Title The Structure of Protease Inhibitors Produced by Streotomyces exfoliatus SMF13
Author Kim, In Seop · Han, Young Tae¹ · Barrow, Kevin D. ¹ · Lee, Kye Joon *
Address Department of Microbiology and Research Center for Molecular Microbiology, Seoul National University; ¹School of Biochemistry and Molecular Genetics, The University of New South Wales
Bibliography Korean Journal of Microbiology, 31(4),326-334, 1993
Key Words Streotomyces exfoliatus, leupeptin, NMR, structure determination
Abstract Streotomyces exfoliatus SMF13 produced extra-cellular protease inhibitors inhibiting thiol protease/trypsin and extra-cellular protease generated by the strain. The inhibitors in culture filtrate were partially purified by adsorption on activated charcoal, butanol extraction, silica gel chromatography, and sequential ion exchange chromatography usin Amberlite CG 50(H^+ form) and Dowex-1(Cl^- form). The inhibitors obtained from the chromatographies consisted of three mixtures of free aldehyde form, hydrate form, and carbibolamie form because of the C-terminal arginal. The inhibitors were oxidized with KMnO_4 for fixing the C-terminal aldehyde to carboxylic acid and separated into individual components by HPLC. Based upon amino acid, IR spectrometry, mass spectrometry, ^1H-NMR, and ^13C-NMR analysis for each of individual peaks, the structure of protease inhibitors were found to ba Acetyl-Leu-Leu Arginal(leupeptin), Acetyl-Ile-Leu-Arginal, and Acetyl-Leu-Ile-Arginal. The latter molecule represents new leupeptin analogue of microbial origin.
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