A simple, fast and commercial method for trypsin purification from bovine pancreas

Editorial

Authors

1 Razi Vaccine and Serum Research Institute

2 Biology Section, Payame noor University

3 Biochemistry Section ov Medicine Faculty, Tehran University

4 Biology Section, Payame Noor University

Abstract

Trypsin is a serine protease found in the digestive system of many vertebrates, where it hydrolyses proteins. Trypsin has been used widely in various biotechnological processes. In this study trypsin was purified using two precipitation cut, 35% and 75% as saturation percent of Ammonium sulfate solutions, TCA precipitation (2.5% W/V), pH precipitation and finally affinity chromatography. This method could be used to obtain 2.8 g purified trypsin with specific activity 155 U/mg and 26% recovery from 1kg bovine pancreas. Therefore, this method is a simple, fast and commercial method for scale-up of trypsin extraction from bovine pancreas.

1- Bond, J.S. (1989) Commercially Available Proteases, Appendix II. In: Proteolytic Enzymes, A Practical Approach. R.J. Beynon and J.S. Bond, eds., IRL Press, Oxford, U.K.
2- Bradford, M.M. (1976) A rapid and sensitive method for the quantification of microgram quantities of proteins utilizing the principle of protein dye binding. Anal. Biochem. 72: 248– 254.
3- Bricteux, S., Gregoire, G. (1966) Study of trypsinogen and trypsin of sheep pancreas. BBA. 127: 277-281.
4- Bricteux, S., Gregoire, G. (1968) Study of trypsinogen and trypsin of goat pancreas. Arch. Intern. Physiol. Biochim. 76: 571-74.
5- Charles, M. (1963) Extraction and characterization of trypsin from pig pancreas. BBA. 69: 115- 121.
6- Erlanager,B., Kokowsky,N., Cohen,W. (1961) The preparation and properties of two new chromogenic stubstrates of trypsin. ArchBiochem. Biophys. 95: 271-77.
7- Harvis, I., Hofmann, T. (1969) Isolation and characterization of trypsin from horse pancreas. Biochim. J. 114: 82 P.
8- Haynes, R., Winter, W.P. (1968) Purification and characterizationof gar (Dog- Fish) trypsin. BBA. Personal communication.
9- Jellouli,K., Bougatef,A., Daassi,D., Balti,R., Barkia,A., Nasri,M. (2009) New alkaline trypsin from the intestine of Grey triggerfish (Balistes capriscus) with high activity at low temperature: Purification and characterization. Food Chemistry 116: 644–650.
10- Kunitz,M., and Northrop,J. (1936) Isolation from beef pancreas of crystalline trypsinogen, trypsin, a trypsin inhibitor and an inhibitor- trypsin compound. J Gen Physiol. 19: 991-98.
11- Laemmli U.K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680-85.
12- Layne, E. (1957) Spectrophotometric and Turbidimetric Methods for Measuring Proteins. Methods in Enzymology 3: 447-455.
13- Northrop,J., Kunitz,M. (1932) Crystalline trypsin. I. Isolation and tests of Purity. J Gen Physiol. 16: 267-75.
14- Polg.r,L. (2005) The catalytic triad of serine peptidases. Cell. Mol. Life Sci. 62: 2161–72.
15- Rawlings ND, Barrett A.J. (1994) Families of serine peptidases”. Meth. Enzymol. 244: 19–61.
16- Rice,R.H. (1977) Stabilization of bovine trypsin by reductive methylation. Biochem. Biophys. Acta. 492: 316–21.
17- Stoscheck, CM. (1990) Quantitation of Protein. Methods inEnzymology 182: 50-69.
18- Tub G., Nerli B., Pic´G. (2007) Partitioning features of bovine trypsin and chymotrypsin in polyethyleneglycol-sodium citrate aqueous two-phase systems. Journal of Chromatography B. 852: 244–249.
19- Walker J.M. (1996) SDS polyacrylamide gel electrophoresis ofproteins. In the protein protocols hand book. Humana Press. Oxford, U.K, pp28-47.
20- Wilimowska,A., (1978) A simple method for purification trypsin, Analytical Biochemistry. 90: 816-20.
21- Wilkinson, J.M. (1986) Fragmentation of Polypeptides by EnzymicMethods. I Practical Protein Chemistry: A Handbook. A. Darbre, ed., John Wiley and Sons, New York, N.Y.
22- Woodard,S., Mayor,J., Bailey,M., Barker,D., Love,R.J. (2003) Applications of Proteolytic Enzymes in industrials. Biotechnol. Appl.Biochem. 38: 123-139.