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Partial purification and characterization of an extracellular proteinase from Aeromonas hydrophila strain A4

Published online by Cambridge University Press:  01 June 2009

Efstathios Alichanidis
Efstathios Alichanidis
Affiliation:
Department of Dairy Technology, University of Thessaloniki, Thessaloniki, Greece
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Summary

An extracellular metalloproteinase from Aeromonas hydrophila strain A4, isolated from milk, was purified by a factor of 300 by chromatogrpahy on DEAE-cellulose and Sephadex G-150. The enzyme had a mol. wt of 43000 and contained 2 g atom Ca/mol. It was active over a pH range 4·8–9·5 and had optimum activity on casein at pH 7·0 with Km = 0·17 mM. It was strongly inactivated by metal chelators and the apoenzyme was fully reactivated with Ca2+, Mn2+ or Co2+. Heavy metal ions such as Ag+, Hg2+, Fe2+, Zn2+, Cd2+, Ni2+ and Cu2+ totally or partly inactivated the enzymic activity at 5 mM concentration. The enzyme was not inactivated by diisopropylfluorophosphate, soyabean trypsin inhibitor or sulphydryl group reagents. It was optimally active at 45 °C; above 50 °C activity declined rapidly, but significant activity persisted at 4 °C. It was heat labile in phosphate or Tris-maleate buffer but exogenous Ca2+ afforded protection.

Type
Original articles
Information
Journal of Dairy Research , Volume 55 , Issue 1 , February 1988 , pp. 97 - 107
Copyright
Copyright © Proprietors of Journal of Dairy Research 1988

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