Mineralisation of urea and urea derivatives in anaerobic soils

Summary The rates of mineralisation of urea and urea derivatives were studied in a laboratory anaerobic incubation experiment. Urea, urea phosphate and sulphur coated urea were hydrolysed rapidly and, even at the highest level of application, had disappeared in just over 8 days. The presence of the phosphate anion depressed pH in the early stages. Hydrolysis of the less soluble organic derivatives of urea, isobutylidine diurea, ureaform and glycoluril was very much slower and in the case of glycoluril a lag period of 8 to 16 days occurred before hydrolysis began.In the initial stages the system was anaerobic but between days 8 and 16 a change to partial aerobic conditions occurred. At this stage nitrification commenced and at day 16 nitrite was detected. Reduction of Fe(III) increased with time, reaching a maximum at day 32. More Fe(II) was produced in the presence of organic derivatives of urea than with the other fertilizers, possibly due to stabilisation by organic ligands. From day 16 nitrification, denitrification and reduction of Fe(III) proceeded together even through E_h values indicated that oxidation of Fe(II) would be expected. This did not occur until after day 32. Once nitrification began denitrification quickly followed so that for all six fertilisers, except at the highest level of application, virtually all the mineralised-N had been lost by denitrification at the end of the experiment.     

Molecular evolution of urea amidolyase and urea carboxylase in fungi

Background  

Urea amidolyase breaks down urea into ammonia and carbon dioxide in a two-step process, while another enzyme, urease, does this in a one step-process. Urea amidolyase has been found only in some fungal species among eukaryotes. It contains two major domains: the amidase and urea carboxylase domains. A shorter form of urea amidolyase is known as urea carboxylase and has no amidase domain. Eukaryotic urea carboxylase has been found only in several fungal species and green algae. In order to elucidate the evolutionary origin of urea amidolyase and urea carboxylase, we studied the distribution of urea amidolyase, urea carboxylase, as well as other proteins including urease, across kingdoms.     

Comparative transport efficiencies of urea analogues through urea transporter UT-B

Expression of urea transporter UT-B confers high urea permeability to mammalian erythrocytes. Erythrocyte membranes also permeate various urea analogues, suggesting common transport pathways for urea and structurally similar solutes. In this study, we examined UT-B-facilitated passage of urea analogues and other neutral small solutes by comparing transport properties of wildtype to UT-B-deficient mouse erythrocytes. Stopped-flow light-scattering measurements indicated high UT-B permeability to urea and chemical analogues formamide, acetamide, methylurea, methylformamide, ammonium carbamate, and acrylamide, each with P_s > 5.0 × 10^− 6 cm/s at 10 °C. UT-B genetic knockout and phloretin treatment of wildtype erythrocytes similarly reduced urea analogue permeabilities. Strong temperature dependencies of formamide, acetamide, acrylamide and butyramide transport across UT-B-null membranes (E_a > 10 kcal/mol) suggested efficient diffusion of these amides across lipid bilayers. Urea analogues dimethylurea, acryalmide, methylurea, thiourea and methylformamide inhibited UT-B-mediated urea transport by > 60% in the absence of transmembrane analogue gradients, supporting a pore-blocking mechanism of UT-B inhibition. Differential transport efficiencies of urea and its analogues through UT-B provide insight into chemical interactions between neutral solutes and the UT-B pore.     

Comparative transport efficiencies of urea analogues through urea transporter UT-B

