Development of an Amine Dehydrogenase for Synthesis of Chiral Amines

Abrahamson, Michael J.; Vazquez-Figueroa, Eduardo; Woodall, Nicholas B.; Moore, Jeffrey C.; Bommarius, Andreas S.

doi:10.1002/anie.201107813

Cited by 247 publications

(190 citation statements)

References 36 publications

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“…For ammonium chloride buffers, pH values <8.5 cannot be attained and hence ammonium formate buffer was investigated at various pH values (28). At pH 8 to 8.5, the amination of (R)-1a (20 mM) was achieved in 93% conversion and >99% ee ( The hydrogen-borrowing cascade was initially tested on a limited number of 1-phenyl-2-propanol derivatives 1a-1e (Table 1) for amination with inversion of configuration (entries 1-5), retention of configuration (entries [15][16][17][18][19] and asymmetric amination of racemic alcohols (entry 29-33). Conversion varied from moderate to excellent, whereas the ee was excellent in almost all cases.…”

Section: Europe Pmc Funders Author Manuscriptsmentioning

confidence: 99%

“…The cascade requires only catalytic quantities of a nicotinamide coenzyme that shuttles hydride from the oxidative step to the reductive step. The method has been successfully applied to: i) amination of optically active secondary alcohols with inversion of configuration; ii) amination of the corresponding enantiomeric secondary alcohols with retention of configuration; iii) asymmetric amination of racemic secondary alcohols; iv) amination of primary alcohols.Initially we examined the catalytic activity of the amine dehydrogenase variant that was recently obtained by protein engineering of the wild-type phenylalanine dehydrogenase from Bacillus badius (Ph-AmDH) (18,19). The substrate scope of the Ph-AmDH variant K78S -N277L for the conversion of a broad range of ketone substrates has not been reported; only the reductive amination of para-fluoro-phenylacetone (2b) was previously described using glucose and glucose dehydrogenase (GDH) for cofactor regeneration and very recently three other ketones were also tested (20).…”

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confidence: 99%

“…Initially we examined the catalytic activity of the amine dehydrogenase variant that was recently obtained by protein engineering of the wild-type phenylalanine dehydrogenase from Bacillus badius (Ph-AmDH) (18,19). The substrate scope of the Ph-AmDH variant K78S -N277L for the conversion of a broad range of ketone substrates has not been reported; only the reductive amination of para-fluoro-phenylacetone (2b) was previously described using glucose and glucose dehydrogenase (GDH) for cofactor regeneration and very recently three other ketones were also tested (20).…”

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Conversion of alcohols to enantiopure amines through dual-enzyme hydrogen-borrowing cascades

Mutti

Knaus

Scrutton

et al. 2015

Science

377

277

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Abstractα-Chiral amines are key intermediates for the synthesis of a plethora of chemical compounds on industrial scale. Here we present a biocatalytic hydrogen-borrowing amination of primary and secondary alcohols that allows for the efficient and environmentally benign production of enantiopure amines. The method relies on the combination of an alcohol dehydrogenase (ADHs from Aromatoleum sp., Lactobacillus sp. and Bacillus sp.) enzyme operating in tandem with an amine dehydrogenase (AmDHs engineered from Bacillus sp.) to aminate a structurally diverse range of aromatic and aliphatic alcohols (up to 96% conversion and 99% enantiomeric excess). Furthermore, primary alcohols are aminated with high conversion (up to 99%). This redox selfsufficient network possesses high atom efficiency, sourcing nitrogen from ammonium and generating water as the sole by-product.Amines are amongst the most frequently used chemical intermediates for the production of APIs (active pharmaceutical ingredients), fine chemicals, agrochemicals, polymers, dyestuffs, pigments, emulsifiers and plasticizing agents (1). However, the requisite amines are scarce in nature and their industrial production mainly relies upon the metal-catalysed hydrogenation of enamides (i.e. obtained from related ketone precursors), a process requiring transition metal complexes, which are expensive and increasingly unsustainable (2). Moreover, the asymmetric synthesis of amines from ketone precursors requires protection and deprotection steps that generate copious amounts of waste. As a consequence, various chemical processes for the direct conversion of alcohols into amines have been developed during the last decade. The intrinsic advantage of the direct amination of an Europe PMC Funders Author ManuscriptsEurope PMC Funders Author Manuscripts alcohol is that the reagent and the product are in the same oxidation state and therefore, theoretically, additional redox equivalents are not required.However, many of these methods have low efficiency and high environmental impact (e.g. Mitsunobu reaction) (3). The amination of simple alcohols such as methanol and ethanol, via heterogeneous catalysis, requires harsh conditions (>200 °C) and more structurally diverse alcohols are either converted with extremely low chemoselectivity or not converted at all (4). Furthermore, most of the work in this field involves non-chiral substrates whereas 40% of the commercial optically active drugs are chiral amines (2). Increasingly, biocatalytic methods are applied for the production of optically active amines, e.g. the lipase catalysed resolution of racemic mixtures of amines or the ω-transaminase process with a most recent example employing an engineered enzyme applied to the industrial manufacture of the diabetes medication Januvia® (sitagliptin) (5,6,7).Multi-step chemical reactions in one pot avoid the need for isolation of intermediates and purification steps. This approach leads to economic as well as environmental benefits since time-consuming intermediate work-ups are not...

