Publisher : Korean Society for Food Science of Animal Resources
DOI : 10.5851/kosfa.2014.34.2.151
Title & Authors
Effects of Concentration and Reaction Time of Trypsin, Pepsin, and Chymotrypsin on the Hydrolysis Efficiency of Porcine Placenta Jung, Kyung-Hun; Choi, Ye-Chul; Chun, Ji-Yeon; Min, Sang-Gi; Hong, Geun-Pyo;
This study investigated the effects of three proteases (trypsin, pepsin and chymotrypsin) on the hydrolysis efficiency of porcine placenta and the molecular weight (Mw) distributions of the placental hydrolysates. Because placenta was made up of insoluble collagen, the placenta was gelatinized by applying thermal treatment at for 1 h and used as the sample. The placental hydrolyzing activities of the enzymes at varying concentrations and incubation times were determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and gel permeation chromatography (GPC). Based on the SDS-PAGE, the best placental hydrolysis efficiency was observed in trypsin treatments where all peptide bands disappeared after 1 h of incubation as compared to 6 h of chymotrypsin. Pepsin hardly hydrolyzed the placenta as compared to the other two enzymes. The Mw distribution revealed that the trypsin produced placental peptides with Mw of 106 and 500 Da. Peptides produced by chymotrypsin exhibited broad ranges of Mw distribution (1-20 kDa), while the pepsin treatment showed Mw greater than 7 kDa. For comparisons of pre-treatments, the subcritical water processing (37.5 MPa and ) of raw placenta improved the efficiency of tryptic digestions to a greater level than that of a preheating treatment ( for 1 h). Consequently, subcritical water processing followed by enzymatic digestions has the potential of an advanced collagen hydrolysis technique.
Subcritical Water Processing of Proteins: An Alternative to Enzymatic Digestion?, Analytical Chemistry, 2016, 88, 12, 6425
Effect of Sub- and Super-critical Water Treatment on Physicochemical Properties of Porcine Skin, Korean Journal for Food Science of Animal Resources, 2015, 35, 1, 35
Effect of Soy Protein Hydrolysates Prepared by Subcritical Water Processing on the Physicochemical Properties of Pork Patty during Chilled Storage, Korean Journal for Food Science of Animal Resources, 2015, 35, 4, 557
Ahmad, M., Benjakul, S., and Nalinanon, S. (2010) Compositional and physicochemical characteristics of acid solubilized collagen extracted from the skin of unicorn leatherjacket (Aluterus monoceros). Food Hydrocolloid. 24, 588-594.
AOAC (1990) Official methods of analysis. 15th ed, Association of Official Analytical Chemists, Washington, DC.
Appel, W. (1986) Chymotrypsin: molecular and catalytic properties. Clin. Biochem. 19, 317-322.
Brunner, G. (2009) Near critical and supercritical water. Part I. Hydrolytic and hydrothermal processes. J. Supercritical Fluid. 47, 373-381.
Chai, H. J., Li, J. H., Huang, H. N., Li, T. L., Chan, Y. L., Shiau, C. Y., and Wu, C. J. (2010) Effects of sizes and conformations of fish-scale collagen peptides on facial skin qualities and transdermal penetration efficiency. J. Biomed. Biotechnol. 2010, 1-9.
Denis, A., Brambati, N., Dessauvages, B., Guedj, S., Ridoux, C., Meffre, N., and Autier, C. (2008) Molecular weight determination of hydrolyzed collagens. Food Hydrocolloid. 22, 989-994.
Fruton, J. S. (1970) The specificity and mechanism of pepsin action. Adv. Enzymol. Relat. Areas Mol. Bio. 33, 401-443.
Gomez-Guillen, M. C., Gimenez, B., Lopez-Caballero, M. E., and Montero, M. P. (2011) Functional and bioactive properties of collagen and gelatin from alternative sources: A review. Food Hydrocolloid. 25, 1813-1827.
