Urease action

Kinetics of formation of the dinuclear iron(III) complex [(tpa)Fe (p-0)(p-urea)Fe(tpa)]s+ tpa = tris(2-pyridylmethyl)amine were investigated in relation to the suggestion that urease action in vivo involves an intermediate containing Ni (p - O H) (p -ur e a) Ni. The mechanism of formation of the di-iron species from [(tpa)(H20)Fe(p-0)Fe(0H)(tpa)]3+ is proposed to involve three reversible steps (350). Three kinetically distinct steps are also involved in the deposition of FeO(OH) in... 

The mechanism of urease action is probably related to those of metalloproteases such as carboxypeptidase A (Fig. 12-16) and of the zinc-dependent carbonic... 

The chief sources of this important enzyme are (a) the jack bean (Canavalia ensiformis). (b) the soy (or soja) bean (Glycine hispida). The enzyme is of great value in identifying and estimating urea. The action of urease on urea is specific, the reaction catalysed being ... 

The contents of B, which act as a control, are treated with mercuric chloride in order to inhibit the action of the enzyme, and then 10 ml. of urease solution are added. The solution is diluted with water and ammonium chloride added (in order to balance the ammonium chloride subsequently formed in A). Meth) l-red is then added and the solution is titrated with Mj 10 HCl from a second burette B until a bright red colour is obtained. 

Enzymes are proteins of high molecular weight and possess exceptionally high catalytic properties. These are important to plant and animal life processes. An enzyme, E, is a protein or protein-like substance with catalytic properties. A substrate, S, is the substance that is chemically transformed at an accelerated rate because of the action of the enzyme on it. Most enzymes are normally named in terms of the reactions they catalyze. In practice, a suffice -ase is added to the substrate on which die enzyme acts. Eor example, die enzyme dial catalyzes die decomposition of urea is urease, the enzyme dial acts on uric acid is uricase, and die enzyme present in die micro-organism dial converts glucose to gluconolactone is glucose oxidase. The diree major types of enzyme reaction are ... 

Recently, the old alkaline phenol method has been revived, and is being widely used in clinical laboratories, without protein preclpltatlon(27). In this procedure, the serum is added to an alkaline phenol reagent, and the ammonia generated from urea is determined either after the action of urease or after strong alkaline treatment of the serum. The objection to this procedure is first, that all urease is rich in ammonia, and second, the color produced with alkaline phenol is not specific for ammonia. It will react with other compounds, especially for those that liberate ammonia. By this procedure one obtains a useful number from the point of view of determining whether the patient has nitrogen retention, but a value which is somewhere between a urea and an N.P.N. determination. 

Enzymes are usually named in terms of the reaction that is catalyzed, commonly adding the suffix -ase to the name of the stoichiometrically converted reactant or substrate. For instance, an enzyme that catalyses the hydrolysis of urea is urease. Enzyme names can only be applied to single enzymes, especially those with termination -ase. For systems that involve the action of two or more enzymes the use of the term should be avoided and the word system should be included. 

Both dietary and endogenous ammoniagenic substrates are removed from the intestinal lumen by the osmotic cathartic action of nonabsorbable disaccharides such as lactulose and lactitol. These compounds are currently the main therapeutic agents for chronic HE. The efficacy of oral lactulose for the treatment of HE has been established in controlled trials [41-43]. Besides having a cathartic effect, lactulose lowers the colonic pH as a result of the production of organic acids by bacterial fermentation. The decrease in pH creates an environment that is hostile to the survival of urease-producing intestinal bac-... 

Kinetic evidence obtained for intramolecular proton transfer between nickel and coordinated thiolate, in a tetrahedral complex containing the bulky triphos ligand (Pl PCE CE PPh to prevent interference from binuclear p-thiolate species, is important with respect to the mechanisms of action of a number of metalloenzymes, of nickel (cf. urease, Section VII. B.4) and of other metals (289). 

The enzyme urease was discovered in soybeans by Takeuchi in 1909 it catalyzed the conversion of urea to ammonium carbonate. Jack beans were another excellent source of the enzyme. Jack bean powder could be stored for considerable periods and very active, soluble, urease extracted. After the action of urease, the ammonia could be estimated colorimetrically by Nesslerisation or titrimetrically. The Conway diffusion apparatus was specially developed for the estimation of urea titrimetrically and remained in use into the 1950s. 

