A polarimetric method for collagenase activity measurement

Brüning, Adrian Rudolf Nicolaus Ernst (1992) A polarimetric method for collagenase activity measurement. Masters thesis, Rhodes University.

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Abstract

A polarimetric method for monitoring the rate of soluble collagen breakdown by collagenase enzyme action has been developed. The method represents an extension of previous physicochemical techniques based on viscometry, but is simpler and easier to carry out, particularly in the case of reaction rate studies. The method was developed arising from reports of collagenase activity measurement on inappropriate substrates such as gelatin, modified collagens and synthetic polypeptides. The optical method depends on measurement of the loss in optical rotation in solutions of soluble calfskin collagen resulting from initial enzymic cleavage of the collagen trip1e-helix, followed by spontaneous unwinding of the resultant unstable helical fragments. Specific assay conditions were chosen to ensure that the loss in optical rotation following enzymic cleavage was rapid and complete. The method is specific since in the absence of collagenase, non-specific proteinases produce only a limited decrease in solution optical activity. The method has also been compared with established physicochemical assay techniques and compares favourably with both viscometric and titrimetric collagenase assays. The availability of a rapid, sensitive and quantitative procedure for measurement of collagenase activity provides a convenient means for detecting the presence of collagenase in solution and examination of hide bacterial cultures for collagenase production. In addition, a study of biocidal compounds of potential interest in hide preservation for possible inhibitory effects on collagenase is conveniently carried out with the method. Fundamental research into synergistic action in enzymic hydrolysis of collagen is now possible, providing valuable insight into the mechanism of raw hide biodeterioration.

Item Type:Thesis (Masters)
Uncontrolled Keywords:Hides, Skins, Preservation, Collagenases, Enzyme action, Synthetic polypeptides, Gelatin, Biodeterioration
Subjects:Q Science > QD Chemistry > QD241 Organic chemistry > QD415 Biochemistry
Divisions:Faculty > Faculty of Science > Biochemistry, Microbiology & Biotechnology
Supervisors:Russell, Allan E. and Duncan, John R.
ID Code:4136
Deposited By: Philip Clarke
Deposited On:26 Nov 2012 12:15
Last Modified:26 Nov 2012 12:15
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