Functional Analysis of a Gene Cluster from Chitinophaga pinensis Involved in Biosynthesis of the Pyrrolidine Azasugar DAB-1
| العنوان: | Functional Analysis of a Gene Cluster from Chitinophaga pinensis Involved in Biosynthesis of the Pyrrolidine Azasugar DAB-1 |
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| المؤلفون: | Nicole A. Horenstein, Claribel Nuñez |
| المصدر: | Journal of Natural Products. 82:3401-3409 |
| بيانات النشر: | American Chemical Society (ACS), 2019. |
| سنة النشر: | 2019 |
| مصطلحات موضوعية: | Pharmacology, chemistry.chemical_classification, Bacillus amyloliquefaciens, biology, Organic Chemistry, Pharmaceutical Science, biology.organism_classification, Pyrrolidine, Analytical Chemistry, chemistry.chemical_compound, Enzyme, Complementary and alternative medicine, chemistry, Biosynthesis, Biochemistry, Drug Discovery, biology.protein, Molecular Medicine, Monosaccharide, Heterologous expression, Maltase, Alcohol dehydrogenase |
| الوصف: | Azasugars, "nitrogen in the ring" analogues of monosaccharides, are known to be distributed in select plant, fungal. and bacterial species. We identify Chitinophaga pinensis DSM 2588 as the first bacterial source of the plant pyrrolidine azasugar 1,4-dideoxy-1,4-aminoarabinitol (DAB-1). Comparative sequence analyses identified C. pinensis as a putative azasugar producer, via observation of a three-gene cluster coding for putative aminotransferase, alcohol dehydrogenase, and sugar phosphatase enzymes, similar to the previously reported azasugar biosynthetic signature identified in Bacillus amyloliquefaciens FZB42. Multistep fractionation of C. pinensis culture media guided by a maltase inhibition assay yielded a component with a mass consistent with the structure of DAB-1. Heterologous expression of the three-gene cluster in E. coli, a non-azasugar producer, led to the isolation of nectrisine, a biosynthetic precursor to DAB-1, which displayed potent slow tight binding inhibition of maltase. Reduction of nectrisine with NaBH4 removed the slow tight binding inhibition kinetics, and MS analysis provided evidence for the production of a compound matching that of the isolated DAB-1 from C. pinensis. 1H NMR analysis of the nectrisine produced in E. coli after NaBD4 reduction produced a spectrum consistent with DAB-1 deuterated at C-1, primarily at the pro-S position. These results support the idea that the azasugar three-gene cluster represents a general biosynthetic path leading to several different compounds, which may prove useful for the identification of other azasugar-producing organisms. |
| تدمد: | 1520-6025 0163-3864 |
| DOI: | 10.1021/acs.jnatprod.9b00758 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_________::a3b0ce13ab1503a66788a91fe1dc968b https://doi.org/10.1021/acs.jnatprod.9b00758 |
| Rights: | CLOSED |
| رقم الانضمام: | edsair.doi...........a3b0ce13ab1503a66788a91fe1dc968b |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 15206025 01633864 |
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| DOI: | 10.1021/acs.jnatprod.9b00758 |