, Volume 17, Issue 2, pp 147-153

Measurement of protein synthesis by soil bacterial assemblages with the leucine incorporation technique

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The incorporation of [14C]-leucine into protein by soil organisms was measured both in soil slurries and for bacteria extracted from soil by homogenization-centrifugation. The result was compared with thymidine incorporation. Using a soil slurry, 9.1×10-10 mol leucine h-1g-1 dry weight of soil was incorporated into protein, with a calculated leucine: thymidine ratio (mol:mol) of about 34. Non-specific labelling of macromolecules other than protein was observed with both the soil slurry and the homogenization-centrifugation method. With the latter, 46.5% of the total incorporation was found in the protein fraction (hot-acid insoluble). The incorporation of leucine was linear with time for at least 4 h for extracted bacteria. Even at 2000 nM, [14C]-leucine did not saturate incorporation into protein. Isotope dilution plots indicated that with 750 nM leucine, the degree of participation of the labelled substance in protein synthesis was 0.59. With this value, the ratio of leucine:thymidine incorporation into total macromolecules was calculated as 41 for extracted bacteria. On the basis of incorporation into protein (leucine) and incorporation into DNA (thymidine) only, the leucine:thymidine ratio was calculated as 117. The mean turnover time of bacteria at 22°C, calculated using conversion factors from published studies and leucine incorporation into protein of extracted bacteria, was 4.3 days.