Yakup Kolcuoğlu

Publications (3)0.81 Total impact

• Article: AN ESTEROLYTIC ACTIVITY FROM A WILD EDIBLE MUSHROOM, LYCOPERDON PERLATUM

  AHMET COLAK, YASEMIN CAMEDAN, ÖZLEM FAIZ, ERTUGRUL SESLI, YAKUP KOLCUOĞLU
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  ABSTRACT: Lycoperdon perlatum Pers. (Lycoperdaceae, Agaricales, Agaricomycetidae, Agaricomycetes, Basidiomycota, Fungi) was evaluated for its esterolytic potential. Native electrophoresis of the crude extracts showed four bands having Rf values of 0.34, 0.39, 0.52 and 0.59. The esterase showed the highest activity toward a short-chain substrate, p-nitrophenyl acetate. Optimum reaction conditions for L. perlatum crude extract were attained at pH 8.0 and 40C. Esterolytic activity of enzyme extract was stimulated in the presence of Mn2+, Fe2+, Ca2+ and Zn2+ in the reaction mixture. The enzyme activity was stimulated by incubation at pH 6.0 but retained 77% of its original activity at its optimum pH after 24 h. Thermal inactivation was displayed after incubation for 20 min at various temperatures above 30C. At 1 mM final concentration, 2-mercaptoethanol, dithiothreitol, ethylenediamine tetraacetic acid and p-methylphenyl sulfonylfluoride inhibited the esterolytic reaction. These results support that the crude L. perlatum extract possesses an esterolytic activity having properties similar to other esterases.PRACTICAL APPLICATIONSEsterases catalyzing the cleavage and formation of ester bonds are known α/β-hydrolases (EC 3.1.1.X). Esterases are used for the synthesis of flavor esters for the food industry, modification of triglycerides for fat and oil industry and resolution of racemic mixtures used for the synthesis of fine chemicals for the pharmaceutical industry. Therefore, the search for new enzyme sources is important for the development of new enzymes and applications.
  Journal of Food Biochemistry 06/2009; 33(4):482 - 499. · 0.81 Impact Factor
• Article: Cloning, expression and characterization of highly thermo- and pH-stable maltogenic amylase from a thermophilic bacterium Geobacillus caldoxylosilyticus TK4

  Yakup Kolcuoğlu, Ahmet Colak, Ozlem Faiz, Ali Osman Belduz
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  ABSTRACT: Maltogenic amylases (MAases), a subclass of cyclodextrin (CD)-hydrolyzing enzymes, belong to glycoside hydrolase family 13. A gene corresponding to MA in Geobacillus caldoxylosilyticus TK4 (GcaTK4MA) was cloned into pET28a(+) vector and expressed in Escherichia coli with 6xHis-tag at the N-terminus. Herein, we report on the biochemical properties of a new thermo- and pH-stable MA. GcaTK4MA has similar properties those of other MAases in terms of the primary structure, preference for CD over starch and having an extra domain at its N- and C-terminals. The recombinant protein was purified efficiently by using one-step nickel affinity chromatography. The purified enzyme exhibited optimal activity for β-CD hydrolysis at 50 °C and pH 7.0. When the enzyme was separately incubated at 4 °C and 50 °C in the buffer solutions (pH 3.0–9.0) up to 7 days, it was seen that the enzyme had the higher stability at 50 °C than 4 °C. The enzyme retained about 80% of its original activity when it was incubated at 50 °C for 7 days. The enzyme activity was significantly inhibited by SDS and EDTA at the final concentration of 1%. These results suggest that this is the first reported MA having an extremely pH- and thermal stabilities.
  Process Biochemistry.
• Article: Comparative characterization of monophenolase and diphenolase activities from a wild edible mushroom (Macrolepiota mastoidea)

  Yakup Kolcuoğlu, Ahmet Colak, Ertuğrul Sesli, Melike Yildirim, Nagihan Saglam
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  ABSTRACT: In this study, Macrolepiota mastoidea, a wild edible mushroom, was evaluated for its polyphenol oxidase potential. Native electrophoresis, stained by l-dihydroxyphenylalanine, of the crude extracts from this species showed two bands having Rf values of 0.38 (minor) and 0.50 (major), supporting a polyphenol oxidase potential. The crude extracts showed monophenolase activity against 3-(4-hydroxyphenyl)-propionic acid and diphenolase activity against 4-methylcatechol as substrates. Monophenolase and diphenolase activities of enzyme extract prepared from M. mastoidea showed pH optimum values at pH 6.0 and pH 4.0, respectively. The extracts retained about 100% and 60% of their original monophenolase and diphenolase activities at their optimum pH values, respectively. It was estimated from thermodynamic data that M. mastoidea had a thermostable monophenolase activity. Thiourea and ascorbic acid were highly potential inhibitors for monophenolase, and ascorbic acid and sodium metabisulphite for diphenolase activity. It is clear from the present results that the enzyme extracts prepared from M. mastoidea possess polyphenol oxidase activities with interesting properties.
  Food Chemistry.