What question did this study set out to answer?

Isolate, purify, and biochemically characterize the peptide synthetases responsible for activating precursor residues during pristinamycin I biosynthesis.

February 1, 1997Journal of BacteriologyOpen Access

Purification of peptide synthetases involved in pristinamycin I biosynthesis

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Authors

DTD ThibautUniversity of Stuttgart DBD BischSmiths Detection (France)NRN RatetSmiths Detection (France)

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Implication

Biochemical analysis reveals three peptide synthetases activating pristinamycin I precursors in Streptomyces pristinaespiralis, highlighting the multienzyme logic of nonribosomal antibiotic assembly.

Key Points

Isolate, purify, and biochemically characterize the peptide synthetases responsible for activating precursor residues during pristinamycin I biosynthesis.
Monitored enzymatic activities in Streptomyces pristinaespiralis cell extracts using adenylate and thioester formation assays, including ATP-pyrophosphate exchange.
Purified candidate synthetases to near homogeneity and estimated molecular masses using sodium dodecyl sulfate-polyacrylamide gel electrophoresis.
Analyzed primary structure via stepwise Edman degradation of intact polypeptides and internal peptide fragments.
Purified SnbA (monomer, 67 kDa) 200-fold as a 3-hydroxypicolinic acid:AMP ligase that activates the initial pristinamycin I residue.
Purified SnbC (homodimer with 240 kDa subunits) 100-fold, demonstrating covalent thioester binding of L-threonine and activation of L-aminobutyric acid for subsequent epimerization.
Purified SnbD (homodimer with 250–350 kDa subunits) 200-fold to catalyze thioesterification and N-methylation of 4-dimethylamino-L-phenylalanine alongside L-proline activation, and identified SnbE (170 kDa, purified 3,000-fold) as its proteolytic fragment.

Cite This Study

Thibaut et al. (1997) studied this question.

synapsesocial.com/papers/6a952d06b7c083589ab2cd05 https://doi.org/10.1128/jb.179.3.697-704.1997

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Implication

Biochemical analysis reveals three peptide synthetases activating pristinamycin I precursors in Streptomyces pristinaespiralis, highlighting the multienzyme logic of nonribosomal antibiotic assembly.

Purification of peptide synthetases involved in pristinamycin I biosynthesis

Discussion

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Implication

Key Points

Cite This Study

Purification of peptide synthetases involved in pristinamycin I biosynthesis

Discussion

Member takes

Implication

Key Points

Cite This Study

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