A Process Concept for High-Purity Production of Amines by Transaminase-Catalyzed Asymmetric Synthesis: Combining Enzyme Cascade and Membrane-Assisted ISPR

Börner, Tim; Rehn, Gustav; Grey, Carl; Adlercreutz, Patrick

doi:10.1021/acs.oprd.5b00055

Cited by 40 publications

(38 citation statements)

References 42 publications

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“…Further interpretation of the data resulted in a novel process concept for the production of pure amines using ALA as more efficient amine donor (than IPA). Together with a selective extraction unit, a pure and enriched amine product was obtained . The example of a successful process design based on kinetic data validates the usefulness of the automated HPLC platform in terms of significantly reduced labor, time and deviation.…”

Section: Resultsmentioning

confidence: 61%

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Generic HPLC platform for automated enzyme reaction monitoring: Advancing the assay toolbox for transaminases and other PLP‐dependent enzymes

Börner

Grey

Adlercreutz

2016

Biotechnology Journal

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Methods for rapid and direct quantification of enzyme kinetics independent of the substrate stand in high demand for both fundamental research and bioprocess development. This study addresses the need for a generic method by developing an automated, standardizable HPLC platform monitoring reaction progress in near real-time. The method was applied to amine transaminase (ATA) catalyzed reactions intensifying process development for chiral amine synthesis. Autosampler-assisted pipetting facilitates integrated mixing and sampling under controlled temperature. Crude enzyme formulations in high and low substrate concentrations can be employed. Sequential, small (1 µL) sample injections and immediate detection after separation permits fast reaction monitoring with excellent sensitivity, accuracy and reproducibility. Due to its modular design, different chromatographic techniques, e.g. reverse phase and size exclusion chromatography (SEC) can be employed. A novel assay for pyridoxal 5'-phosphate-dependent enzymes is presented using SEC for direct monitoring of enzyme-bound and free reaction intermediates. Time-resolved changes of the different cofactor states, e.g. pyridoxal 5'-phosphate, pyridoxamine 5'-phosphate and the internal aldimine were traced in both half reactions. The combination of the automated HPLC platform with SEC offers a method for substrate-independent screening, which renders a missing piece in the assay and screening toolbox for ATAs and other PLP-dependent enzymes.

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Section: Resultsmentioning

confidence: 61%

“…Experimental times, see Supporting information, Table S3. Data in ( A ), ( E ), ( D ) and the ALA data in ( C ) were adopted from .…”

Section: Resultsmentioning

confidence: 99%

Generic HPLC platform for automated enzyme reaction monitoring: Advancing the assay toolbox for transaminases and other PLP‐dependent enzymes

Börner

Grey

Adlercreutz

2016

Biotechnology Journal

Self Cite

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“…Specifically, membrane‐based three liquid phase (3LP) systems were developed by filling the pores of an hydrophobic hollow fibre membrane contactor with an hydrophobic solvent. This operation allowed to physically separate the reaction and the extraction aqueous solutions by using a supported liquid membrane . The only 3LP system developed without membranes was reported by Yun and Kim, 2008.…”

Section: Figurementioning

confidence: 99%

Three‐liquid‐phase Spinning Reactor for the Transaminase‐catalyzed Synthesis and Recovery of a Chiral Amine

et al. 2020

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A device for the transaminase‐catalysed synthesis combined with continuous recovery of chiral amines was designed. The system enabled the separation of the reaction components in three liquid phases: a reaction phase, an organic solvent phase (where the poorly water soluble ketone substrate was supplied), and an aqueous extraction phase for continuous product recovery. The transaminase‐mediated asymmetric synthesis of (S)‐1‐methyl‐3‐phenylpropylamine was employed as model reaction. Factors influencing the performance of the system, such as reactor geometry, working volumes and operating parameters, were investigated. Specifically, reaction yield and product recovery were enhanced by i) reducing the thickness of the reaction phase, while continuously stirring and ii) reducing the volume of the extraction phase. Under the optimal condition tested, 85 % of the product formed was extracted and a product concentration value of 9 g/L was reached. However, co‐extraction of the unreacted amine donor (17 %) was observed. Advantages and drawbacks of this process compared to existing technologies, as well as possible optimization strategies are discussed.

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“…In another study with the amine transaminase catalyzed synthesis of chiral amines, alanine was investigated as amine donor for the reductive amination of a poorly water-soluble ketone (4-phenyl-2-butanone) in a combined in situ product removal approach using liquid membrane extraction together with an enzyme cascade. This strategy afforded very high product purity (>98%) with an integrated enrichment step and eliminated product as well as co-product inhibition [120].…”

Section: Liquid Membranesmentioning

confidence: 99%

Chiral Separation in Preparative Scale: A Brief Overview of Membranes as Tools for Enantiomeric Separation

2017

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Given the importance of chirality in the biological response, regulators, industries and researchers require chiral compounds in their enantiomeric pure form. Therefore, the approach to separate enantiomers in preparative scale needs to be fast, easy to operate, low cost and allow obtaining the enantiomers at high level of optical purity. A variety of methodologies to separate enantiomers in preparative scale is described, but most of them are expensive or with restricted applicability. However, the use of membranes have been pointed out as a promising methodology for scale-up enantiomeric separation due to the low energy consumption, continuous operability, variety of materials and supports, simplicity, eco-friendly and the possibility to be integrated into other separation processes. Different types of membranes (solid and liquid) have been developed and may provide applicability in multi-milligram and industrial scales. In this brief overview, the different types and chemical nature of membranes are described, showing their advantages and drawbacks. Recent applications of enantiomeric separations of pharmaceuticals, amines and amino acids were reported.

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A Process Concept for High-Purity Production of Amines by Transaminase-Catalyzed Asymmetric Synthesis: Combining Enzyme Cascade and Membrane-Assisted ISPR

Cited by 40 publications

References 42 publications

Generic HPLC platform for automated enzyme reaction monitoring: Advancing the assay toolbox for transaminases and other PLP‐dependent enzymes

Generic HPLC platform for automated enzyme reaction monitoring: Advancing the assay toolbox for transaminases and other PLP‐dependent enzymes

Three‐liquid‐phase Spinning Reactor for the Transaminase‐catalyzed Synthesis and Recovery of a Chiral Amine

Chiral Separation in Preparative Scale: A Brief Overview of Membranes as Tools for Enantiomeric Separation

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