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OPTIMIZATION OF β-1,4—GLUCOSIDASE AND ACID PHOSPHATASE ENZYME METHODS FOR AGRICULTURAL SOIL STUDIES

G. Galieva, S. Selivanovskaya, P. Galitskaya
Wednesday 19 December 2018 by Libadmin2018

ABSTRACT

In our study, we optimized two methods of enzyme activity estimations characterizing C and P cycles (β-1,4-glucosidase and acid phosphatase) for agricultural soil with 2.629% of total C and 0.062 % of total N sampled in Tatarstan (Russia). Concentration of the substrates (4-MUF-β-d-glucopyranoside, 4-MUF-phosphate) in the range from 20 to 900 μmol g-1 soil was used, four levels of buffer pH (3, 6, 8, 11) were compared, and three different dilutions of soil suspension (1%, 2%, 10% w/v) were investigated. Besides, the optimal time of measurement was chosen. For β-1,4-glucosidase, it was revealed that 4-MUB-β-d-glucopyranoside concentration should be 400 μmol g-1 of soil, buffer pH should be 8.0, and soil suspension should be prepared using 1 part of soil and 100 parts of water. The measurements should be done after 60 min of incubation. For acid phosphatase, it was revealed that 4-MUB-phosphate concentration should be 400 μmol g-1 of soil, buffer pH should be 11.0, and soil suspension should be prepared using 1 part of soil and 50 parts of water. The measurements should be done after 30 min of incubation.

Keywords: soil microbial enzyme activities, synthetic fluorescent dye-conjugated substrates, β-1,4-glucosidase, acid phosphatase, buffer pН, soil suspension, incubation time, saturation curve.


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