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Purification and characterization of the extracellular proteinase of Pseudomonas fluorescens NCDO 2085

Published online by Cambridge University Press:  01 June 2009

David J. Fairbairn  and
Barry A. Law
David J. Fairbairn
Affiliation:
AFRC Institute of Food Research, Reading Laboratory (University of Reading), Shinfield, Reading RG2 9 AT, UK
Barry A. Law
Affiliation:
AFRC Institute of Food Research, Reading Laboratory (University of Reading), Shinfield, Reading RG2 9 AT, UK
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Summaey

Pseudomonas fluorescens NCDO 2085 produced a single heat-stable extracellular proteinase in Na caseinate medium at 20 °C and pH 7·0. The proteinase was purified to electrophoretic homogeneity using chromatofocusing, gel filtration and ion-exchange chromatography. The purification procedure resulted in a 158-fold increase in the specific activity and a yield of 3·5% of the original activity. The enzyme is a metalloproteinase containing Zn and Ca, with an isoelectric point at 5·40±0·05 and a mol. wt of 40200±2100. It is heat-stable having D-values at 74 and 140 °C of 1·6 and 1·0 min respectively; 40 and 70% of the original activity remained after HTST (74 °C/17 s) and ultra high temperature (140°C/4 s) treatments respectively. The amino acid composition of the proteinase was determined and compared with those from other Pseudomonas spp.

Type
Original Articles
Information
Journal of Dairy Research , Volume 53 , Issue 3 , August 1986 , pp. 457 - 466
Copyright
Copyright © Proprietors of Journal of Dairy Research 1986

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