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| 裂殖壶菌脂肪酶基因克隆及底物特异性研究 |
| Gene cloning of lipase from Schizochytrium sp.and substrate specificity analysis |
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| DOI： |
| 中文关键词: 裂殖壶菌 脂肪酶基因 毕赤酵母 底物特异性 |
| 英文关键词:Schizochytrium sp. lipase gene Pichia pastoris substrate specificity |
| 基金项目: |
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| 中文摘要: |
| 裂殖壶菌甘油三酯合成和水解的过程中，脂肪酶起着重要的作用。根据同源性分析结果，筛选得到裂殖壶菌脂肪酶基因STGL2。以cDNA为模板获得STGL2脂肪酶基因，将其连接到pPIC9K载体上，得到重组质粒pPIC9K- STGL2，并在毕赤酵母中表达；系统分析了重组脂肪酶（Stgl2p）的底物特异性。结果表明：STGL2脂肪酶基因在毕赤酵母中成功表达；裂殖壶菌脂肪酶Stgl2p的最适底物为对硝基苯酚月桂酸酯（ p-NPL），其最适pH为7.5，最适反应温度为35?℃。综上所述，脂肪酶Stgl2p在裂殖壶菌细胞内对甘油三酯中长碳链脂肪酸的水解起着重要的作用。 |
| 英文摘要: |
| Lipases play an important role in the synthesis and hydrolysis of triglycerides in Schizochytrium sp. According to the results of homology analysis, lipase gene STGL2 from Schizochytrium sp. was screened out. The lipase gene STGL2 was obtained with cDNA as atemplate, then it was inserted into the expression vector pPIC9K to obtain recombinant plasmid pPIC9K- STGL2 and transformed into Pichia pastoris SMD1168. The substrate specificity of recombinant lipase (Stgl2p) was analyzed systematically. The results showed that the lipase gene STGL2 was successfully expressed in Pichia pastoris. The optimal substrate for lipase Stgl2p was p-nitrophenol laurate (C12),and the optimal pH and temperature were 75 and 35?℃ respectively. In conclusion, lipase of Stgl2p played an important role in the hydrolysis of medium-long chain fatty acids in triglyceride in Schizochytrium sp.. |
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