Thermal stability of an extracellular proteinase from Pseudomonas fluorescens AFT 36

The Journal of Dairy Research
L Stepaniak, P F Fox

Abstract

A metalloproteinase, isolated from a shaken milk culture of Pseudomonas fluorescens AFT 36 by chromatography in DEAE and CM-cellulose and Sephadex G-150, was very unstable in 0.1 M-phosphate buffer, pH 6.6, being completely denatured above 70 degrees C in 1 min. It was also unstable in a Ca-containing buffer (synthetic milk salts, SMS) between 50 and 60 degrees C (minimum at 55 degrees C), but stability was very high above 80 degrees C in this buffer. D-values were determined at 10 degrees C intervals in the range 70-150 degrees C in SMS from which a Z value of 31.9 degrees C and an Ea of 8.82 X 10(4) J mol-1 were calculated; the half-life at 150 degrees C was 9 s. Instability at 55 degrees C was due to autolysis as evidenced by gel electrophoresis, gel filtration and increase in 2,4,6-trinitrobenzenesulphonic acid-reactive amino groups. The extent of inactivation experienced at 80 degrees C was inversely related to the rate of heating to 80 degrees C, i.e. length of time spent in the neighbourhood of 55 degrees C. Addition of increasing concentrations of caseinate substrate reduced inactivation of the enzyme at 55 degrees C, presumably due to substrate binding. Attempts to stabilize the enzyme at 55 degrees C by addition of ED...Continue Reading

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Citations

Oct 1, 1984·Zeitschrift Für Lebensmittel-Untersuchung Und -Forschung·S Kroll, H Klostermeyer
May 1, 1993·The Journal of Dairy Research·H KumuraZ Saito
Feb 1, 1987·The Journal of Dairy Research·P DiermayrH Klostermeyer
Feb 1, 1986·The Journal of Dairy Research·D J Fairbairn, B A Law
Feb 1, 1985·The Journal of Dairy Research·L Stepaniak, P F Fox
Apr 1, 1994·The Journal of Applied Bacteriology·V K UplackshR K Malik
Sep 6, 2000·Journal of Applied Microbiology·R Koka, B C Weimer
Oct 26, 1999·Journal of Dairy Science·H KumuraK Shimazaki
Apr 1, 1992·Journal of Dairy Science·M Gobbetti, J Rossi
Jan 1, 1984·Zeitschrift Für Lebensmittel-Untersuchung Und -Forschung·S Kroll, H Klostermeyer

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