Chemoenzymatic oxidizing enzymes

Chemoenzymatic processes involving oxidizing enzymes have been reported particularly for specific chemical syntheses. For example, industrially important amino acids can be deracemized by exploiting the enantioselectivity of amino acid oxidases a commercial process has recently been developed in which efficient... 

The asymmetric hydrolysis of (exo,exo)-7-oxabicyclo[2.2.1]heptane-2,3-dimethanol, diacetate ester (37) to the corresponding chiral monoacetate ester (38) (Fig. 12B) has been demonstrated with lipases [61]. Lipase PS-30 from P. cepacia was most effective in asymmetric hydrolysis to obtain the desired enantiomer of monoacetate ester. The reaction yield of 75 M% and e.e. of >99% were obtained when the reaction was conducted in a biphasic system with 10% toluene at 5 g/liter of the substrate. Lipase PS-30 was immobilized on Accurel PP and the immobilized enzyme was reused (5 cycles) without loss of enzyme activity, productivity, or e.e. of product (38). The reaction process was scaled up to 80 liters (400 g of substrate) and monoacetate ester (38) was isolated in 80 M% yield with 99.3% e.e. The product was isolated in 99.5% chemical purity. The chiral monoacetate ester (38) was oxidized to its corresponding aldehyde and subsequently hydrolyzed to give chiral lactol (33) (Fig. 12B). The chiral lactol (33) obtained by this enzymatic process was used in chemoenzymatic synthesis of thromboxane A2 antagonist (35). 

As a part of ongoing efforts to synthesize a potent, orally active anti-platelet agent, xemilofiban 1 [1], development of an efficient chemoenzymatic process for 2, the chiral yS-amino acid ester synthon (Fig. 1) was proposed. The scheme emphasized the creation of the stereogenic center as the key step. In parallel with the enzymatic approach, chemical synthesis of the / -amino acid ester synthon emphasized formation of a chiral imine, nucleophilic addition of the Reformatsky reagent, and oxidative removal of the chiral auxiliary. This chapter describes a selective amida-tion/amide hydrolysis using the enzyme Penicillin G amidohydrolase from E. coli to synthesize (R)- and (S)-enantiomers of ethyl 3-amino-5-(trimethylsilyl)-4-pen-tynoate in an optically pure form. The design of the experimental approach was applied in order to optimize the critical reaction parameters to control the stereoselectivity of the enzyme Penicillin G amidohydrolase. 

Still, there remain many open problems. It would be efficient to be able to prenylate or ferf-prenylate indole regioselectively at the benzene positions 4, 5 and 6 without having to rely on pre-functionalisation such as halogenation or hydroxylation. Here, deeper investigation of prenyl shifts and of CH functionalisation on indole is required. Enantioselective catalysis has to be explored further towards the synthesis of optically pure 3-prenylated or -tert-prenylated alkaloids. A chiral version of NBS would be helpful. In the case of conformationally flexible starting materials, the diastereoselectivity of oxidative cyclisations of tryptophan-derived diketopiperazines is still not convincing. In the area of chemoenzymatic synthesis, the number and availability of enzymes has to be enhanced and their substrate tolerance has to be elucidated in more detail. 

An example of this technique is shown in Scheme 5.57, where a chemoenzymatic enantiomerization occurs by the involvement of enantiomer-selective oxidation and non-selective hydride reduction [147]. It has been shown that one enantiomer of a racemic sec-amine can be enantiospecifically oxidized by an enzyme, giving an achiral intermediary imine. This is simultaneously reduced to the racemic starting amine in a non-selective manner. Repetition of the sequence leads to an overall chiral inversion of the faster-reacting enantiomer to the slower reacting enantiomer to give the final product in 100% theoretical chemical and optical yields. 

A further exciting development in the field of chemoenzymatic one-pot synthesis is the integration of artificial metalloproteins (which can then be regarded as the chemocatalytic component ) in such processes. Such a concept was successfully realized by HoUmann, Turner, and Ward et al. in the combination of an artificial transfer hydrogenase with various redox biocatalysts, comprising NADH-, FAD, and heme-dependent enzymes [47]. A selected example is shown in Scheme 19.18. Therein, readily available L-lysine is oxidized by an L-amino acid oxidase toward Al-piperidine<arboxylic acid (52), which is then reduced by the iridium complex-containing metalloprotein to racemic pipecohc acid (rac-53). 

A chemoenzymatic way to produce poly(hydroquinone) was achieved by enzymatic oxidative polymerization of 4-hydroxyphenyl benzoate, followed by alkahne hydrolysis of the resulting polymer [45]. HRP and SBP were used as enzymes. The molecular weight of the resulting poly(4-hydroxyphenyl benzoate) varied between 1100 and 2400 g/mol. The structure was said to consist of phenylene and oxyphenylene moieties, which was found by IR analysis and titration of the residual amount of phenolic groups in the polymer. Other phenol polymers have shown their potential for electronic applications as well. Besides hydroquinone, catechol has also been used as substrate for peroxidase-catalyzed polymerization. The molecular weights of the reac-... 

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