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Journal of Biological Sciences

Year: 2002 | Volume: 2 | Issue: 9 | Page No.: 591-595
DOI: 10.3923/jbs.2002.591.595
A Single Step Purification of Gastricsin-Like Proteinase from Atlantic Cod (Gadus morhua)
M. A. Amiza and R.K. Owusu Apenten

Abstract: A gastricsin-like proteinase was purified from gastric mucosa of Atlantic cod by a single step purification scheme on ion-exchange of Amberlite CG-50. The purification was very efficient as the recovery was 205% and the purification factor was 1796-fold. The enzyme preparation was homogeneous as observed by SDS-PAGE and isoelectric focusing. The enzyme has an estimated molecular weight of 34 kDa and the pI of 4.4.

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How to cite this article
M. A. Amiza and R.K. Owusu Apenten, 2002. A Single Step Purification of Gastricsin-Like Proteinase from Atlantic Cod (Gadus morhua). Journal of Biological Sciences, 2: 591-595.

Keywords: Enzyme, purification, cod, cod and protease

REFERENCES
  • Almas, K.A., 1990. Utilization of Marine Biomass for Production of Microbial Growth Media and Biochemicals. In: Advances in Fisheries Technology and Biotechnology for Increased Profitability, Voigt, M.N. and J.R. Botta (Eds.). Technomic Publishing Co. Inc., Lancaster, PA., pp: 509-524

  • Anson, M.L. and A.E. Mirsky, 1932. The estimation of pepsin with hemoglobin. J. Genet. Physiol., 16: 59-63.

  • Arunchalam, K. and N.F. Haard, 1985. Isolation and characterization of pepsin from polar cod (Boreogadus saida). Comp. Biochem. Physiol., 80: 467-473.

  • Asgeirsson, B., J.W. Fox and J.B. Bjarnason, 1989. Purification and characterization of trypsin from the poikilotherm Gadus morhua. Eur. J. Biochem., 180: 85-94.

  • Bjarnason, J.B. and B. Asgeirsson, 1993. Psychrophilic proteolytic enzymes from Atlantic cod: Their characteristics and applications. Genet. Eng. Biotechnol., 13: 31-39.

  • Brewer, P., N. Helbig and N.F. Haard, 1984. Atlantic cod pepsin-characterization and use as a rennet substitute. Can. Inst. Food Sci. Technol., 17: 38-43.

  • Cano-Lopez, A., B.K. Simpson and N.F. Haard, 1987. Extraction of carotenoprotein from shrimp waste with trypsin from Atlantic cod. J. Food Sci., 52: 503-506.

  • Gildberg, A., 1988. Aspartic proteinases in fishes and aquatic invertebrates. Comp. Biochem. Physiol., 91: 425-435.

  • Gildberg, A. and K.A. Almas, 1986. Utilization of Fish Viscera. In: Food Engineering and Processing Application, Le Maguer, M. and P. Jelen (Eds.). Vol. 2, Elsevier Applied Science Publications, London, pp: 383-393

  • Gildberg, A. and J. Raa, 1983. Purification and characterization of pepsins from the arctic fish capelin (Mallotus villosus). Comp. Biochem. Physiol., 75: 337-342.

  • Gildberg, A., R.L. Olsen and J.B. Bjarnason, 1990. Characteristics and composition of pepsins from Atlantic cod (Gadus morhua). Adv. Exp. Med. Biol., 306: 107-110.

  • Gildberg, A., 1992. Recovery of proteinase and protein hydrolysates from fish viscera. Bioresour. Technol., 39: 271-276.

  • Gudmundsdottir, A., S. Oskarsson, A.E. Eakin, C.S. Craik and J.B. Bjarnason, 1994. Atlantic cod cDNA encoding a psychrophilic chymotrypsinogen. Biochim. Biophys. Acta, 1219: 211-214.

  • Gudmundsdottir, A., S. Oskarsson, E. Gudmundsdottir, J.B. Bjarnason, A.K. Eakin and C.S. Craik, 1993. Isolation and characterization of cDNAs from Atlantic cod encoding two different forms of trypsinogen. Eur. J. Biochem., 217: 1091-1097.

  • Haard, N.F., 1986. Atlantic cod gastric proteases. Characterization with casein and milk substrate and influence of sepharose immobilization on salt activation, temperature characteristics and milk clotting reaction. J. Food Sci., 51: 313-316.

  • Haard, N.F., 1992. A review of proteolytic enzymes from marine organisms and their application in the food industry. J. Aquat. Food Prod. Technol., 1: 17-35.

  • Haard, N.F., N. Helbig and L.A.W. Feltham, 1981. The Temperature Characteristics of Pepsin from two Stocks of American Smelt. In: Research in the Labrador Coastal and Offshore Region, Denner, W. (Ed.). Newfoundland Institute of Cold Ocean Science, St. John's, pp: 175-196

  • Herriot, R.M., 1962. Pepsinogen and pepsin. J. Genet. Physiol., 45: 57-67.

