Enhancement of EPA and DHA biosynthesis by over-expression of masu salmon Δ6-desaturase-like gene in zebrafish

Ali, Alimuddin; Yoshizaki, Goro; Kiron, Viswanath; Satoh, Shuichi; Takeuchi, Toshio

doi:10.1007/s11248-004-7435-7

Cited by 59 publications

(37 citation statements)

References 32 publications

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“…There have also been research studies in fish specifically targeting LC-PUFA metabolism. Transgenic zebrafish (Danio rerio) have been produced by transformation with Δ6 and Δ5 fatty acyl desaturases of masu salmon (Oncorhynchus masou) (Alimuddin et al, 2005(Alimuddin et al, , 2007, and both zebrafish (Alimuddin et al, 2008) and the marine teleost, nibe croaker Nibea mitsukurii (Kabeya et al, 2014), have been transformed with fatty acid elongase from masu salmon. Recently, the production of n-3 PUFA, specifically LNA from 18:1n-9, was demonstrated in GM-zebrafish transformed with fat-1 and fat-2 genes (Pang et al, 2014).…”

Section: Enhancing Endogenous Lc-pufa Production and Retentionmentioning

confidence: 99%

Omega-3 long-chain polyunsaturated fatty acids and aquaculture in perspective

2015

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Abbreviations: ARA, arachidonic acid (20:4n-6); CVD, cardiovascular disease; DHA, docosahexaenoic acid (22:6n-3); DPA, docosapentaenoic acid (22:5n-3); EFA, essential fatty acid; Elovl, fatty acid elongase; EPA, eicosapentaenoic acid (20:5n-3); Fads, fatty acyl desaturase; FM, fishmeal; FO, fish oil; LC-PUFA, long-chain polyunsaturated fatty acids (≥ C20 ≥ 3 double bonds); LOA, linoleic acid (18:2n-6); LNA, linolenic acid (18:3n-3); PUFA, polyunsaturated fatty acid; TAG, triacylglycerol; VO, vegetable oil. ABSTRACTIn the 40 years since the essentiality of polyunsaturated fatty acids (PUFA) in fish was first established by determining quantitative requirements for 18:3n-3 and 18:2n-6 in rainbow trout, essential fatty acid (EFA) research has gone through distinct phases. For 20 years the focus was primarily on determining qualitative and quantitative EFA requirements of fish species. Nutritional and biochemical studies showed major differences between fish species based on whether C 18 PUFA or long-chain (LC)-PUFA were required to satisfy requirements. In contrast, in the last 20 years, research emphasis shifted to determining "optimal" levels of EFA to support growth of fish fed diets with increased lipid content and where growth expectations were much higher. This required greater knowledge of the roles and functions of EFA in metabolism and physiology, and how these impacted on fish health and disease. Requirement studies were more focused on early life stages, in particular larval marine fish, defining not only levels, but also balances between different EFA. Finally, a major driver in the last 10-15 years has been the unavoidable replacement of fish oil and fishmeal in feeds and the impacts that this can have on n-3 LC-PUFA contents of diets and farmed fish, and the human consumer. Thus, dietary n-3 in fish feeds can be defined by three levels.Firstly, the minimum level required to satisfy EFA requirements and thus prevent nutritional pathologies. This level is relatively small and easy to supply even with today's current high demand for fish oil. The second level is that required to sustain maximum growth and optimum health in fish being fed modern high-energy diets. The balance between different PUFA and LC-PUFA is important and defining them is more challenging, and so ideal levels and balances are still not well understood, particularly in relation to fish health. The third level is currently driving much research; how can we supply sufficient n-3 LC-PUFA to maintain the nutritional quality of farmed fish at the same level as 20 years ago, and similar or better than in wild fish? This level far exceeds the biological requirements of the fish itself and to satisfy it we require entirely new sources of n-3 LC-PUFA. We cannot rely on the finite and limited marine resources that we can sustainably harvest or 3 efficiently recycle. We need to produce n-3 LC-PUFA de novo and all possible options should be considered.Keywords: Fish oil; essential fatty acids; nutrition; health; metabolism; sustainability 4 Highl...

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Section: Enhancing Endogenous Lc-pufa Production and Retentionmentioning

confidence: 99%

Omega-3 long-chain polyunsaturated fatty acids and aquaculture in perspective

2015

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“…Chemical analysis showed that the muscles of transgenic fish contained considerably higher levels of EPA and DHA compared with control fish (Alimuddin et al, 2008). Summing up the findings of this series of model studies, it ought to be stressed that masu salmon desaturases and elongase transgenes were functional in zebrafish and were able to beneficially alter the species' fatty-acid metabolic pathway, suggesting that this technology might be used to modify farmed fish to produce nutritionally improved, healthy meat for human consumption (Alimuddin et al, 2005(Alimuddin et al, , 2007(Alimuddin et al, and 2008.…”

Section: Modifications Of the Composition Of Milk Lipidsmentioning

confidence: 88%

“…Overexpression of this transgene in modified zebrafish highly elevated body EPA (by a factor of 1.4) and DHA concentrations (by a factor of 2.1). Simultaneously, the content of ALA, being a substrate of Δ6 desaturase, decreased, while total lipids were not changed (Alimuddin et al, 2005). The authors indicated that their results demonstrated the possibility of beneficial modification of the fatty-acid metabolic pathway in fish through the transgenic technique.…”

