Efficient Synthesis of Purine Nucleoside Analogs by a New Trimeric Purine Nucleoside Phosphorylase from <i>Aneurinibacillus migulanus</i> AM007
Purine nucleoside phosphorylases (PNPs) are promising biocatalysts for the synthesis of purine nucleoside analogs. Although a number of PNPs have been reported, the development of highly efficient enzymes for industrial applications is still in high demand. Herein, a new trimeric purine nucleoside p...
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MDPI AG
2019-12-01
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| Series: | Molecules |
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| Online Access: | https://www.mdpi.com/1420-3049/25/1/100 |
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| author | Gaofei Liu Tiantong Cheng Jianlin Chu Sui Li Bingfang He |
| author_facet | Gaofei Liu Tiantong Cheng Jianlin Chu Sui Li Bingfang He |
| author_sort | Gaofei Liu |
| collection | DOAJ |
| description | Purine nucleoside phosphorylases (PNPs) are promising biocatalysts for the synthesis of purine nucleoside analogs. Although a number of PNPs have been reported, the development of highly efficient enzymes for industrial applications is still in high demand. Herein, a new trimeric purine nucleoside phosphorylase (<i>Am</i>PNP) from <i>Aneurinibacillus migulanus</i> AM007 was cloned and heterologously expressed in <i>Escherichia coli</i> BL21(DE3). The <i>Am</i>PNP showed good thermostability and a broad range of pH stability. The enzyme was thermostable below 55 °C for 12 h (retaining nearly 100% of its initial activity), and retained nearly 100% of the initial activity in alkaline buffer systems (pH 7.0−9.0) at 60 °C for 2 h. Then, a one-pot, two-enzyme mode of transglycosylation reaction was successfully constructed by combining pyrimidine nucleoside phosphorylase (<i>Bb</i>PyNP) derived from <i>Brevibacillus borstelensis</i> LK01 and <i>Am</i>PNP for the production of purine nucleoside analogs. Conversions of 2,6-diaminopurine ribonucleoside (<b>1</b>), 2-amino-6-chloropurine ribonucleoside (<b>2</b>), and 6-thioguanine ribonucleoside (<b>3</b>) synthesized still reached >90% on the higher concentrations of substrates (pentofuranosyl donor: purine base; 20:10 mM) with a low enzyme ratio of <i>Bb</i>PyNP: <i>Am</i>PNP (2:20 μg/mL). Thus, the new trimeric <i>Am</i>PNP is a promising biocatalyst for industrial production of purine nucleoside analogs. |
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| issn | 1420-3049 |
| language | English |
| last_indexed | 2024-12-21T21:44:44Z |
| publishDate | 2019-12-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Molecules |
| spelling | doaj.art-37900f3be52647fb97b57b8f93194c0c2022-12-21T18:49:15ZengMDPI AGMolecules1420-30492019-12-0125110010.3390/molecules25010100molecules25010100Efficient Synthesis of Purine Nucleoside Analogs by a New Trimeric Purine Nucleoside Phosphorylase from <i>Aneurinibacillus migulanus</i> AM007Gaofei Liu0Tiantong Cheng1Jianlin Chu2Sui Li3Bingfang He4College of Biotechnology and Pharmaceutical Engineering, Nanjing Tech University, Nanjing 211800, ChinaCollege of Biotechnology and Pharmaceutical Engineering, Nanjing Tech University, Nanjing 211800, ChinaSchool of Pharmaceutical Sciences, Nanjing Tech University, Nanjing 211800, ChinaSchool of Pharmaceutical Sciences, Nanjing Tech University, Nanjing 211800, ChinaSchool of Pharmaceutical Sciences, Nanjing Tech University, Nanjing 211800, ChinaPurine nucleoside phosphorylases (PNPs) are promising biocatalysts for the synthesis of purine nucleoside analogs. Although a number of PNPs have been reported, the development of highly efficient enzymes for industrial applications is still in high demand. Herein, a new trimeric purine nucleoside phosphorylase (<i>Am</i>PNP) from <i>Aneurinibacillus migulanus</i> AM007 was cloned and heterologously expressed in <i>Escherichia coli</i> BL21(DE3). The <i>Am</i>PNP showed good thermostability and a broad range of pH stability. The enzyme was thermostable below 55 °C for 12 h (retaining nearly 100% of its initial activity), and retained nearly 100% of the initial activity in alkaline buffer systems (pH 7.0−9.0) at 60 °C for 2 h. Then, a one-pot, two-enzyme mode of transglycosylation reaction was successfully constructed by combining pyrimidine nucleoside phosphorylase (<i>Bb</i>PyNP) derived from <i>Brevibacillus borstelensis</i> LK01 and <i>Am</i>PNP for the production of purine nucleoside analogs. Conversions of 2,6-diaminopurine ribonucleoside (<b>1</b>), 2-amino-6-chloropurine ribonucleoside (<b>2</b>), and 6-thioguanine ribonucleoside (<b>3</b>) synthesized still reached >90% on the higher concentrations of substrates (pentofuranosyl donor: purine base; 20:10 mM) with a low enzyme ratio of <i>Bb</i>PyNP: <i>Am</i>PNP (2:20 μg/mL). Thus, the new trimeric <i>Am</i>PNP is a promising biocatalyst for industrial production of purine nucleoside analogs.https://www.mdpi.com/1420-3049/25/1/100trimeric nucleoside phosphorylasethermostabilitypurine nucleoside analogsenzyme ratio |
| spellingShingle | Gaofei Liu Tiantong Cheng Jianlin Chu Sui Li Bingfang He Efficient Synthesis of Purine Nucleoside Analogs by a New Trimeric Purine Nucleoside Phosphorylase from <i>Aneurinibacillus migulanus</i> AM007 Molecules trimeric nucleoside phosphorylase thermostability purine nucleoside analogs enzyme ratio |
| title | Efficient Synthesis of Purine Nucleoside Analogs by a New Trimeric Purine Nucleoside Phosphorylase from <i>Aneurinibacillus migulanus</i> AM007 |
| title_full | Efficient Synthesis of Purine Nucleoside Analogs by a New Trimeric Purine Nucleoside Phosphorylase from <i>Aneurinibacillus migulanus</i> AM007 |
| title_fullStr | Efficient Synthesis of Purine Nucleoside Analogs by a New Trimeric Purine Nucleoside Phosphorylase from <i>Aneurinibacillus migulanus</i> AM007 |
| title_full_unstemmed | Efficient Synthesis of Purine Nucleoside Analogs by a New Trimeric Purine Nucleoside Phosphorylase from <i>Aneurinibacillus migulanus</i> AM007 |
| title_short | Efficient Synthesis of Purine Nucleoside Analogs by a New Trimeric Purine Nucleoside Phosphorylase from <i>Aneurinibacillus migulanus</i> AM007 |
| title_sort | efficient synthesis of purine nucleoside analogs by a new trimeric purine nucleoside phosphorylase from i aneurinibacillus migulanus i am007 |
| topic | trimeric nucleoside phosphorylase thermostability purine nucleoside analogs enzyme ratio |
| url | https://www.mdpi.com/1420-3049/25/1/100 |
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