| 摘要: |
| 以目前报道油脂产量最高的解脂耶氏酵母菌株（Yarrowia lipolytica） ATCC 30162为对象，采用逆转录PCR扩增到脂肪酶编码基因Yllip1和Yllip2，编码产物分别为816和549个氨基酸。保守结构域预测表明，Yllip1包含Patatin类磷脂酶和功能未知的DUF3336结构域，而Yllip2包含lipase_3类脂肪酶结构域，且这两个蛋白都具有1~4个跨膜区域。与不同物种来源的脂肪酶同源蛋白的多序列比对表明Yllip1和Yllip2分别包含8和6个保守区域，这些生物信息学分析表明这两个来源于解脂耶氏酵母的脂肪酶作用底物可能分别为细胞内膜磷脂和酰基甘油酯。荧光定量PCR分析表明：培养基中添加油酸在短期内(6 h)诱导了这两个脂肪酶基因Yllip1和Yllip2的显著上调表达，表明它们可能参与了酵母分解利用油酸的生化过程。 |
| 关键词: 解脂耶氏酵母 脂肪酶 保守结构域 多序列比对 |
| DOI：10.14188/j.ajsh.2019.02.011 |
| 分类号:Q939.9 |
| 基金项目:中国农业科学院科技创新工程项目CAAS-ASTIP-2013-OCRI中国农业科学院科技创新工程项目(CAAS-ASTIP-2013-OCRI) |
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| Cloning of two lipase genes Yllip1 and Yllip2 from Yarrowia lipolytica ATCC 30162 and their protein sequence analysis |
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YOU Lingjie1,YE Yunyao1,2,WANG Dan1,DAI Nijie1,GONG Yangmin1
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1.Oil Crops Research Institute, Chinese Academy of Agriculture Science, Wuhan 430062, Hubei, China;2.Xiaogan High School, Xiaogan 432000, Hubei, China
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| Abstract: |
| Reverse transcription PCR was conducted to amplify two lipase genes from the yeast strain Yarrowia lipolytica ATCC 30162, which was reported to produce highest lipid yield till now. The sizes of the encoding products of these two lipase genes, Yllip1 and Yllip2, are 816 and 549 amino acids, respectively. Prediction of conserved domain suggests that Yllip1 contains a Patatin-like phospholipase and a DUF3336 conserved domain with unknown function, whereas Yllip2 contains a lipase_3 class domain. Both of these two lipase proteins have 1~4 transmembrane regions. Multiple sequence alignment of Yllip1, Yllip2 and their homologues shows that they contain 8 and 6 conserved regions, respectively. These bioinformatics analyses suggest that these two lipase proteins from the yeast Yarrowia lipolytica ATCC 30162 may have different substrates with Yllip1 using intracellular membrane phospholipids as substrate while Yllip2 using acyl glycerides as substrate. Quantitative RT-PCR analysis shows that supplementation of oleic acid in the media within 6 h induces the up-regulation of both Yllip1 and Yllip2, suggesting that these two lipases possibly participate in the biochemical process of the utilization of oleic acid. |
| Key words: Yarrowia lipolytica lipase conserved domain multiple sequence alignment |
