Glutamine sensor

While selectivity in certain instances has been excellent (i.e., the above glutamine sensor), most tissue and bacterial electrodes are plagued by simultaneous response to several biochemical species. This is because there are often several enzymes present in a given cell which liberate the electrode-detectable product. Consequently, in situ type measurements with such... 

The typical behavior of a potentiometric based enzyme sensors is shown in Fig. 10 adopted from Rechnitz. In this design of a glutamine sensor, an organism was chosen that was particularly rich in glutamic deaminase. The reaction is... 

Blankenstein G., Preuschoff F., Spohn U., Mohr K.H., Kula M.R., Determination of L-glutamate and L-glutamine by flow-injection analysis and chemiluminescence detection comparison of an enzyme column and enzyme membrane sensor, Anal. Chim. Acta 1993 271 231-237. 

Iida T., Kawabe T., Noguchi F., Mitamura T., Nagata K., Tomita K., ISFET-type -glutamate sensor using thermophilic glutamine-synthetase from a thermophile, Nippon-Kagaku-Kaishi 1987 10 1817-1821. 

Figure 5 Five-channel enzyme sensor for the simultaneous determination of glucose, lactate, glutamate, glutamine, and ammonium. MFM, microfiltration module WV, valves P, pumps DC, dialysis cell B, blank reactors MC, reactor D, biosensor flow cell. (Adapted with permission from Ref. 34.)...

Bacterial electrodes [11, 31, 33, 46, 48, 49, 60] In this type of electrode, a suspension of suitable bacteria is placed between the sensor proper and a dialysis membrane that prevents passage of high-molecular substances (see fig. 8.3). The sensor is usually a gas probe. In the simple types of bacterial electrode, the determinand is converted by a suitable strain of bacteria into a product sensed by the gas probe. Thus it is possible to determine arginine [46], glutamine [48],/.-aspartic acid [31],/.-histidine [60] and nitrate [33]. Hybrid bacterial - enzyme electrodes contain both a bacterial strain and a suitable enzyme. For example, an extract from ivingas Neurospora chossa can be used as a source of NAD nucleosidase and an Escherichia coli culture as a source of nicotinamide deaminase, so that the electrode responds to NAD [49] as a result of the series of reactions... 

Tissue electrodes [2, 3, 4, 5, 45,57], In these biosensors, a thin layer of tissue is attached to the internal sensor. The enzymic reactions taking place in the tissue liberate products sensed by the internal sensor. In the glutamine electrode [5, 45], a thick layer (about 0.05 mm) of porcine liver is used and in the adenosine-5 -monophosphate electrode [4], a layer of rabbit muscle tissue. In both cases, the ammonia gas probe is the indicator electrode. Various types of enzyme, bacterial and tissue electrodes were compared [2]. In an adenosine electrode a mixture of cells obtained from the outer (mucosal) side of a mouse small intestine was used [3j. The stability of all these electrodes increases in the presence of sodium azide in the solution that prevents bacterial decomposition of the tissue. In an electrode specific for the antidiuretic hormone [57], toad bladder is placed over the membrane of a sodium-sensitive glass electrode. In the presence of the antidiuretic hormone, sodium ions are transported through the bladder and the sodium electrode response depends on the hormone concentration. 

Fe/S clusters in regulatory enzymes have been proposed to act as sensors in such a manner that, upon detection of a measurand, the cluster disintegrates and activity stops. Putative examples are NO sensing by the [2Fe-2S] cluster in the terminal enzyme of heme synthesis, ferrochelatase [8], and 02 sensing by the [4Fe-4S] cluster in the regulatory enzyme of purine nucleotide biosynthesis, glutamine 5-phosphoribosyl-l-pyrophosphate amidotransferase [9], This is of course not a catalytic activity, since the cluster is destroyed in the action. 

Biosensors. Sensors are required to adequately monitor bioreactor performance. Ideally, one would like to have online sensors to minimize the number of samples to be taken from the bioreactor and to automate the bioreactor process. Most bioreactors have autoclavable pH and dissolved oxygen (D.O.) electrodes as online sensors, and use offline detectors to measure other critical parameters such as glucose and glutamine concentration, cell density, and carbon dioxide partial pressure (pC02). An online fiber-optic-based pC02 sensor is commercially available and appears to be robust.37 Probes are also commercially available that determine viable cell density by measuring the capacitance of a cell suspension. Data from perfusion and batch cultures indicate that these probes are reasonably accurate at cell concentrations greater than 0.5 X 106 cells/mL.38,39... 

Substitution of other oxoreductase enzymes for glucose oxidase allows amperometric biosensors for other substrates of clinical interest to be constructed. Practical sensors with commercial application in critical care analyzers for blood lactate have been realized. Other amperometric biosensors reported include cholesterol, pyruvate, alanine, glutamate, and glutamine. By using the multiple enzyme cascade shown in the reactions below, an amperometric biosensor for creatinine is also possible. Electrochemical oxidation of H2O2 is the detection mechanism. 

Decarboxylases of phenylalanine, tyrosine, and lysine and ammonia lyases of histidine, glutamine, and asparagine are also highly selective. Guilbault et al. (1988) described a potentiometric enzyme sensor for the determination of the artificial sweetener aspartame (L-aspartyl-L-phen-ylalanine methylester) based on L-aspartase (EC 4.3.1.1). The ammonia liberated in the enzyme reaction created a slope of 30 mV/decade for the enzyme-covered ammonia sensitive electrode. The specificity of the sensor was excellent however, the measuring time of 40 min per sample appears not to be acceptable. The measuring time has been decreased to about 20 min by coimmobilizing carboxypeptidase A with L-aspartase (Fatibello-Filho et al., 1988). 

Lei Y, Chen W, Mulchandani A (2006) Microbial biosensors. Anal Chim Acta 568(l-2) 200-220 Ligler FS (2006) Biosensors for detection of bioterrorist threats. In Baldini F et al (eds) Optical chemical sensors NATO science series II, vol 224. Springer, Berlin, pp 437 55 Moser I et al (1995) Miniaturized thin film glutamate and glutamine biosensors. Biosens Bioelectron 10(6-7) 527-532... 

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