Advanced SearchSearch Tips
Protease Inhibitors in Porcine Colostrum: Potency Assessment and Initial Characterization
facebook(new window)  Pirnt(new window) E-mail(new window) Excel Download
 Title & Authors
Protease Inhibitors in Porcine Colostrum: Potency Assessment and Initial Characterization
Zhou, Q.; He, R.G.; Li, X.; Liao, S.R.;
  PDF(new window)
Porcine colostrum and milk were separated into the acid-soluble and casein fractions by acidification followed by centrifuge. The acid-soluble fraction of porcine colostrum was further separated by liquid chromatography and anisotropic membrane filtration. Trypsin and chymotrypsin inhibitory capacity in porcine colostrum, milk and their components was determined by incubating bovine trypsin or chymotrypsin in a medium containing their corresponding substrates with or without addition of various amounts of porcine colostrum, porcine milk or their components. The inhibition of insulin-like growth factor I (IGF-I) and epidermal growth factor (EGF) degradation in pig small intestinal contents by porcine colostrum was measured by incubating iodinated IGF-I or EGF with the intestinal contents with or without addition of porcine colostrum. Degradation of labeled IGF-I or EGF was determined by monitoring the generation of radioactivity soluble in 30% trichloroacetic acid (TCA). The results showed that porcine colostrum had high levels of trypsin and chymotrypsin inhibitory activity and increased the stability of IGF-I and EGF in pig intestinal contents. The inhibitory activity declined rapidly during lactation. It was also found that trypsin and chymotrypsin inhibitory activity and the inhibition on IGF-I and EGF degradation in the acid-soluble fraction were higher than that in the casein fraction. Heat-resistance study indicated that trypsin inhibitors in porcine colostrum survived heat treatments of water bath for up to 10 min, but exposure to boiling water bath for 30 min significantly decreased the inhibitory activity. Compared with the trypsin inhibitors, the chymotrypsin inhibitors were more heatsensitive. Separation of the acid-soluble fraction of porcine colostrum by liquid chromatography and anisotropic membrane filtration revealed that the trypsin and chymotrypsin inhibitory capacity was mainly due to a group of small proteins with molecular weight of 10,000-50,000. In conclusion, the present study confirmed the existence of high levels of protease inhibitors in porcine colostrum, and the inhibition of porcine colostrum on degradation of milk-borne growth factors in the pig small intestinal tract was demonstrated for the first time.
Porcine Colostrum;Protease Inhibitors;Growth Factors;GI Tract;
 Cited by
Dietary Insulin Affects Leucine Aminopeptidase, Growth Hormone, Insulin-Like Growth Factor I and Insulin Receptors in the Intestinal Mucosa of Neonatal Pigs, Neonatology, 2006, 89, 4, 265  crossref(new windwow)
Carlsson, L. C. T. and B. W. Karlsson. 1972. Trypsin inhibiting capacity in the serum, digestive contents, urine and organs of the developing pig. Enzymologia. 43:89-103.

Carlsson, L. C. T., I. S. S. Bergelin and B. W. Karlsson. 1974. Trypsin inhibition in urine of developing neonatal pigs and in sow’s colostrum. Enzyme. 18:176-188.

Carlsson, L. C. T., B. W. Karlsson and B. R. Westrom. 1975. Trypsin inhibition in serum and urine of neonatal and lactating rats and in rat colostrum and milk. Int. J. Biochem. 6:173-180.

Carlsson, L. C. T., B. R. Westrom and B. W. Karlsson. 1980. Intestinal absorption of proteins by the neonatal piglet fed on sow’s colostrum with either natural or experimentally eliminated trypsin-inhibitory activity. Biol. Neonate. 38:309-320.

Fox, P. F. 1992. Advanced Dairy Chemistry-1: Proteins. London: Elsevier Applied Science.

Freund, R. J. and W. J. Wilson. 1993. Statistical Methods. Academic Press, New York, 237-240.

Geiger, R. 1984. Chymotrypsin. In: (Ed. H. U. Bergmeyer), Methods of Enzymatic Analysis, 3rd edition, Verlag Chemie, Weinhein, German, 5:99-107.

Geiger, R. and F. Hans. 1984. Trypsin. In: (Ed. H. U. Bergmeyer), Methods of Enzymatic Analysis, 3rd edition, Verlag Chemie, Weinhein, German, 5:119-124.

Greenwood, F. C., W. M. Hunter and J. S. Glover. 1963. The preparation of 131-I-labeled human growth hormone of high specific radioactivity. Biochem. J. 89:114-123.

Jensen, P. T. and K. B. Pedersen. 1979. Studies on immunoglobulins and trypsin inhibitor in colostrum and milk from sows and in serum of their piglets. Acta Vet. Scand. 20:60-72.

Koldovsky, O. 1996. The potential physiological significance of milk-borne hormonally active substances for the neonate. J. Mammary Gland Biology and Neoplasia 1:317-323.

