Lyases benzaldehyde lyase

A biochemically related benzaldehyde lyase (BAL) (EC 4.1.2.38) catalyzes the same carboligation reactions, but with opposite (J )-selectivity (mf-110) [178]. All these enzymes seem to display a rather useful substrate tolerance for variously substituted aldehyde precursors. 

R)-Benzoins and (/ )-2-hydroxypropiophcnonc derivatives are formed on a preparative scale by benzaldehyde lyase (BAL)-catalyzed C-C bond formation from aromatic aldehydes and acetaldehyde in aqueous buffer/DMSO solution with remarkable ease in high chemical yield and high optical purity (Eq. 8.112).303 Less-stable mixed benzoins were also generated via reductive coupling of benzoyl cyanide and carbonyl compounds by aqueous titanium(III) ions.304... 

Starting from enantiomerically pure 4-methylsulfanyl-mandelonitrile, thiamphenicol and florfenicol have been enantioselectively synthesized (Figure 5.14). The enantiomerically pure 4-methylsulfanyl-mandelonitrile was obtained by hydrocyanation reaction of 4-methy lsulfany 1-benzaldehyde catalyzed by (M)-hydroxynitrile lyase of Badamu (almond from Xinjiang, China) (Prunus communis L. var. dulcis Borkh), which, after an extensive screening, was found to be a highly effective bio-catalyst for this reaction [85]. 

A new hydroxynitrile lyase (HNL) was isolated from the seed of Japanese apricot Prunus mume). It accepts benzaldehyde and a large number of unnatural substrates for the addition of HCN to produce the corresponding (7 )-cyanohydrins in excellent optical and chemical yields. A new high-performance liquid chromatography (HPLC)-based enantioselective assay technique was developed for the enzyme, which promotes the addition of KCN to benzaldehyde in a buffered solution (pH 4.0). Asymmetric synthesis of (7 )-cyanohydrins by a new HNL is described (Figure 8.4). ... 

Miiller and co-workers have developed an enantioselective enzymatic crossbenzoin reaction (Table 2) [43, 44], This is the first example of an enantioselective cross-benzoin reaction and takes advantage of the donor-acceptor concept. This transformation is catalyzed by thiamin diphosphate (ThDP) 23 in the presence of benzaldehyde lyase (BAL) or benzoylformate decarboxylase (BFD). Under these enzymatic reaction conditions the donor aldehyde 24 is the one that forms the acyl anion equivalent and subsequently attacks the acceptor aldehyde 25 to provide a variety of a-hydroxyketones 26 in good yield and excellent enantiomeric excesses without contamination of the other cross-benzoin products 27. The authors chose 2-chlorobenzaldehyde 25 as the acceptor because of its inability to form a homodimer under enzymatic reaction conditions. 

This enzyme [EC 4.1.2.10], also known as hydroxynitrile lyase and (i )-oxynitrilase, catalyzes the conversion of mandelonitrile to cyanide and benzaldehyde. 

Benzoylformate decarboxylase (BFD EC 4.1.1.7) belongs to the class of thiamine diphosphate (ThDP)-dependent enzymes. ThDP is the cofactor for a large number of enzymes, including pyruvate decarboxylase (PDC), benzaldehyde lyase (BAL), cyclohexane-1,2-dione hydrolase (CDH), acetohydroxyacid synthase (AHAS), and (lR,6] )-2-succinyl-6-hydroxy-2,4-cyclohexadiene-l-carboxylate synthase (SHCHC), which all catalyze the cleavage and formation of C-C bonds [1]. The underlying catalytic mechanism is summarized elsewhere [2] (see also Chapter 2.2.3). 

In the framework of SFB380, two projects dealt extensively with acyloin-con-densing ThDP-dependent enzymes such as pyruvate decarboxylase (PDC), ben-zoylformate decarboxylase (BFD), or benzaldehyde lyase (BAL) (see Chapters 2.2.3 and 2.2.7). Another ThDP-dependent decarboxylase, phosphonopyruvate decarboxylase (PPD) from Streptomyces viridochromogenes, became available only recently and was studied in project B21. We wanted to find out whether this PDC-related enzyme could be a valuable tool in the provision of acyloin condensations involving C-P bonds (see Section 2.2.2.23). 

