Alcohol dehydrogenase immobilization

Choi J.W., Bae J.Y., Min J., Cho K.S., Lee W.H., Fiber-optic ethanol sensor using alcohol dehydrogenase-immobilized Langmuir-Blodgett film, Sensor Mater. 1996 8 493-504. 

Addo et al. (2011) investigated a rechargeable alcohol biobattery. The anode compartment of this battery consisted of NAD-dependent alcohol dehydrogenase immobilized into a carbon composite paste. Ferrocene was added to the electrolyte to shuttle electrons to and from the electrode surface. In discharge mode the bioanode catalyzed the oxidation of ethanol to acetaldehyde. The cathode... 

There have been a number of reports of electrocatalysis of alcohol oxidation using immobilized PQQ-dependent alcohol dehydrogenases or flavin-containing alcohol dehydrogenases or oxidases with dissolved mediators in solution. Co-immobihzing the mediator with the enzyme is advantageous, as set out in Section 17.1, and several such strategies have been employed for electrocatalytic alcohol oxidation. 

Alcohol oxidase was used to generate H202 followed by its reaction with luminol in the presence of K3[Fe(CN)6] as a catalyst [53], The luminescence was transmitted from the flow cell to the detector via optical fibers. Ethanol can be determined in the 3-750-pmol/L concentration range, with a detection limit of 3 pmol/L. Also, using an immobilized alcohol dehydrogenase reactor in glass beads, a FIA sensor for a reduced form of NADH was constructed by the ECL using the above-mentioned ruthenium tris(2,2 -biryridine) complex. The sensor was satisfactorily applied to the determination of ethanol concentration [54],... 

Benzyl alcohol can be produced from benzaldehyde (S) by a dehydrogenation reaction catalyzed by yeast alcohol dehydrogenase (YADH). Nikolova et al. (1995) obtained initial-rate data for this reaction using immobilized YADH immersed in iso-octane with 1% v/v water. The following data were obtained ... 

The amperometric dehydrogenase sensor for ethanol consists of a platinum electrode on the surface of which alcohol dehydrogenase (ADH), Meldra blue (MB) and NAD are immobilized with a conductive polypyrrole membrane as schematically illustrated in Fig.24. 

K. F. Gu and T. M. S. Chang, Conversion of ammonia or urea into essential amino acids, using artificial cells containing an immobilized multienzyme system and dextran-NAD+ 2. Yeast alcohol dehydrogenase for coenzyme recycling, Biotechnol. Appl. Biochem., 12, 227-236 (1990). 

All the enzymes used in the work described above are quite stable at room temperature and can be used in a free form. They can also be used in an immobilized form to improve the stability and to facilitate the recovery. Many immobilization techniques are available today (25). The recent procedure developed by Whitesides et al using water-insoluble, cross-linked poly(aerylamide-acryloxysuccinimide) appears to be very useful and applicable to many enzymes (37). We have found that the non-crosslinked polymer can be used directly for immobilization in the absence of the diamine cross-linking reagent. Reaction of an enzyme with the reactive polymer produces an immobilized enzyme which is soluble in aqueous solutions but insoluble in organic solvents. Many enzymes have been immobilized by this way and the stability of each enzyme is enhanced by a factor of greater than 100. Horse liver alcohol dehydrogenase and FDP aldolase, for example, have been successfully immobilized and showed a marked increase in stability. 

The activity of immobilized alcohol dehydrogenase was probed and modified by changing the local pH [133], The tip was positioned close to the enzyme film and used to increase the local pH by reducing water and producing hydroxide ions. A significant increase in enzyme activity was observed at higher pH. The opposite effect, that is, local inactivation of the immobilized enzyme (diaphorase) by chlorine or bromine species electrogenerated at the tip, also has been reported [134]. 

Immobilied Cibacron Blue 3G-A has been found very efficient for the isolation of human serum albumin interferons produced in tissue cultures phosphogly-cerate kinase from Saccharomyces cerevisiae and alcohol dehydrogenase from equine liver. Immobilized Procion Red HE-3B and Prcxaon Blue MX-4GD have been successfully applied for the isolation of 3-hydroxybutyrate and malate dehydrogenases... 

Formate dehydrogenase in conjunction with polyethyleneglycol-immobilized nicotinamide adenine dinudeotide has been used to good effect as a cofactor recycle system (39). The alcohol dehydrogenase from Thermoanaerobium hrockii catalyzed the reduction of ketones independently when driven by the cooxidation of isopropanol (40,41). 

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