Characterization of GMPP from Dendrobium huoshanense yielding GDP-D-mannose
Dendrobium huoshanense has been used for centuries in China and its polysaccharides are the main active components in treating loss of body fluids resulting from fever and asthenic symptoms. However, the biosynthetic pathway of polysaccharides in D. huoshanense remains to be elucidated. In this stud...
| Main Authors: | , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
De Gruyter
2021-02-01
|
| Series: | Open Life Sciences |
| Subjects: | |
| Online Access: | https://doi.org/10.1515/biol-2021-0015 |
| _version_ | 1828108164269080576 |
|---|---|
| author | Yi Yuqi Liu Lulu Zhou Wenyan Peng Daiyin Han Rongchun Yu Nianjun |
| author_facet | Yi Yuqi Liu Lulu Zhou Wenyan Peng Daiyin Han Rongchun Yu Nianjun |
| author_sort | Yi Yuqi |
| collection | DOAJ |
| description | Dendrobium huoshanense has been used for centuries in China and its polysaccharides are the main active components in treating loss of body fluids resulting from fever and asthenic symptoms. However, the biosynthetic pathway of polysaccharides in D. huoshanense remains to be elucidated. In this study, we obtained a guanosine diphosphate (GDP)-mannose pyrophosphorylase (DhGMPP) from D. huoshanense and characterized its function to catalyze the conversion of α-D-mannose-phosphate to GDP-D-mannose involved in the production of polysaccharides. DhGMPP, with the open reading frame of 1,245 bp, was isolated from RNA-Seq data of D. huoshanense. Phylogenetic analysis as well as sequence characterization suggested its involvement in the biosynthesis of GDP-D-mannose. In vitro enzyme assay demonstrated that GMPP encoded a pyrophosphorylase that converted α-D-mannose-phosphate and GTP into GDP-D-mannose. Identification of DhGMPP could provide more insights into the mechanism concerning polysaccharide biosynthesis in D. huoshanense and be utilized for enhancing polysaccharide accumulation through metabolic engineering. |
| first_indexed | 2024-04-11T10:43:27Z |
| format | Article |
| id | doaj.art-8abe55b1c8624feaa63a18f3e369bffd |
| institution | Directory Open Access Journal |
| issn | 2391-5412 |
| language | English |
| last_indexed | 2024-04-11T10:43:27Z |
| publishDate | 2021-02-01 |
| publisher | De Gruyter |
| record_format | Article |
| series | Open Life Sciences |
| spelling | doaj.art-8abe55b1c8624feaa63a18f3e369bffd2022-12-22T04:29:08ZengDe GruyterOpen Life Sciences2391-54122021-02-0116110210710.1515/biol-2021-0015Characterization of GMPP from Dendrobium huoshanense yielding GDP-D-mannoseYi Yuqi0Liu Lulu1Zhou Wenyan2Peng Daiyin3Han Rongchun4Yu Nianjun5School of Pharmacy, Anhui University of Chinese Medicine, No. 1, Qianjiang Road, Yaohai District, Hefei 230012, ChinaDepartment of Research and Development, Shanghai Zenith Pharmaceutical Technology Co. Ltd.; Shanghai 201199, ChinaDepartment of Research and Development, Hefei Yifan Biopharmaceutical Co. Ltd.; Hefei 230061, ChinaSchool of Pharmacy, Anhui University of Chinese Medicine, No. 1, Qianjiang Road, Yaohai District, Hefei 230012, ChinaSchool of Pharmacy, Anhui University of Chinese Medicine, No. 1, Qianjiang Road, Yaohai District, Hefei 230012, ChinaSchool of Pharmacy, Anhui University of Chinese Medicine, No. 1, Qianjiang Road, Yaohai District, Hefei 230012, ChinaDendrobium huoshanense has been used for centuries in China and its polysaccharides are the main active components in treating loss of body fluids resulting from fever and asthenic symptoms. However, the biosynthetic pathway of polysaccharides in D. huoshanense remains to be elucidated. In this study, we obtained a guanosine diphosphate (GDP)-mannose pyrophosphorylase (DhGMPP) from D. huoshanense and characterized its function to catalyze the conversion of α-D-mannose-phosphate to GDP-D-mannose involved in the production of polysaccharides. DhGMPP, with the open reading frame of 1,245 bp, was isolated from RNA-Seq data of D. huoshanense. Phylogenetic analysis as well as sequence characterization suggested its involvement in the biosynthesis of GDP-D-mannose. In vitro enzyme assay demonstrated that GMPP encoded a pyrophosphorylase that converted α-D-mannose-phosphate and GTP into GDP-D-mannose. Identification of DhGMPP could provide more insights into the mechanism concerning polysaccharide biosynthesis in D. huoshanense and be utilized for enhancing polysaccharide accumulation through metabolic engineering.https://doi.org/10.1515/biol-2021-0015gdp-mannose pyrophosphorylasepolysaccharide biosynthesisguanosine-5-triphosphoric acid |
| spellingShingle | Yi Yuqi Liu Lulu Zhou Wenyan Peng Daiyin Han Rongchun Yu Nianjun Characterization of GMPP from Dendrobium huoshanense yielding GDP-D-mannose Open Life Sciences gdp-mannose pyrophosphorylase polysaccharide biosynthesis guanosine-5-triphosphoric acid |
| title | Characterization of GMPP from Dendrobium huoshanense yielding GDP-D-mannose |
| title_full | Characterization of GMPP from Dendrobium huoshanense yielding GDP-D-mannose |
| title_fullStr | Characterization of GMPP from Dendrobium huoshanense yielding GDP-D-mannose |
| title_full_unstemmed | Characterization of GMPP from Dendrobium huoshanense yielding GDP-D-mannose |
| title_short | Characterization of GMPP from Dendrobium huoshanense yielding GDP-D-mannose |
| title_sort | characterization of gmpp from dendrobium huoshanense yielding gdp d mannose |
| topic | gdp-mannose pyrophosphorylase polysaccharide biosynthesis guanosine-5-triphosphoric acid |
| url | https://doi.org/10.1515/biol-2021-0015 |
| work_keys_str_mv | AT yiyuqi characterizationofgmppfromdendrobiumhuoshanenseyieldinggdpdmannose AT liululu characterizationofgmppfromdendrobiumhuoshanenseyieldinggdpdmannose AT zhouwenyan characterizationofgmppfromdendrobiumhuoshanenseyieldinggdpdmannose AT pengdaiyin characterizationofgmppfromdendrobiumhuoshanenseyieldinggdpdmannose AT hanrongchun characterizationofgmppfromdendrobiumhuoshanenseyieldinggdpdmannose AT yunianjun characterizationofgmppfromdendrobiumhuoshanenseyieldinggdpdmannose |