Enzymes chirality

Catalysts and enzymes Chiral synthesis BINAP" Novozym 388 2,2 -Bis (diphenylphosphino) l,l -binaphthyl 1,3-Specific lipase Rhodia, France Solvias, Switz. Novozymes, Denmark... 

The use of various catalysts in the asymmetric addition of HCN (or an equivalent reagent) to achiral aldehydes and ketones, giving optically active cyanohydrins has been discussed. The catalysts include enzymes, chiral polymers, boron, and chiral titanium complexes.1908... 

We applied a similar approach under kinetic resolution conditions (Figure 5) (19). First a mixture of 2 and racemic 4-methylcaprolactone (4-MeCL) was reacted until all (S)-4-MeCL had been converted, i.e., to about 50 % total monomer conversion. Subsequently, oxygen was removed from the reaction medium by several consecutive freeze-pump-thaw cycles. The ATRP was initiated by adding MMA and Ni(PPh3)2Br2 and raising the reaction temperature to 80 C. In this case the nickel complex acts as both ATRP initiator and enzyme inhibitor thereby preventing any side reactions caused by the enzyme. Chiral block copolymers were obtained by this approach as evident from SEC analysis. 

As an interesting illustration of this enzyme chirality principle, Stephen B. Kent, then at The Scripps Research Institute in La JoUa, CaMomia, synthesized the enantiomer of a natural enzyme by using the enantiomers of the amino acids normally found in proteins as building blocks. As expected, this synthetic enzyme enantiomer only catalyzed a reaction with a substrate that was the enantiomer of the chiral substrate utilized by the natural enzyme. 

In addition to the homogeneous and heterogeneous TM catalysts discussed above, other catalysts/methods such as uano catalysts, carbon materials, enzymes, chiral catalysts and continuous flow techuiques have also been developed successfully and applied in /-alkylation reactions of amines/amides with alcohols. 

The method of analysis developed is for acetic acid and involves the use of two enzymes. Chiral acetic acid [as 1.26)] is irreversibly condensed as its CoA-derivative with glyoxylic acid (/.27), using the enzyme malate synthase, to give malic acid 1.28). The condensation occurs with loss of hydrogen isotope by a primary kinetic isotope effect (A h kj). This means that loss of H is favoured over loss of D which is in turn favoured over loss of T. The result is a high retention of tritium. 

This example illustrates a subtle control of a chemical reaction by a delicate manipulation of tire stereochemical environment around a metal centre dictated by tire selection of tire ligands. This example hints at tire subtlety of nature s catalysts, tire enzymes, which are also typically stereochemically selective. Chiral catalysis is important in biology and in tire manufacture of chemicals to regulate biological functions, i.e., phannaceuticals. 

Application of the CCM to small sets (n < 6) of enzyme inhibitors revealed correlations between the inhibitory activity and the chirality measure of the inhibitors, calculated by Eq. (26) for the entire structure or for the substructure that interacts with the enzyme (pharmacophore) [41], This was done for arylammonium inhibitors of trypsin, Di-dopamine receptor inhibitors, and organophosphate inhibitors of trypsin, acetylcholine esterase, and butyrylcholine esterase. Because the CCM values are equal for opposite enantiomers, the method had to be applied separately to the two families of enantiomers (R- and S-enantiomers). 

Open-chain 1,5-polyenes (e.g. squalene) and some oxygenated derivatives are the biochemical precursors of cyclic terpenoids (e.g. steroids, carotenoids). The enzymic cyclization of squalene 2,3-oxide, which has one chiral carbon atom, to produce lanosterol introduces seven chiral centres in one totally stereoselective reaction. As a result, organic chemists have tried to ascertain, whether squalene or related olefinic systems could be induced to undergo similar stereoselective cyclizations in the absence of enzymes (W.S. Johnson, 1968, 1976). 

Optically inactive starting materials can give optically active products only if they are treated with an optically active reagent or if the reaction is catalyzed by an optically active substance The best examples are found m biochemical processes Most bio chemical reactions are catalyzed by enzymes Enzymes are chiral and enantiomerically homogeneous they provide an asymmetric environment m which chemical reaction can take place Ordinarily enzyme catalyzed reactions occur with such a high level of stereo selectivity that one enantiomer of a substance is formed exclusively even when the sub strate is achiral The enzyme fumarase for example catalyzes hydration of the double bond of fumaric acid to malic acid m apples and other fruits Only the S enantiomer of malic acid is formed m this reaction... 

The enzyme is a single enantiomer of a chiral molecule and binds the coenzyme and substrate m such a way that hydride is transferred exclusively to the face of the carbonyl group that leads to (5) (+) lactic acid Reduction of pyruvic acid m the absence of an enzyme however say with sodium borohydride also gives lactic acid but as a racemic mixture containing equal quantities of the R and S enantiomers... 

FIGURE 17 14 (a) Binding sites of enzyme discriminate between prochiral faces of substrate One prochiral face can bind to the enzyme better than the other (b) Reaction attaches fourth group to substrate producing only one enantiomer of chiral product... 

