化 毕赤酵母 分泌表达 脱墨 |
Expression of Bacillus subtilis Alkaline Lipase Gene in Pichia pastoris |
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| Dai Su
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Zhao Xiaolan
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Lin Ying
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Han Shuangyan.Expression of Bacillus subtilis Alkaline Lipase Gene in Pichia pastoris[J].Biotechnology Bulletin,2012,0(3):135-140. |
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| Authors: | Dai Su Zhao Xiaolan Lin Ying Han Shuangyan |
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| Institution: | Dai Su Zhao Xiaolan Lin Ying Han Shuangyan (School of Bioscience and Bioengineering,South China University of Technology,Guangzhou 510006) |
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| Abstract: | The aim of the present study was to express and evaluate a previously cloned lipase gene.The alkaline lipase gene was optimized and subcloned into the secreted expression vector pPICZαA to construction a recombinant plasmid pPICZαA-bsl,then the recombinant plasmid was transformated into the Pichia pastoris X33.The recombinant yeast x33/pPICZαA-bsl could secrete alkaline lipase,and the maximum activity of the culture supernatant was 4.78 U/mL when the recombinant strain was cultured on a rotary shaker.The properties of the recombinant lipase was be tentatively researched,the results showed that the optimal temperature and pH of this lipase were 40-60℃ and 9.0,and the lipase was extreme alkaline-resistant.The recombinant lipase showed obvious deinking ability in old news paper deinking test. |
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| Keywords: | Bacillus subtilis Alkaline lipase Codon optimization Pichia pastoris Secreted expression Deinking |
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