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Characterization of rennet preparations produced by genetic engineering in comparison to calf rennet


Characterization of rennet preparations produced by genetic engineering in comparison to calf rennet



Milchwissenschaft 42(12): 787-789



ISSN/ISBN: 0026-3788

For a comparative analysis of calf rennet and rennet preparations produced by genetic engineering different methods for the purity and identity test as well as for measuring the proteolytic activity are described. The anion exchange column chromatography was used for the pre-purification of the chymosin and the pepsin fraction, resp. By the fine structure of UV spectra in derivative spectroscopy (2nd and 4th derivative) it was possible to differentiate pepsin and chymosin. The immunochemical detection of chymosin and pepsin was done by radial agar gel double diffusion. By its high resolution the ultrathin-layer isoelectric focusing (UDIEF) was adapted for the identification of genetic variants of chymosin. For a differentiation of the proteolytic enzymes from the concomitant rennet proteins in the electropherogram the substrate gel technique was used. A skim-milk powder agarose diffusion test served as a micromethod for the quantitative determination of the proteolytic enzymes. The proteolytic activity of rennet enzymes could be characterized by renneting strength determination and by the HPLC peptide pattern after the proteolysis of .alpha.S1-casein. The content of phosphotungstic acid soluble amino-N which could be determined by o-phthaldialdehyde (OPA) in aqueous cheese extracts was an appropriate measure of proteolysis in cheese.

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Accession: 004931299

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