Novel lysophospholipid acyltransferase PLAT1 of Aurantiochytrium limacinum F26-b responsible for generation of palmitate-docosahexaenoate- phosphatidylcholine and phosphatidylethanolamine

Eriko Abe, Kazutaka Ikeda, Eri Nutahara, Masahiro Hayashi, Atsushi Yamashita, Ryo Taguchi, Kosaku Doi, Daiske Honda, Nozomu Okino, Makoto Ito

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

N-3 polyunsaturated fatty acids (PUFA), such as docosahexaenoic acid (DHA, 22:6n-3), have been reported to play roles in preventing cardiovascular diseases. The major source of DHA is fish oils but a recent increase in the global demand of DHA and decrease in fish stocks require a substitute. Thraustochytrids, unicellular marine protists belonging to the Chromista kingdom, can synthesize large amounts of DHA, and, thus, are expected to be an alternative to fish oils. DHA is found in the acyl chain(s) of phospholipids as well as triacylglycerols in thraustochytrids; however, how thraustochytrids incorporate DHA into phospholipids remains unknown. We report here a novel lysophospholipid acyltransferase (PLAT1), which is responsible for the generation of DHA-containing phosphatidylcholine and phosphatidylethanolamine in thraustochytrids. The PLAT1 gene, which was isolated from the genomic DNA of Aurantiochytrium limacinum F26-b, was expressed in Saccharomyces cerevisiae, and the FLAG-tagged recombinant enzyme was characterized after purification with anti-FLAG affinity gel. PLAT1 shows wide specificity for donor substrates as well as acceptor substrates in vitro, i.e, the enzyme can adopt lysophosphatidylcholine, lysophosphatidylethanolamine, lysophosphatidylserine and lysophosphatidylinositol as acceptor substrates, and 15:0/16:0-CoA and DHA-CoA as donor substrates. In contrast to the in vitro experiment, only lysophosphatidylcholine acyltransferase and lysophosphatidylethanolamine acyltransferase activities were decreased in plat1-knockout mutants, resulting in a decrease of 16:0-DHA-phosphatidylcholine (PC) [PC(38:6)] and 16:0-DHA-phosphatidylethanolamine (PE) [PE(38:6)], which are two major DHA-containing phospholipids in A. limacinum F26-b. However, the amounts of other phospholipid species including DHA-DHA-PC [PC(44:12)] and DHA-DHA-PE [PE(44:12)] were almost the same in plat-knockout mutants and the wild-type. These results indicate that PLAT1 is the enzyme responsible for the generation of 16:0-DHA-PC and 16:0-DHA-PE in the thraustochytrid.

Original languageEnglish
Article numbere102377
JournalPloS one
Volume9
Issue number8
DOIs
Publication statusPublished - Aug 4 2014

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1-Acylglycerophosphocholine O-Acyltransferase
acyltransferases
Docosahexaenoic Acids
Palmitates
palmitates
phosphatidylethanolamines
Phosphatidylcholines
phosphatidylcholines
Phospholipids
phospholipids
lysophosphatidylcholine
knockout mutants
Fish Oils
Substrates
Coenzyme A
fish oils
Enzymes
enzymes
Lysophosphatidylcholines
Omega-3 Fatty Acids

All Science Journal Classification (ASJC) codes

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)
  • General

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Novel lysophospholipid acyltransferase PLAT1 of Aurantiochytrium limacinum F26-b responsible for generation of palmitate-docosahexaenoate- phosphatidylcholine and phosphatidylethanolamine. / Abe, Eriko; Ikeda, Kazutaka; Nutahara, Eri; Hayashi, Masahiro; Yamashita, Atsushi; Taguchi, Ryo; Doi, Kosaku; Honda, Daiske; Okino, Nozomu; Ito, Makoto.

In: PloS one, Vol. 9, No. 8, e102377, 04.08.2014.

Research output: Contribution to journalArticle

Abe, Eriko ; Ikeda, Kazutaka ; Nutahara, Eri ; Hayashi, Masahiro ; Yamashita, Atsushi ; Taguchi, Ryo ; Doi, Kosaku ; Honda, Daiske ; Okino, Nozomu ; Ito, Makoto. / Novel lysophospholipid acyltransferase PLAT1 of Aurantiochytrium limacinum F26-b responsible for generation of palmitate-docosahexaenoate- phosphatidylcholine and phosphatidylethanolamine. In: PloS one. 2014 ; Vol. 9, No. 8.
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abstract = "N-3 polyunsaturated fatty acids (PUFA), such as docosahexaenoic acid (DHA, 22:6n-3), have been reported to play roles in preventing cardiovascular diseases. The major source of DHA is fish oils but a recent increase in the global demand of DHA and decrease in fish stocks require a substitute. Thraustochytrids, unicellular marine protists belonging to the Chromista kingdom, can synthesize large amounts of DHA, and, thus, are expected to be an alternative to fish oils. DHA is found in the acyl chain(s) of phospholipids as well as triacylglycerols in thraustochytrids; however, how thraustochytrids incorporate DHA into phospholipids remains unknown. We report here a novel lysophospholipid acyltransferase (PLAT1), which is responsible for the generation of DHA-containing phosphatidylcholine and phosphatidylethanolamine in thraustochytrids. The PLAT1 gene, which was isolated from the genomic DNA of Aurantiochytrium limacinum F26-b, was expressed in Saccharomyces cerevisiae, and the FLAG-tagged recombinant enzyme was characterized after purification with anti-FLAG affinity gel. PLAT1 shows wide specificity for donor substrates as well as acceptor substrates in vitro, i.e, the enzyme can adopt lysophosphatidylcholine, lysophosphatidylethanolamine, lysophosphatidylserine and lysophosphatidylinositol as acceptor substrates, and 15:0/16:0-CoA and DHA-CoA as donor substrates. In contrast to the in vitro experiment, only lysophosphatidylcholine acyltransferase and lysophosphatidylethanolamine acyltransferase activities were decreased in plat1-knockout mutants, resulting in a decrease of 16:0-DHA-phosphatidylcholine (PC) [PC(38:6)] and 16:0-DHA-phosphatidylethanolamine (PE) [PE(38:6)], which are two major DHA-containing phospholipids in A. limacinum F26-b. However, the amounts of other phospholipid species including DHA-DHA-PC [PC(44:12)] and DHA-DHA-PE [PE(44:12)] were almost the same in plat-knockout mutants and the wild-type. These results indicate that PLAT1 is the enzyme responsible for the generation of 16:0-DHA-PC and 16:0-DHA-PE in the thraustochytrid.",
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AU - Ikeda, Kazutaka

AU - Nutahara, Eri

AU - Hayashi, Masahiro

AU - Yamashita, Atsushi

AU - Taguchi, Ryo

AU - Doi, Kosaku

AU - Honda, Daiske

AU - Okino, Nozomu

AU - Ito, Makoto

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