Organic chemists often use enantiomencally homogeneous starting materials for the synthe SIS of complex molecules (see Chiral Drugs p 296) A novel preparation of the S enantiomer of compound B has been descnbed using a bacterial cyclohexanone monooxygenase enzyme system... 

Before leaving this biosynthetic scheme notice that PGE2 has four chirality cen ters Even though arachidomc acid is achiral only the stereoisomer shown m the equa tion IS formed Moreover it is formed as a single enantiomer The stereochemistry is controlled by the interaction of the substrate with the enzymes that act on it Enzymes offer a chiral environment m which biochemical transformations occur and enzyme catalyzed reactions almost always lead to a single stereoisomer Many more examples will be seen m this chapter... 

Ammonia reacts with the ketone carbonyl group to give an mine (C=NH) which is then reduced to the amine function of the a ammo acid Both mine formation and reduc tion are enzyme catalyzed The reduced form of nicotinamide adenine diphosphonu cleotide (NADPH) is a coenzyme and acts as a reducing agent The step m which the mine is reduced is the one m which the chirality center is introduced and gives only L glutamic acid... 

The primary disadvantage of the conjugate addition approach is the necessity of performing two chiral operations (resolution or asymmetric synthesis) ia order to obtain exclusively the stereochemicaHy desired end product. However, the advent of enzymatic resolutions and stereoselective reduciag agents has resulted ia new methods to efficiently produce chiral enones and CO-chain synthons, respectively (see Enzymes, industrial Enzymes in ORGANIC synthesis). Eor example, treatment of the racemic hydroxy enone (70) with commercially available porciae pancreatic Hpase (PPL) ia vinyl acetate gave a separable mixture of (5)-hydroxyenone (71) and (R)-acetate (72) with enantiomeric excess (ee) of 90% or better (204). 

It is generally beheved that selectivity of hydrolytic enzymes strongly depends on the proximity of the chiral center to the reacting carbonyl group, and only a few examples of successful resolutions exist for compounds that have the chiral center removed by more than three bonds. A noticeable exception to this rule is the enantioselective hydrolysis by Pseudomonasfluorescens Hpase (PEL) of racemic dithioacetal (5) that has a prochiral center four bonds away from the reactive carboxylate (24). The monoester (6) is obtained in 89% yield and 98% ee. 

Alcohol dehydrogenase-catalyzed reduction of ketones is a convenient method for the production of chiral alcohols. HLAD, the most thoroughly studied enzyme, has a broad substrate specificity and accommodates a variety of substrates (Table 11). It efficiendy reduces all simple four- to nine-membered cycHc ketones and also symmetrical and racemic cis- and trans-decalindiones (167). Asymmetric reduction of aUphatic acycHc ketones (C-4—C-10) (103,104) can be efficiendy achieved by alcohol dehydrogenase isolated from Thermoanaerohium hrockii (TBADH) (168). The enzyme is remarkably stable at temperatures up to 85°C and exhibits high tolerance toward organic solvents. Alcohol dehydrogenases from horse Hver and T. hrockii... 

The enzyme-catalyzed interconversion of acetaldehyde and ethanol serves to illustrate a second important feature of prochiral relationships, that ofprochiral faces. Addition of a fourth ligand, different from the three already present, to the carbonyl carbon of acetaldehyde will produce a chiral molecule. The original molecule presents to the approaching reagent two faces which bear a mirror-image relationship to one another and are therefore enantiotopic. The two faces may be classified as re (from rectus) or si (from sinister), according to the sequence rule. If the substituents viewed from a particular face appear clockwise in order of decreasing priority, then that face is re if coimter-clockwise, then si. The re and si faces of acetaldehyde are shown below. 

Most enzyme-catalyzed processes, such as the examples just discussed, are highly enantioselective, leading to products of high enantiomeric purity. Reactions with other chiral reagents exhibit a wide range of enantioselectivity. A fiequent objective of the smdy... 

Forskolin is an activator of the enzyme adenylate cyclase which has therapeutic utility. Outlined below are stereocontrolled routes to racemic and natural chiral forms of forskolin derived by multistrategic retrosynthedc analysis. 

In the early work on the synthesis of prostaglandins, zinc borohydride was used for the reduction of the 15-ketone function and a 1 1 mixture of epimeric 15(S)- and 15(/ )-alcohols was generally obtained. Subsequent studies led to reaction conditions for highly selective reduction to the desired 15(S)-alcohol. Some of the results are summarized in the following table. The most practical method is E which utilizes borane as the stoichiometric reductant and a chiral, enzyme-like catalyst which is shown. 

As in the previous categories in this section, there are numerous compounds which have been prepared based on a sugar subunit. Examples may be found in Refs. 7,35,42-45, 57, 82-85, 117—121,175,176,193 and 208. Much of the work in these references has been reported by Stoddart and his coworkers, who have pioneered this field. As with the compounds prepared by Cram, the goal was to prepare a chiral receptor for ammonium ions which could be utilized in enzyme model studies. 

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