Surface biocatalysis

The protein-containing colloidal solutions of water-in-organic solvents are optically transparent. Hence, absorption spectroscopy, circular dichroism spectroscopy and fluorescence spectroscopy are found to be convenient for studying biocatalysis [53]. The reversed micelles are interesting models for studying bioconversion, since the majority of the enzymes in vivo act inside or on the surface of biological membranes. 

Because enzymes are insoluble in organic solvent, mass-transfer limitations apply as with any heterogeneous catalyst. Water-soluble enzymes (which represent the majority of enzymes currently used in biocatalysis) have hydrophilic surfaces and so tend to form aggregates or stick to reaction vessel walls rather than form the fine dispersions that are required for optimum efficiency. This can be overcome by enzyme immobilization, as discussed in Section 1.5. 

Bioreactions. The use of supercritical fluids, and in particular C02, as a reaction media for enzymatic catalysis is growing. High diffusivities, low surface tensions, solubility control, low toxicity, and minimal problems with solvent residues all make SCFs attractive. In addition, other advantages for using enzymes in SCFs instead of water include reactions where water is a product, which can be driven to completion increased solubilities of hydrophobic materials increased biomolecular thermostability and the potential to integrate both the reaction and separation bioprocesses into one step (98). There have been a number of biocatalysis reactions in SCFs reported (99—101). The use of lipases shows perhaps the most commercial promise, but there are a number of issues remaining unresolved, such as solvent—enzyme interactions and the influence of the reaction environment. A potential area for increased research is the synthesis of monodisperse biopolymers in supercritical fluids (102). 

Before any catalysis can occur, at least one of the substrates must coordinate to the catalyst. This means that the catalyst must have a vacant active site. In homogeneous metal complex catalysis and biocatalysis, this will be a vacant coordination site at the metal atom. In heterogeneous catalysis, the vacant site could be a metal crystallite or an ion on the surface. For the latter, we speak of desorption and adsorption instead of dissociation and coordination. Remember that our reactions are not in vacuum, so there is no vacant site . Thus, before any chemical species can coordinate to the metal complex (or to the active site in heterogeneous catalysis or biocatalysis) the species already occupying this space must first vacate it. This happens constantly, as the system is dynamic (Figure 3.3) [15]. At any given moment... 

In some cases, substrates and enzymes are not soluble in the same solvent. To achieve efficient substrate conversion, a large interface between the immiscible fluids has to be established, by the formation of microemulsions or multiple-phase flow that can be conveniently obtained in microfluidic devices. Until now only a couple of examples are published in which a two-phase flow is used for biocatalysis. Goto and coworkers [431] were first to study an enzymatic reaction in a two-phase flow in a microfluidic device, in which the oxidation ofp-chlorophenol by the enzyme laccase (lignin peroxidase) was analyzed (Scheme 4.106). The surface-active enzyme was solubilized in a succinic acid aqueous buffer and the substrate (p-chlorophenol) was dissolved in isooctane. The transformation ofp-chlorophenol occurred mainly at... 

Fischer-Colbrie, G., Herrmaim, M., Henmann, S., Pnolakka, A., Wirth, A., Cavaco-Panlo, A., and Gnebitz, G.M. 2006. Surface modification of polyacrylonitrile with nitrile hydratase and amidase from Agrobacterium tumefaciens. Biocatalysis and Biotransformation, 24 419-25. 

Here, we will report on the design, synthesis, characterization, and applications of template-synthesized nanotube membranes. Then, we will briefly review the synthesis of the template-synthesized nanotube membranes. Some details of differential-surface chemistry on nanombes, and nanombes for bioextraction and biocatalysis are presented. We discuss in detail the drug detoxification using functionalized nanotubes [2], and epoenzyme-, enzyme- and antibody-immobilized nanotubes for enantiomeric separations, biocatalysis, and bioextractions [3-5]. We also describe our recent results on DNA-functionalized nanombe membranes with single-nucleotide mismatch selectivity [6], and the fabrication of artificial ion-channel using single-conical nanombe membrane [7]. 

Adsorption on a solid catalyst surface, complex formation in homogeneous catalysis with metallo-organic complexes and in biocatalysis with enzymes share the same principle, i.e. the total number of sites is constant. Therefore, the rate expressions for reactions on heterogeneous, homogeneous and biocatalysts have a similar form. The constant number of active sites results in rate expressions that differ from homogeneous gas phase kinetics. Partial pressures are usually used in rate expressions for gas-phase reactions, while concentrations are used when the reactions take place in the liquid phase. It appears that definitions and nomenclature of particular kinetics constants in the different sub-communities differ sometimes. In the following sections the expressions used by the different subdisciplines will be compared and their conceptual basis outlined. 

In biocatalysis /C2 is the rate measured when all enzyme molecules are complexed with reactant divided by the total concentration of enz)one present. This is the Turn-Over Number according to biochemists definition. Note that this differs from the Turn-Over Frequency as defined in heterogeneous catalysis where it is simply the rate normalised to the total number of surface sites present. In the latter case it is a function of the gas phase composition. 

The form of the resulting expression differs from the gas-phase reaction rate expressions due to the presence of a denominator representing the reduction in rate due to adsorption phenomena. The individual terms of this denominator respresent the distribution of the active sites among the possible surface complexes and vacancies. Expressions of this type are termed the Langmuir-Hinshel-wood-Hougen-Watson (LHHW) rate expressions in heterogeneous catalysis and Michaelis-Menten expressions in biocatalysis. 

Miniaturization in biocatalysis and fermentation is another necessary step. This will allow continuous processes with the benefits that could derive in terms of process intensification and reduction of waste. Miniature (less than 10 mL) stirred reactors and microtiter plates (MTP) have been introduced mainly with the idea of allowing high-throughput screening to speed up bioprocess development, even though they are available now also for production uses [172-174]. Notably, problems emerge with these miniature bioreactors (MBRs), such as evaporation and surface tension, which determine the performances, but which are masked in larger bioreactors. 

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