Urease surface

Bacterial catabolism of oral food residue is probably responsible for a higher [NHj] in the oral cavity than in the rest of the respiratory tract.Ammonia, the by-product of oral bacterial protein catabolism and subsequent ureolysis, desorbs from the fluid lining the oral cavity to the airstream.. Saliva, gingival crevicular fluids, and dental plaque supply urea to oral bacteria and may themselves be sites of bacterial NH3 production, based on the presence of urease in each of these materials.Consequently, oral cavity fNTi3)4 is controlled by factors that influence bacterial protein catabolism and ureolysis. Such factors may include the pH of the surface lining fluid, bacterial nutrient sources (food residue on teeth or on buccal surfaces), saliva production, saliva pH, and the effects of oral surface temperature on bacterial metabolism and wall blood flow. The role of teeth, as structures that facilitate bacterial colonization and food entrapment, in augmenting [NH3J4 is unknown. 

Quartz surfaces 16-Carbon spacer chain, urease [119]... 

Jackbean urease was immobilized on kaolinite and montmorillonite [98]. The amounts of urease required for maximum immobilization were 70 and 90 mg g 1 of kaolinite and montmorillonite, respectively. The Km values of immobilized urease (25.1-60.8 mM) were of the same order of magnitude as that of free urease (29.4 mM) but one order of magnitude higher than those of soil urease (1.77-2.90 mM). Immobilization of urease on clay surfaces leads to increases in the kinetic constants. 

Figure 15.4(A) shows the effect of the R = Zn2+/Al3+ ratio, which determines the charge density of the LDH layer, on the Freundlich adsorption isotherms. K values are far higher than those measured for smectite or other inorganic matrices. The increase in Kf with the charge density (Kf= 215, 228, 325mg/g, respectively, for R = 4, 3 and 2) is supported by a mechanism of adsorption based on an anion exchange reaction. The desorption isotherms confirm that urease is chemically adsorbed by the LDH surface. The aggregation of the LDH platelets can affect noticeably their adsorption capacity for enzymes and the preparation of LDH adsorbant appears to be a determinant step for the immobilization efficiency. [ZnRAl]-urease hybrid LDH was also prepared by coprecipitation with R = 2, 3 and 4 and Q= urease/ZnRAl from 1 /3 up to 2.5. For Q < 1.0,100 % of the urease is retained by the LDH matrix whatever the R value while for higher Q values an increase in the enzyme/LDH weight ratio leads to a decrease in the percentage of the immobilized amount.

An enzyme deposited on the LAPS surface allows one to observe the spatial distribution of a specific substrate. In a urea-selective sensor urease was immobilized on a pH-selective LAPS [75],... 

Tor [7] developed a new method for the preparation of thin, uniform, self-mounted enzyme membrane, directly coating the surface of glass pH electrodes. The enzyme was dissolved in a solution containing synthetic prepolymers. The electrode was dipped in the solution, dried, and drained carefully. The backbone polymer was then cross-linked under controlled conditions to generate a thin enzyme membrane. The method was demonstrated and characterized by the determination of acetylcholine by an acetylcholine esterase electrode, urea by a urease electrode, and penicillin G by a penicillinase electrode. Linear response in a wide range of substrate concentrations and high storage and operational stability were recorded for all the enzymes tested. 

In 1989, to the best of our knowledge, the first report appeared on the use of an avidin-biotin system for the immobilization of enzymes in the preparation of biosensors. Walt et al. have immobilized biotin-modified enzymes (urease, esterase, and penicillinase) on the surface of biotin-modified optical fiber using avidin as a binder (Figure 5a). They have demonstrated the general use of this procedure in immobilizing several types of enzymes. At nearly the same time, Gunaratna and Wilson used an enzyme column in which choline esterase and choline oxidase were immobilized through avidin-biotin complexation for the determination of acetyl-... 

