Chorismate biosynthesis

By use of synthetic medium the formation of 3,4-trans-CH D was maintained for a 36-h cultivation period, resulting in accumulation of up to 790 mg L 1 3,4-trans-CHD 1 [11]. With this strain, however, it is necessary to separate the growth and production phases, because the substances for which E. coli strain BN117 is known to be auxotrophic, i.e. tryptophan, tyrosine, phenylalanine, proline, arginine, histidine, and p-aminobenzoate, also partially inhibit entry to the chorismate biosynthesis pathway (feedback inhibition of DAHP-synthase). 

II CH 1 FIGURE 1 4.1 Chorismate Biosynthesis. Chorismate is an intermediate in the shikimate pathway. The formation... 

Plants synthesize all 20 common amino acids de novo. Glyphosate, a weed killer sold under the trade name Roundup, is an analog of phosphoenolpyruvate that specifically inhibits 3-enolpyruvylshikimate 5-phosphate synthase, a key enzyme of the pathway for chorismate biosynthesis. This compound is a very effective plant herbicide, but has virtually no effect on mammals. Why ... 

Animals caimot synthesize the naphthoquinone ring of vitamin K, but necessary quantities are obtained by ingestion and from manufacture by intestinal flora. In plants and bacteria, the desired naphthoquinone ring is synthesized from 2-oxoglutaric acid (12) and shikimic acid (13) (71,72). Chorismic acid (14) reacts with a putative succinic semialdehyde TPP anion to form o-succinyl benzoic acid (73,74). In a second step, ortho-succmY benzoic acid is converted to the key intermediate, l,4-dihydroxy-2-naphthoic acid. Prenylation with phytyl pyrophosphate is followed by decarboxylation and methylation to complete the biosynthesis (75). 

Biological examples of pericyclic reactions are relatively rare, although one much-studied example occurs during biosynthesis in bacteria of the essential amino acid phenylalanine. Phenylalanine arises from the precursor chorismate,... 

Figure 30.14 Pathway for the bacterial biosynthesis of phenylalanine from chorismate, involving a Claisen rearrangement.

The shikimate pathway is the major route in the biosynthesis of ubiquinone, menaquinone, phyloquinone, plastoquinone, and various colored naphthoquinones. The early steps of this process are common with the steps involved in the biosynthesis of phenols, flavonoids, and aromatic amino acids. Shikimic acid is formed in several steps from precursors of carbohydrate metabolism. The key intermediate in quinone biosynthesis via the shikimate pathway is the chorismate. In the case of ubiquinones, the chorismate is converted to para-hydoxybenzoate and then, depending on the organism, the process continues with prenylation, decarboxylation, three hydroxy-lations, and three methylation steps. - ... 

The Shikimate pathway is responsible for biosynthesis of aromatic amino acids in bacteria, fungi and plants [28], and the absence of this pathway in mammals makes it an interesting target for designing novel antibiotics, fungicides and herbicides. After the production of chorismate the pathway branches and, via specific internal pathways, the chorismate intermediate is converted to the three aromatic amino acids, in addition to a number of other aromatic compounds [29], The enzyme chorismate mutase (CM) is a key enzyme responsible for the Claisen rearrangement of chorismate to prephenate (Scheme 1-1), the first step in the branch that ultimately leads to production of tyrosine and phenylalanine. 

Schultz and coworkers (Jackson et a ., 1988) have generated an antibody which exhibits behaviour similar to the enzyme chorismate mutase. The enzyme catalyses the conversion of chorismate [49] to prephenate [50] as part of the shikimate pathway for the biosynthesis of aromatic amino acids in plants and micro-organisms (Haslam, 1974 Dixon and Webb, 1979). It is unusual for an enzyme in that it does not seem to employ acid-base chemistry, nucleophilic or electrophilic catalysis, metal ions, or redox chemistry. Rather, it binds the substrate and forces it into the appropriate conformation for reaction and stabilizes the transition state, without using distinct catalytic groups. 

Aryl side chain containing L-a-amino acids, such as phenylalanine (Phe), tyrosine (Tyr), and tryptophan (Trp), are derived through the shikimate pathway. The enzymatic transformation of phosphoenolpyr-uvate (PEP) and erythro-4-phosphate, through a series of reactions, yields shikimate (Scheme 2). Although shikimate is an important biosynthetic intermediate for a number of secondary metabolites, this chapter only describes the conversion of shikimate to amino acids containing aryl side chains. In the second part of the biosynthesis, shikimate is converted into chorismate by the addition of PEP to the hydroxyl group at the C5 position. Chorismate is then transformed into prephenate by the enzyme chorismate mutase (Scheme 3). 

During the biosynthetic transformation, chorismate is the point of divergence for the biosynthesis of Phe, Tyr, Trp, and other amino acids containing aromatic groups. For example, the biosynthesis of Trp begins with the conversion of chorismate to anthranilate (Scheme 4(a)). A sequence of amination and aromatization reactions produces anthranilate, which is then condensed with phosphoribosylpyrophosphate. The intermediate is carried through a series of reactions to yield Trp (Scheme 4(b)). 

