Three diacylglycerol acyltransferases contribute to oil biosynthesis and normal growth in Yarrowia lipolytica
Three diacylglycerol acyltransferases contribute to oil biosynthesis and normal growth in Yarrowia lipolytica
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DOI：
10.1002/yea.1914
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发表时间：
2012-01
期刊：
影响因子：
2.6
通讯作者：
Hongxiang Zhang;H. Damude;N. Yadav
中科院分区：
文献类型：
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作者：
Hongxiang Zhang;H. Damude;N. Yadav
甘油二酯酰基转移酶（DAG）催化甘油三酯生物合成的最后一步。真核生物通常含有多达三种不同类别的DAG酰基转移酶：酰基-CoA：二酰基甘油酰基转移酶1（DGAT 1）、酰基-CoA：二酰基甘油酰基转移酶2（DGAT 2）和磷脂：二酰基甘油酰基转移酶（PDAT）。非传统的产油酵母，解脂耶氏酵母，含有每类的至少一个同源物，并作为一个很好的模型来理解不同的DAG酰基转移酶在油的生物合成中的作用，油是一种关键的细胞组分，作为储存分子以及游离脂肪酸的缓冲液。我们在Y染色体上使用了基因突变。通过在lipolytica和体外酶测定中进行比较，以确认编码所有三种DAG酰基转移酶的基因的身份，并证明它们一起解释几乎所有的油生物合成，并且所有三种都显著地促进其油生物合成。耶氏酵母中对于解脂菌ATCC 20362菌株，在pdat、dgat 1、dgat 2、dgat 1/dgat 2双突变体和dgat 1/dgat 2/pdat三突变体中，作为野生型菌株的百分比的总脂质%干细胞重量（DCW）分别为70%、57%、36%、18%和13%。三重突变体在增加的滞后期和较慢的生长速率中显示出显著的生长缺陷，表明油生物合成有助于该菌株的正常生长。版权所有© 2011约翰威利父子有限公司.
Diacylglycerol (DAG) acyltransferase catalyses the final and committed step of triacylglycerol biosynthesis. Eukaryotes commonly contain up to three distinct classes of DAG acyltransferases: acyl‐CoA:diacylglycerol acyltransferase 1 (DGAT1), acyl‐CoA:diacylglycerol acyltransferase 2 (DGAT2), and phospholipid:diacylglycerol acyltransferase (PDAT). The non‐conventional oleaginous yeast, Yarrowia lipolytica, contains at least one homologue of each class and serves as a good model to understand the role of different DAG acyltransferases in the biosynthesis of oil, a critical cellular component that serves as a storage molecule as well as a buffer for free fatty acids. We used gene disruptions in Y. lipolytica and in vitro enzyme assays to confirm the identity of genes encoding all three DAG acyltransferases and demonstrate that together they account for almost all oil biosynthesis and that all three contribute significantly to its oil biosynthesis. In Y. lipolytica ATCC 20362 strain, the total lipid% dry cell weight (DCW) as a percentage of the wild‐type strain in pdat, dgat1, dgat2, dgat1/dgat2 double mutant and dgat1/dgat2/pdat triple mutant was 70%, 57%, 36%, 18% and 13%, respectively.This is the first example of DGAT1 contributing significantly to oil biosynthesis in a microorganism. The triple mutant shows significant growth defect in both increased lag phase and slower growth rate, suggesting that oil biosynthesis contributes to normal growth in this strain. Copyright © 2011 John Wiley & Sons, Ltd.