Summary
Purification of soil phosphatase-, urease-, casein- and benzoylarginamide-hydrolysing proteases was obtained by exhaustively ultrafiltering a soil extract using 0.1M pyrophosphate solution at pH 7.1, separating the retained material into fractions of molecular weight higher (AI) and lower (AII) than 105 and eluting the fractions on gel chromatography.
Three peaks of phosphatase and urease activity were obtained after gel chromatography of fraction AI on Sephadex G200 using 0.1M pyrophosphate solution as eluant. Only one distinct activity peak was observed when casein- and benzoylarginamide-hydrolysing proteases were assayed in the eluted fractions. Elution diagrams obtained by gel chromatography of fraction AII on Sephadex G100, using a water as eluant, were characterized by one peak each of phosphatase-, casein- and benzoylarginamide-hydrolysing activity and by two peaks of urease activity.
Gel chromatography of both AI and AII, generally, but not always, increased specific activity on a C and N basis of derivative fractions. Both proteases showed the highest increase in specific activity due to a marked decrease in organic C and N and an increase in total activity.
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Nannipieri, P., Ceccanti, B., Bianchi, D. et al. Fractionation of hydrolase-humus complexes by gel chromatography. Biol Fert Soils 1, 25–29 (1985). https://doi.org/10.1007/BF00710967
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DOI: https://doi.org/10.1007/BF00710967