Characterization of a hyperthermophilic phosphatase from Archaeoglobus fulgidus and its application in in vitro synthetic enzymatic biosystem
Abstract Background Haloacid dehalogenase (HAD)-like hydrolases represent the largest superfamily of phosphatases, which release inorganic phosphate from phosphate containing compounds, such as sugar phosphates. The HAD-like phosphatases with highly substrate specificity, which perform irreversible...
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| Format: | Article |
| Language: | English |
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SpringerOpen
2019-06-01
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| Series: | Bioresources and Bioprocessing |
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| Online Access: | http://link.springer.com/article/10.1186/s40643-019-0257-5 |
| _version_ | 1811313890526494720 |
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| author | Wei Wang Dongdong Meng Qiangzi Li Zhimin Li Chun You |
| author_facet | Wei Wang Dongdong Meng Qiangzi Li Zhimin Li Chun You |
| author_sort | Wei Wang |
| collection | DOAJ |
| description | Abstract Background Haloacid dehalogenase (HAD)-like hydrolases represent the largest superfamily of phosphatases, which release inorganic phosphate from phosphate containing compounds, such as sugar phosphates. The HAD-like phosphatases with highly substrate specificity, which perform irreversible dephosphorylation, are always integrated into in vitro synthetic enzymatic biosystems as the last enzymatic step for the cost-efficient production of biochemicals. Therefore, identification and characterization of substrate specificity of HAD-like phosphatases are important for exploring their application. Results In this study, a hyperthermophilic HAD-like phosphatase from Archaeoglobus fulgidus (AfPase) was cloned, expressed, and characterized. AfPase was identified as a type I Mg2+-dependent HAD-like phosphatase with high optimal temperature and thermostability. Among the tested phosphate containing compounds, AfPase exhibited the highest catalytic activity on p-nitrophenyl phosphate, followed by dihydroxyacetone phosphate (DHAP). On the basis of the high catalytic activity of AfPase to generate 1,3-dihydroxyacetone (DHA) from DHAP, an in vitro synthetic enzymatic biosystem containing this phosphatase and other five enzymes was constructed for the biosynthesis of DHA from inexpensive maltodextrin in one pot. About 14 mM (1.26 g/L) DHA was produced from 10 g/L maltodextrin. Conclusions A hyperthermophilic HAD-like phosphatase from Archaeoglobus fulgidus was characterized carefully, and the success of an in vitro synthetic enzymatic biosystem containing this phosphatase provided a promising approach for DHA production from maltodextrin. |
| first_indexed | 2024-04-13T11:02:39Z |
| format | Article |
| id | doaj.art-c7e5c725e6204704a7ba8c97d01173c3 |
| institution | Directory Open Access Journal |
| issn | 2197-4365 |
| language | English |
| last_indexed | 2024-04-13T11:02:39Z |
| publishDate | 2019-06-01 |
| publisher | SpringerOpen |
| record_format | Article |
| series | Bioresources and Bioprocessing |
| spelling | doaj.art-c7e5c725e6204704a7ba8c97d01173c32022-12-22T02:49:21ZengSpringerOpenBioresources and Bioprocessing2197-43652019-06-016111110.1186/s40643-019-0257-5Characterization of a hyperthermophilic phosphatase from Archaeoglobus fulgidus and its application in in vitro synthetic enzymatic biosystemWei Wang0Dongdong Meng1Qiangzi Li2Zhimin Li3Chun You4State Key Laboratory of Bioreactor Engineering, East China University of Science and TechnologyTianjin Institute of Industrial Biotechnology, Chinese Academy of SciencesTianjin Institute of Industrial Biotechnology, Chinese Academy of SciencesState Key Laboratory of Bioreactor Engineering, East China University of Science and TechnologyTianjin Institute of Industrial Biotechnology, Chinese Academy of SciencesAbstract Background Haloacid dehalogenase (HAD)-like hydrolases represent the largest superfamily of phosphatases, which release inorganic phosphate from phosphate containing compounds, such as sugar phosphates. The HAD-like phosphatases with highly substrate specificity, which perform irreversible dephosphorylation, are always integrated into in vitro synthetic enzymatic biosystems as the last enzymatic step for the cost-efficient production of biochemicals. Therefore, identification and characterization of substrate specificity of HAD-like phosphatases are important for exploring their application. Results In this study, a hyperthermophilic HAD-like phosphatase from Archaeoglobus fulgidus (AfPase) was cloned, expressed, and characterized. AfPase was identified as a type I Mg2+-dependent HAD-like phosphatase with high optimal temperature and thermostability. Among the tested phosphate containing compounds, AfPase exhibited the highest catalytic activity on p-nitrophenyl phosphate, followed by dihydroxyacetone phosphate (DHAP). On the basis of the high catalytic activity of AfPase to generate 1,3-dihydroxyacetone (DHA) from DHAP, an in vitro synthetic enzymatic biosystem containing this phosphatase and other five enzymes was constructed for the biosynthesis of DHA from inexpensive maltodextrin in one pot. About 14 mM (1.26 g/L) DHA was produced from 10 g/L maltodextrin. Conclusions A hyperthermophilic HAD-like phosphatase from Archaeoglobus fulgidus was characterized carefully, and the success of an in vitro synthetic enzymatic biosystem containing this phosphatase provided a promising approach for DHA production from maltodextrin.http://link.springer.com/article/10.1186/s40643-019-0257-5HAD-like hydrolasePhosphataseEnzyme promiscuityIn vitro synthetic enzymatic biosystem1,3-Dihydroxyacetone |
| spellingShingle | Wei Wang Dongdong Meng Qiangzi Li Zhimin Li Chun You Characterization of a hyperthermophilic phosphatase from Archaeoglobus fulgidus and its application in in vitro synthetic enzymatic biosystem Bioresources and Bioprocessing HAD-like hydrolase Phosphatase Enzyme promiscuity In vitro synthetic enzymatic biosystem 1,3-Dihydroxyacetone |
| title | Characterization of a hyperthermophilic phosphatase from Archaeoglobus fulgidus and its application in in vitro synthetic enzymatic biosystem |
| title_full | Characterization of a hyperthermophilic phosphatase from Archaeoglobus fulgidus and its application in in vitro synthetic enzymatic biosystem |
| title_fullStr | Characterization of a hyperthermophilic phosphatase from Archaeoglobus fulgidus and its application in in vitro synthetic enzymatic biosystem |
| title_full_unstemmed | Characterization of a hyperthermophilic phosphatase from Archaeoglobus fulgidus and its application in in vitro synthetic enzymatic biosystem |
| title_short | Characterization of a hyperthermophilic phosphatase from Archaeoglobus fulgidus and its application in in vitro synthetic enzymatic biosystem |
| title_sort | characterization of a hyperthermophilic phosphatase from archaeoglobus fulgidus and its application in in vitro synthetic enzymatic biosystem |
| topic | HAD-like hydrolase Phosphatase Enzyme promiscuity In vitro synthetic enzymatic biosystem 1,3-Dihydroxyacetone |
| url | http://link.springer.com/article/10.1186/s40643-019-0257-5 |
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