Exploring Bacterial Carboxylate Reductases for the Reduction of Bifunctional Carboxylic Acids

Khusnutdinova, Anna N.; Flick, Robert; Popović, Ana; Brown, Greg; Tchigvintsev, Anatoli; Nocek, B.; Correia, Kevin; Joo, Jeong Chan; Mahadevan, Radhakrishnan; Yakunin, Alexander F.

doi:10.1002/biot.201600751

Cited by 77 publications

(102 citation statements)

References 52 publications

(122 reference statements)

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“…They were confirmed as true carboxylic acid reductases. Of the enzymes tested, the MavCAR displayed the highest catalytic activity, being nearly six times more active than the previously characterized MmCAR and slightly more active than the best previously reported CAR, from Mycobacterium abscessus …”

Section: Resultsmentioning

confidence: 74%

Characterization of Carboxylic Acid Reductases for Biocatalytic Synthesis of Industrial Chemicals

et al. 2018

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Carboxylic acid reductases (CARs) catalyze the reduction of a broad range of carboxylic acids into aldehydes, which can serve as common biosynthetic precursors to many industrial chemicals. This work presents the systematic biochemical characterization of five carboxylic acid reductases from different microorganisms, including two known and three new ones, by using a panel of short-chain dicarboxylic acids and hydroxy acids, which are common cellular metabolites. All enzymes displayed broad substrate specificities. Higher catalytic efficiencies were observed when the carbon chain length, either of the dicarboxylates or of the terminal hydroxy acids, was increased from C to C . In addition, when substrates of the same carbon chain length are compared, carboxylic acid reductases favor hydroxy acids over dicarboxylates as their substrates. Whole-cell bioconversions of eleven carboxylic acid substrates into the corresponding alcohols were investigated by coupling the CAR activity with that of an aldehyde reductase in Escherichia coli hosts. Alcohol products were obtained in yields ranging from 0.5 % to 71 %. The de novo stereospecific biosynthesis of propane-1,2-diol enantiomer was successfully demonstrated with use of CARs as the key pathway enzymes. E. coli strains accumulated 7.0 mm (R)-1,2-PDO (1.0 % yield) or 9.6 mm (S)-1,2-PDO (1.4 % yield) from glucose. This study consolidates carboxylic acid reductases as promising enzymes for sustainable synthesis of industrial chemicals.

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Section: Resultsmentioning

confidence: 74%

Characterization of Carboxylic Acid Reductases for Biocatalytic Synthesis of Industrial Chemicals

et al. 2018

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“…The carboxylic acid reductases (CARs) are enzymes that have increasing interest as biocatalysts . CARs catalyze the reduction of carboxylic acids to aldehydes in mild conditions, and can connect many types of enzyme reaction allowing the construction of novel multi‐enzyme pathways (Figure A) . Previously, we have shown CARs to be a fairly promiscuous enzyme class, catalyzing the reduction of both aliphatic and aromatic carboxylic acids.…”

Section: Introductionmentioning

confidence: 99%

Engineering a Seven Enzyme Biotransformation using Mathematical Modelling and Characterized Enzyme Parts

et al. 2019

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Multi‐step enzyme reactions offer considerable cost and productivity benefits. Process models offer a route to understanding the complexity of these reactions, and allow for their optimization. Despite the increasing prevalence of multi‐step biotransformations, there are few examples of process models for enzyme reactions. From a toolbox of characterized enzyme parts, we demonstrate the construction of a process model for a seven enzyme, three step biotransformation using isolated enzymes. Enzymes for cofactor regeneration were employed to make this in vitro reaction economical. Good modelling practice was critical in evaluating the impact of approximations and experimental error. We show that the use and validation of process models was instrumental in realizing and removing process bottlenecks, identifying divergent behavior, and for the optimization of the entire reaction using a genetic algorithm. We validated the optimized reaction to demonstrate that complex multi‐step reactions with cofactor recycling involving at least seven enzymes can be reliably modelled and optimized.

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“…NADPH could be regenerated by mutant formate dehydrogenase (FDH m ) from Pseudomonas sp. 101, using formate [Scheme A].…”

Section: Methodsmentioning

confidence: 99%

Glutamate as an Efficient Amine Donor for the Synthesis of Chiral β‐ and γ‐Amino Acids Using Transaminase

et al. 2019

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A recyclable glutamate amine donor system employing transaminase (TA), glutamate dehydrogenase (GluDH) and mutant formate dehydrogenase (FDHm) was developed, wherein amine donor Glu was regenerated using GluDH and thereby circumvented the inhibition of TA by α‐ketoglutarate. Various enantiopure β‐, γ‐amino acids, and amines were successfully synthesized with high conversions and excellent enantiomeric excess using this system.

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Exploring Bacterial Carboxylate Reductases for the Reduction of Bifunctional Carboxylic Acids

Cited by 77 publications

References 52 publications

Characterization of Carboxylic Acid Reductases for Biocatalytic Synthesis of Industrial Chemicals

Characterization of Carboxylic Acid Reductases for Biocatalytic Synthesis of Industrial Chemicals

Engineering a Seven Enzyme Biotransformation using Mathematical Modelling and Characterized Enzyme Parts

Glutamate as an Efficient Amine Donor for the Synthesis of Chiral β‐ and γ‐Amino Acids Using Transaminase

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