Gu, R. Z., Li, C. Y., Liu, W. Y., Yi, W. X. and Cai, M. Y. (2011) Angiotensin I-converting enzyme inhibitory activity of low-molecular-weight peptides from Atlantic salmon (Salmo salar L.) skin. Food Res. Int. 44, 1536-1540.
He, S., Franco, C., and Zhang, W. (2013) Functions, applications and production of protein hydrolysates from fish processing co-products (FPCP). Food Res. Int. 50, 289-297.
Johnston, N., Dettmar, P. W., Bishwokarma, B., Lively, M. O., and Koufman, J. A. (2007) Activity/stability of human pepsin: Implications for reflux attributed laryngeal disease. Laryngoscope 117, 1036-1039.
Kageyama, T. (2004) Role of S'1 loop residues in the substrate specificities of pepsin A and chymosin. Biochem. 43, 15122-15130.
Klomklao, S., Benjakul, S., Visessanguan, W., Kishimura, H., and Simpson, B. K. (2006) Proteolytic degradation of sardine (Sardinella gibbosa) proteins by trypsin from skipjack tuna (Katsuwonus pelamis) spleen. Food Chem. 98, 14-22.
Laemmli, U. K. (1970) Cleavage of structural proteins during assembly of head of bacteriophage T4. Nature 227, 680-685.
Lee, M. Y., Choi, Y. C., Chun, J. Y., Min, S. G., and Hong, G. P. (2013) Effects of high pressure/high temperature processing on the recovery and characteristics of porcine placenta hydrolysates. Korean J. Food Sci. An. 33, 474-480.
Liu, D., Liang, L., Regenstein, J. M. and Zhou, P. (2012) Extraction and characterization of pepsin-solubilised collagen from fins, scales, skins, bones and swim bladders of bighead carp (Hypophthalmichthys nobilis). Food Chem. 133, 1441-1448.
Ma, W., Tang, C., and Lai, L. (2005) Specificity of trypsin and chymotrypsin: Loop-motion-controlled dynamic correlation as a determinant. Biophys. J. 89, 1183-1193.
Miller, E. J. (1988) Collagen types: Structure, distribution, and functions. In: Collagen. Nimni, M. E. (ed) Boca Raton, CRC Press, Boca Raton, vol. 1, pp. 139-156.
Olsen, J. V., Ong, S., and Mann, M. (2004) Trypsin cleaves exclusively C-terminal to arginine and lysine residues. Mol. Cell Proteomics 3, 608-614.
Rodriguez, J., Gupta, N., Smith, R. D., and Prevner, P. A. (2008) Does trypsin cut before proline? J. Proteome Res. 7, 300-305.
Sipos, T. and Merkel, J. R. (1970) An effect of calcium ions on the activity, heat stability, and structure of trypsin. Biochem. 9, 2766-2775.
Vajda, T. and Szabo, T. (1976) Specificity of trypsin and alpha-chymotrypsin towards neutral substrates. Acta Biochim. Biophys. Acad. Sic. Hung. 11, 287-294.
Watchararuji, K., Goto, M., Sasaki, M., and Shotiprunk, A. (2008) Value-added subcritical water hydrolysate from rice bran and soybean meal. Bioresour. Technol. 99, 6207-6213.
Yang, Y., Hawthorne, S. B., and Miller, D. J. (1997) Classselective extraction of polar, moderately polar, and nonpolar organics from hydrocarbon wastes using subcritical water. Environ. Sci. Technol. 31, 430-437.
Yorgancioglu, A. and Bayramoglu, E. E. (2013) Production of cosmetic purpose collagen containing antimicrobial emulsion with certain essential oils. Ind. Crop. Prod. 44, 378-382.
Zhang, Z., Li, G., and Shi, B. (2006) Physicochemical properties of collagen, gelatin and collagen hydrolysate derived from bovine limed split wastes. J. Society Leather Technol. Chem. 90, 23-28.