Partial purification and characterization of alkaline proteases from the Black Sea anchovy (Engraulis encrasicholus) digestive tract
Alkaline proteases from the digestive tract of anchovy were partially purified by ammonium sulfate fractionation, dialysis and Sephadex G-75 gel filtration. The purification fold and yield were 6.23 and 4.49%, respectively. The optimum activities of partially purified alkaline proteases were observed at 60°C and at pH 11.0. The alkaline proteases were stable within the temperature range of 40 to 50°C and pH range of 9.0 to 11.0. They were inhibited by the serine-protease inhibitor phenylmethylsulfonyl fluoride (PMSF) and trypsin specific inhibitor benzamidine, but were not inhibited by the β-mercaptoethanol. The enzymes were slightly activated by metal ions such as Na+ and Ba2+ and inhibited by Cu2+, Zn2+, K+ and Mn2+ at different degrees. The molecular weight of the partially purified enzyme was 24 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE).
Key words: Alkaline proteases, Engraulis encrasicholus, purification, characterization, digestive tract.