A co-expression system to shift the equilibrium of transamination reactions toward the synthesis of enantiomerically pure amines

Telzerow, Aline; Hobisch, Markus; Müller, Monika; Schürmann, Martin; Schwab, Helmut; Steiner, Kerstin

doi:10.1016/j.mcat.2019.04.006

Cited by 9 publications

(8 citation statements)

References 26 publications

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“…Single whole-cell biocatalysts co-expressing multiple enzymes are advantageous in terms of convenient handling and enhanced mass transfer efficiency (Tufvesson et al, 2014;Iglesias et al, 2017;Telzerow et al, 2019). In a previous study, we reported that ratio of esterase to TA should be balanced for fine-tuning of the activity (TAIC-Est PS).…”

Section: Application Of Promoter Engineering Strategy For the Synthesis Of Sitagliptin Intermediatementioning

confidence: 99%

Synthesis of Sitagliptin Intermediate by a Multi-Enzymatic Cascade System Using Lipase and Transaminase With Benzylamine as an Amino Donor

Khobragade

Sarak

Pagar

et al. 2021

Front. Bioeng. Biotechnol.

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Herein, we report the development of a multi-enzyme cascade using transaminase (TA), esterase, aldehyde reductase (AHR), and formate dehydrogenase (FDH), using benzylamine as an amino donor to synthesize the industrially important compound sitagliptin intermediate. A panel of 16 TAs was screened using ethyl 3-oxo-4-(2,4,5-trifluorophenyl) butanoate as a substrate (1). Amongst these enzymes, TA from Roseomonas deserti (TARO) was found to be the most suitable, showing the highest activity towards benzylamine (∼70%). The inhibitory effect of benzaldehyde was resolved by using AHR from Synechocystis sp. and FDH from Pseudomonas sp., which catalyzed the conversion of benzaldehyde to benzyl alcohol at the expense of NAD(P)H. Reaction parameters, such as pH, buffer system, and concentration of amino donor, were optimized. A single whole-cell system was developed for co-expressing TARO and esterase, and the promoter engineering strategy was adopted to control the expression level of each biocatalyst. The whole-cell reactions were performed with varying substrate concentrations (10–100 mM), resulting in excellent conversions (ranging from 72 to 91%) into the desired product. Finally, the applicability of this cascade was highlighted on Gram scale, indicating production of 70% of the sitagliptin intermediate with 61% isolated yield. The protocol reported herein may be considered an alternative to existing methods with respect to the use of cheaper amine donors as well as improved synthesis of (R) and (S) enantiomers with the use of non-chiral amino donors.

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Section: Application Of Promoter Engineering Strategy For the Synthesis Of Sitagliptin Intermediatementioning

confidence: 99%

Synthesis of Sitagliptin Intermediate by a Multi-Enzymatic Cascade System Using Lipase and Transaminase With Benzylamine as an Amino Donor

Khobragade

Sarak

Pagar

et al. 2021

Front. Bioeng. Biotechnol.

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“…A very common approach is the removal of unwanted co-products or regeneration of cofactors and/ or co-substrates (to drive equilibria towards the desired product; e.g. [31,32]). Cofactor regeneration may also be achieved by using a parallel cascade.…”

Section: Uses and Categorisation Of Cascade Reactionsmentioning

confidence: 99%

Using enzyme cascades in biocatalysis: Highlight on transaminases and carboxylic acid reductases

Cutlan

Rose²,

Isupov³

et al. 2020

Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics

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This is a PDF file of an article that has undergone enhancements after acceptance, such as the addition of a cover page and metadata, and formatting for readability, but it is not yet the definitive version of record. This version will undergo additional copyediting, typesetting and review before it is published in its final form, but we are providing this version to give early visibility of the article. Please note that, during the production process, errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.

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“…A small selection of ion exchange materials was mentioned in the past within biocatalytic processes, but their applications are still somehow limited with a few noticeable exceptions. [47][48][49][50] In this study, the ion exchange resins are applied to the IRED-reaction system to selectively remove the reactants from the reaction solution without any significant changes to the aqueous reaction medium, which eventually minimizes the processing steps during a potential DSP. In addition, this strategy opens the possibility of simply reusing the biocatalyst without any potentially harmful biocatalyst immobilization steps.…”

Section: Introductionmentioning

confidence: 99%

“…This study aims at presenting the utilization of ion exchange resins as an easy option to perform the product isolation from IRED‐catalyzed reaction systems, including its potential usage on a preparative scale. A small selection of ion exchange materials was mentioned in the past within biocatalytic processes, but their applications are still somehow limited with a few noticeable exceptions 47‐50 . In this study, the ion exchange resins are applied to the IRED‐reaction system to selectively remove the reactants from the reaction solution without any significant changes to the aqueous reaction medium, which eventually minimizes the processing steps during a potential DSP.…”

Section: Introductionmentioning

confidence: 99%

Integration of ion exchange resin materials for a downstream‐processing approach of an imine reductase‐catalyzed reaction

Meyer

Brundiek²,

Langermann

2020

Biotechnology Progress

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In this study, an ion exchange resin-based downstream-processing concept for imine reductase (IRED)-catalyzed reactions was investigated. As a model reaction, 2-methylpyrroline was converted to its corresponding product (S)-2-methylpyrrolidine with >99% of conversion by the (S)-selective IRED from Paenibacillus elgii B69. Under optimized reaction conditions full conversion was achieved using a substrate concentration of 150 and 500 mmol/L of D-glucose. Seven commercially available cation-and anion-exchange resins were studied with respect to their ability to recover the product from the reaction solution. Without any pretreatment, cation-exchange resins Amberlite IR-120(H), IRN-150, Dowex Monosphere 650C, and Dowex Marathon MSC showed high recovery capacities (up to >90%). A 150-ml preparative scale reaction was performed yielding 1 g hydrochloride salt product with >99% purity. Any further purification steps, for example, by column chromatography or recrystallization, were not required.

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A co-expression system to shift the equilibrium of transamination reactions toward the synthesis of enantiomerically pure amines

Cited by 9 publications

References 26 publications

Synthesis of Sitagliptin Intermediate by a Multi-Enzymatic Cascade System Using Lipase and Transaminase With Benzylamine as an Amino Donor

Synthesis of Sitagliptin Intermediate by a Multi-Enzymatic Cascade System Using Lipase and Transaminase With Benzylamine as an Amino Donor

Using enzyme cascades in biocatalysis: Highlight on transaminases and carboxylic acid reductases

Integration of ion exchange resin materials for a downstream‐processing approach of an imine reductase‐catalyzed reaction

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