Method of isolation and purification of trypsin from pronase protease and use thereof
View Patent ↗The present invention provides methods of isolation and purification of Streptomyces griseus trypsin (SGT) from PRONASE protease mixture in a single affinity chromatography step and uses of the purified SGT.
1. A method of isolating Streptomyces griseus trypsin (SGT) from Streptomyces griseus Pronase, the method comprising contacting said Pronase with an immobilized affinity moiety selected from the group consisting of an amidine, a guanadine, or an amine containing species and eluting said SGT selectively from said immobilized affinity moiety with an eluant comprising arginine, wherein the isolated SGT further comprises arginine and has a specific activity of at least 25×10 3 U/mg protein.
2. The method of claim 1 , wherein said arginine is in a concentration between 0.5-1.2 M.
3. The method according to claim 1 , wherein said eluant has a pH in the range of between about pH 4.0. and about 9.0.
4. The method according to claim 1 , wherein said affinity moiety is benzamidine.
5. The method according to claim 1 , wherein said eluant comprises an inorganic salt in a concentration between about 0.1 and about 1 M.
6. The method according to claim 1 , wherein said SGT is obtained in a solution comprising 0.5 to 1.2 M arginine and has a purity of at least 95%.
7. The method according to claim 1 , further comprising removing the eluting agent from the purified SGT.