共轭亚油酸合成相关基因在耶氏解脂酵母中异源表达及活性研究
Heterologous expression and activity of the genes related to CLA synthesis in Yarrowia lipolytica
  
DOI:
中文关键词:  亚油酸  共轭亚油酸  耶氏解脂酵母  异源表达
英文关键词:linoleic acid  conjugated linoleic acid  Yarrowia lipolytica  heterologous expression
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Author NameAffiliation
LIU Yunyun, YANG Bo, ZHANG Hao,etc  
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中文摘要:
      为了探究植物乳杆菌(Lactobacillus plantarum)中与共轭亚油酸(CLA)生物合成相关的3个基因:(亚)油酸水合酶基因(mcra)、短链脱氢酶/氧化还原酶基因(dh)、乙酰乙酸脱羧酶基因(dc)在耶氏解脂酵母(Yarrowia lipolytica,Y. lipolytica)中异源表达后能否具有活性,利用两个耶氏解脂酵母整合表达质粒(pINA 1269和pINA 1312),将3个基因分别导入耶氏解脂酵母营养缺陷型宿主菌Polf(Ura-,Leu-)中,构建了重组菌株。在不同重组菌中添加相应的底物:亚油酸(LA)和10-羟基-顺12-十八碳烯酸(10-HOE),然后对反应体系进行脂肪酸检测,得到基因对应的不同产物:10-HOE和10-氧代-反11-十八碳烯酸(10-oxo-trans 11-octadecenoic acid),证明mcra、dh、dc在耶氏解脂酵母中进行了异源表达并且具有活性。
英文摘要:
      To explore whether three genes involved in conjugated linoleic acid (CLA) biosynthesis in Lactobacillus plantarum(Myosin cross reactive antigen(mcra), Short-chain dehydrogenase/oxidoreductase(dh) and Acetoacetate decarboxylase(dc)) were active after heterologously expressed in Yarrowia lipolytica (Y. lipolytica), with two Y. lipolytica expression plasmids (pINA 1269 and pINA 1312), the three genes were introduced into Y. lipolytica auxotrophic host Polf (Ura-, Leu-), and finally the recombinant strains were constructed. The corresponding substrates linoleic acid (LA) and 10-hydroxy-cis-12-octadecenoic acid (10-HOE) were added into different recombinant strains, and the fatty acids were detected in the reaction system to obtain different products corresponding to the genes: 10-HOE and 10-oxo-trans 11-octadecenoic acid, demonstrating that mcra, dh and dc were heterologously expressed and were active in Y. lipolytica.
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