Kinetic resolution of α-methylbenzylamine by recombinant Pichia pastoris expressing ω-transaminase

Bea, Han-Seop; Seo, Young-Man; Ho, Min; Kim, Byung‐Gee; Yun, Hyungdon

doi:10.1007/s12257-009-3093-1

Cited by 23 publications

(24 citation statements)

References 15 publications

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“…3C). These results are consistent with our previous studies [10][11][12][13][14] and indicated that the removal of acetophenone would be a key factor in the successful enzymatic production of (R)-3-fluoroalanine using ω-TA.…”

Section: Substrate and Product Inhibition Of ω-Ta For Asymmetric Syntsupporting

confidence: 93%

“…The kinetic resolution of various amines by ω-TA have been typically carried out using pyruvate as an amino acceptor to accept the amino group from the amine, which is the main target compound [9][10][11][12][13][14][15][16]. However, in this study F-pyruvate was used as a target substrate and (S)-α-MBA was used as a co-substrate to donate the amino group to Fpyruvate.…”

Section: Substrate and Product Inhibition Of ω-Ta For Asymmetric Syntmentioning

confidence: 99%

“…TAs belonging to subgroup III are called ω-TAs, and include ω-amino acid:pyruvate TA, ornithine TA, 4-aminobutyrate TA, and others [10]. Unlike α-TA, ω-TA can transfer an amino group from a non-α position amino acid (such as 4-aminobutyrate) or an amine compound with no carboxylic group (such as α-MBA) to an amino acceptor (such as 2-ketoglutarate or pyruvate) [10,11].…”

Section: Introductionmentioning

confidence: 99%

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Asymmetric synthesis of (R)-3-fluoroalanine from 3-fluoropyruvate using omega-transaminase

Bea

Lee

Yun

2011

Biotechnol Bioproc E

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In this study, (R)-3-fluoroalanine was asymmetrically synthesized from 3-fluoropyruvate (F-pyruvate) and (S)-α-methylbenzylamine (MBA) using recombinant ω-transaminase (TA) from Vibrio fluvialis JS17. The reaction was severely inhibited by acetophenone (deaminated product of α-MBA). In the presence of 5 mM acetophenone, the reactivity of the enzyme towards F-pyruvate decreased by 78%. To overcome the product inhibition by acetophenone, a biphasic reaction was successfully used. The conversion of F-pyruvate into (R)-3-fluoroalanine (enatiomeric exess (e.e.) > 99%) was about 95% in the biphasic system (75 mM F-pyruvate, 100 mM (S)-α-MBA, and 3.0 U/mL), whereas 31% was obtained without product extraction. The use of racemic α-MBA as an amino donor instead of (S)-α-MBA can reduce the reaction cost and also produce chiral amines through kinetic resolution. When the kinetic resolution of racemic α-MBA (40 mM) was carried out with F-pyruvate (30 mM) and ω-TA (3.0 U/mL) in 100 mM phosphate buffer (pH 7.0), the e.e. of (R)-α-MBA reached 98.4% with 52.2% conversion for 10 h and 21 mM (R)-3-fluoroalanine was produced with 70% conversion and an e.e. > 99%.

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Section: Substrate and Product Inhibition Of ω-Ta For Asymmetric Syntsupporting

confidence: 93%

Section: Substrate and Product Inhibition Of ω-Ta For Asymmetric Syntmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Asymmetric synthesis of (R)-3-fluoroalanine from 3-fluoropyruvate using omega-transaminase

Bea

Lee

Yun

2011

Biotechnol Bioproc E

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“…Another advantage of a whole-cell reaction system is that cofactor-dependent enzymatic reactions can be effectively performed by using endogenous cofactors without adding costly cofactors [30]. Sometimes, a whole cell reaction is less susceptible to enzyme inhibition by substrate and product due to the diffusion barrier of the cell membrane [31].…”

Section: Kinetic Resolution Of Amines Using E Coli Co-expressing Amdmentioning

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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“…The wild-type microorganism containing the desired transaminase may be used, but the more common approach is to clone the desired transaminase into a host vector. For example the use of recombinant E. coli (Ingram et al, 2007;Koszelewski et al, 2009) or Pichia pastoris (Bea et al, 2010) expressing v-transaminase, optionally following a similar approach as seen for cascades, creating so called cassettes over-expressing the production of the enzymes involved in the degradation or recycling of the co-product. Nevertheless, the number of available v-transaminases with a known gene sequence is still rather limited (Clay et al, 2010;Koszelewski et al, 2010b).…”

Section: Whole-cell Biocatalysismentioning

confidence: 99%

Process considerations for the asymmetric synthesis of chiral amines using transaminases

Tufvesson

Lima-Ramos

Jensen

et al. 2011

Biotech & Bioengineering

228

183

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Biocatalytic transamination is being established as key tool for the production of chiral amine pharmaceuticals and precursors due to its excellent enantioselectivity as well as green credentials. Recent examples demonstrate the potential for developing economically competitive processes using a combination of modern biotechnological tools for improving the biocatalyst alongside using process engineering and integrated separation techniques for improving productivities. However, many challenges remain in order for the technology to be more widely applicable, such as technologies for obtaining high yields and productivities when the equilibrium of the desired reaction is unfavorable. This review summarizes both the process challenges and the strategies used to overcome them, and endeavors to describe these and explain their applicability based on physiochemical principles. This article also points to the interaction between the solutions and the need for a process development strategy based on fundamental principles.

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Kinetic resolution of α-methylbenzylamine by recombinant Pichia pastoris expressing ω-transaminase

Cited by 23 publications

References 15 publications

Asymmetric synthesis of (R)-3-fluoroalanine from 3-fluoropyruvate using omega-transaminase

Asymmetric synthesis of (R)-3-fluoroalanine from 3-fluoropyruvate using omega-transaminase

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

Process considerations for the asymmetric synthesis of chiral amines using transaminases

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