  • Kristjansson, M.M., S. Gudmundsdottir, J.W. Fox and J.B. Bjarnson, 1995. Characterization of a collagenolytic serine proteinase from the Atlantic cod (Gadus morhua). Comp. Biochem. Physiol., 110: 707-717.
    PubMed    Direct Link    

  • Laemmli, U.K., 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, 227: 680-685.
    CrossRef    Direct Link    

  • Martinez, A. and R.L. Olsen, 1989. Characterization of pepsins from cod. US Biochem. Corp., 16: 22-23.

  • Merrett, T.G., E. Bar-Eli and H. Van Vunakis, 1969. Pepsinogens A, C and D from smooth dogfish. Biochemistry, 8: 3696-3702.

  • Noda, M. and K. Murakami, 1981. Studies on proteinases from the digestive organs of sardine. II. Purification and characterization of two acid proteinases from the stomach. Biochim. Biophys. Acta, 658: 27-34.

  • Norris, E.R. and D.W.J. Ellam, 1940. Preparation and properties of crystalline salmon pepsin. J. Biol. Chem., 134: 443-454.

  • Owen, T.G. and A.J. Wiggs, 1971. Thermal compensation in the stomach of the Brook trout (Salvelinus fontinalis mitchill). Comp. Biochem. Physiol., 40: 465-473.

  • Peterson, G.L., 1977. A simplification of the protein assay method of Lowry et al. which is more generally applicable. Anal. Biochem., 83: 346-356.
    CrossRef    PubMed    Direct Link    

  • Raa, J., 1990. Biotechnology in Aquaculture and the Fish Processing Industry: A Success Story in Norway. In: Advances in Fisheries Technology and Biotechnology for Increased Profitability, Voigt, M.N. and J.R. Botta (Eds.). Technomic Publishing Co. Inc., Lancaster, PA., pp: 509-524

  • Richmond, V., J. Tang, S. Wolf, R.E. Trucco and R. Caputto, 1958. Chromatographic isolation of gastricsin, the proteolytic enzymes from gastric juice with pH optimum 3.2. Biochem. Biophys. Acta, 29: 453-453.

  • Sanchez-Chiang, L., E. Cisternas and O. Ponce, 1987. Partial purification of pepsin from adult and juvenile salmon fish, Oncorhynchus keta. Effect of NaCl on proteolytic activity. Comp. Biochem. Physiol., 87: 793-797.

  • Sanchez-Chiang, L. and O. Ponce, 1981. Gastricsinogens and gastricsins from Merluccius gayi, purification and properties. Comp. Biochem. Physiol., 68: 251-257.

  • Sikorski, Z.E., B.S. Pan and F. Shahidi, 1994. Sea Food Protein. Chapman and Hall, New York

  • Simpson, B.K. and N.F. Haard, 1984. Trypsin from Greenland cod, Gadus ogac. Isolation and comparative properties. Comp. Biochem. Physiol., 79: 613-622.

  • Simpson, B.K. and N.F. Haard, 1985. Characterization of the trypsin fraction from cunner, Tautogolabrus adspersus. Comp. Biochem. Physiol., 80: 475-480.

  • Smalas, A.O., E.S. Heimstad, A. Hordvik, P. Willasen and R. Male, 1994. Cold adaptation of enzymes: Structural comparison between salmon and bovine trypsins. Protein Struct. Funct. Genet., 20: 149-166.

  • Squires, J., N.F. Haard and L.A.W. Feltham, 1986. Gastric proteases of the Greenland cod Gadus ogac. I. Isolation and kinetic properties. Biochem. Cell Biol., 64: 205-214.

  • Squires, J., N.F. Haard and L.A.W. Feltham, 1986. Gastric proteases of the Greenland cod (Gadus ogac). II. Comparison of structural properties. Biochem. Cell Biol., 64: 215-222.

  • Stefansson, G. and U. Steingrimsdottir, 1990. Application of Enzymes for Fish Processing in Iceland-Present and Future Aspects. In: Advances in Fisheries Technology and Biotechnology for Increased Profitability, Voigt, M.N. and Botta, J.R. (Eds.). Technomic Publishing Co. Inc., Lancaster, PA., pp: 237-250

  • Suparno and A. Poernomi, 1992. Fish waste utilization in Indonesia. ASEAN Food J., 7: 67-72.
    Direct Link    

  • Tang, J., 1970. Gastricsin and Pepsin. In: Methods in Enzymology, Perlman, G.E. and K. Lorand (Eds.). Vol. 9, Academic Press, New York and London, pp: 406-420

  • Tanji, M., T. Kageyama and K. Takahashi, 1988. Tuna pepsinogens and pepsins. Purification, characterization and amino-acid sequences. Eur. J. Biochem., 177: 2511-2591.

  • Twining, S.S., P.A. Alexander, K. Huibregtse and D.M. Glick, 1983. A pepsinogen from rainbow trout. Comp. Biochem. Physiol., 75: 109-112.

  • Xu, R.A., R.J. Wong, M.L. Rogers and G.C. Fletcher, 1996. Purification and characterization of acidic proteases from the stomach of the deep water finfish orange roughy (Hoplostethus atlanticus). J. Food Biochem., 20: 31-48.

  • Reece, P., 1988. Recovery of proteases from fish waste. Process Biochem., 33: 62-66.

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