Biocatalytic <i>N</i>-Alkylation of Amines Using Either Primary Alcohols or Carboxylic Acids via Reductive Aminase Cascades

Ramsden, Jeremy I.; Heath, Rachel S.; Derrington, Sasha R.; Montgomery, Sarah L.; Mangas‐Sánchez, Juan; Mulholland, Keith R.; Turner, Nicholas J.

doi:10.1021/jacs.8b11561

Cited by 98 publications

(82 citation statements)

References 38 publications

(51 reference statements)

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“…In an alternative approach, adenylate kinase (which catalyzes the reversible phosphorylation of AMP by ATP) was used in place of PPK. Coupling of this enzyme with PAP pushes the equilibrium towards ATP regeneration (Figure B) …”

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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Section: Introductionmentioning

confidence: 99%

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

et al. 2019

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“…[8] In vitro systems on the other hand require the imitation of all relevant reaction steps by applying appropriate enzymes. [9] They provide advantages in testing and optimizing the reaction conditions, ideally allowing reducing-if not eliminating-side reactions of the startingmaterials and aldehyde products. On the otherh and, possible incompatibilities with reagents required to regeneratet he co-factors and pH shifts during the reactionc ourse that would have played no role in aw hole-cellt ransformation need to be considered.…”

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confidence: 99%

Enzymatic One‐Step Reduction of Carboxylates to Aldehydes with Cell‐Free Regeneration of ATP and NADPH

Strohmeier

Eiteljörg

Schwarz

et al. 2019

Chemistry A European J

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The direct generation of aldehydes from carboxylic acids is often a challenging synthetic task but undoubtedly attractive in view of abundant supply of such feedstocks from nature. Though long known, biocatalytic carboxylate reductions are at an early stage of development, presumably because of their co‐factor requirement. To establish an alternative to whole‐cell‐based carboxylate reductions which are limited by side reactions, we developed an in vitro multi‐enzyme system that allows for quantitative reductions of various carboxylic acids with full recycling of all cofactors and prevention of undesired over‐reductions. Regeneration of adenosine 5′‐triphosphate is achieved through the simultaneous action of polyphosphate kinases from Meiothermus ruber and Sinorhizobium meliloti and β‐nicotinamide adenine dinucleotide 2′‐phosphate is reduced by a glucose dehydrogenase. Under these conditions and in the presence of the carboxylate reductases from Neurospora crassa or Nocardia iowensis , various aromatic, heterocyclic and aliphatic carboxylic acids were quantitatively reduced to the respective aldehydes.

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“…Additionally, enzymes can be engineered to change their properties for novel catalysis. [12][13][14][15][16] Oxidoreductases are one of the largest enzyme classes and are vital to the pharmaceutical industry; their requirement for nicotinamide cofactors as a hydride source/sink [17][18][19][20][21][22][23] makes them perfect candidates for use in the electrochemical leaf technology. Given their expense and requirement in stoichiometric amounts, industry must recycle nicotinamide cofactors in situ to allow reactions to progress.…”

Section: Giorgio Morello Holds a Master's Degree (2015)mentioning

confidence: 99%

Electrified Nanoconfined Biocatalysis with Rapid Cofactor Recycling

et al. 2019

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In living cells, the overall rates of catalytic reaction chains (cascades) are massively enhanced by nanoconfinement of enzymes in tiny enclosed volumes: presented in such a way, interdependent catalysts are highly concentrated, and distances (active site‐to‐active site) across which intermediates and cofactors must diffuse, may be tiny. In a parallel technology exploiting this principle, enzyme cascades are powered, amplified, and monitored in real time as they work in concert, being nanoconfined within the pores of an electrically conductive metal oxide electrode. The technology, nicknamed the electrochemical leaf, mimics chloroplast biosynthesis by exploiting nicotinamide adenine dinucleotide (NADP(H)) recycling catalysed by the flavoenzyme, ferredoxin NADP+ reductase (FNR). Adsorbed on the inner walls of the nanopores, FNR rapidly transfers electrons between the electrode and NADP(H). This activity is coupled to an oxidoreductase enzyme, also nanoconfined within the pores, which recycles the cofactor and selectively synthesises a desired product or senses an analyte. Use of catalyst and cofactor is very efficient. The technology is simple, inexpensive and adaptable, and scalable to both microscopic levels (diagnostics) and macroscopic levels (organic synthesis). Whereas native FNR is specific for NADP(H), the technology can be extended by genetic engineering to include NAD(H) as recycling cofactor.

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Biocatalytic N-Alkylation of Amines Using Either Primary Alcohols or Carboxylic Acids via Reductive Aminase Cascades

Cited by 98 publications

References 38 publications

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

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

Enzymatic One‐Step Reduction of Carboxylates to Aldehydes with Cell‐Free Regeneration of ATP and NADPH

Electrified Nanoconfined Biocatalysis with Rapid Cofactor Recycling

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