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Characterization of urease derived from Citrullus lanatus (watermelon) seeds to estimate total Kjeldahl nitrogen in human urine

Lim, Kahui, Leverenz, Harold
International journal of environmental analytical chemistry 2019 v.99 no.5 pp. 486-499
Citrullus lanatus, ammonium nitrogen, enzyme kinetics, field methods, liquids, nitrogen, pH, refrigeration, seeds, slurries, total Kjeldahl nitrogen, urea, urease, urine, watermelons, wet digestion method
A urease extract prepared by decanting liquid from a suspension of finely ground Citrullus lanatus (watermelon) seeds was characterized and applied to dilute urine samples to demonstrate a low-cost field method to estimate total Kjeldahl nitrogen (TKN) concentrations in human urine. The extract exhibited a Michaelis-Menten constant, Kₘ, of 3.00 mM urea and a specific activity of up to 12.2 U/mg protein at an optimum pH of 8.1. A statistical F-test on 54 samples demonstrated that TKN can be estimated as the total ammonium-nitrogen recovered upon addition of urease in dilute fresh and stale urine samples. The total ammonium-nitrogen in urine samples determined after treatment with watermelon seed urease was consistent with that determined using traditional acid digestion techniques. The extract retained 85% of its initial capacity after three months of refrigeration. The effectiveness of this method to assay nitrogen in unbuffered urine samples will be useful in nitrogen analyses in nutrient recovery and urine or slurry storage contexts. Accordingly, this study is useful in understanding the kinetics of a plant-derived urease acting in dilute urine.