Diversity‐Oriented Production of Metabolites Derived from Chorismate and Their Use in Organic Synthesis

Bongaerts, Johannes; Esser, Simon; Lorbach, Volker; Al-Momani, Lo’ay; Franke, Dirk; Grondal, Christoph; Kurutsch, Anja; Bujnicki, Robert; Raeven, Leon; Wubbolts, Marcel; Bovenberg, Roel A. L.; Nieger, Martin; Schürmann, Melanie; Trachtmann, Natalie; Kozak, Stefan; Sprenger, Georg A.; Müller, Michael

doi:10.1002/ange.201103261

Cited by 11 publications

(7 citation statements)

References 45 publications

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“…The compound with t R =3.9 min was purified and analyzed by NMR spectroscopy. The signals agree with the data reported for the regioisomer of 8 , iso‐SDHCHC (iso‐ 8 ; Scheme ) …”

Section: Methodssupporting

confidence: 89%

“…The shikimate pathway has been genetically engineered for the production of known natural products, for example, (5 S ,6 S )‐5,6‐dihydroxycyclohexa‐1,3‐diene‐1‐carboxylate (2,3‐ trans ‐CHD; 4 ) and (5 S ,6 S )‐6‐amino‐5‐hydroxycyclohexa‐1,3‐diene‐1‐carboxylate (2,3‐ trans ‐CHA; 5 ), themselves chiral building blocks useful for stereoselective synthesis . Both compounds are known from a biosynthetic context and have been produced by microbial fermentation with Escherichia coli strains (Scheme ) …”

Section: Methodsmentioning

confidence: 99%

“…With the same strategy as used for the in vivo conversion of 4 , a 2,3‐ trans ‐CHA ( 5 ) overproducer strain was used for the expression of MenD. Following the approach used by Bongaerts et al., E. coli F68/ pC35‐2 was obtained by expressing two genes ( phzD ‐ phzE ) from Pseudomonas aeruginosa encoding ADIC synthase, which forms 3 that is hydrolyzed by the isochorismatase PhzD to 5 (Scheme ) . Genes encoding wild‐type MenD and MenD Arg107Ile were cloned into this strain, resulting in yields of 10 and 64 mg L −1 SAHCHC ( 10 ), respectively.…”

Section: Methodsmentioning

confidence: 99%

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Alteration of the Route to Menaquinone towards Isochorismate‐Derived Metabolites

et al. 2019

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Chorismate and isochorismate constitute branch‐point intermediates in the biosynthesis of many aromatic metabolites in microorganisms and plants. To obtain unnatural compounds, we modified the route to menaquinone in Escherichia coli . We propose a model for the binding of isochorismate to the active site of MenD ((1 R ,2 S , 5 S ,6 S )‐2‐succinyl‐5‐enolpyruvyl‐6‐hydroxycyclohex‐3‐ene‐1‐carboxylate (SEPHCHC) synthase) that explains the outcome of the native reaction with α‐ketoglutarate. We have rationally designed variants of MenD for the conversion of several isochorismate analogues. The double‐variant Asn117Arg–Leu478Thr preferentially converts (5 S ,6 S )‐5,6‐dihydroxycyclohexa‐1,3‐diene‐1‐carboxylate (2,3‐ trans ‐CHD), the hydrolysis product of isochorismate, with a >70‐fold higher ratio than that for the wild type. The single‐variant Arg107Ile uses (5 S ,6 S )‐6‐amino‐5‐hydroxycyclohexa‐1,3‐diene‐1‐carboxylate (2,3‐ trans ‐CHA) as substrate with >6‐fold conversion compared to wild‐type MenD. The novel compounds have been made accessible in vivo (up to 5.3 g L −1 ). Unexpectedly, as the identified residues such as Arg107 are highly conserved (>94 %), some of the designed variations can be found in wild‐type SEPHCHC synthases from other bacteria (Arg107Lys, 0.3 %). This raises the question for the possible natural occurrence of as yet unexplored branches of the shikimate pathway.

