Cloning and functional characterization of PmΔ5FAD in pearl oyster Pinctada fucata martensii
Pinctada fucata martensii produces high-quality pearls. However, excess immune and inflammatory response after grafting operation will lead to nucleus rejection, pearl sac formation failure, and death of the host pearl oyster. Polyunsaturated fatty acids (PUFAs) are critically important for its regu...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2022-04-01
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| Series: | Aquaculture Reports |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2352513422000321 |
| _version_ | 1818356212155023360 |
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| author | Chuangye Yang Ruijuan Hao Chengzhang He Yuewen Deng Qingheng Wang |
| author_facet | Chuangye Yang Ruijuan Hao Chengzhang He Yuewen Deng Qingheng Wang |
| author_sort | Chuangye Yang |
| collection | DOAJ |
| description | Pinctada fucata martensii produces high-quality pearls. However, excess immune and inflammatory response after grafting operation will lead to nucleus rejection, pearl sac formation failure, and death of the host pearl oyster. Polyunsaturated fatty acids (PUFAs) are critically important for its regulation function in inflammation and their synthesis process was limited by the related enzymes including fatty acid desaturase (FAD). In this study, FAD gene in the pearl oyster was characterized in P. f. martensii (PmΔ5FAD). We established that the full-length sequence of PmΔ5FAD contained a 1302-bp open reading frame that encoded a sequence of 433 amino acids. PmΔ5FAD domain analysis revealed the presence of a distinctive Cyt-b5 domain and a FA-desaturase domain that were highly similar to the Δ5FAD protein sequences identified in Haliotis discus hannai (69.25%) and Mimachlamys nobilis (62.96%). PmΔ5FAD expressed in all detected tissues. Functional analysis of PmΔ5FAD in recombinant Saccharomyces cerevisiae yeast showed the existence of Δ5-desaturation activity, which towards PUFA substrates and it efficiently desaturated exogenous PUFA C20:3n-6 to C20:4n-6 (ARA) with a desaturation conversion rate of 19.49%. Furthermore, PmΔ5FAD showed a significantly higher expression level on day 1 after the grafting operation, which may upregulate ARA synthesis in response to inflammatory and immune. These results provide important information concerning the function of FADs during pearl oyster PUFA biosynthesis and grafting operation. |
| first_indexed | 2024-12-13T19:53:37Z |
| format | Article |
| id | doaj.art-186561ae94e744c59eb1f54defa716a7 |
| institution | Directory Open Access Journal |
| issn | 2352-5134 |
| language | English |
| last_indexed | 2024-12-13T19:53:37Z |
| publishDate | 2022-04-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Aquaculture Reports |
| spelling | doaj.art-186561ae94e744c59eb1f54defa716a72022-12-21T23:33:22ZengElsevierAquaculture Reports2352-51342022-04-0123101036Cloning and functional characterization of PmΔ5FAD in pearl oyster Pinctada fucata martensiiChuangye Yang0Ruijuan Hao1Chengzhang He2Yuewen Deng3Qingheng Wang4Fisheries College, Guangdong Ocean University, Zhanjiang 524088, China; Guangdong Science and Innovation Center for Pearl Culture, Zhanjiang 524088, China; Guangdong Provincial Engineering Laboratory for Mariculture Organism Breeding, Zhanjiang 524088, ChinaFisheries College, Guangdong Ocean University, Zhanjiang 524088, China; Development and Research Center for Biological Marine Resources, Southern Marine Science and Engineering Guangdong Laboratory (Zhanjiang), Zhanjiang 524006, ChinaFisheries College, Guangdong Ocean University, Zhanjiang 524088, ChinaFisheries College, Guangdong Ocean University, Zhanjiang 524088, China; Development and Research Center for Biological Marine Resources, Southern Marine Science and Engineering Guangdong Laboratory (Zhanjiang), Zhanjiang 524006, China; Guangdong Science and Innovation Center for Pearl Culture, Zhanjiang 524088, China; Guangdong Provincial Engineering Laboratory for Mariculture Organism Breeding, Zhanjiang 524088, China; Corresponding author at: Fisheries College, Guangdong Ocean University, Zhanjiang 524088, China.Fisheries College, Guangdong Ocean University, Zhanjiang 524088, China; Guangdong Science and Innovation Center for Pearl Culture, Zhanjiang 524088, China; Guangdong Provincial Engineering Laboratory for Mariculture Organism Breeding, Zhanjiang 524088, ChinaPinctada fucata martensii produces high-quality pearls. However, excess immune and inflammatory response after grafting operation will lead to nucleus rejection, pearl sac formation failure, and death of the host pearl oyster. Polyunsaturated fatty acids (PUFAs) are critically important for its regulation function in inflammation and their synthesis process was limited by the related enzymes including fatty acid desaturase (FAD). In this study, FAD gene in the pearl oyster was characterized in P. f. martensii (PmΔ5FAD). We established that the full-length sequence of PmΔ5FAD contained a 1302-bp open reading frame that encoded a sequence of 433 amino acids. PmΔ5FAD domain analysis revealed the presence of a distinctive Cyt-b5 domain and a FA-desaturase domain that were highly similar to the Δ5FAD protein sequences identified in Haliotis discus hannai (69.25%) and Mimachlamys nobilis (62.96%). PmΔ5FAD expressed in all detected tissues. Functional analysis of PmΔ5FAD in recombinant Saccharomyces cerevisiae yeast showed the existence of Δ5-desaturation activity, which towards PUFA substrates and it efficiently desaturated exogenous PUFA C20:3n-6 to C20:4n-6 (ARA) with a desaturation conversion rate of 19.49%. Furthermore, PmΔ5FAD showed a significantly higher expression level on day 1 after the grafting operation, which may upregulate ARA synthesis in response to inflammatory and immune. These results provide important information concerning the function of FADs during pearl oyster PUFA biosynthesis and grafting operation.http://www.sciencedirect.com/science/article/pii/S2352513422000321PmΔ5FADPUFA biosynthesisPearl oysterPinctada fucata martensiiGrafting operation |
| spellingShingle | Chuangye Yang Ruijuan Hao Chengzhang He Yuewen Deng Qingheng Wang Cloning and functional characterization of PmΔ5FAD in pearl oyster Pinctada fucata martensii Aquaculture Reports PmΔ5FAD PUFA biosynthesis Pearl oyster Pinctada fucata martensii Grafting operation |
| title | Cloning and functional characterization of PmΔ5FAD in pearl oyster Pinctada fucata martensii |
| title_full | Cloning and functional characterization of PmΔ5FAD in pearl oyster Pinctada fucata martensii |
| title_fullStr | Cloning and functional characterization of PmΔ5FAD in pearl oyster Pinctada fucata martensii |
| title_full_unstemmed | Cloning and functional characterization of PmΔ5FAD in pearl oyster Pinctada fucata martensii |
| title_short | Cloning and functional characterization of PmΔ5FAD in pearl oyster Pinctada fucata martensii |
| title_sort | cloning and functional characterization of pmδ5fad in pearl oyster pinctada fucata martensii |
| topic | PmΔ5FAD PUFA biosynthesis Pearl oyster Pinctada fucata martensii Grafting operation |
| url | http://www.sciencedirect.com/science/article/pii/S2352513422000321 |
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