Kress, L. F, S. R. Martin and M. Sr. Laskowski. 1971. Isolation of isoinhibitors of trypsin from porcine colostrum. Biochem. Biophys. Acta. 299:836-841.

K, Kita and J. Okumura. 2001. Chicken Insulin-Like Growth Factor-I Stimulates Protein Synthesis of Chicken Embryo Myoblasts Cultured in Serum-Free Medium. Asian-Aust. J. Anim. Sci. 14:17-20.

K, Kita, T. Shibata, K. Nagao, H. Hwangbo and J. Okumura. 2002. Effects of refeeding with a protein-free diets supplemented with various essential amino acids on the plasma insulin-like growth factor-I concentration in fasting young chickens. Asian-Aust. J. Anim. Sci. 15:406-409.

Laskowski, M., Jr. and M. Laskowski. 1951. Crystalline trypsin inhibitor from colostrum. J. Biol. Chem. 190:563-573.

Laskowski, M., B. Kassell and G. Hagerty. 1957. A crystalline trypsin inhibitor from swine colostrum. Biochem. Biophys. Acta. 24:300-305.

Lehninger, A. L. 1982. Principles in Biochemistry. World Publishers, Inc., New York.

Lindberg, T. 1979. Protease inhibitors in human milk. Pediatr. Res. 13:969-972.

Lindberg, T., K. Ohlsson and B. Westrom. 1982. Protease inhibitors and their relation to protease activity in human milk. Pediatr. Res. 16:479-483.

Lowry, O. H., N. J. Rosebrough, A. L. Farr and R. J. Randall. 1951. Protein measurement with the Folin phenol reagent. J. Biol. Chem. 193:265-275.

Odle, J., R. T. Zijlstra and S. M. Donovan. 1996. Intestinal effects of milk-borne growth factors in neonates of agricultural importance. J. Anim. Sci. 74:2509-2522.

Ohlsson, B. G., B. R. Westrom and B. W. Karlsson. 1982. In vitro interaction of porcine serum and colostrum protease inhibitors with pancreatic trypsin, chymotrypsin and elastase. Biochimica et Biophysica Acta. 705:357-365.

Ohlsson, B. 1987. Characterization and Physiological Function of Proteinases of Pancreatic Origin and Proteinase Inhibitors in the Developing Pig. PhD thesis.

Rao, R. K., O. Koldovsky and T. P. Davis. 1990. Inhibition of intestinal degradation of somatostatin by rat milk. Am. J. Physiol. 258:G426-431.

Rao, R. K., K. Lam, A. F. Philipps, C. Williams, M. Lake and O. Koldovsky. 1993. Presence of multiple forms of peptidase inhibitors in rat milk. J. Pediatr. Gastroenterol. Nutr. 17:414-420.

Telemo, E., B. R. Westrom and B. W. Karlsson. 1987. Proteolytic activity as a regulator of the transmission of orally fed proteins from the gut to the blood serum in the suckling rat. Biol. Neonate. 41:85-93.

Udall, J. N., K. J. Bloch, G. Vachino and W. A. Walker. 1984. Development of gastrointestinal mucosal barrier. IV. The effect of inhibition of proteolysis on the uptake of macromolecules by the intestine of the newborn. Biol. Neonate. 45:289-295.

Weber, B. A. and S. S. Nielsen. 1991. Isolation and partial characterization of a native serine-type protease inhibitor from bovine milk. J. Dairy Sci. 74:764-771.

Westrom, B. R. and L. C. T. Carlsson. 1976. Trypsin inhibitors in serum of adult and suckling rats and in rat milk. Int. J. Biochem. 7:41-47.

Westrom, B. R, L. C. T. Carlsson and B. W. Karlsson. 1979. The importance of colostral trypsin inhibitors for the absorption of proteins by the neonatal piglet. In: (Ed. W. A. Hemmings), Protein Transmission through Living Membranes, Elsevier/North-Holland Biomedical Press, 225-232.

Westrom, B. R., J. Svendsen and B. W. Karlsson. 1982. Protease inhibitor levels in porcine mammary secretions. Biol. Neonate. 42:185-194.

Westrom, B. R., B. G. Ohlsson, J. Svendsen, C. Tagesson and B. W. Karlsson. 1985. Intestinal transmission of macromolecules (BSA and FTIC-dextran) in the neonatal pig: enhancing effect of colostrum, proteins and proteinase inhibitors. Biol. Neonate. 47:359-366. crossref(new window)

Xian, C. J., C. A. Shoubridge and L. C. Read. 1995. Degradation of IGF-I in the adult rat gastrointestinal tract is limited by a specific antiserum or the dietary protein casein. J. Endocrinol. 146:215-225.

Xu, R. J. 1996. Development of the newborn GI tract and its relation to colostrum/milk intake: a review. Reprod. Fertil. Dev. 8:35-48.

Xu, R. J., Y. L. Mao and M. Y. W. Tso. 1996. Stability of gastrin in the gastrointestinal lumen of suckling, weaning and adult pigs. Biol. Neonate. 70:60-68.