Cloning, characterization and transcription analysis of fatty acyl desaturase ( Δ6Fad‐like ) and elongase ( Elovl4‐like , Elove5‐like ) during embryos development of Strongylocentrotus intermedius

2019 ◽  
Vol 50 (12) ◽  
pp. 3483-3492
Author(s):  
Jun Ding ◽  
Dan Yang ◽  
Heng Wang ◽  
Rantao Zuo ◽  
Shoubing Qi ◽  
...  
Keyword(s):  
Fatty Acyl ◽  
Strongylocentrotus Intermedius ◽  
Transcription Analysis ◽  
Fatty Acyl Desaturase

scholarly journals Characterization of type -2 diacylglycerol acyltransferases in Haematococcus lacustris reveals their functions and engineering potential in triacylglycerol biosynthesis

2020 ◽  
Author(s):  
Hongli Cui ◽  
Chunchao Zhao ◽  
Wenxin Xu ◽  
Hongjiang Zhang ◽  
Wei Hang ◽  
...  
Keyword(s):  
Heterologous Expression ◽  
Feeding Strategy ◽  
Expression Patterns ◽  
Fatty Acyl ◽  
Functional Complementation ◽  
Yeast Cells ◽  
Transcription Analysis ◽  
Haematococcus Lacustris ◽  
Tag Biosynthesis

Abstract Background: Haematococcus lacustris is an ideal source of astaxanthin (AST), which is stored in oil bodies containing esterified AST (EAST) and triacylglycerol (TAG). Diacylglycerol acyltransferases (DGATs) catalyze the last step of acyl-CoA-dependent TAG biosynthesis and are also considered as crucial enzymes involved in EAST biosynthesis in H. lacustris. Previous studies have identified four putative DGAT2-encoding genes in H. lacustris, and only HpDGAT2D allowed the recovery of TAG biosynthesis, but the engineering potential of HpDGAT2s in TAG biosynthesis remains ambiguous.Results: Five putative DGAT2 genes (HpDGAT2A, HpDGAT2B, HpDGAT2C, HpDGAT2D, and HpDGAT2E) were identified in H. lacustris. Transcription analysis showed that the expression levels of the HpDGAT2A, HpDGAT2D, and HpDGAT2E genes markedly increased under high light and nitrogen deficient conditions with distinct patterns, which led to significant TAG and EAST accumulation. Functional complementation demonstrated that HpDGAT2A, HpDGAT2B, HpDGAT2D, and HpDGAT2E had the capacity to restore TAG synthesis in a TAG-deficient yeast strain (H1246) showing a large difference in enzymatic activity. Fatty acid (FA) profile assays revealed that HpDGAT2A, HpDGAT2D, and HpDGAT2E, but not HpDGAT2B, preferred monounsaturated fatty acyl-CoAs (MUFAs) for TAG synthesis in yeast cells, and showed a preference for polyunsaturated fatty acyl-CoAs (PUFAs) based on their feeding strategy. The heterologous expression of HpDGAT2D in Arabidopsis thaliana and Chlamydomonas reinhardtii significantly increased the TAG content and obviously promoted the MUFAs and PUFAs contents.Conclusions: Our study represents pioneering work on the characterization of HpDGAT2s by systematically integrating expression patterns, AST/TAG accumulation, functional complementation, and heterologous expression in yeast, plants, and algae. These results (1) update the gene models of HpDGAT2s, (2) prove the TAG biosynthesis capacity of HpDGAT2s, (3) show the strong preference for MUFAs and PUFAs, and (4) offer target genes to modulate TAG biosynthesis by using genetic engineering methods.

scholarly journals Characterization of type-2 diacylglycerol acyltransferases in Haematococcus lacustris reveals their functions and engineering potential in triacylglycerol biosynthesis

