Three-enzyme process

A two-step three-enzyme process for atorvastatin intermediate. 

An example of three-enzyme process is given in Fig. 8.20. The biotransformation of D-glucose to fructose-1,6-diphosphate with glucose-6-phosphate and fructose-6-phosphate as intermediate products involves, respectively, the following enzymes glu-cokinase, phosphoglucoisomerase, and phosphofructokinase. 

Figure 2.13 Reactions and enzymes involved in the production of L-amino acids from racemic hydantoins by the three-enzyme hydantoinase process [55],...

In many cases, the racemization of a substrate required for DKR is difficult As an example, the production of optically pure cc-amino acids, which are used as intermediates for pharmaceuticals, cosmetics, and as chiral synfhons in organic chemistry [31], may be discussed. One of the important methods of the synthesis of amino acids is the hydrolysis of the appropriate hydantoins. Racemic 5-substituted hydantoins 15 are easily available from aldehydes using a commonly known synthetic procedure (Scheme 5.10) [32]. In the next step, they are enantioselectively hydrolyzed by d- or L-specific hydantoinase and the resulting N-carbamoyl amino acids 16 are hydrolyzed to optically pure a-amino acid 17 by other enzymes, namely, L- or D-specific carbamoylase. This process was introduced in the 1970s for the production of L-amino acids 17 [33]. For many substrates, the racemization process is too slow and in order to increase its rate enzymes called racemases are used. In processes the three enzymes, racemase, hydantoinase, and carbamoylase, can be used simultaneously this enables the production of a-amino acids without isolation of intermediates and increases the yield and productivity. Unfortunately, the commercial application of this process is limited because it is based on L-selective hydantoin-hydrolyzing enzymes [34, 35]. For production of D-amino acid the enzymes of opposite stereoselectivity are required. A recent study indicates that the inversion of enantioselectivity of hydantoinase, the key enzyme in the... 

Hematological Effects. The effects of lead on the hematopoietic system have been well documented. These effects, which are seen in both humans and animals, include increased urinary porphyrins, coproporphyrins, ALA, EP, FEP, ZPP, and anemia. The process of heme biosynthesis is outlined in Figure 2-10. Lead interferes with heme biosynthesis by altering the activity of three enzymes ALAS,... 

Although the present situation and the way ahead appear uncertain, it is clear that enzyme treatment alone does not fulfil the technical requirements for shrink-resist finishing. Even with enzyme treatment, some degree of chlorination (with the attendant AOX problems) and/or application of a resin will still be required. Two-stage or even three-stage processes have been proposed [116] ... 

The immobilization of the white rot fungus F. trogii in Na-ALG beads allowed the decolorization of the dye Acid Black 52 in a stirred tank reactor operated in batch [55]. Three enzymes, laccase, MnP, LiP, secreted by fungus were reported during decolorization process. Results showed that laccase enzyme activity increased with increasing alginate concentration from 0 to 4%. Cell growth at immobilized cultivation was maintained more stably than suspended cultivation. Total amount of removed dye was reported to be 469 mg/L for immobilized cultures and 440 mg/L for suspended cultures. 

Chymotrypsin catalysis takes place through a three-step process, equation (11), where ES is an enzyme substrate complex which breaks down to give an acylated enzyme intermediate, ES and Pj,... 

An elegant four-enzyme cascade process was described by Nakajima et al. [28] for the deracemization of an a-amino acid (Scheme 6.13). It involved amine oxidase-catalyzed, (i )-selective oxidation of the amino acid to afford the ammonium salt of the a-keto acid and the unreacted (S)-enantiomer of the substrate. The keto acid then undergoes reductive amination, catalyzed by leucine dehydrogenase, to afford the (S)-amino acid. NADH cofactor regeneration is achieved with formate/FDH. The overall process affords the (S)-enantiomer in 95% yield and 99% e.e. from racemic starting material, formate and molecular oxygen, and the help of three enzymes in concert. A fourth enzyme, catalase, is added to decompose the hydrogen peroxide formed in the first step which otherwise would have a detrimental effect on the enzymes. 

Pyridoxal phosphate is the coenzyme for the enzymic processes of transamination, racemization and decarboxylation of amino-acids, and for several other processes, such as the dehydration of serine and the synthesis of tryptophan that involve amino-acids (Braunstein, 1960). Pyridoxal itself is one of the three active forms of vitamin B6 (Rosenberg, 1945), and its biochemistry was established by 1939, in considerable part by the work of A. E. Braunstein and coworkers in Moscow (Braunstein and Kritzmann, 1947a,b,c Konikova et al 1947). Further, the requirement for the coenzyme by many of the enzymes of amino-acid metabolism had been confirmed by 1945. In addition, at that time, E. E. Snell demonstrated a model reaction (1) for transamination between pyridoxal [1] and glutamic acid, work which certainly carried with it the implication of mechanism (Snell, 1945). 

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