Penicillin enzyme electrode

Papariello G, Mukherji A and Shearer C 1973 Penicillin enzyme electrode Anal. 

Mifflin, T. E. Andriano, K. M. Robbins, W. B. Determination of Penicillin Using an Immobilized Enzyme Electrode, /. Chem. Educ. 1984, 61, 638-639. 

Urea in kidney dialysate can be determined by immobilizing urease (via silylation or with glutaraldehyde as binder) on commercially available acid-base cellulose pads the process has to be modified slightly in order not to alter the dye contained in the pads [57]. The stopped-flow technique assures the required sensitivity for the enzymatic reaction, which takes 30-60 s. Synchronization of the peristaltic pumps PI and P2 in the valveless impulse-response flow injection manifold depicted in Fig. 5.19.B by means of a timer enables kinetic measurements [62]. Following a comprehensive study of the effect of hydrodynamic and (bio)chemical variables, the sensor was optimized for monitoring urea in real biological samples. A similar system was used for the determination of penicillin by penicillinase-catalysed hydrolysis. The enzyme was immobilized on acid-base cellulose strips via bovine serum albumin similarly as in enzyme electrodes [63], even though the above-described procedure would have been equally effective. 

These reactions are coupled with a signal transducer using mainly the pH decrease and the reaction enthalpy. Whereas (3-lactamase has been combined with various transducer types, penicillin amidase has been applied only in one enzyme electrode for penicillin (Table 9). 

Fig. 75. Construction of a sterilizable enzyme electrode for the determination of penicillin. A flat glass electrode, B reaction chamber containing enzyme, C dialysis membrane, D holder. (Redrawn from Enfors, 1987).

Enzyme electrodes have been described for the majority of the low-molecular substances of interest, such as amino acids, sugars, phosphates, penicillin and gluconic acid. In contrast, fundamental problems... 

Nielsen PM, Olsen HS (2002) Enzymic modification of food protein. In Whitehurst RJ, Law BA (eds). Enzymes in food technology. CRC Press, Boca Raton, pp 109-143 Nilsson H, Mosbach K, Enfors SO et al. (1978) An enzyme electrode for measurement of penicillin in fermentation broth a step toward the application of enzyme electrodes in fermentation control. Biotechnol Bioeng 20(4) 527-539... 

Investigation of design and response characteristics of an enzyme electrode for the determination of penicillin Investigation of the kinetic properties and stability of the active immobilized enzyme Living immobilized cells for the production of penicillin G from D-glucose... 

Enzyme Electrode. The determination of penicillin is possible by immobilising penicillinase onto glass electrodes. The principle of operation of the sensor is based on the changes in the H " concentration resulting from the enzymatic hydrolysis of penicillin by penicillinase. 

In process control and the food industry enzyme electrodes are employed mainly for the determination of carbohydrates, ascorbic acid, amino acids, and penicillin. However, the application of such sensors in situ in bioreactors is associated with significant difficulties ... 

Immobilized enzymes are used in the industrial production of many compounds. They are used in the manufacture of amino acids, sugar products such as glucose and high fructose com syrup, and many types of pharmaceutical compounds including antibiotics such as penicillin (1-4). The enzyme is used in these cases for enantioselective synthesis, to produce the desired enantiomer or isomer. Enzymes are currently being explored for their use in the manufacture of biofuels (5). Enzymes are used in analysis in both free and immobilized states. In both states, they are able to determine, with great sensitivity, the concentration of substrates in complex matrices. Immobilized enzymes can be deployed in several different form factors, such as reactor tubes, microwell plates, and as enzyme electrodes in biosensor devices. 

Other, practically useful, enzyme electrodes include those based on urease and penicillinase. The penicillin electrode utilizes a pH electrode with immobilized penicillinase. It is widely used to monitor the penidllin content of fermentation reaction mixtures ... 

The most commonly used biomedical sensors are the dissolved oxygen p02 and pH electrodes, although enzyme electrodes for glucose and penicillin are becoming more widely accepted. 

An amygdalin-sensitive enzyme electrode is obtained by simply immobilizing dte enzyme galacrosidase on a pCN electrode [127]. Similarly, penicillin is determined by cleaving die amide bond of the P-lactame ring with penicillinase 10 produce poiicilloic acid ... 

Microbial electrodes measure antibiotics in a very different way to enzyme electrodes. The penicillinase electrode transforms penicillin into penicilloic acid [108], which is detectable by the transducer. The microbial sensor for antibiotics makes use of the inhibitory effect of the antibiotic nystatin on the respiration of the yeast, Streptomyces species [63]. The enzyme sensor measures the number of moles of penicillin whereas the microbial sensor gives the unit activity of nystatin. 

Enfors S.O. and Nilson H. (1979) Design and response characteristics of an enzyme electrode for measurement of penicillin in fermentation broth. Enzyme Microb. Technol., 1, 260-264. 

Directions for preparing a potentiometric biosensor for penicillin are provided in this experiment. The enzyme penicillinase is immobilized in a polyacrylamide polymer formed on the surface of a glass pH electrode. The electrode shows a linear response to penicillin G over a concentration range of 10 M to 10 M. 

Mifflin and associates described a membrane electrode for the quantitative analysis of penicillin in which the enzyme penicillinase is immobilized in a polyacrylamide gel that is coated on a glass pH electrode. The following data were collected for a series of penicillin standards. 

Penicillin can likewise be determined by using the enzyme penicillinase to destroy the penicillin with production of hydrogen ions which can be determined using a normal glass pH electrode. Many other organic materials can be determined by similar procedures.34,35... 

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. 

Electrodes with enzymes (e.g., urease) incorporated into a gel membrane. They are selective for biochemical compounds such as urea, glucose, L-amino acids, and penicillin (see - biosensors). 

Electrodes based on the immobilization of enzymes at the electrode surface have been reported for a few pharmaceutical applications. Seegopaul and Rechnitz developed a CO2 electrode that responds to methotrexate due to the inhibition of dihydrofolate reductase by the analyte. Additional applications have included the detection of penicillin consumption by penicillinase as well as urea by urease with a pH electrode and analysis of L-ascorbic acid by ascorbate oxidase.f ... 

Meier, H. Tran Minh, C. Determination of penicillin-V in standard solution and in fermentation broth by flow-injection analysis using fast-responding enzyme glass electrodes in different detection cells. Anal. Chim. Acta 1992,264, 13-22. 

Potentiometric penicillin sensors are mainly based on glass electrodes working with free enzyme or with enzyme included in an electrode cover. In the first penicillin electrode, (5-lactamase was directly photopolymer-ized in acrylamide on the electrode (Papariello et al., 1973). Enfors and Nilsson (1979) designed a sterilizable penicillin sensor where a solution containing (5-lactamase was pumped into a reaction chamber in front of a flat glass electrode, after sterilization of the electrode (Fig. 75). 

Several methods for enzyme immobilization can be found in literature. In our laboratory, we have developed a new enzyme immobilization technique making possible response time of the biosensor much inferior to any of the response times so far reported for a penicillin sensor the combined pH electrode to be coated with the enzyme was left... 

Many potentiometric en2yme electrodes for widely different analyte species can be prepared merely by choosing the appropriate ISE transducer and immobilized enzymatic reagent. Table 5 summarizes the enzymes and ISEs/gas sensors used to construct biosensors for a number of important biomolecules, ranging from urea and creatinine to amino acids, nucleotides, and even glucose and penicillin. In the... 

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