Diatoms and Plants Acyl-CoA:lysophosphatidylcholine Acyltransferases (LPCATs) Exhibit Diverse Substrate Specificity and Biochemical Properties
The search of the <i>Phaeodactylum tricornutum</i> genome database revealed the existence of six genes potentially encoding lysophospholipid acyltransferases. One of these genes, Phatr3_J20460, after introduction to yeast <i>ale1</i> mutant disrupted in the LPCAT gene, produc...
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2021-08-01
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| author | Ada Połońska Katarzyna Jasieniecka-Gazarkiewicz Lingjie You Xiahui Hao Sylwia Klińska Yangmin Gong Antoni Banaś |
| author_facet | Ada Połońska Katarzyna Jasieniecka-Gazarkiewicz Lingjie You Xiahui Hao Sylwia Klińska Yangmin Gong Antoni Banaś |
| author_sort | Ada Połońska |
| collection | DOAJ |
| description | The search of the <i>Phaeodactylum tricornutum</i> genome database revealed the existence of six genes potentially encoding lysophospholipid acyltransferases. One of these genes, Phatr3_J20460, after introduction to yeast <i>ale1</i> mutant disrupted in the LPCAT gene, produced a very active acyl-CoA:lysophosphatidylcholine (LPCAT) enzyme. Using in vitro assays applying different radioactive and non-radioactive substrates and microsomal fractions from such yeast, we have characterized the biochemical properties and substrate specificities of this <i>Pt</i>LPCAT1. We have found that the substrate specificity of this enzyme indicates that it can completely supply phosphatidylcholine (PC) with all fatty acids connected with a biosynthetic pathway of very long-chain polyunsaturated fatty acids (VLC-PUFAs) used further for the desaturation process. Additionally, we have shown that biochemical properties of the <i>Pt</i>LPCAT1 in comparison to plant LPCATs are in some cases similar (such as the dependency of its activity on pH value), differ moderately (such as in response to temperature changes), or express completely different properties (such as in reaction to calcium and magnesium ions or toward some acyl-CoA with 20C polyunsaturated fatty acids). Moreover, the obtained results suggest that cloned “Phatr3_J20460” gene can be useful in oilseeds plant engineering toward efficient production of VLC-PUFA as LPCAT it encodes can (contrary to plant LPCATs) introduce 20:4-CoA (n-3) to PC for further desaturation to 20:5 (EPA, eicosapentaenoic acid). |
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| last_indexed | 2024-03-10T08:43:51Z |
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| spelling | doaj.art-5c6f54a61e664e0b900e40cedcf135ca2023-11-22T08:04:47ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672021-08-012216905610.3390/ijms22169056Diatoms and Plants Acyl-CoA:lysophosphatidylcholine Acyltransferases (LPCATs) Exhibit Diverse Substrate Specificity and Biochemical PropertiesAda Połońska0Katarzyna Jasieniecka-Gazarkiewicz1Lingjie You2Xiahui Hao3Sylwia Klińska4Yangmin Gong5Antoni Banaś6Intercollegiate Faculty of Biotechnology, University of Gdansk and Medical University of Gdańsk, 80-307 Gdansk, PolandIntercollegiate Faculty of Biotechnology, University of Gdansk and Medical University of Gdańsk, 80-307 Gdansk, PolandOil Crops Research Institute, Chinese Academy of Agricultural Sciences, Wuhan 430062, ChinaOil Crops Research Institute, Chinese Academy of Agricultural Sciences, Wuhan 430062, ChinaIntercollegiate Faculty of Biotechnology, University of Gdansk and Medical University of Gdańsk, 80-307 Gdansk, PolandOil Crops Research Institute, Chinese Academy of Agricultural Sciences, Wuhan 430062, ChinaIntercollegiate Faculty of Biotechnology, University of Gdansk and Medical University of Gdańsk, 80-307 Gdansk, PolandThe search of the <i>Phaeodactylum tricornutum</i> genome database revealed the existence of six genes potentially encoding lysophospholipid acyltransferases. One of these genes, Phatr3_J20460, after introduction to yeast <i>ale1</i> mutant disrupted in the LPCAT gene, produced a very active acyl-CoA:lysophosphatidylcholine (LPCAT) enzyme. Using in vitro assays applying different radioactive and non-radioactive substrates and microsomal fractions from such yeast, we have characterized the biochemical properties and substrate specificities of this <i>Pt</i>LPCAT1. We have found that the substrate specificity of this enzyme indicates that it can completely supply phosphatidylcholine (PC) with all fatty acids connected with a biosynthetic pathway of very long-chain polyunsaturated fatty acids (VLC-PUFAs) used further for the desaturation process. Additionally, we have shown that biochemical properties of the <i>Pt</i>LPCAT1 in comparison to plant LPCATs are in some cases similar (such as the dependency of its activity on pH value), differ moderately (such as in response to temperature changes), or express completely different properties (such as in reaction to calcium and magnesium ions or toward some acyl-CoA with 20C polyunsaturated fatty acids). Moreover, the obtained results suggest that cloned “Phatr3_J20460” gene can be useful in oilseeds plant engineering toward efficient production of VLC-PUFA as LPCAT it encodes can (contrary to plant LPCATs) introduce 20:4-CoA (n-3) to PC for further desaturation to 20:5 (EPA, eicosapentaenoic acid).https://www.mdpi.com/1422-0067/22/16/9056LPCATVLC-PUFAeicosapentaenoic acideicosatetraenoic aciddiatoms<i>Phaeodactylum tricornutum</i> |
| spellingShingle | Ada Połońska Katarzyna Jasieniecka-Gazarkiewicz Lingjie You Xiahui Hao Sylwia Klińska Yangmin Gong Antoni Banaś Diatoms and Plants Acyl-CoA:lysophosphatidylcholine Acyltransferases (LPCATs) Exhibit Diverse Substrate Specificity and Biochemical Properties International Journal of Molecular Sciences LPCAT VLC-PUFA eicosapentaenoic acid eicosatetraenoic acid diatoms <i>Phaeodactylum tricornutum</i> |
| title | Diatoms and Plants Acyl-CoA:lysophosphatidylcholine Acyltransferases (LPCATs) Exhibit Diverse Substrate Specificity and Biochemical Properties |
| title_full | Diatoms and Plants Acyl-CoA:lysophosphatidylcholine Acyltransferases (LPCATs) Exhibit Diverse Substrate Specificity and Biochemical Properties |
| title_fullStr | Diatoms and Plants Acyl-CoA:lysophosphatidylcholine Acyltransferases (LPCATs) Exhibit Diverse Substrate Specificity and Biochemical Properties |
| title_full_unstemmed | Diatoms and Plants Acyl-CoA:lysophosphatidylcholine Acyltransferases (LPCATs) Exhibit Diverse Substrate Specificity and Biochemical Properties |
| title_short | Diatoms and Plants Acyl-CoA:lysophosphatidylcholine Acyltransferases (LPCATs) Exhibit Diverse Substrate Specificity and Biochemical Properties |
| title_sort | diatoms and plants acyl coa lysophosphatidylcholine acyltransferases lpcats exhibit diverse substrate specificity and biochemical properties |
| topic | LPCAT VLC-PUFA eicosapentaenoic acid eicosatetraenoic acid diatoms <i>Phaeodactylum tricornutum</i> |
| url | https://www.mdpi.com/1422-0067/22/16/9056 |
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