Most commonly used enzyme names have the suffix "-ase" attached to the substrate of the reaction (for example, glucosi-dase, urease, sucrase), or to a description of the action performed (for example, lactate dehydrogenase and adenylyl cyclase). [Note Some enzymes retain their original trivial names, which give no hint of the associated enzymic reaction, for example, trypsin and pepsin.]... 

Fate of urea Urea diffuses from the liver, and is transported in the blood to the kidneys, where it is filtered and excreted in the urine. A portion of the urea diffuses from the blood into the intestine, and is cleaved to CO2 and NH3 by bacterial urease. This ammonia is partly lost in the feces, and is partly reabsorbed into the blood. In patients with kidney failure, plasma urea levels are elevated, promoting a greater transfer of urea from blood into the gut. The intestinal action of urease on this urea becomes a clinically important source of ammonia, contributing to the hyperam-... 

From bacterial action in the intestine Ammonia is formed fan urea by the action of bacterial urease in the lumen of the htes-tine. This ammonia is absorbed from the intestine by way of tie portal vein and is almost quantitatively removed by the liver ve conversion to urea. 

Enzymes are usually impressively specific in their action. The specificity toward substrate is sometimes almost absolute. For many years urea was believed to be the only substrate for the enzyme urease and succinate the only substrate for succinate dehydrogenase. Even after much searching for other substrates, only... 

A possible mechanism for the action of urease is pictured in Fig. 16-25 and Eq. 16-47. Carbamate is thought to be one intermediate. Can you suggest an alternative possibility for the initial nickel ion-dependent steps. See Barrios and Lippard.476... 

UREASE. Enzyme present in low-percentages in jackbean and soybean water soluble, its action is inhibited by heavy-metal ions. Its principal use is in the determination of urea in urine, blood, and other body fluids it splits urea into ammonia and carbon dioxide or ammonium carbonate. 

The isoelectric point, determined by Sumner and Hand (50) to have a value of 5.0-5.1, has been redetermined by the electrofocusing technique (43). The value obtained was 4.8. The solubility is extremely small at this pH, but the urease can be located by its enzymic action. It is a point of interest that the solubility of the isoelectric species increases spectacularly if the sulfhydryl groups are substituted with A-ethylmaleimide (NEM) (43). 

The Enzyme Commission catalog (EC 3.5.1.5) lists the urease reaction as urea + 2 H20 = C02 + 2 NH3. Since two C-N bonds are broken it is evident that the stoichiometric relation above is the result of two component reactions. Any conjecture concerning the mechanisms of these reactions and the nature of the intermediates must encompass the action of inhibitors and the spectrum of substrates. Some of the organic inhibitors that have been reported are shown in Table I. The substrates that have been shown to be hydrolyzed are listed in Table II. 

Thomas, C. R. and P. Dunnill, "Action of Shear on Enzymes Studies with Catalase and Urease," Biotechnol Bioeng. 21 (1979) 2279 - 2302. 

Roughly 30% of enzymes are metalloenzymes or require metal ions for activity and the present chapter will concentrate on the chemisty and structure of the plant metalloenzymes. As analytical methods have improved it has been possible to establish a metal ion requirement for a variety of enzymes which were initially considered to be pure proteins. A dramatic example is provided by the enzyme urease isolated from Jack beans and first crystallised by Sumner (1926) (the first enzyme to be crystallised). Sumner defined an enzyme as a pure protein with catalytic activity, however, Zerner and his coworkers (Dixon et al., 1975) established that urease is in fact a nickel metalloenzyme. Jack bean urease contains two moles of nickel(II) per mole of active sites and at least one of these metal ions is implicated in its mechanism of action. 

Dixon et al. (1980) have proposed a mechanism for the action of urease (Fig. 5-3). The mechanism involves nucleophilic attack by hydroxide coordinated to one nickel, on urea which is coordinated to the other nickel(II) via the carbonyl oxygen. 

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