Hostname: page-component-669899f699-tpknm Total loading time: 0 Render date: 2025-04-28T05:53:00.085Z Has data issue: false hasContentIssue false
  • English
  • Français

Idendification of sulphur-rich proteins which resist rumen degradation and are hydrolysed rapidly by intestinal proteases

Published online by Cambridge University Press:  09 March 2007

Kerrie R. Hancock ,
Paul M. Ealing  and
Derek W.R. White
Kerrie R. Hancock
Affiliation:
Plant Molecular Genetics Laboratory, New Zealand Pastoral Agriculture Resarch Institute, Palmerston North, New Zealand
Paul M. Ealing
Affiliation:
Plant Molecular Genetics Laboratory, New Zealand Pastoral Agriculture Resarch Institute, Palmerston North, New Zealand
Derek W.R. White
Affiliation:
Plant Molecular Genetics Laboratory, New Zealand Pastoral Agriculture Resarch Institute, Palmerston North, New Zealand
Rights & Permissions [Opens in a new window]

Abstract

Core share and HTML view are not available for this content. However, as you have access to this content, a full PDF is available via the ‘Save PDF’ action button.

Several proteins with high proportions of S-containing essential amino acids were incubated in sheep rumen fluid in vitro and their rate of digestion was examined by sodium dodecyl sulphate-polyacrylamide-gel electrophoresis. The S-rich proteins rice prolamin (10 kDa), maize zein (10 kDa) and the 3·2 kDa pumpkin (Cucurbita maxima L.) trypsin inhibitor-1 (CMTI-1) were highly resistant to rumen fluid degradation, relative to control proteins of known degradation rate (casein, bovine serum albumin (BSA) and pea (Pisum sativum) albumin-1 (PA1)). Comparison of PA1 and a recombinant N-terminal epitope-tagged PA1 indicated that addition of the epitope caused a slight increase in resistance to rumen degradation. The proteins were also incubated with a mixture of trypsin (EC 3·4·21·4) and chymotrypsin (EC 3·4·21.1). PA1, BSA and casein were hydrolysed less rapidly than rice prolamin, maize zein and CMTI-1. Digestion by these intestinal proteases appeared to be complete. Thus, the prolamin, zein and CMTI-1 proteins are suitable candidates for expression as foreign proteins in pasture plants to increase throughput and uptake of essential amino acids in sheep.

Keywords

Sulphur-rich proteinsRumen proteolysisRuminant nutrition
Type
Suiphur-rich Proteins Which resist dagaration in the rumen
Information
British Journal of Nutrition , Volume 72 , Issue 6 , December 1994 , pp. 855 - 863
Copyright
Copyright © The Nutrition Society 1994