In another kinetics study, Huang and Chen immobilized jack bean urease in the form of a thin film on the surface of a reticulated polyurethane foam. The residual apparent activity of the urease after immobilization was about 50%. The good hydrodynamic properties and flexibility of the support were retained in solution after immobilization. Urea hydrolysis was examined in both a batch squeezer and circulated flow reactor. The results suggest potential for practical applications in various reactors. 

Figure 5.8 depicts a typical immunoassay-based LAPS. The reaction of an antibody to its antigen will immobilise the species and after washing, only those species that have been successfully bound will be left onto the LAPS surface. Here, a conjugated enzyme, e.g., urease, will change the pH value of the test sample by enzymatic catalysis after injection of urea. 

I.G. Mourzina, T. Yoshinobu, Y.E. Ermolenko, Y.G. Vlasov, M.J. Schoning and H. Iwasaki, Immobilization of urease and cholinesterase on the surface of semiconductor transducer for the development of light-addressable potentiometric sensors, Microchim. Acta, 144(1-3) (2004) 41-50. 

The objective is to describe a new non-enzymatic urea sensor based on catalytic chemical reaction. The sensor consists of screen-printed transducer (IVA, Ekaterinburg, Russia) and catalytic system which is immobilized on the transducer surface as a mixture with carbon ink. The sensor is used for measuring concentration of urea in blood serum, dialysis liquid. Detection limit is 0.007 mM, while the correlation coefficient is 0.99. Some analysis data of serum samples using the proposed sensor and urease-containing sensor (Vitros BUN/UREA Slide, Johnson Johnson Clinical Diagnostics, Inc.) are presented. 

Continuous-flow microreactors were successfully fabricated by etching channels in silicon and immobilizing urease onto channel surfaces by a layer-by-layer self-assembly technique. Preliminary results show urea conversion. The potential advantages of this surface-coating technique in microreactors warrant continued investigation. 

The gas-sensing electrodes also are used for the potentiometric measurement of biologically important species. An enzyme is immobilized at or near the gas probe. The gas sensor measures the amount of characteristic gas produced by the reaction of the analyzed substance with the enzyme. For example, an enzyme electrode for urea [NH2C(0)NH2] determination is constructed by the immobilization of urease onto the surface of an ammonia-selective electrode. When the electrode is inserted into a solution that contains urea, the enzyme catalyzes its conversion to ammonia ... 

Because the urea-urease interaction leads to a pH increase, a polymer that increases erosion rate with increasing pH is needed. A useful polymer for this application is a partially esterified copolymer of methyl vinyl ether and maleic anhydride. This copolymer undergoes surface erosion with an erosion rate that is extraordinarily pH-dependent (J). The polymer dissolves by ionization of the carboxylic acid groups as shown below ... 

Urea undergoes microbial hydrolysis catalyzed by urease, leading to loss of as much as 30% of its nitrogen from ammonia volatilization. The reduced nitrogen availability in the soils appears particularly when urea is surface broadcast on soils. The factors that influence ammonia volatilization include levels of urease activity, moisture availability, nitrification rate, and soil texture (Bernard et al., 2009). 

Chemically binding enzymes to nylon net is very simple and gives strong mechanically resistant membranes (135). The nylon net is first activated by methylation and then quickly treated with lysine. Finally, the enzyme is chemically bound with GA. The immobilized disks are fixed direcdy to the sensor surface or stored in a phosphate buffer. GOD, ascorbate oxidase, cholesterol oxidase, galactose oxidase, urease, alcohol oxidase (135), and lactate oxidase (142) have been immobilized by this procedure and the respective enzyme electrode performance has been established. 

Another less-utilized transduction mechanism for biosensors involves the acoustoelectric effect. In principle, any biochemical process that produces a change in the electrical properties of the solution, can be monitored by observing changes in the frequency and/or attenuation of the device if its surface is not metallized. For example, a SH-SAW device has been reported for the detection of pH changes associated with the enzyme-catalyzed hydrolysis of urea [235]. Using an immobilized urease membrane on the sensor surface, it was anticipated that urea concentrations as small as 3 /u.M could be reliably detected. 

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