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Response of phosphomonoesterases in soils to chloroform fumigation
Authors:Susanne Klose  M Ali Tabatabai
Abstract:The chloroform fumigation technique has been successfully employed to quantify intracellular and extracellular urease and arylsulfatase activities in soil. In this study, the same approach was evaluated for its ability to differentiate between various pools of phosphomonoesterase activities in soils and reference proteins purified from plant and microbial sources. The activities of acid and alkaline phosphatases were assayed in 10 surface soils and reference proteins at their optimal pH values before and after chloroform fumigation and in the presence and absence of toluene. Chloroform fumigation decreased the activities of acid and alkaline phosphatases in soils, on average, by 6 and 8%, respectively. Similarly, the activities of two purified reference enzyme proteins were decreased after fumigation, with acid and alkaline phosphatase activities exhibiting a reduction of 17 and 8%, respectively. Toluene treatment caused an increase in the activities of acid and alkaline phosphatases by 8 to 18% in nonfumigated soils, but showed no effect in the fumigated soils. Average enzyme protein concentrations, calculated for the 10 soils based on the activity values of the soils and the specific activity of the purified enzymes (i.e., activity values per mg protein), were 22.5 and 2.1 mg protein (kg soil)—1 for acid and alkaline phosphatase, respectively. The decrease in enzyme activity by the fumigant was either by direct denaturing of the periplasmic and extracellular portion of the particular protein after lysis of the microbial cell membrane, by absorption and/or inhibition of the released phosphomonoesterases by organic and inorganic constituents or by degradation of the protein by soil proteases. The ratios of acid phosphatase protein concentrations relative to organic C in six soils were significantly, but negatively correlated with soil organic C, suggesting differences in organic C quality. Comparison of the activity values of soil phosphatases with those of the protein concentrations present in soils indicated that alkaline phosphatase has greater catalytic efficiency than does acid phosphatase.
Keywords:intracellular and extracellular enzyme pools  chloroform fumigation  P cycling in soils  phosphomonoesterases  enzyme protein concentrations
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