Ion exchange chromatography methods which have been
developed in recent years appear to offer a sensitive technique that
can be utilized very advantageously in studies on various proteins
and their properties. The application of such a procedure for the
successful fractionation of bovine sarcoplasmic proteins should stimulate
interest and research in characterizing the changes occurring
in beef muscle during the post-mortem aging period.
The research described herein pertains to the development and
application of a DEAE-cellulose ion exchange chromatography procedure
for the fractionation of bovine sarcoplasmic proteins. Results
of preliminary experiments indicated that columns packed under
pressure possessed superior fractionational qualities than did columns
packed without pressure. Also in the preliminary experiments,
a Tris buffer system (a starting buffer of 0.04 M Tris phosphate, pH 9.0, and a limiting buffer of 0.5 M Tris H₂PO₄, pH 3.6) and a
concave gradient elution procedure were developed which were found
to separate the sarcoplasmic proteins satisfactorily.
At least 16 components were recognized to be fractionated in the
chromatography of beef sarcoplasmic proteins extracted two hours
post-mortem. Duplication of the chromatographic results was found
to be quite good.
Some changes that had occurred in the sarcoplasmic proteins
during a post-mortem aging period of 10 days were detected by the
chromatographic technique. The changes observed were the appearance
of new components, and disappearance of some fractions while
others diminished.
Data obtained from experiments on the pre-chromatographic
treatment of the samples showed that any deviation in procedure
always resulted in chromatographic differences. Hence, strict
uniformity must be maintained throughout the chromatographic
procedure in order to obtain reproducible results.
Although the experimental evidence obtained in this study indicates
that further research must be completed on the chromatographic
procedure, the method does offer a sensitive technique for gaining
new information on the properties of the sarcoplasmic proteins. / Graduation date: 1965
| Identifer | oai:union.ndltd.org:ORGSU/oai:ir.library.oregonstate.edu:1957/26778 |
| Date | 01 September 1964 |
| Creators | Rampton, James Henry |
| Contributors | Anglemier, Allen F. |
| Source Sets | Oregon State University |
| Language | en_US |
| Detected Language | English |
| Type | Thesis/Dissertation |
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