2020 ◽  
Author(s):  
Hongli Cui ◽  
Chunchao Zhao ◽  
Wenxin Xu ◽  
Hongjiang Zhang ◽  
Wei Hang ◽  
...  
Keyword(s):  
Heterologous Expression ◽  
Feeding Strategy ◽  
Expression Patterns ◽  
Fatty Acyl ◽  
Functional Complementation ◽  
Yeast Cells ◽  
Transcription Analysis ◽  
Haematococcus Lacustris ◽  
Tag Biosynthesis

Abstract Background: Haematococcus lacustris is an ideal source of astaxanthin (AST), which is stored in oil bodies containing esterified AST (EAST) and triacylglycerol (TAG). Diacylglycerol acyltransferases (DGATs) catalyze the last step of acyl-CoA-dependent TAG biosynthesis and are also considered as crucial enzymes involved in EAST biosynthesis in H. lacustris. Previous studies have identified four putative DGAT2-encoding genes in H. lacustris, and only HpDGAT2D allowed the recovery of TAG biosynthesis, but the engineering potential of HpDGAT2s in TAG biosynthesis remains ambiguous.Results: Five putative DGAT2 genes (HpDGAT2A, HpDGAT2B, HpDGAT2C, HpDGAT2D, and HpDGAT2E) were identified in H. lacustris. Transcription analysis showed that the expression levels of the HpDGAT2A, HpDGAT2D, and HpDGAT2E genes markedly increased under high light and nitrogen deficient conditions with distinct patterns, which led to significant TAG and EAST accumulation. Functional complementation demonstrated that HpDGAT2A, HpDGAT2B, HpDGAT2D, and HpDGAT2E had the capacity to restore TAG synthesis in a TAG-deficient yeast strain (H1246) showing a large difference in enzymatic activity. Fatty acid (FA) profile assays revealed that HpDGAT2A, HpDGAT2D, and HpDGAT2E, but not HpDGAT2B, preferred monounsaturated fatty acyl-CoAs (MUFAs) for TAG synthesis in yeast cells, and showed a preference for polyunsaturated fatty acyl-CoAs (PUFAs) based on their feeding strategy. The heterologous expression of HpDGAT2D in Arabidopsis thaliana and Chlamydomonas reinhardtii significantly increased the TAG content and obviously promoted the MUFAs and PUFAs contents.Conclusions: Our study represents systematic work on the characterization of HpDGAT2s by integrating expression patterns, AST/TAG accumulation, functional complementation, and heterologous expression in yeast, plants, and algae. These results (1) update the gene models of HpDGAT2s, (2) prove the TAG biosynthesis capacity of HpDGAT2s, (3) show the strong preference for MUFAs and PUFAs, and (4) offer target genes to modulate TAG biosynthesis by using genetic engineering methods.

scholarly journals Characterization of type-2 diacylglycerol acyltransferases in Haematococcus lacustris reveals their functions and engineering potential in triacylglycerol biosynthesis and possible roles in astaxanthin esterification

2020 ◽  
Author(s):  
Hongli Cui ◽  
Chunchao Zhao ◽  
Wenxin Xu ◽  
Hongjiang Zhang ◽  
Wei Hang ◽  
...  
Keyword(s):  
Molecular Docking ◽  
Feeding Strategy ◽  
Fatty Acyl ◽  
Functional Complementation ◽  
Yeast Cells ◽  
Transcription Analysis ◽  
Over Expression ◽  
Haematococcus Lacustris ◽  
Tag Biosynthesis

