geoscience.net logo
+ Resolve Article
+ Follow Us
Follow on FacebookFollow on Facebook
Follow on TwitterFollow on Twitter

+ Translate
+ Subscribe to Site Feed
GeoScience Most Shared ContentMost Shared Content

Purification and characterization of trypsin from the pyloric ceca of the new zealand hoki fish (macruronus novaezealandiae)


, : Purification and characterization of trypsin from the pyloric ceca of the new zealand hoki fish (macruronus novaezealandiae). Journal of food biochemistry 31(6): 772-796

Trypsin was purified from the pyloric ceca of hoki fish by (NH)SO fractionation, acetone precipitation and affinity chromatography. The purified trypsin migrated as a single band in sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and the isoelectric point was determined as 6.5. The molecular weight was determined as 26,000 Da by SDS-PAGE and as 23,791 Da by matrix-assisted laser desorption/ionization time-of-flight mass spectroscopy. The enzyme was stable within the pH range of 6.0-11.0, and exhibited optimum activity for the hydrolysis of N-l-benzoyl- dl-arginine-p-nitroanilide ( dl-BAPNA) at pH 9.0. The enzyme was stable up to 40C, and its optimum temperature for dl-BAPNA hydrolysis was 60C, and was inhibited by soybean trypsin inhibitor, aprotinin, phenyl methyl sulfonyl fluoride and benzamidine. The apparent Michaelis-Menten constant (Km') and substrate turnover number (kcat) for the dl-BAPNA hydrolase reaction was 0.06 mM and 0.33 sp#, respectively, while the corresponding values for tosyl arginine methyl ester hydrolysis were 2.08 mM and 19.0 sp#, respectively. The N-terminal 20 amino acid residues of hoki trypsin, IVGGQECVPNSQPFMASLNY, displayed considerable homology with other fish trypsins. Trypsins from various sources catalyze the hydrolysis of peptide bonds on the carboxyl sides of arginine and lysine. Thus, it is expected that like other trypsins, hoki trypsin would also be useful in biomedical, food and beverage industry applications. Examples of the practical applications of trypsins include their use in mammalian cell culture to disaggregate adherent cells for research and the production of recombinant proteins; in cancer therapy to break down tumors and cancer cells; and in wound care, for treatment of acne and other skin disorders. Other uses are as a digestive aid, for treating pain, inflammations, arthritis, tendonitis, ulcers, hemorrhoids and navel infections in newborns. Trypsins are also used in diabetes diagnosis and therapy; in detergents and bating of leather; and in the food industry for the production of protein hydrolysates, as meat tenderizer and as an aid to improve yields in oil extractions.

US$19.90

DOI: 10.1111/j.1745-4514.2007.00141.x


Other references

Reisner, H.M.; Price, W.A.; Blatt, P.M.; Barrow, E.S.; Graham, J.B., 1980: Factor VIII coagulant antigen in hemophilic plasma: a comparison of five alloantibodies. Five independent alloantibodies directed to factor VIII coagulant antigen (VIII:CAg) were assessed against normal, von Willebrand's disease, and severe hemophilia A plasmas. Immunoradiometric assays (IRMAs) were developed for each antibody, o...

Bates, J., 2008: Unfriendly. NlmCategory="UNASSIGNED">'Your friend is waiting for you just round the corner,' I informed a patient. 'He is my husband!' she replied indignantly. Quite how indignant was emerging in the animated conversation the...

Fujii, Y.; Takeuchi, S.; Sasaki, O.; Minakawa, T.; Koike, T.; Tanaka, R., 1997: Serial changes of hemostasis in aneurysmal subarachnoid hemorrhage with special reference to delayed ischemic neurological deficits. This study was undertaken to elucidate comprehensively the serial changes occurring in hemostatic systems after aneurysmal subarachnoid hemorrhage (SAH) and thereby to ascertain whether the examination of the integrity of these systems is helpful...

Abdul-Wahab, A.; Robson, A.; Agar, N., 2013: An asymptomatic nodule on the upper thigh. Clinical and Experimental Dermatology 37(8): 926-927

Droukas, V., 1989: Sounds from the TM joint. Etiology and pathological significance. The irregularities of the disc in relation to the condyle, as the production of pathological sounds, from the temporomandibular joints, is one of the simultaneous and very difficult topics that occupied the physiology of the stomatognathic system,...

Kenney, G.; Yee, J., 2007: SCHIP at a crossroads: experiences to date and challenges ahead. As reauthorization of the State Children's Health Insurance Program (SCHIP) looms, we examine the program's first decade and identify changes needed so that SCHIP can better serve its target population. We conclude that by many objective...

Gupta, M.K.; Bogner, R.H.; Goldman, D.; Tseng, Y-Chao., 2002: Mechanism for further enhancement in drug dissolution from solid-dispersion granules upon storage. The present study was performed to investigate the further increase in drug dissolution on storage of ternary solid-dispersion granules containing poorly water-soluble drugs. Ternary solid-dispersion granules of the drug, a dispersion carrier, and...

Wilgosh L.; Mulcahy R.; Watters B., 1986: Assessing intellectual performance of culturally different inuit children with the wisc r. This paper examines problems in the assessment of intelligence using standardized intelligence tests with culturally different children, for a sample of Canadian Inuit (Eskimo) children whose WISC-R scores, using the original WISC-R norms, would f...

Gur-Yosef, J.A.; Leske, G.S.; Ripa, L.W., 1987: A student-to-student dental outreach program. Journal of American College Health 36(3): 185-187

García Ribes, M., 2016: 2006-2016: Ten years of immersion in the world of rare diseases. Atencion Primaria 48(4): 217-218