Chromatographic separation and partial characterization of microbial low-molecular-weight proteinase inhibitors. 1988

I Kourteva
Institute of Microbiology, Bulgarian Academy of Sciences, Sofia.

A crude preparation, inhibiting trypsin (T), chymotrypsin (C) and papain (P), was isolated from the culture filtrate of Actinomyces sp. 9 by butanol extraction. Ion-exchange high-performance liquid chromatography (HPLC) of this preparation on a Mono S column resulted in the separation of two inhibitory fractions: one active against T (TI-9) and the other active against C and P (CPI-9). These two fractions, having the same inhibitory specificity, were also obtained from the crude extract by size-exclusion HPLC on a Superose 12 column. This method proved to be better in terms of the purity of the fractionated inhibitors than ion-exchange chromatography. Further purification of TI-9 and CPI-9 was achieved by using reversed-phase HPLC on a PEP RPC column. In this case, TI-9 was obtained as an apparently homogeneous peak. Studies on the physicochemical properties of the purified inhibitors showed them to be small molecules, similar in hydrophobicity, pH and thermal stability, but differing in solubility. Amino acid and spectral analyses provided the peptidic structure of TI-9 and gave a molecular weight of 1643, while parallel analyses of CPI-9 showed that it lacks the common amino acids.

UI MeSH Term Description Entries
D011480 Protease Inhibitors Compounds which inhibit or antagonize biosynthesis or actions of proteases (ENDOPEPTIDASES). Antiprotease,Endopeptidase Inhibitor,Endopeptidase Inhibitors,Peptidase Inhibitor,Peptidase Inhibitors,Peptide Hydrolase Inhibitor,Peptide Hydrolase Inhibitors,Peptide Peptidohydrolase Inhibitor,Peptide Peptidohydrolase Inhibitors,Protease Antagonist,Protease Antagonists,Antiproteases,Protease Inhibitor,Antagonist, Protease,Antagonists, Protease,Hydrolase Inhibitor, Peptide,Hydrolase Inhibitors, Peptide,Inhibitor, Endopeptidase,Inhibitor, Peptidase,Inhibitor, Peptide Hydrolase,Inhibitor, Peptide Peptidohydrolase,Inhibitor, Protease,Inhibitors, Endopeptidase,Inhibitors, Peptidase,Inhibitors, Peptide Hydrolase,Inhibitors, Peptide Peptidohydrolase,Inhibitors, Protease,Peptidohydrolase Inhibitor, Peptide,Peptidohydrolase Inhibitors, Peptide
D002851 Chromatography, High Pressure Liquid Liquid chromatographic techniques which feature high inlet pressures, high sensitivity, and high speed. Chromatography, High Performance Liquid,Chromatography, High Speed Liquid,Chromatography, Liquid, High Pressure,HPLC,High Performance Liquid Chromatography,High-Performance Liquid Chromatography,UPLC,Ultra Performance Liquid Chromatography,Chromatography, High-Performance Liquid,High-Performance Liquid Chromatographies,Liquid Chromatography, High-Performance
D002855 Chromatography, Thin Layer Chromatography on thin layers of adsorbents rather than in columns. The adsorbent can be alumina, silica gel, silicates, charcoals, or cellulose. (McGraw-Hill Dictionary of Scientific and Technical Terms, 4th ed) Chromatography, Thin-Layer,Thin Layer Chromatography,Chromatographies, Thin Layer,Chromatographies, Thin-Layer,Thin Layer Chromatographies,Thin-Layer Chromatographies,Thin-Layer Chromatography
D000190 Actinomyces A genus of gram-positive, rod-shaped bacteria whose organisms are nonmotile. Filaments that may be present in certain species are either straight or wavy and may have swollen or clubbed heads.
D000596 Amino Acids Organic compounds that generally contain an amino (-NH2) and a carboxyl (-COOH) group. Twenty alpha-amino acids are the subunits which are polymerized to form proteins. Amino Acid,Acid, Amino,Acids, Amino

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