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Highly efficient production of chiral amines in batch and continuous flow by immobilized ω-transaminases on controlled porosity glass metal-ion affinity carrier

In this study, two stereocomplementary ω-transaminases from Arthrobacter sp. (AsR-ωTA) and Chromobacterium violaceum (Cv-ωTA) were immobilized via iron cation affinity binding onto polymer-coated controlled porosity glass beads (EziG™). The immobilization procedure was studied with different types o...

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Autores principales: Böhmer, Wesley, Knaus, Tanja, Volkov, Alexey, Slot, Thierry K., Shiju, N. Raveendran, Engelmark Cassimjee, Karim, Mutti, Francesco G.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7116800/
https://www.ncbi.nlm.nih.gov/pubmed/30550957
http://dx.doi.org/10.1016/j.jbiotec.2018.12.001
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author Böhmer, Wesley
Knaus, Tanja
Volkov, Alexey
Slot, Thierry K.
Shiju, N. Raveendran
Engelmark Cassimjee, Karim
Mutti, Francesco G.
author_facet Böhmer, Wesley
Knaus, Tanja
Volkov, Alexey
Slot, Thierry K.
Shiju, N. Raveendran
Engelmark Cassimjee, Karim
Mutti, Francesco G.
author_sort Böhmer, Wesley
collection PubMed
description In this study, two stereocomplementary ω-transaminases from Arthrobacter sp. (AsR-ωTA) and Chromobacterium violaceum (Cv-ωTA) were immobilized via iron cation affinity binding onto polymer-coated controlled porosity glass beads (EziG™). The immobilization procedure was studied with different types of carrier materials and immobilization buffers of varying compositions, concentrations, pHs and cofactor (PLP) concentrations. Notably, concentrations of PLP above 0.1 mM were correlated with a dramatic decrease of the immobilization yield. The highest catalytic activity, along with quantitative immobilization, was obtained in MOPS buffer (100mM, pH 8.0, PLP 0.1 mM, incubation time 2 h). Leaching of the immobilized enzyme was not observed within 3 days of incubation. EziG-immobilized AsR-ωTA and Cv-ωTA retained elevated activity when tested for the kinetic resolution of rac-α-methylbenzylamine (rac-α-MBA) in single batch experiments. Recycling studies demonstrated that immobilized EziG(3)-AsR-ωTA could be recycled for at least 16 consecutive cycles (15min per cycle) and always affording quantitative conversion (TON ca. 14,400). Finally, the kinetic resolution of rac-α-MBA with EziG(3)-AsR-ωTA was tested in a continuous flow packed-bed reactor (157 µL reactor volume), which produced more than 5g of (S)-α-MBA (> 49% conversion, > 99% ee) in 96 h with no detectable loss of catalytic activity. The calculated TON was more than 110,000 along with a space-time yield of 335 g L(−1)h(−1).
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spelling pubmed-71168002021-02-22 Highly efficient production of chiral amines in batch and continuous flow by immobilized ω-transaminases on controlled porosity glass metal-ion affinity carrier Böhmer, Wesley Knaus, Tanja Volkov, Alexey Slot, Thierry K. Shiju, N. Raveendran Engelmark Cassimjee, Karim Mutti, Francesco G. J Biotechnol Article In this study, two stereocomplementary ω-transaminases from Arthrobacter sp. (AsR-ωTA) and Chromobacterium violaceum (Cv-ωTA) were immobilized via iron cation affinity binding onto polymer-coated controlled porosity glass beads (EziG™). The immobilization procedure was studied with different types of carrier materials and immobilization buffers of varying compositions, concentrations, pHs and cofactor (PLP) concentrations. Notably, concentrations of PLP above 0.1 mM were correlated with a dramatic decrease of the immobilization yield. The highest catalytic activity, along with quantitative immobilization, was obtained in MOPS buffer (100mM, pH 8.0, PLP 0.1 mM, incubation time 2 h). Leaching of the immobilized enzyme was not observed within 3 days of incubation. EziG-immobilized AsR-ωTA and Cv-ωTA retained elevated activity when tested for the kinetic resolution of rac-α-methylbenzylamine (rac-α-MBA) in single batch experiments. Recycling studies demonstrated that immobilized EziG(3)-AsR-ωTA could be recycled for at least 16 consecutive cycles (15min per cycle) and always affording quantitative conversion (TON ca. 14,400). Finally, the kinetic resolution of rac-α-MBA with EziG(3)-AsR-ωTA was tested in a continuous flow packed-bed reactor (157 µL reactor volume), which produced more than 5g of (S)-α-MBA (> 49% conversion, > 99% ee) in 96 h with no detectable loss of catalytic activity. The calculated TON was more than 110,000 along with a space-time yield of 335 g L(−1)h(−1). 2019-02-10 2018-12-11 /pmc/articles/PMC7116800/ /pubmed/30550957 http://dx.doi.org/10.1016/j.jbiotec.2018.12.001 Text en https://creativecommons.org/licenses/BY/4.0/ This is an open access article under the CC BY license (https://creativecommons.org/licenses/BY/4.0/).
spellingShingle Article
Böhmer, Wesley
Knaus, Tanja
Volkov, Alexey
Slot, Thierry K.
Shiju, N. Raveendran
Engelmark Cassimjee, Karim
Mutti, Francesco G.
Highly efficient production of chiral amines in batch and continuous flow by immobilized ω-transaminases on controlled porosity glass metal-ion affinity carrier
title Highly efficient production of chiral amines in batch and continuous flow by immobilized ω-transaminases on controlled porosity glass metal-ion affinity carrier
title_full Highly efficient production of chiral amines in batch and continuous flow by immobilized ω-transaminases on controlled porosity glass metal-ion affinity carrier
title_fullStr Highly efficient production of chiral amines in batch and continuous flow by immobilized ω-transaminases on controlled porosity glass metal-ion affinity carrier
title_full_unstemmed Highly efficient production of chiral amines in batch and continuous flow by immobilized ω-transaminases on controlled porosity glass metal-ion affinity carrier
title_short Highly efficient production of chiral amines in batch and continuous flow by immobilized ω-transaminases on controlled porosity glass metal-ion affinity carrier
title_sort highly efficient production of chiral amines in batch and continuous flow by immobilized ω-transaminases on controlled porosity glass metal-ion affinity carrier
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7116800/
https://www.ncbi.nlm.nih.gov/pubmed/30550957
http://dx.doi.org/10.1016/j.jbiotec.2018.12.001
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