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Section: Europe Pmc Funders Author Manuscriptsmentioning

confidence: 99%

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confidence: 99%

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confidence: 99%

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Conversion of alcohols to enantiopure amines through dual-enzyme hydrogen-borrowing cascades

Mutti

Knaus

Scrutton

et al. 2015

Science

377

277

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“…Recently, AmDHs have been created through protein engineering using the existing L-amino acid dehydrogenase. First, AmDH was engineered by the Bommarius group using leucine dehydrogenase (LeuDH) from Bacillus stearothomophilus to accept ketone substrates through the introduction of 2-4 point mutations [16]. Subsequently, a second amine dehydrogenase was developed based on the scaffold of phenylalanine dehydrogenase (PheDH) from Bacillus badius by the same group using a similar approach [17].…”

Section: Introductionmentioning

confidence: 99%

The Kinetic Resolution of Racemic Amines Using a Whole-Cell Biocatalyst Co-Expressing Amine Dehydrogenase and NADH Oxidase

et al. 2017

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Amine dehydrogenase (AmDH) possesses tremendous potential for the synthesis of chiral amines because AmDH catalyzes the asymmetric reductive amination of ketone with high enatioselectivity. Although a reductive application of AmDH is favored in practice, the oxidative route is interesting as well for the preparation of chiral amines. Here, the kinetic resolution of racemic amines using AmDH was first extensively studied, and the AmDH reaction was combined with an NADH oxidase (Nox) to regenerate NAD + and to drive the reaction forward. When the kinetic resolution was carried out with 10 mM rac-2-aminoheptane and 5 mM rac-α-methylbenzylamine (α-MBA) using purified enzymes, the enantiomeric excess (ee) values were less than 26% due to the product inhibition of AmDH by ketone and the inhibition of Nox by the substrate amine. The use of a whole-cell biocatalyst co-expressing AmDH and Nox apparently reduces the substrate and product inhibition, and/or it increases the stability of the enzymes. Fifty millimoles (50 mM) rac-2-aminoheptane and 20 mM rac-α-MBA were successfully resolved into the (S)-form with >99% ee using whole cells. The present study demonstrates the potential of a whole-cell biocatalyst co-expressing AmDH and Nox for the kinetic resolution of racemic amines.

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“…2 Enzymes offer highly sustainable and efficient synthetic alternatives to current existing chemical methods, 3 providing access to enantiopure amines and derivatives with excellent selectivity. 4 Traditionally, hydrolases have been the biocatalysts of choice for the preparation of optically active amines through classical or dynamic kinetic resolutions of the corresponding racemates, 5 but the development of other classes of enzymes such as amine oxidases, 6 transaminases, 7 berberine bridge enzymes, 8 amine dehydrogenases 9 and more recently imine reductases 10 presents biotransformations as versatile tools for the development of highly efficient asymmetric transformations to prepare these compounds.…”

Section: Introductionmentioning

confidence: 99%

Biocatalytic Transamination for the Asymmetric Synthesis of Pyridylalkylamines. Structural and Activity Features in the Reactivity of Transaminases

et al. 2016

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ABSTRACT. A set of transaminases has been investigated for the biocatalytic amination of 1-(4-chloropyridin-2-yl)alkan-1-ones. The influence of the chain length of the n-1-alkanone at the C-2 position of the pyridine has been studied in the reaction with different (R)-and (S)-selective transaminases. Thus, enantiopure amines were isolated with high purity starting from a wide selection of prochiral ketones. On the one hand excellent yields (97->99% conversion, up to 93% isolated yield) and stereoselectivity values (>99% ee for both amine enantiomers) were found for n-1-alkanone linear short chain substituents such as ethanone or propanone. On the other hand, more hindered substrates were accepted only when using evolved enzymes such as an evolved variant of (R)-Arthrobacter (ArRmut11-TA). An initial common structural feature was the presence of a chlorine atom on the C-4 position of the pyridine core, which was found to increase the reactivity of the starting ketone, giving extra versatility for the introduction of other chemical functionalities towards more complex and applicable organic molecules. In order to gain a deeper understanding about the substrate specificity of different transaminases, additional structural features were considered by variation of the acetyl group position on the pyridine ring and the use of related acetophenone derivatives.

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Development of an Amine Dehydrogenase for Synthesis of Chiral Amines

Cited by 247 publications

References 36 publications

Conversion of alcohols to enantiopure amines through dual-enzyme hydrogen-borrowing cascades

Conversion of alcohols to enantiopure amines through dual-enzyme hydrogen-borrowing cascades

The Kinetic Resolution of Racemic Amines Using a Whole-Cell Biocatalyst Co-Expressing Amine Dehydrogenase and NADH Oxidase

Biocatalytic Transamination for the Asymmetric Synthesis of Pyridylalkylamines. Structural and Activity Features in the Reactivity of Transaminases

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