The extracellular p-I-type proteinase of Lactococcus lactis hydrolyzes beta-casein into more than one hundred different oligopeptides
| Autor: | V. Juillard, Harry Laan, Wn Konings, C. M. Jeronimus-Stratingh, A. P. Bruins, Edmund R.S. Kunji |
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| Přispěvatelé: | Bactéries Lactiques et Pathogènes Opportunistes (UBLO), Institut National de la Recherche Agronomique (INRA) |
| Jazyk: | angličtina |
| Rok vydání: | 1995 |
| Předmět: |
Molecular Sequence Data
Peptide Tripeptide Oligopeptide transport Microbiology Mass Spectrometry 03 medical and health sciences Bacterial Proteins Endopeptidases LACTOCOCCUS LACTIS Amino Acid Sequence Molecular Biology Peptide sequence Chromatography High Pressure Liquid ComputingMilieux_MISCELLANEOUS 030304 developmental biology chemistry.chemical_classification 0303 health sciences Oligopeptide Chromatography biology 030306 microbiology Hydrolysis Lactococcus lactis Serine Endopeptidases Proteolytic enzymes Caseins Biological Transport biology.organism_classification Amino acid [SDV.MP]Life Sciences [q-bio]/Microbiology and Parasitology chemistry Biochemistry Oligopeptides Research Article Chromatography Liquid |
| Zdroj: | Journal of Bacteriology Journal of Bacteriology, American Society for Microbiology, 1995, 177 (12), pp.3472-3478 |
| ISSN: | 0021-9193 1098-5530 |
| Popis: | The peptides released from beta-casein by the action of PI-type proteinase (PrtP) from Lactococcus lactis subsp. cremoris Wg2 have been identified by on-line coupling of liquid chromatography to mass spectrometry. After 24 h of incubation of beta-casein with purified PrtP, a stable mixture of peptides was obtained. The trifluoroacetic acid-soluble peptides of this beta-casein hydrolysate were fractionated by high-performance liquid chromatography and introduced into the liquid chromatography-ion spray mass spectrometry interface. Multiply charged ions were generated from trifluoroacetic acid-soluble peptides under low nozzle voltage conditions, yielding the MH+ mass of each eluted peptide. All peptides corresponding to each of the MH+ calculated masses were determined. In those cases in which different peptides were possible, further identification was achieved by collision-induced dissociation under higher nozzle voltage conditions. Hydrolysis of beta-casein by PrtP was observed to proceed much further than reported previously. More than 40% of the peptide bonds are cleaved by PrtP, resulting in the formation of more than 100 different oligopeptides. With the exception of Phe, significant release of amino acids or di- and tripeptides could not be observed. Interestingly, one-fifth of the identified oligopeptides are small enough to be taken up by the oligopeptide transport system. Uptake of these peptides could supply L. lactis with all amino acids, including the essential ones, indicating that growth of L. lactis might be possible on peptides released from beta-casein by proteinase only. |
| Databáze: | OpenAIRE |
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