Identification of glycine betaine as a host-derived molecule required for the vegetative proliferation of the protozoan parasite Perkinsus olseni
Perkinsus olseni is an industrially significant protozoan parasite of Manila clam, Ruditapes philippinarum. So far, various media, based on Dulbecco's Modified Eagle Medium and Ham's F-12 nutrient mixture with supplementation of fetal bovine serum (FBS), have been developed to proliferate...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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Cambridge University Press
2023-09-01
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| Series: | Parasitology |
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| Online Access: | https://www.cambridge.org/core/product/identifier/S0031182023000768/type/journal_article |
| _version_ | 1797659549889462272 |
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| author | Yuqi Liu Akihiro Ninomiya Tomoyoshi Yoshinaga Naoki Itoh |
| author_facet | Yuqi Liu Akihiro Ninomiya Tomoyoshi Yoshinaga Naoki Itoh |
| author_sort | Yuqi Liu |
| collection | DOAJ |
| description | Perkinsus olseni is an industrially significant protozoan parasite of Manila clam, Ruditapes philippinarum. So far, various media, based on Dulbecco's Modified Eagle Medium and Ham's F-12 nutrient mixture with supplementation of fetal bovine serum (FBS), have been developed to proliferate the parasitizing trophozoite stage of P. olseni. The present study showed that P. olseni did not proliferate in FBS-deficient Perkinsus broth medium (PBMΔF), but proliferated well in PBMΔF supplemented with tissue extract of host Manila clams, indicating that FBS and Manila clam tissue contained molecule(s) required for P. olseni proliferation. Preliminary characterization suggested that the host-derived molecule(s) was a heat-stable molecule(s) with a molecular weight of less than 3 kDa, and finally a single molecule required for the proliferation was purified by high-performance liquid chromatography processes. High-resolution electrospray ionization mass spectrometry and nuclear magnetic resonance analyses identified this molecule as glycine betaine (=trimethylglycine), and the requirement of this molecule for P. olsseni proliferation was confirmed by an assay using chemically synthesized, standard glycine betaine. Although glycine betaine was required for the proliferation of all examined Perkinsus species, supplementation of glycine betaine precursors, such as choline and betaine aldehyde, enhanced the proliferation of 4 Perkinsus species (P. marinus, P. chesapeaki, P. mediterraneus and P. honshuensis), but not of 2 others (P. olseni and P. beihaiensis). Thus, it was concluded that the ability to biosynthesise glycine betaine from its precursors varied among Perkinsus species, and that P. olseni and P. beihaiensis lack the ability required to biosynthesize glycine betaine for proliferation. |
| first_indexed | 2024-03-11T18:16:44Z |
| format | Article |
| id | doaj.art-f7afe5c6f30a492cb65a2f5c61b24f9a |
| institution | Directory Open Access Journal |
| issn | 0031-1820 1469-8161 |
| language | English |
| last_indexed | 2024-03-11T18:16:44Z |
| publishDate | 2023-09-01 |
| publisher | Cambridge University Press |
| record_format | Article |
| series | Parasitology |
| spelling | doaj.art-f7afe5c6f30a492cb65a2f5c61b24f9a2023-10-16T06:57:14ZengCambridge University PressParasitology0031-18201469-81612023-09-0115093994910.1017/S0031182023000768Identification of glycine betaine as a host-derived molecule required for the vegetative proliferation of the protozoan parasite Perkinsus olseniYuqi Liu0Akihiro Ninomiya1Tomoyoshi Yoshinaga2Naoki Itoh3https://orcid.org/0000-0003-1944-4253Laboratory of Fish Diseases, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Tokyo, JapanLaboratory of Aquatic Natural Products Chemistry, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Tokyo, JapanLaboratory of Fish Diseases, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Tokyo, JapanLaboratory of Fish Diseases, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Tokyo, JapanPerkinsus olseni is an industrially significant protozoan parasite of Manila clam, Ruditapes philippinarum. So far, various media, based on Dulbecco's Modified Eagle Medium and Ham's F-12 nutrient mixture with supplementation of fetal bovine serum (FBS), have been developed to proliferate the parasitizing trophozoite stage of P. olseni. The present study showed that P. olseni did not proliferate in FBS-deficient Perkinsus broth medium (PBMΔF), but proliferated well in PBMΔF supplemented with tissue extract of host Manila clams, indicating that FBS and Manila clam tissue contained molecule(s) required for P. olseni proliferation. Preliminary characterization suggested that the host-derived molecule(s) was a heat-stable molecule(s) with a molecular weight of less than 3 kDa, and finally a single molecule required for the proliferation was purified by high-performance liquid chromatography processes. High-resolution electrospray ionization mass spectrometry and nuclear magnetic resonance analyses identified this molecule as glycine betaine (=trimethylglycine), and the requirement of this molecule for P. olsseni proliferation was confirmed by an assay using chemically synthesized, standard glycine betaine. Although glycine betaine was required for the proliferation of all examined Perkinsus species, supplementation of glycine betaine precursors, such as choline and betaine aldehyde, enhanced the proliferation of 4 Perkinsus species (P. marinus, P. chesapeaki, P. mediterraneus and P. honshuensis), but not of 2 others (P. olseni and P. beihaiensis). Thus, it was concluded that the ability to biosynthesise glycine betaine from its precursors varied among Perkinsus species, and that P. olseni and P. beihaiensis lack the ability required to biosynthesize glycine betaine for proliferation.https://www.cambridge.org/core/product/identifier/S0031182023000768/type/journal_articleBetaine aldehydebiosynthesischolinehost–parasite interactionmollusksPerkinsozoaschizogonytrimethylglycinetrophozoite |
| spellingShingle | Yuqi Liu Akihiro Ninomiya Tomoyoshi Yoshinaga Naoki Itoh Identification of glycine betaine as a host-derived molecule required for the vegetative proliferation of the protozoan parasite Perkinsus olseni Parasitology Betaine aldehyde biosynthesis choline host–parasite interaction mollusks Perkinsozoa schizogony trimethylglycine trophozoite |
| title | Identification of glycine betaine as a host-derived molecule required for the vegetative proliferation of the protozoan parasite Perkinsus olseni |
| title_full | Identification of glycine betaine as a host-derived molecule required for the vegetative proliferation of the protozoan parasite Perkinsus olseni |
| title_fullStr | Identification of glycine betaine as a host-derived molecule required for the vegetative proliferation of the protozoan parasite Perkinsus olseni |
| title_full_unstemmed | Identification of glycine betaine as a host-derived molecule required for the vegetative proliferation of the protozoan parasite Perkinsus olseni |
| title_short | Identification of glycine betaine as a host-derived molecule required for the vegetative proliferation of the protozoan parasite Perkinsus olseni |
| title_sort | identification of glycine betaine as a host derived molecule required for the vegetative proliferation of the protozoan parasite perkinsus olseni |
| topic | Betaine aldehyde biosynthesis choline host–parasite interaction mollusks Perkinsozoa schizogony trimethylglycine trophozoite |
| url | https://www.cambridge.org/core/product/identifier/S0031182023000768/type/journal_article |
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