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. 2014;34(2):151-7.
doi: 10.5851/kosfa.2014.34.2.151. Epub 2014 Apr 30.

Effects of Concentration and Reaction Time of Trypsin, Pepsin, and Chymotrypsin on the Hydrolysis Efficiency of Porcine Placenta

Kyung-Hun Jung  1 , Ye-Chul Choi  1 , Ji-Yeon Chun  1 , Sang-Gi Min  1 , Geun-Pyo Hong  2
Affiliations

Effects of Concentration and Reaction Time of Trypsin, Pepsin, and Chymotrypsin on the Hydrolysis Efficiency of Porcine Placenta

Kyung-Hun Jung et al. Korean J Food Sci Anim Resour. 2014.

Abstract

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 90 ℃ 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 200 ℃ of raw placenta improved the efficiency of tryptic digestions to a greater level than that of a preheating treatment (90 ℃ for 1 h). Consequently, subcritical water processing followed by enzymatic digestions has the potential of an advanced collagen hydrolysis technique.

Keywords: collagen peptide; efficiency; hydrolysis; porcine placenta; protease.

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Figures

Fig. 1.
Fig. 1.. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis patterns of raw and preheated (90℃ for 1 h) porcine placenta after a 24 h incubation with various concentrations of trypsin.
Fig. 2.
Fig. 2.. Effects of trypsin concentration and incubation time on (A) sodium dodecyl sulfate-polyacrylamide gel electrophoresis pattern and (B) molecular weight distribution of preheated (90℃ for 1 h) porcine placenta hydrolysates. The placenta and enzyme mixture was incubated at 37℃.
Fig. 3.
Fig. 3.. Effects of pepsin concentration and incubation time on (A) sodium dodecyl sulfate-polyacrylamide gel electrophoresis pattern and (B) molecular weight distribution of preheated (90℃ for 1 h) porcine placenta hydrolysates. The placenta and enzyme mixture was incubated at 37℃.
Fig. 4.
Fig. 4.. Effects of chymotrypsin concentration and incubation time on (A) sodium dodecyl sulfate-polyacrylamide gel electrophoresis pattern and (B) molecular weight distribution of preheated (90℃ for 1 h) porcine placenta hydrolysates. The placenta and enzyme mixture was incubated at 37℃.
Fig. 5.
Fig. 5.. Gel permeation chromatography pattern of preheated (90℃ for 1 h) and subcritical water processed (37.5MPa and 200℃) porcine placenta hydrolysates incubated with 25.0 BAEE unit/mg trypsin at 37℃ for 24 h.
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References

    1. Ahmad M., Benjakul S., Nalinanon S. Compositional and physicochemical characteristics of acid solubilized collagen extracted from the skin of unicorn leatherjacket (Aluterus monoceros) Food Hydrocolloid. (2010);24:588–594. doi: 10.1016/j.foodhyd.2010.03.001. - DOI
    1. AOAC. Official methods of analysis. 15th ed Association of Official Analytical Chemists; Washington, DC: (1990).
    1. Appel W. Chymotrypsin: molecular and catalytic properties. Clin. Biochem. (1986);19:317–322. doi: 10.1016/S0009-9120(86)80002-9. - DOI - PubMed
    1. Brunner G. Near critical and supercritical water. Part I. Hydrolytic and hydrothermal processes. J. Supercritical Fluid. (2009);47:373–381. doi: 10.1016/j.supflu.2008.09.002. - DOI
    1. Chai H. J., Li J. H., Huang H. N., Li T. L., Chan Y. L., Shiau C. Y., Wu C. J. Effects of sizes and conformations of fish-scale collagen peptides on facial skin qualities and transdermal penetration efficiency. J. Biomed. Biotechnol. (2010);2010:1–9. - PMC - PubMed