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“…The compound with t R =3.9 min was purified and analyzed by NMR spectroscopy. The signals agree with the data reported for the regioisomer of 8 , iso‐SDHCHC (iso‐ 8 ; Scheme ) …”

Section: Methodssupporting

confidence: 89%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Alteration of the Route to Menaquinone towards Isochorismate‐Derived Metabolites

et al. 2019

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“…Moreover, many enzymes that catalyse reactions starting from chorismate ( Fig. 1) are useful tools in the production of structurally diverse valuable, often chiral building blocks [4][5][6]. Knowledge of the three-dimensional structure of an enzyme usually greatly aids the understanding of the enzyme's catalytic mechanism.…”

Section: Introductionmentioning

confidence: 99%

Mechanistic Implications for the Chorismatase FkbO Based on the Crystal Structure

Juneja

Hubrich

Diederichs

et al. 2014

Journal of Molecular Biology

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Chorismate-converting enzymes are involved in many biosynthetic pathways leading to natural products and can often be used as tools for the synthesis of chemical building blocks. Chorismatases such as FkbO from Streptomyces species catalyse the hydrolysis of chorismate yielding (dihydro)benzoic acid derivatives. In contrast to many other chorismate-converting enzymes, the structure and catalytic mechanism of a chorismatase had not been previously elucidated. Here we present the crystal structure of the chorismatase FkbO in complex with a competitive inhibitor at 1.08Å resolution. FkbO is a monomer in solution and exhibits pseudo-3-fold symmetry; the structure of the individual domains indicates a possible connection to the trimeric RidA/YjgF family and related enzymes. The co-crystallised inhibitor led to the identification of FkbO's active site in the cleft between the central and the C-terminal domains. A mechanism for FkbO is proposed based on both interactions between the inhibitor and the surrounding amino acids and an FkbO structure with chorismate modelled in the active site. We suggest that the methylene group of the chorismate enol ether takes up a proton from an active-site glutamic acid residue, thereby initiating chorismate hydrolysis. A similar chemistry has been described for isochorismatases, albeit implemented in an entirely different protein scaffold. This reaction model is supported by kinetic data from active-site variants of FkbO derived by site-directed mutagenesis.

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“…Several other products are accessible from the biotransformations of chorismate catalysed by different enzymes, including isochorismate synthase (MenF or EntC), salicylate synthase (MbtI), anthranilate synthase (TrpE) and aminodeoxychorismate synthase (PabB) to yield isochorismate, salicylate, anthranilate and p ‐aminodeoxychorismate, respectively . Using a diversity‐oriented synthesis approach, microbial production of a number of chorismate transformation products was achieved and the application of these metabolites as synthetic building blocks was demonstrated …”

Section: Substrate Specificities and Biocatalytic Applications Of Isomentioning

confidence: 99%

Biocatalytic Potential of Enzymes Involved in the Biosynthesis of Isoprenoid Quinones

2017

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Naturally occurring isoprenoid quinones mediate electron transfer in respiratory or photosynthetic chains of living organisms. Tremendous progress has been made in the elucidation of biosynthetic pathways of prenylated quinones and a number of enzymes that catalyze specific transformation steps with remarkably high regio‐ and stereoselectivity have been characterized. Interestingly, some of these enzymes possess broad substrate scope towards synthetic analogues, thereby enabling their application as synthetic tools. The availability of mechanistic, structural and mutagenesis information for many of the pathway enzymes presents opportunities for protein engineering, a powerful approach that may improve the suitability of these biocatalysts for industrial processes.

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Diversity‐Oriented Production of Metabolites Derived from Chorismate and Their Use in Organic Synthesis

Cited by 11 publications

References 45 publications

Alteration of the Route to Menaquinone towards Isochorismate‐Derived Metabolites

Alteration of the Route to Menaquinone towards Isochorismate‐Derived Metabolites

Mechanistic Implications for the Chorismatase FkbO Based on the Crystal Structure

Biocatalytic Potential of Enzymes Involved in the Biosynthesis of Isoprenoid Quinones

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