Expression of urea transporter UT-B confers high urea permeability to mammalian erythrocytes. Erythrocyte membranes also permeate various urea analogues, suggesting common transport pathways for urea and structurally similar solutes. In this study, we examined UT-B-facilitated passage of urea analogues and other neutral small solutes by comparing transport properties of wildtype to UT-B-deficient mouse erythrocytes. Stopped-flow light-scattering measurements indicated high UT-B permeability to urea and chemical analogues formamide, acetamide, methylurea, methylformamide, ammonium carbamate, and acrylamide, each with P(s)>5.0 x 10(-6) cm/s at 10 degrees C. UT-B genetic knockout and phloretin treatment of wildtype erythrocytes similarly reduced urea analogue permeabilities. Strong temperature dependencies of formamide, acetamide, acrylamide and butyramide transport across UT-B-null membranes (E(a)>10 kcal/mol) suggested efficient diffusion of these amides across lipid bilayers. Urea analogues dimethylurea, acryalmide, methylurea, thiourea and methylformamide inhibited UT-B-mediated urea transport by >60% in the absence of transmembrane analogue gradients, supporting a pore-blocking mechanism of UT-B inhibition. Differential transport efficiencies of urea and its analogues through UT-B provide insight into chemical interactions between neutral solutes and the UT-B pore.     

土壤盐渍化对尿素与磷酸脲氨挥发的影响

氨挥发是肥料氮素损失的重要途径之一,肥料类型、土壤类型、肥料用量以及土壤全盐量均影响氨挥发损失率及挥发特征。本文采用通气法测定了磷酸脲和尿素两种肥料六个施肥量处理分别施入六个不同盐渍化程度（1.7、9.9、16.4、23.2、29.1、37.9 g/kg）的土壤后氨挥发累积状况和动力学特性,以及土壤氨挥发累积量与土壤电导值之间的相关性。结果表明：（1）在土壤总盐介于1.66 -37.9 g/kg的范围内,随着土壤含盐量增加,尿素与磷酸脲处理的氨挥发累积量显著增加;土壤含盐量对氨挥发速率有显著的促进作用。（2）各处理二次线性函数拟合的二项式系数a均为负值,表明：在不同盐渍化条件下肥料的挥发速率是随着时间增长而降低的;一次线性函数和Elovich 方程的斜率a随土壤含盐量增加而增大,表明：土壤盐渍化将加剧土壤的氨挥发速率。（3）土壤氨挥发累积量与电导值拟合结果符合logistic方程（︱R︱分别为0.9732,0.9815,0.965,0.9182,0.9817,0.9971︱R︱>r0.01=0.9172, n=6）,氨挥发累积量随土壤电导值呈“S”型增长。     

The urea carboxylase and allophanate hydrolase activities of urea amidolyase are functionally independent

Urea amidolyase (UAL) is a multifunctional biotin‐dependent enzyme that contributes to both bacterial and fungal pathogenicity by catalyzing the ATP‐dependent cleavage of urea into ammonia and CO₂. UAL is comprised of two enzymatic components: urea carboxylase (UC) and allophanate hydrolase (AH). These enzyme activities are encoded on separate but proximally related genes in prokaryotes while, in most fungi, they are encoded by a single gene that produces a fusion enzyme on a single polypeptide chain. It is unclear whether the UC and AH activities are connected through substrate channeling or other forms of direct communication. Here, we use multiple biochemical approaches to demonstrate that there is no substrate channeling or interdomain/intersubunit communication between UC and AH. Neither stable nor transient interactions can be detected between prokaryotic UC and AH and the catalytic efficiencies of UC and AH are independent of one another. Furthermore, an artificial fusion of UC and AH does not significantly alter the AH enzyme activity or catalytic efficiency. These results support the surprising functional independence of AH from UC in both the prokaryotic and fungal UAL enzymes and serve as an important reminder that the evolution of multifunctional enzymes through gene fusion events does not always correlate with enhanced catalytic function.     

Treatment of urea cycle disorders

Recent advances in the treatment of inborn errors of urea synthesis have significantly decreased mortality. Treatment has included combining a high-quality low-protein diet with supplements of deficient metabolites and stimulation of alternate pathways of waste nitrogen excretion. Long-term alternate pathway therapy, using sodium benzoate and sodium phenylacetate, has generally been unassociated with signs of toxicity. However, acute intoxications have simulated hyperammonemic crises. Neurologic outcome appears to be primarily a function of duration of neonatal hyperammonemic coma, although ongoing accumulation of urea cycle intermediates may also play a role. Early recognition and treatment are critical if a good outcome is to be possible.