Hostname: page-component-586b7cd67f-gb8f7 Total loading time: 0 Render date: 2024-11-30T05:20:07.618Z Has data issue: false hasContentIssue false
  • English
  • Français

Large-scale isolation of fraction 1 leaf protein (18S) from lucerne (Medicago sativa L)

Published online by Cambridge University Press:  27 March 2009

W. T. Jones  and
J. L. Mangan
W. T. Jones
Affiliation:
Biochemistry Department, Institute of Animal Physiology, Agricultural Research Council, Babraham, Cambridge, England
J. L. Mangan
Affiliation:
Biochemistry Department, Institute of Animal Physiology, Agricultural Research Council, Babraham, Cambridge, England
Get access Rights & Permissions [Opens in a new window]

Summary

Fraction 1 protein (18S) can be isolated in large quantities (order 100 g) in a soluble state by heating lucerne juice, adjusted to pH 6·7 to 6·9, from a Pirie extractor to 63 °C for 10 min. Low speed oentrifugation (2500 g) removed coagulated chloroplast fragments and most of the heat-denatured Fraction 2 proteins. Fraction 1 (18S) protein (> 95% pure) was purified from low molecular weight materials (sugars, phenolics etc.) by ammonium sulphate fractionation and gel filtration on Sephadex G-75 gels.

Type
Research Article
Information
The Journal of Agricultural Science , Volume 86 , Issue 3 , June 1976 , pp. 495 - 501
Copyright
Copyright © Cambridge University Press 1976

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Arnon, D. I. (1949). Copper enzymes in isolated chloroplasts. Polyphenoloxidase in Beta Vulgaris. Plant Physiology 24, 1–15.CrossRefGoogle ScholarPubMed
de Fremery, D., Miller, R. E., Edwards, R. H., Knuckles, B. E., Bickoff, E. M. & Kohler, G. O. (1973). Centrifugal separation of white and green protein fractions from alfalfa juice following controlled heating. Journal of Agriculture and Food Chemistry 21, 886–9.CrossRefGoogle ScholarPubMed
Jones, W. T. (1972). Ph.D. thesis ‘Studies on the Foaming Properties of Proteins’, Massey University, Palmerston North, New Zealand.Google Scholar
Jones, W. T. & Lyttleton, J. W. (1972). Further studies of the foaming properties of leaf proteins. New Zealand Journal of Agricultural Research 15, 267–78.CrossRefGoogle Scholar
Jones, W. T. & Lyttleton, J. W. (1972). The importance of inhibiting polyphenol oxidase in the extraction of Fraction 1 leaf protein. Phytochemistry 11, 1595–6.CrossRefGoogle Scholar
Kohler, G. O. & Bickoff, E. M. (1971). In Leaf Protein – its Agronomy, Preparation, Quality and Use. I.B.P. Handbook No. 20, Oxford: Blackwell Scientific Publications, 69–77.Google Scholar
Lyttleton, J. W. & T'so, P. O. P. (1958). The localization of Fraction 1 protein of green leaves in chloroplasts. Archives of Biochemistry and Biophysics 73, 120–6.CrossRefGoogle ScholarPubMed
Mangan, J. L. (1972). Quantitative studies on nitrogen metabolism in the bovine rumen. British Journal of Nutrition 27, 261–83.CrossRefGoogle ScholarPubMed
Mayaudon, J. (1957). Study of association between the main nucleoprotein of green leaves and carboxydismutase. Enzymologia 18, 343–54.Google ScholarPubMed
MoArthur, J. E. & Miltimore, J. E. (1966). Pasture bloat and the role of 18 S protein. Proceedings of the Tenth International Grassland Congress, 518.Google Scholar
Pirie, N. W. (1971). In Leaf Protein – its Agronomy, Preparation, Quality and Use. I.B.P. Handbook No. 20, Oxford: Blackwell Scientific Publications. 53–62.Google Scholar
Reid, C. S. W. & Clarke, R. T. J. (1974). Foamy bloat of cattle. A Review. Journal of Dairy Science 57, 753–85.Google Scholar
Reid, M. S. & Bieleski, R. L. (1968). A simple apparatus for vertical flat sheet polyacrylamide gel electrophoresis. Analytical Biochemistry 22, 374–81.CrossRefGoogle ScholarPubMed
Singer, S. J., Eggman, L., Campbell, J. M. & Wildman, S. (1952). The proteins of green leaves. Journal of Biological Chemistry 197, 233–9.CrossRefGoogle ScholarPubMed
Stifel, F. B., Vetter, R. L. & Allen, R. S. (1968). Relationship between calcium and magnesium binding to Fraction 1 chloroplast protein and bloat. Agriculture dk Food Chemistry 16, 500–4.CrossRefGoogle Scholar
Trown, P. W. (1965). An improved method for the isolation of carboxydismutase. Probable identity with Fraction 1 protein and the protein moiety of photochlorophyll holochrome. Biochemistry 4, 908–18.CrossRefGoogle Scholar