Section: Modifications Of the Composition Of Milk Lipidsmentioning

confidence: 91%

“…Model studies on fish transgenesis for the improvement of fatty-acid profiles of fish body lipids were carried out in Japan (Alimuddin et al, 2005 and. In the first experiment, a gene construct containing the masu salmon Δ6 desaturaselike gene was introduced by microinjection to the genome of zebrafish (Danio rerio).…”

Section: Modifications Of the Composition Of Milk Lipidsmentioning

confidence: 99%

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The use of genetic engineering techniques to improve the lipid composition in meat, milk and fish products: a review

Świątkiewicz

Arczewska-Włosek

et al. 2015

Animal

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The health-promoting properties of dietary long-chain n-3 polyunsaturated fatty acids (n-3 LCPUFAs) for humans are well-known. Products of animal-origin enriched with n-3 LCPUFAs can be a good example of functional food, that is food that besides traditionally understood nutritional value may have a beneficial influence on the metabolism and health of consumers, thus reducing the risk of various lifestyle diseases such as atherosclerosis and coronary artery disease. The traditional method of enriching meat, milk or eggs with n-3 LCPUFA is the manipulation of the composition of animal diets. Huge progress in the development of genetic engineering techniques, for example transgenesis, has enabled the generation of many kinds of genetically modified animals. In recent years, one of the aims of animal transgenesis has been the modification of the lipid composition of meat and milk in order to improve the dietetic value of animal-origin products. This article reviews and discusses the data in the literature concerning studies where techniques of genetic engineering were used to create animal-origin products modified to contain health-promoting lipids. These studies are still at the laboratory stage, but their results have demonstrated that the transgenesis of pigs, cows, goats and fishes can be used in the future as efficient methods of production of healthy animal-origin food of high dietetic value. However, due to high costs and a low level of public acceptance, the introduction of this technology to commercial animal production and markets seems to be a distant prospect.

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“…Total lipid was extracted from a pool of 3-4 adults in triplicate for each transgenic and non-transgenic sibling using the chloroform-methanol (2:1) method as described by Folch et al (1957). Fatty acid methyl esters were prepared and analyzed by gas liquid chromatography as described by Alimuddin et al (2005).…”

Section: Analysis Of Epa/dha Contentmentioning

confidence: 99%

Over-Expression of Gene Encoding Fatty Acid Metabolic Enzymes in Fish

Alimuddin

Yoshizaki

Takeuchi

et al. 2008

Indonesian Aquaculture Journal

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Eicosapentaenoic acid (EPA, and docosahexaenoic acid (DHA, 22:6n-3) have important nutritional benefits in humans. EPA and DHA are mainly derived from fish, but the decline in the stocks of major marine capture fishes could result in these fatty acids being consumed less. Farmed fish could serve as promising sources of EPA and DHA, but they need these fatty acids in their diets. Generation of fish strains that are capable of synthesizing enough amounts of EPA/DHA from the conversion of α-linolenic acid (LNA, 18:3n-3) rich oils can supply a new EPA/DHA source. This may be achieved by over-expression of genes encoding enzymes involved in HUFA biosynthesis. In aquaculture, the successful of this technique would open the possibility to reduce the enrichment of live food with fish oils for marine fish larvae, and to completely substitute fish oils with plant oils without reducing the quality of flesh in terms of EPA and DHA contents. Here, three genes, i.e. Δ6-desaturase-like (OmΔ6FAD), Δ5-desaturase-like (OmΔ5FAD) and elongase-like (MELO) encoding EPA/ DHA metabolic enzymes derived from masu salmon (Oncorhynchus masou) were individually transferred into zebrafish (Danio rerio) as a model to increase its ability for synthesizing EPA and DHA. Fatty acid analysis showed that EPA content in whole body of the second transgenic fish generation over-expressing OmΔ6FAD gene was 1.4 fold and that of DHA was 2.1 fold higher (P<0.05) than those in non-transgenic fish. The EPA content in whole body of transgenic fish over-expressing OmΔ5FAD gene was 1.21-fold, and that of DHA was 1.24-fold higher (P<0.05) than those in nontransgenic fish. The same patterns were obtained in transgenic fish over-expressing MELO gene. EPA content was increased by 1.30-fold and DHA content by 1.33-fold higher (P<0.05) than those in non-transgenic fish. The results of studies demonstrated that fatty acid content of fish can be enhanced by over-expressing gene encoding enzymes involved in fatty acid biosynthesis, and perhaps this could be applied to tailor farmed fish as even better sources of valuable human food.

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Enhancement of EPA and DHA biosynthesis by over-expression of masu salmon Δ6-desaturase-like gene in zebrafish

Cited by 59 publications

References 32 publications

Omega-3 long-chain polyunsaturated fatty acids and aquaculture in perspective

Omega-3 long-chain polyunsaturated fatty acids and aquaculture in perspective

The use of genetic engineering techniques to improve the lipid composition in meat, milk and fish products: a review

Over-Expression of Gene Encoding Fatty Acid Metabolic Enzymes in Fish

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