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A qualitative screening procedure for the detection of casein hydrolysis by bacteria, using sodium dodecyl sulphate polyacrylamide gel electrophoresis

Published online by Cambridge University Press:  01 June 2009

Susan H. A. Hill  and
Michael J. Gasson
Susan H. A. Hill
Affiliation:
AFRC Institute of Food Research, Reading Laboratory (University of Reading), Shinfield, Reading RG2 9 AT, UK
Michael J. Gasson
Affiliation:
AFRC Institute of Food Research, Reading Laboratory (University of Reading), Shinfield, Reading RG2 9 AT, UK
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Summary

Incubation of bacteria with casein at 30 °C (pH 7·2) resulted in the formation of a number of peptide fragments. The peptides were separable when subjected to electrophoresis. A proteinase-positive strain of Streptococcus lactis preferentially hydrolysed the β-casein moiety of whole casein, and after incubation for 5 min produced a characteristic pattern of four peptides on sodium dodecyl sulphate (SDS) polyacrylamide gels. After 30 min incubation only one peptide remained. This single polypeptide was then further hydrolysed to produce one (60 min) and eventually two (120 min) lower molecular weight peptides. Hydrolysis was generally complete after about 480 min. Two proteinase-negative variants of this strain did not hydrolyse casein. Crude proteinase preparations gave the same characteristic peptide patterns. SDS-polyacrylamide gel electrophoresis was used to screen more proteinase-positive strains of Str. Lactis for casein hydrolysing activity. Of those so far tested all produced the same peptide patterns from β-casein. Proteinase negative variants of all these strains did not hydrolyse casein.

Type
Original Articles
Information
Journal of Dairy Research , Volume 53 , Issue 4 , November 1986 , pp. 625 - 629
Copyright
Copyright © Proprietors of Journal of Dairy Research 1986

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References

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