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Variant Amino Acid Residues Alter the Enzyme Activity of Peanut Type 2 Diacylglycerol Acyltransferases

文献类型： 外文期刊

作者： Zheng, Ling 1 ; Shockey, Jay; Bian, Fei 2 ; Chen, Gao 2 ; Shan, Lei 2 ; Li, Xinguo 2 ; Wan, Shubo 1 ; Peng, Zhenying 1 ;

作者机构： 1.Shandong Univ, Coll Life Sci, Jinan, Shandong, Peoples R China

2.Shandong Acad Agr Sci, Dept Biotech Res Ctr, Shandong Prov Key Lab Genet Improvement Ecol & Ph, Jinan, Shandong, Peoples R China

3.Shandong Acad Agr Sci, Dept Biotech Res Ctr, Shandon

关键词： diacylglycerol acyltransferase;triacylglycerol;lipid;transformation;peanut oil

期刊名称：FRONTIERS IN PLANT SCIENCE （ 影响因子：5.753； 五年影响因子：6.612 ）

ISSN： 1664-462X

年卷期： 2017 年 8 卷

页码：

收录情况： SCI

摘要： Diacylglycerol acyltransferase (DGAT) catalyzes the final step in triacylglycerol (TAG) biosynthesis via the acyl-CoA-dependent acylation of diacylglycerol. This reaction is a major control point in the Kennedy pathway for biosynthesis of TAG, which is the most important form of stored metabolic energy in most oil-producing plants. In this study, Arachis hypogaea type 2 DGAT (AhDGAT2) genes were cloned from the peanut cultivar 'Luhua 14.' Sequence analysis of 11 different peanut cultivars revealed a gene family of 8 peanut DGAT2 genes (designated AhDGAT2a-h). Sequence alignments revealed 21 nucleotide differences between the eight ORFs, but only six differences result in changes to the predicted amino acid (AA) sequences. A representative fulllength cDNA clone (AhDGAT2a) was characterized in detail. The biochemical effects of altering the AhDGAT2a sequence to include single variable AA residues were tested by mutagenesis and functional complementation assays in transgenic yeast systems. All six mutant variants retained enzyme activity and produced lipid droplets in vivo. The N6D and A26P mutants also displayed increased enzyme activity and/or total cellular fatty acid (FA) content. N6D mutant mainly increased the content of palmitoleic acid, and A26P mutant mainly increased the content of palmitic acid. The A26P mutant grew well both in the presence of oleic and C18: 2, but the other mutants grew better in the presence of C18: 2. AhDGAT2 is expressed in all peanut organs analyzed, with high transcript levels in leaves and flowers. These levels are comparable to that found in immature seeds, where DGAT2 expression is most abundant in other plants. Over-expression of AhDGAT2a in tobacco substantially increased the FA content of transformed tobacco seeds. Expression of AhDGAT2a also altered transcription levels of endogenous tobacco lipid metabolic genes in transgenic tobacco, apparently creating a larger carbon 'sink' that supports increased FA levels.

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[1]Discovery of a new mechanism for regulation of plant triacylglycerol metabolism: The peanut diacylglycerol acyltransferase-1 gene family transcriptome is highly enriched in alternative splicing variants. Zheng, Ling,Guo, Feng,Shi, Lingmin,Li, Xinguo,Shan, Lei,Wan, Shubo,Peng, Zhenying,Shockey, Jay.

[2]Development and application of KASP marker for high throughput detection of AhFAD2 mutation in peanut. Zhao, Shuzhen,Li, Aiqin,Li, Changsheng,Xia, Han,Zhao, Chuanzhi,Zhang, Ye,Hou, Lei,Wang, Xingjun. 2017

[3]Effect of Diacylglycerol on Postprandial Serum Triacylglycerol Concentration: A Meta-analysis. Xu, Tongcheng,Zhang, Zhiguo,Li, Duo,Li, Xia,Xu, Tongcheng,Ma, Xiaohang,Zhang, Tiansong. 2009

[4]Effect of diacylglycerol on body weight: a meta-analysis. Xu, Tongcheng,Zhang, Zhiguo,Li, Duo,Xu, Tongcheng,Li, Xia,Ma, Xiaohang. 2008

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