07 خرداد 1403
عباس زماني

عباس زمانی

مرتبه علمی: دانشیار
نشانی: ملایر - کیلومتر 4 جاده اراک - دانشگاه ملایر - دانشکده منابع طبیعی و محیط زیست - گروه علوم و مهندسی شیلات
تحصیلات: دکترای تخصصی / مهندسی منابع طبیعی - شیلات
تلفن: +988132355330
دانشکده: دانشکده منابع طبیعی و محیط زیست

مشخصات پژوهش

عنوان
Trypsin Enzyme from Viscera of Common Kilka (Clupeonella Cultriventris Caspia): Purification, Characterization, and Its Compatibility with Oxidants and Surfactants
نوع پژوهش مقاله چاپ شده
کلیدواژه‌ها
characterization, Clupeonella cultriventris caspia, purification, trypsin
سال
2014
مجله Journal of Aquatic Food Product Technology
پژوهشگران عباس زمانی

چکیده

The purification of trypsin from the common kilka (Clupeonella cultriventris caspia) viscera (pyloric caeca) resulted in a 28.3-fold increase and 12% yield by ammonium sulfate precipitation (30-60%), Sephadex G-75, and DEAE–cellulose chromatography. Trypsin showed a molecular weight of 23.2 kDa and appeared as a single band on sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE), native-PAGE, and zymography. The trypsin had optimal activity at pH 8.0 and 60 °C for the hydrolysis of α-N-benzoyl-DL-arginine-ρ-nitroanilide hydrochloride (BAPNA) substrate. Trypsin was stable up to 50 °C and at pH range of 7.0-10.0. Activity was significantly inhibited by soybean trypsin inhibitor (SBTI) and N-ρ-tosyl-L-lysinechloromethylketone (TLCK) inhibitors (P < 0.05). The enzyme was relatively stable towards oxidizing agents, retaining 59.7 and 98.0% of its initial activity after 1 h incubation in the presence of 15% H2O2 and 1% sodium perborate, respectively. Trypsin was significantly activated by surfactants and Ca2+, Mg2+, and Mn2+ and inactivated by Fe2+, Zn2+, Cu2+, Al3+, Ba2+, and Co2+ (P < 0.05). Nevertheless, Na+ and K+ had no significant effect on trypsin activity (P > 0.05). The purified trypsin showed significantly higher catalytic efficiency (kcat/Km) than porcine pancreatic trypsin against BAPNA and N-α-p-Tosyl-L-arginine methyl ester hydrochloride (TAME) substrates (P < 0.05).