LB-ADH was used for the synthesis of vic-diols. Starting from benzaldehyde and acetaldehyde, (lS,2S)-l-phenylpropane-l,2-diol (de = 98%) and (lS,2R)-l-phenyl-propane-l,2-diol (de = 99%), respectively, could be produced in a stereoselective two-step enzymatic synthesis using benzaldehyde lyase (BAL) and accordingly benzoylformate decarboxylase (BFD) as well as LB-ADH [8] (Scheme 2.2.4.3). 

Table 3.1.2 Stability of benzaldehyde lyase under process conditions.

Benzoin Condensation with Entrapped Benzaldehyde Lyase... 

Enzymatic reactions forming new carbon-carbon bonds are a further important field of biotransformations in natural product synthesis. The construction of new, often complex carbon frameworks or their decomposition is performed by nature under catalysis of a set of enzymes. For organic chemists some of these enzymes, belonging to the enzyme class of lyases, such as aldolases, decarboxylases, hydro-xynitrile lyases (HNLs), or benzaldehyde lyases (BALs), have been proven to represent versatile amendments to their synthetic toolbox. 

Enantiopure, bifunctional acyloins (a-hydroxy ketones) are versatile intermediates in natural product synthesis (also see Sect. 2.3, Fig. 11). In nature, the formation of a-hydroxy ketones is efficiently catalyzed by thiamine diphosphate-dependent enzymes transketolases, decarboxylases, and other lyases, such as BALs. A great portfolio of biotransformations, especially with benzaldehyde derivatives as starting materials, were realized [204]. 

Fig. 43 (a) Benzaldehyde lyase-catalyzed acyloin formation, (b) Application of acyloins in natural product synthesis... 

Demir AS, Sesenoglu O et al (2003) Benzaldehyde lyase-catalyzed enantioselective carbo-ligation of aromatic aldehydes with mono- and dimethoxy acetaldehyde. Org Lett 5 2047-2050... 

The bifunctional nature and the presence of a stereocenter make a-hydroxyketones (acyloins) amenable to further synthetic transformations. There are two classical chemical syntheses for these a-hydroxyketones the acyloin condensation and the benzoin condensation. In the acyloin condensation a new carbon-carbon bond is formed by a reduction, for instance with sodium. In the benzoin condensation the new carbon-carbon bond is formed with the help of an umpolung, induced by the formation of a cyanohydrin. A number of enzymes catalyze this type of reaction, and as might be expected, the reaction conditions are considerably milder [2-4, 26, 27]. In addition the enzymes such as benzaldehyde lyase (BAL) catalyze the formation of a new carbon-carbon bond enantioselectively. Transketolases (TK)... 

L-Phe can be prepared via the enantioselective hydrolysis of N-acetyl-D,L-Phe and microbial reductive amination of phenylpyruvate (see Fig. 8.14) [87]. The stoichiometric yields of these processes were high but the precursors required 3-4 synthetic steps from the basic starting materials in most cases. The phenyl-ammonia lyase route, in contrast [89], provided L-Phe in only two steps from the basic chemicals benzaldehyde and acetic anhydride [90]. The enzymatic step... 

Another approach towards immobilization via ionic interactions is the use of tethered metal ions, such as Cu, Co, or Ni, in order to bind the enzyme [53, 54]. This is particularly so when the enzyme contains an easily accessible imidazole residue from histidine [55] or a His tag (Figure 2.6) [56-59], that is a short tag with six histidines. This tag can be readily introduced by genetically modifying the enzyme. Little influence of the tag on the catalytic performance has been noticed. When benzaldehyde lyase [60] was immobilized on an Ni -containing polyvinylpyrrolidinone-based matrix, it could be reused several times for the formation of benzoin (12) (Scheme 2.4) [58]. 

Lrthreonine aldolase (L-threonine acetaldehyde-lyase) catalyzes the reversible condensation of acetaldehyde and glycine to form L-threonine. The enzyme has been shown to be an activity distinct from serine hydroxy-methyltransferase that also catalyzes the above reaction (85,86). The substrate specifically of the adolase has been demonstrated to be flexible with respect to the aldehyde involved. The enzyme has been shown to form phenylserine derivatives from substituted benzaldehydes and glycine (86). 

Benzaldehyde lyase Reversible benzoin condensation, a carboligase/lyase C6H5C0CH(0H)C6H5 2C6H5CHO None... 

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