Abstract Background: Haematococcus lacustris is an ideal source of astaxanthin (AST) which is stored at oil bodies containing esterified AST (EAST) and triacylglycerol (TAG). Diacylglycerol acyltransferases (DGATs) catalyze the last step of the acyl-CoA-dependent TAG biosynthesis and are also considered as the crucial enzymes involving in EAST biosynthesis in H. lacustris. Previous studies have identified four putative DGAT2 encoding genes in H. lacustris and only the HpDGAT2D allowed the recovery of TAG biosynthesis, but the engineering potential of HpDGAT2s in TAG biosynthesis, especially possible roles in AST esterification, remains ambiguous.Results: Five putative DGAT2 genes (HpDGAT2A, HpDGAT2B, HpDGAT2C, HpDGAT2D, and HpDGAT2E) were identified in H. lacustris. Transcription analysis showed that the expression levels of HpDGAT2A, HpDGAT2D, and HpDGAT2E genes markedly increased under high light and nitrogen deficient conditions with distinct patterns, which led to significant TAG and EAST accumulation. Functional complementation demonstrated HpDGAT2A, HpDGAT2B, HpDGAT2D, and HpDGAT2E had the capability to restore TAG synthesis in a TAG-deficient yeast strain (H1246) with the large difference in enzymatic activity. Fatty acids (FAs) profiles assays revealed that HpDGAT2A, HpDGAT2D, and HpDGAT2E except for HpDGAT2B preferred monounsaturated fatty acyl-CoAs (MUFAs) for TAG synthesis in yeast cells, and also showed polyunsaturated fatty acyl-CoAs (PUFAs) preference by feeding strategy. The over-expression of HpDGAT2D in Arabidopsis thaliana and Chlamydomonas reinhardtii significantly increased the TAG content and obviously promoted the MUFAs and PUFAs contents. Interestingly, molecular docking analysis implied that HpDGAT2s structures contained AST binding sites, which provides strong evidence for AST esterification function in H. lacustris.Conclusions: Our study represents a pioneering work on the characterization of HpDGAT2s by systematically integrating expression pattern, AST/TAG accumulation, functional complementation, molecular docking, and over-expression in yeast, plants, and algae. These results (1) update the gene models of HpDGAT2s, (2) prove TAG biosynthesis capacity of HpDGAT2s, (3) show the strong preference for MUFAs and PUFAs, (4) offer target genes to modulate TAG biosynthesis by genetic engineering method, and (5) provide new evidence for HpDGAT2s roles in AST esterification.

Molecular cloning and functional characterization of elongase (elovl5) and fatty acyl desaturase (fads2) in sciaenid, Nibea diacanthus (Lacepède, 1802)

Gene ◽  
2019 ◽  
Vol 695 ◽  
pp. 1-11 ◽  
Author(s):  
Weiguang Zou ◽  
Zhideng Lin ◽  
Yisheng Huang ◽  
Samwel Mchele Limbu ◽  
Xiaobo Wen
Keyword(s):  
Molecular Cloning ◽  
Functional Characterization ◽  
Fatty Acyl ◽  
Fatty Acyl Desaturase

FATTY ACID COMPOSITION OF PORCINE DEPOT FAT AS RELATED TO THE EFFECT OF SUPPLEMENTAL DIETARY COPPER ON THE SPECIFIC ACTIVITIES OF FATTY ACYL DESATURASE SYSTEMS

1974 ◽  
Vol 54 (1) ◽  
pp. 23-28 ◽  
Author(s):  
S. K. HO ◽  
J. I. ELLIOT
Keyword(s):  
Fatty Acid Composition ◽  
Fatty Acid ◽  
Acid Composition ◽  
Unsaturated Fatty Acids ◽  
Average Daily Gain ◽  
Fatty Acyl ◽  
Dietary Copper ◽  
Liver Copper ◽  
Fatty Acyl Desaturase ◽  
Specific Activities

Copper supplementation of a swine diet (barley-wheat-soybean meal) at a level of 250 ppm was associated with improved average daily gain, reduction in the melting point of the depot fat, increase in liver copper, decreased proportions of stearic and palmitic acids and concomitant increases in proportion of the medium-chain myristic acid as well as those of the major long-chain unsaturated fatty acids in the depot fat, and increases in the capacities of hepatic and adipose microsomes to desaturate 1-14C-oleate (in the presence of coenzyme A, ATP, Mg2+) and 1-14C-palmitoyl-CoA. There were neither sex differences nor copper × sex interactions. It is suggested that supplemental dietary copper enhances the specific activities of the fatty acyl desaturase systems, and that this enhancement contributes to the observed changes in fatty acid composition of the depot fat.

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