Soluble enzyme batch membrane

Figures 7.21 and 7.22 show typical results for the two cases-with enzymatic gel layer formation and when soluble enzymes are confined only near the membrane surface. Comprehensive models for an immobilized enzyme batch membrane reactor (IEMR) and for a soluble enzyme batch membrane reactor (SEMR) are proposed in References 33 and 30, respectively, for a flat slab membrane configuration.

A plug flow reactor may be realized using immobilized enzymes within a column reactor or using soluble enzymes within a cascade of membrane reactors. A batch or a repetitive batch process with soluble enzymes (see below) has the same productivity as the plug flow reactor. 

Glucuronides have been synthesized batch-wise or in a hollow fiber system using microsomal or soluble enzyme preparations (5-5). Furthermore, they have been prepared with enzymes immobilized to polymeric supports (6). Here we describe the continuous synthesis of glucuronide conjugates in a 10-mL membrane reactor (7). 

A first application using ferroceneboronic acid as mediator [45] was described for the transformation of p-hydroxy toluene to p-hydroxy benzaldehyde which is catalyzed by the enzyme p-cresolmethyl hydroxylase (PCMH) from Pseudomonas putida. This enzyme is a flavocytochrome containing two FAD and two cytochrome c prosthetic groups. To develop a continuous process using ultrafiltration membranes to retain the enzyme and the mediator, water soluble polymer-bound ferrocenes [50] such as compounds 3-7 have been applied as redox catalysts for the application in batch electrolyses (Fig. 12) or in combination with an electrochemical enzyme membrane reactor (Fig. 13) [46, 50] with excellent results. 

Since most enzymes are globular protein, they are soluble in water. Therefore, it is very difficult or impractical to separate the enzyme for reuse in a batch process. Enzymes can be immobilized on the surface of or inside of an insoluble matrix either by chemical or physical methods. They can be also immobilized in their soluble forms by retaining them ivith a semipermeable membrane. 

Another favorable aspect of stirred batch reactors is the fact that they are compatible with most forms of a biocatalyst. The biocatalyst may be soluble, immobilized, or a whole-cell preparation in the latter case a bioconversion might be performed in the same vessel used to culture the organism. Recovery of the biocatalyst is sometimes possible, typically when the enzyme is immobilized or confined within a semi-permeable membrane. The latter configuration is often referred to as a membrane reactor. An example is the hollow fiber reactor where enzymes or whole cells are partitioned within permeable fibers that allow the passage of substrates and products but retain the catalyst. A hollow-fiber reactor can be operated in conjunction with the stirred tank and operated in batch or... 

The mechanisms of CO2 toxicity at near-atmospheric pressures are amplified at the near-critical and supercritical pressures used to achieve sterilization. For example, Spilimbergo et al. (9) examined the mechanism of inactivation of Pseudomonas aeruginosa and B. subtilis at 38-54°C and 5.8-20 MPa in a batch process. Total inactivation of the bacteria was observed when exposed to SCF CO2 at 38°C and 7.4 MPa for 150 s. The mechanism of inactivation is suggested to involve (a) the diffusion of SCF CO2 into the cells leading to a drop in pH and a subsequent loss of activity of key enzymes and (b) the extraction by SCF CO2 of intracellular substances, including phospholipids. Further analysis determined a high solubility of CO2 in model cell membrane phospholipids, suggesting that the enhanced permeability of the membrane in the presence of CO2 contributed to the inactivation of the cells. 

The catalytic behavior of enzymes in immobilized form may dramatically differ from that of soluble homogeneous enzymes. In particular, mass transport effects (the transport of a substrate to the catalyst and diffusion of reaction products away from the catalyst matrix) may result in the reduction of the overall activity. Mass transport effects are usually divided into two categories - external and internal. External effects stem from the fact that substrates must be transported from the bulk solution to the surface of an immobilized enzyme. Internal diffusional limitations occur when a substrate penetrates inside the immobilized enzyme particle, such as porous carriers, polymeric microspheres, membranes, etc. The classical treatment of mass transfer in heterogeneous catalysis has been successfully applied to immobilized enzymes I27l There are several simple experimental criteria or tests that allow one to determine whether a reaction is limited by external diffusion. For example, if a reaction is completely limited by external diffusion, the rate of the process should not depend on pH or enzyme concentration. At the same time the rate of reaction will depend on the stirring in the batch reactor or on the flow rate of a substrate in the column reactor. 

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