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Inhibition by chelating agents of the formation of active extracellular proteinase by Pseudomonas fluorescens 32A

Published online by Cambridge University Press:  01 June 2009

Robin C. Mckeller  and
Hilaire Cholette
Robin C. Mckeller
Affiliation:
Food Research Institute, Researcg Branch, Agriculture Canada, Ottawa, Ontario K1A 0C6, Canada
Hilaire Cholette
Affiliation:
Food Research Institute, Researcg Branch, Agriculture Canada, Ottawa, Ontario K1A 0C6, Canada
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Summary

The effect of chelating agents on extracellular proteinase production by Pseudomonas fluorescens 32A was examined. Increasing concentrations of orthophosphate slightly stimulated growth while inhibiting proteinase synthesis. Fifty percent inhibition was found at 35 and 28 mM-orthophosphate at 5 and 20 °C respectively. Extracellular protein concentration was reduced by 30% when cells were grown with 100 mM-orthophosphate. Polyacrylamide gel electrophoresis of the cell-free supernatants suggested that reduced enzyme synthesis had taken place as evidenced by the decrease in staining intensity of the protein band corresponding to the proteinase. Other phosphate compounds could replace orthophosphate as an inhibitor. Extent of inhibition was related to chain length; polyphosphates with 4–6 or 13–18 phosphorus atoms were the most effective inhibitors. EDTA (0·5 mM) completely inhibited proteinase synthesis. This inhibition could be partly reversed by Ca2+ and, to a lesser extent, Mn2+. Proteinase production at 5 °C in skim milk was completely inhibited by phosphate glass (P13–P18). Control experiments showed that loss of activity with chelators was not due to inhibition of preformed enzyme. The results suggest a possible role for polyphosphates in controlling proteinase production in stored milk.

Type
Original Articles
Information
Journal of Dairy Research , Volume 52 , Issue 1 , February 1985 , pp. 91 - 100
Copyright
Copyright © Proprietors of Journal of Dairy Research 1985

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