The SGNH-hydrolase of Streptomyces coelicolor has (aryl)esterase and a true lipase activity. (CROSBI ID 146067)
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Podaci o odgovornosti
Bielen, Ana ; Ćetković, Helena ; Long, Paul F. ; Schwab, Helmuth ; Abramić, Marija ; Vujaklija, Dušica
engleski
The SGNH-hydrolase of Streptomyces coelicolor has (aryl)esterase and a true lipase activity.
The Streptomyces coelicolor A3(2) gene SCI11.14c was overexpressed and purified as a His-tagged protein from heterologous host, Streptomyces lividans. The purification procedure resulted in 34.1-fold increase in specific activity with an overall yield of 21.4%. Biochemical and physical properties of the purified enzyme were investigated and it was shown that it possesses (aryl)esterase and a true lipase activity. The enzyme was able to hydrolyze p-nitrophenyl-, alpha- and beta-naphthyl esters and poly(oxyethylene) sorbitan monoesters (Tween 20-80). It showed pronounced activity towards p-nitrophenyl and alpha- and beta-naphthyl esters of C(12)-C(16). Higher activity was observed with alpha-naphthyl esters. The enzyme hydrolyzed triolein (specific activity: 91.9U/mg) and a wide range of oils with a preference for those having higher content of linoleic or oleic acid (C18:2 ; C18:1, cis). The active-site serine specific inhibitor 3, 4-dichloroisocoumarin (DCI) strongly inhibited the enzyme, while tetrahydrofurane and 1, 4-dioxane significantly increased (2- and 4- fold, respectively) hydrolytic activity of lipase towards p-nitrophenyl caprylate. The enzyme exhibited relatively high temperature optimum (55 degrees C) and thermal stability. CD analysis revealed predominance of alpha-helical structure (54% alpha-helix, 21% beta-sheet) and a T(m) value at 66 degrees C. Systematic bioinformatic analysis of deduced amino acid sequence of S. coelicolor enzyme placed it to the SGNH-hydrolase family. Phylogenetic analysis of the predicted protein homologues to the S. coelicolor SGNH-hydrolase generated three distinct groups consisting of proteins from Actinomycetales, Ascomycota and nematodes. At present it seems that these enzymes are most conserved among soil inhabiting organisms.
Streptomyces coelicolor ; GDSL/SGNH porodica ; Lipaza ; (Aril)esteraza
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