References

RERERENCES

Ako, H., Foster, R. J. & Ryan, C. A. (1972) The preparation of anhydrotrypsin and its reactivity with naturally occurring proteinase inhibitors. Biochemical and Biophysical Research Communications 47, 1402–1407.CrossRefGoogle ScholarPubMed
Barry, T. N. (1981) Protein metabolism in growing lambs fed on fresh ryegrass (Lolium perenne), clover (Trifolium repens) pasture ad lib. 1. Protein and energy deposition in response to abomasal infusion of casein and methionine. British Journal of Nutrition 46, 521–532.CrossRefGoogle ScholarPubMed
Broderick, G. & Craig, W. M. (1989) Metabolism of peptides and amino acids during in vitro protein degradation by mixed rumen organisms. Journal of Dairy Science 72, 254C–2548.CrossRefGoogle ScholarPubMed
Broderick, G. A., Wallace, R. J. & Ørskov, E. R. (1991) Control of rate and extent of protein degradation. In Physiological Aspects of Digestion and Metabolism in Ruminants. Proceedings of the Seventh International Symposium on Ruminant Physiology, pp. 541–582 [Tsuda, T., Sasaki, Y. and Kawashima, R., editors]. San Diego: Academic Press.CrossRefGoogle Scholar
Field, J., Nikawa, J., Brock, D., McDonald, B., Rodgers, L., Wilson, I. A., Lerner, R. A. & Wigler, M. (1988) Purification of a RAS-responsive adenyl-cychdse complex from Saccharomyces cerevisiae by use of an epitope addition method. Molecular and Cell Biology 8, 2159–2165.Google ScholarPubMed
Higgins, T. J. V., Chandler, P. M., Randall, P. J., Spencer, D., Beach, L. R., Blugrove, R. J., Kortt, A. A. & Inglis, A. S. (1986) Gene structure, protein structure and regulation of the synthesis of a sulphur-rich protein in pea seeds. Journal of Biologicul Chemistry 261, 11124–11130.Google Scholar
Ludevid, M. D., Martinez-Izquierdo, J. A., Armengol, M., Torrent, M., Puigdoménech, P. & Palau, J. (1985) Immunological relations between glutelin-2 and low molecular weight zein-2 proteins from maize (Zea mays L.) endosperm. Plant Science 41, 41–48.CrossRefGoogle Scholar
McDonald, I. W. & Hall, R. J. (1957) The conversion of casein into microbial proteins in the rumen. Biochemical Journal 67, 400–405.Google Scholar
McDougall, E. I. (1948) Studies of ruminant saliva. 1. The composition and output of sheep's saliva. Biochemical Journal 43, 99–109.Google Scholar
Mahadevan, S., Erfle, J. D. & Sauer, F. D. (1980) Degradation of soluble and insoluble proteins by Bacteroides amylophilus protease and by rumen organisms. Journal of Animal Science 50, 723–728.Google Scholar
Mangan, J. L. (1972) Quantitative studies on nitrogen metabolism in the bovine rumen. British Journal of Nutrition 27, 261–283.CrossRefGoogle ScholarPubMed
Masumura, T., Shibata, D., Hibino, T., Kato, T., Kawabe, K., Takeba, G., Tanaka, K. & Fujii, S. (1989) cDNA cloning of an mRNA encoding a sulfur-rich 10 kDa prolamin I polypeptide in rice seeds. Plant Molecular Biology 12, 123–130.Google Scholar
Nugent, J. H. A. & Mangan, J. L. (1981) Characteristics of the rumen proteolysis of fraction I (18S) leaf protein from lucerne (Medicago sativa L). British Journal of Nutrition 46, 39–58.CrossRefGoogle ScholarPubMed
Ørskov, E. R. & Chen, X. B. (1989) Assessment of amino acid requirement in ruminants. In The Rumen Ecosystem, pp. 161–167 [Hoshino, S., Onodera, R., Minato, H. and Habashi, H., editors]. Berlin: Springer-Verlag.Google Scholar
Reis, P. J. & Colebrook, W. F. (1972) The utilisation of abomasal supplements of proteins and amino acids by sheep with special reference to wool growth. Australian Journal of Biological Science 25, 1057–1063.CrossRefGoogle ScholarPubMed
Riggs, P. (1990) Analysis of proteins. In Current Protocols in Molecular Biology, pp. 330–334 [Ausubel, F.M., Brent, R., Kingston, R. E., Moore, D. D., Seidman, J. G., Smith, J. A. and Struhl, K., editors]. New York: Greene Associates/Wiley Interscience.Google Scholar
Rulquin, H. & Verite, R. (1993) Amino acid nutrition of dairy cows: productive effects and animal requirements. In Recent Advances in Animal Nutrition, pp. 55–77 [Garnworthy, P.C. and Cole, D. J. A., editors]. Nottingham: Nottinghan University Press.Google Scholar
Sodek, L. & Wilson, C. M. (1971) Amino acid composition of proteins isolated from normal, opaque-2, and floury-2 corn endosperms by a modified Osborne procedure. Journal of Agricultural Food Chemistry 19, 1144–1150.CrossRefGoogle Scholar
Spencer, D., Higgins, T. J. V., Freer, M., Dove, H. & Coombe, J. B. (1988) Monitoring the fate of dietary proteins in rumen fluid using gel electrophoresis. British Journal of Nutrition 60, 241–247.CrossRefGoogle ScholarPubMed
Sugimoto, T., Tanaka, K. & Kasai, Z. (1986) Improved extraction of rice prolamin. Agricultural and Biological Chemistry 50, 2409–2411.Google Scholar
Ulyatt, M. J., Thomson, D. J., Beever, D. E., Evans, R. T. & Haines, M. J. (1988) The digestion of perennial ryegrass (Lolium perenne cv. Melle) and white clover (Trifolium repens cv. Blanca) by grazing cattle. British Journal of Nurrition 60, 137–149.Google Scholar
Wandelt, C. I., Khan, M. R. I., Craig, S., Schroeder, H. E., Spencer, D. & Higgins, T. J. V. (1992) Vicilin with carboxy-terminal KDEL is retained in the endoplasmic reticulum and accumulates to high levels in the leaves of transgenic plants. The Plant Journal 2, 181–192.Google Scholar