Direct pathway cloning and expression of the radiosumin biosynthetic gene cluster
Autor: | Xiaodan Ouyang, Paul M. D'Agostino, Matti Wahlsten, Endrews Delbaje, Jouni Jokela, Perttu Permi, Greta Giaini, Antti Poso, Piia Bartos, Tobias A. M. Gulder, Hannu Koistinen, David Fewer |
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Přispěvatelé: | Department of Food and Nutrition, Department of Microbiology, Cyanobacteria research, Research Programs Unit, Department of Clinical Chemistry and Hematology, HUS Helsinki and Uusimaa Hospital District, Helsinki Institute of Sustainability Science (HELSUS), Microbial Natural Products |
Jazyk: | angličtina |
Rok vydání: | 2023 |
Předmět: |
11832 Microbiology and virology
Identification Diversity Organic Chemistry Bacillus-subtilis 116 Chemical sciences Fresh-water DNA Protease inhibitors Cyanobacteria Biochemistry Quality Nonribosomal peptide geneettinen monimuotoisuus Natural-products Trypsin-inhibitor Physical and Theoretical Chemistry syanobakteerit |
Popis: | Radiosumins are a structurally diverse family of low molecular weight natural products that are produced by cyanobacteria and exhibit potent serine protease inhibition. Members of this family are dipeptides characterized by the presence of two similar non-proteinogenic amino acids. Here we used a comparative bioinformatic analysis to identify radiosumin biosynthetic gene clusters from the genomes of 13 filamentous cyanobacteria. We used direct pathway cloning to capture and express the entire 16.8 kb radiosumin biosynthetic gene cluster from Dolichospermum planctonicum UHCC 0167 in Escherichia coli. Bioinformatic analysis demonstrates that radiosumins represent a new group of chorismate-derived non-aromatic secondary metabolites. High-resolution liquid chromatography-mass spectrometry, nuclear magnetic resonance spectroscopy and chemical degradation analysis revealed that cyanobacteria produce a cocktail of novel radiosumins. We report the chemical structure of radiosumin D, an N-methyl dipeptide, containing a special Aayp (2-amino-3-(4-amino-2-cyclohexen-1-ylidene) propionic acid) with R configuration that differs from radiosumin A-C, an N-Me derivative of Aayp (Amyp) and two acetyl groups. Radiosumin C inhibits all three human trypsin isoforms at micromolar concentrations with preference for trypsin-1 and -3 (IC50 values from 1.7 mu M to >7.2 mu M). These results provide a biosynthetic logic to explore the genetic and chemical diversity of the radiosumin family and suggest that these natural products may be a source of drug leads for selective human serine proteases inhibitors. |
Databáze: | OpenAIRE |
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