Trypsin from zebra blenny (Salaria basilisca) viscera: Purification,characterisation and potential application as a detergent additive |
| |
Authors: | Naourez Ktari Hayet Ben Khaled Rim Nasri Kemel Jellouli Sofiane Ghorbel Moncef Nasri |
| |
Affiliation: | Laboratoire de Génie Enzymatique et de Microbiologie, Université de Sfax, Ecole Nationale d’Ingénieurs de Sfax, B.P. 1173-3038 Sfax, Tunisia |
| |
Abstract: | An alkaline trypsin was purified from the viscera of zebra blenny (Salaria basilisca) by ammonium sulphate (40?80% saturation) precipitation, Sephadex G-100, Mono Q-Sepharose and ultrafiltration. A yield of 12% with a purification-fold of 4.2 was obtained. The trypsin had an apparent molecular weight of 27 kDa. Soybean trypsin inhibitor and phenylmethylsulfonyl fluoride showed a strong inhibitory effect on the purified trypsin. Trypsin had maximal activity at pH 9.5 and 60 °C for the hydrolysis of Nα-benzoyl-dl-arginine-p-nitroanilide (BAPNA). It was stable at low temperatures and in the pH range of 7.0?12.0. The N-terminal amino acid sequence of the first 12 amino acids of the purified protease was IVGGRECTEPSQ. S. basilisca trypsin, which showed high homology with other fish trypsins, had a charged Arg residue at position 5, where Tyr is common in marine vertebrates and mammalian trypsins. The trypsin kinetic constants, Km and kcat for BAPNA, were 0.6 mM and 1.38 s?1, respectively. |
| |
Keywords: | |
本文献已被 ScienceDirect 等数据库收录! |
|