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Hydrolysis of ¹⁴C-Biuret by In Vitro Rumen Fermentation and Crude Biuretase Preparations

The Dow Chemical Company, Midland, Michigan 48823

Abstract

The hydrolysis of ¹⁴C- biuret by in vitro fermentation using sheep and cattle rumen fluid was studied. Evolution of 14CO₂ or loss of acid-stable ¹⁴C activity in solution were used as measures of biuret hydrolysis. Prior exposure of the source animal to biuret was essential, and most of the biuretolytic activity resided with the microorganisms closely associated with the plant debris. Adding energy source nutrients to the rumen fluid had little effect on the hydrolysis of biuret, but appeared to increase the utilization of the products of biuret hydrolysis, CO₂ and NH₃. The ¹⁴C activity of biuret was transformed to ¹⁴CO₂, ¹⁴CH₄ and ¹⁴C incorporated into bacterial mass.

Cell-free extracts of ruptured rumen microorganisms, especially those associated with the plant debris, were shown to possess biuretase activity. The enzyme is distinct from urease in distribution, action and response to inhibitors, but urease may participate in the complete hydrolysis of biuret to CO₂ and NH₃. The biuretase enzyme contains essential sulhydryl groups and may be prepared in dry form by acetone dehydration.

¹ Agricultural Department, The Dow Chemical Company, Midland, Michigan.