Biocatalytic

In a biocatalytic biosensor the molecular recognition component is an enzyme. Enzymes, macromolecular catalysts that are manufactured by plants and animals, affect the rates of biochemical reactions. Virtually all of the millions of chemical reactions involved in Hfe processes have associated enzymes controlling the rates. CoUectively, there are several thousand enzymes known and perhaps many thousand more yet to be discovered. 

Fig. 2. Schematic of an electrochemical biocatalytic biosensor showing enzyme E immobilized onto the electrode where S is the substrate (target) and P is...

The simple cases where one enzyme is employed afford a limited scope of potential targets. Usually two or more enzyme reactions are coupled, as exemplified by the development of a piezoelectricaHy-transduced biocatalytic biosensor that couples two enzyme reactions to detect glucose [492-62-6] ... 

Fig. 3. Detail of enzyme-modified quartz radio crystal used in a piezoelectric biocatalytic biosensor.

Several other biosensors have been developed usiag this oxygen-quenched fluorescence approach. Target species iaclude ethanol [64-17-5] hydrogen peroxide [7722-84-17, H2O2, lactate, and xanthine [69-89-6] C H4N402, usiag alcohol oxidase, catalase [9001-05-2] lactate oxidase, and xanthine oxidase, respectively. An additional technique for biocatalytic biosensors iavolves the firefly chemiluminescent reaction (17) ... 

Biocatalytic ledox reactions offer great synthetic utility to organic chemists. The majority of oxidase-catalyzed preparative bioconversions are still performed using a whole-ceU technique, despite the fact that the presence of more than one oxidoreductase in cells often leads to product degradation and lower selectivity. Fortunately, several efficient cofactor regeneration systems have been developed (160), making some cell-free enzymatic bioconversions economically feasible (161,162). 

It is apparent that the use of enzymatic catalysis continues to grow Greater availabiUty of enzymes, development of new methodologies for thek utilization, investigation of enzymatic behavior in nonconventional environments, and the design and synthesis of new biocatalysts with altered selectivity and increased stabiUty are essential for the successhil development of this field. As more is learned about selectivity of enzymes toward unnatural substrates, the choice of an enzyme for a particular transformation will become easier to predict. It should simplify a search for an appropriate catalyst and help to estabhsh biocatalytic procedures as a usehil supplement to classical organic synthesis. 

In this chapter, we try to summarize the work so far reported in this field. We first give a short introduction into the different forms of biocatalytic reactions, highlighting some special properties of biocatalysts. 

Generally, there are three ways to use organic solvents or ionic liquids in a biocatalytic process ... 

Thanks to their special properties and potential advantages, ionic liquids may be interesting solvents for biocatalytic reactions to solve some of the problems discussed above. After initial trials more than 15 years ago, in which ethylammonium nitrate was used in salt/water mixtures [29], results from the use of ionic liquids as pure solvent, as co-solvent, or for biphasic systems have recently been reported. The reaction systems are summarized in Tables 8.3-1 and 8.3-2, below. Table 8.3-1 compiles all biocatalytic systems except lipases, which are shown separately in 8.3-2. Some of the entries are discussed in more detail below. 

Biotechnology has attracted enormous interest and high expectations over the past decade. However, the implementation of new technologies into industrial processes has been slower than initially predicted. Although biocatalytic methods hold great industrial potential, there are relatively few commercial applications of biocatalysts in organic chemical synthesis. The main factors that limit the application of biocatalysts are ... 

Once there is an appreciable amount of cells and they are growing very rapidly, the cell number exponentially increases. The optical cell density of a culture can then be easily detected that phase is known as the exponential growth phase. The rate of cell synthesis sharply increases the linear increase is shown in the semi-log graph with a constant slope representing a constant rate of cell population. At this stage carbon sources are utilised and products are formed. Finally, rapid utilisation of substrate and accumulation of products may lead to stationary phase where the cell density remains constant. In this phase, cell may start to die as the cell growth rate balances the death rate. It is well known that the biocatalytic activities of the cell may gradually decrease as they age, and finally autolysis may take place. The dead cells and cell metabolites in the fermentation broth may create... 

Willner, I and Willner, B. Artifical Photosynthetic Model Systems Using Light-Induced Electron Transfer Reactions in Catalytic and Biocatalytic Assemblies. 159, 153-218... 

FIGURE 6-1 Enzyme electrode based on a biocatalytic layer immobilized on an electrode transducer. 

Enzyme electrodes for other substrates of analytical significance have been developed. Representative examples are listed in Table 6-1. Further advances in enzyme technology, and particularly the isolation of new and more stable enzymes, should enhance the development of new biocatalytic sensors. New opportunities (particularly assays of new environments or monitoring of hydrophobic analytes) derive from the finding that enzymes can maintain then biocatalytic activity in organic solvents (31,32). 

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