PMID: 9531495Jun 11, 1998Paper

Characterization of the leupeptin-inactivating enzyme from Streptomyces exfoliatus SMF13 which produces leupeptin

The Biochemical Journal
I S KimK J Lee

Abstract

Leupeptin-inactivating enzyme (LIE) was purified from Streptomyces exfoliatus SMF13 by ammonium sulphate fractionation of cell-free culture broth, ultrafiltration, anion-exchange chromatography on DEAE-Sephadex A-50 and gel filtration chromatography on Sephadex G-75. The molecular mass of the purified enzyme was measured as 34700 Da and the N-terminal amino acid sequence was APTPPDIPLANVPA. Acetyl-leucine, leucine and argininal were identified as the products of leupeptin inactivated by the LIE, indicating that leupeptin is inactivated by hydrolysis of peptide bond between leucine and leucine and between leucine and argininal of leupeptin (acetyl-leucine-leucine-argininal). Synthetic-peptide substrates specificity of LIE showed that LIE has absolute specificity for peptide bonds with leucine in the P1 position, suggesting that LIE is a leucine-specific protease. The optimum pH and temperature were pH 9.0 and 45 degrees C, respectively. LIE activity was inhibited by metalloprotease inhibitors such as EDTA, EGTA, o-phenanthroline and bestatin, but activated by Mg2+ and Ca2+, suggesting that the enzyme is a metalloprotease. Aerial-mycelium growth and aerial spore formation of S. exfoliatus SMF13 were inhibited by the addition of b...Continue Reading

Citations

Apr 7, 2016·Medical Microbiology and Immunology·Mateusz KędziorJan Gutowicz
Jan 22, 2010·FEMS Microbiology Reviews·Keith F ChaterHildgund Schrempf
Jan 2, 2016·International Journal of Biological Macromolecules·Zhibin SunZhongli Cui
Feb 4, 2009·Applied Biochemistry and Biotechnology·Bo WuBin Yao
Jul 19, 2003·European Journal of Biochemistry·J ZotzelH-L Fuchsbauer
Dec 17, 2004·Journal of Bacteriology·Jun-Ya KatoSueharu Horinouchi
Jul 2, 2020·Angewandte Chemie·Jhe-Hao LiJason M Crawford
Apr 9, 2020·Frontiers in Microbiology·Maria Petronella HumanBridget Genevieve Crampton
Oct 10, 1998·Journal of Bacteriology·I NagyR De Mot

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