This enzyme [EC 4.1.3.27] catalyzes the reaction of chorismate with glutamine to generate anthranilate, pyruvate, and glutamate. In certain species, this enzyme is part of a multifunctional protein together with one or more other components of the system for the biosynthesis of tryptophan (Le., indole-3-glycerol-phosphate synthase, anthranilate phosphoribosyltransferase, tryptophan synthase, and phosphoribosylanthranilate isomerase). The anthranilate synthase that is present in these complexes has been reported to be able to utilize either glutamine or ammonia as the nitrogen source. However, it has also been reported that when anthranilate synthase is separated from this complex, only ammonia can serve as a substrate. 

Claisen rearrangement plays an important part in the biosynthesis of several natural products. For example, the chorismate ion is rearranged to the prephenate ion by the Claisen rearrangement, which is catalysed by the enzyme chorismate mutase. This prephenate ion is a key intermediate in the shikimic acid pathway for the biosynthesis of phenylalanine, tyrosine and many other biologically important natural products. 

HGURE 22-16 Biosynthesis of chorismate, an intermediate in the synthesis of aromatic amino acids in bacteria and plants. 

FIGURE 22-17 Biosynthesis of tryptophan from chorismate in bacteria and plants. In E. coli, enzymes catalyzing steps (T) and are subunits of a single complex. 

FIGURE 22-19 Biosynthesis of phenylalanine and tyrosine from chorismate in bacteria and plants. Conversion of chorismate to prephenate is a rare biological example of a Claisen rearrangement. 

The enzyme is organized into two structural domains,201 one of which binds FAD and the other NADP+. Similar single-electron transfers through flavoproteins also occur in many other enzymes. Chorismate mutase, an important enzyme in biosynthesis of aromatic rings (Chapter 25), contains bound FMN. Its function is unclear but involves formation of a neutral flavin radical.276 277... 

Figure 25-2 The biosynthesis of tryptophan from chorismate and related synthetic reactions-...

The biosynthesis of tryptophan. Tryptophan is synthesized in five steps from chorismate. Each step requires a specific enzyme activity. The four intermediates between chorismate and tryptophan serve no function other than as precursors of tryptophan. The detailed steps of each reaction in this sequence are described in chapter 21. 

Aromatic amino acid biosynthesis proceeds via a long series of reactions, most of them concerned with the formation of the aromatic ring before branching into the specific routes to phenylalanine, tyrosine, and tryptophan. Chorismate, the common intermediate of the three aromatic amino acids, (see fig. 21.1) is derived in eight steps from erythrose-4-phosphate and phosphoenolpyruvate. We focus on the biosynthesis of tryptophan, which has been intensively studied by both geneticists and biochemists. 

The biosynthesis of tryptophan from the branchpoint compound, chorismate in E. coli The first step involves the conversion of chorismate to the aromatic compound anthranilate. The anthranilate is transferred to a ribose phosphate chain. The product is cyclized to indoleglycerol phosphate by the removal of water and loss of the ring carboxyl by indoleglycerol phosphate synthase. Finally, in a... 

The first specific step in tryptophan biosynthesis is the glutamine-dependent conversion of chorismate to the simple aromatic compound anthranilate. Like most other glutamine-dependent reactions, the reaction can also occur with ammonia as the source of the amino group. However, high concentrations of ammonia are required. Thus far, almost all the anthranilate synthases examined have the glutamine amidotransferase activity (component II) and the choris-mate-to-anthranilate activity (component I) on separate proteins. 

The shikimate pathway results in the biosynthesis of chorismate, which can subsequently serve as a recursor for the biosynthesis of the aromatic amino acids tryptophan, phenylalanine and tyrosine. The biochemistry of... 

Figure 3-5. Biosynthesis of salicylic acid. The enzymes involved in this pathway are (a) chorismate mutase (E.C. 5.4.99.5), (b) prephenate aminotransferase (E.C. 2.6.1.78 and E.C. 2.6.1.79), (c) arogenate dehydratase (E.C. 4.2.1.91), (d) phenylalanine ammonia lyase (E.C. 4.3.1.5), (e) presumed P-oxidation by a yet to be identified enzyme, (f) benzoic acid 2-hydroxylase, (g) isochorismate synthase (E. C. 5.4.4.2), and (h) a putative plant pyruvate lyase.

Chorismate mutase (CM) catalyzes conversion of chorismate into prephenate in a Claisen rearrangement prephenate is a precursor in the biosynthesis of both i-phenylalanine (i-Phe) and i-tyrosine (i-Ttyr). CM occurs as a dimer for structural studies, the monomer was needed (MacBeath, 1998). The assay for CM activity was based on growth of the colonies in the absence of i-Phe and L-Tyr and thus was based on selection, not screening. One resulting mutant was found to be a monomer and to contain a somewhat polar Ala-Arg-Trp-Pro-Trp-Ala sequence. 

Chloramphenicol.—A synthetase which converts chorismic acid into p-amino-phenylalanine (an intermediate in chloramphenicol biosynthesis) has been partly characterized it requires an aminotransferase and pyridoxal phosphate for activity.58... 

Fig. 6 Four enzymes (one salicylate-AMP ligase YbtE/PchD, two NRPS and/or NRPS/PKS enzymes HMWPl/PchE and HMWP2/PchF, and one reductase YbtU/PchG) are required for the in vitro biosynthesis of (a) yersiniabactin (24) and (b) pyochelin (25). (c) The initial stages of 24 and 25 biosynthesis proceed via a similar mechanism from chorismic acid 26 to the salicylate-bisthiazole intermediate 33...

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