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

ABSTRACT: In this study, two stereocomplementary ?-transaminases from Arthrobacter sp. (AsR-?TA) and Chromobacterium violaceum (Cv-?TA) 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 (100?mM, 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-?TA and Cv-?TA retained elevated activity when tested for the kinetic resolution of rac-?-methylbenzylamine (rac-?-MBA) in single batch experiments. Recycling studies demonstrated that immobilized EziG³-AsR-?TA could be recycled for at least 16 consecutive cycles (15?min per cycle) and always affording quantitative conversion (TON ca. 14,400). Finally, the kinetic resolution of rac-?-MBA with EziG³-AsR-?TA was tested in a continuous flow packed-bed reactor (157??L reactor volume), which produced more than 5?g of (S)-?-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.

SUBMITTER: Bohmer W

PROVIDER: S-EPMC7116800 | biostudies-literature | 2019 Feb

REPOSITORIES: biostudies-literature

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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.

Böhmer Wesley W Knaus Tanja T Volkov Alexey A Slot Thierry K TK Shiju N Raveendran NR Engelmark Cassimjee Karim K Mutti Francesco G FG

Journal of biotechnology 20181211

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<sup>™</sup>). 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 wi ...[more]

PMID: 30550957

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Project description:The current investigation is taken up with the aim of studying repeated batch and continuous degradation of Endosulfan, using Ca-alginate immobilized cells of Pseudomonas aeruginosa isolated from an agricultural soil. The work involves the study of genes and enzymes involved in the degradation of the pesticide and was carried out with an objective of reducing the toxicity of Endosulfan by degrading it to less toxic metabolites. The long-term stability of Endosulfan degradation was studied during its repeated batch degradation, carried out over a period of 35 days. Immobilized cells of Ps. aeruginosa were able to show 60 % degradation of Endosulfan at the end of the 35th cycle with a cell leakage of 642 × 104 Cfu/mL. During continuous treatment, with 2 % concentration of Endosulfan, 100 % degradation was recorded up to 100 mL/h flow rate and with 10 % concentration of the Endosulfan, and 100 and 85 % degradation was recorded at 20 mL/h flow rate and 100 mL/h flow rate, respectively. After degradation of Endosulfan, products were extracted from a large amount of spent medium using two volumes of ethyl acetate and subjected to the LC-MS analysis. Endosulfan lactone and Endosulfan ether were the products of degradation detected by the LCMS analysis. Plasmid curing experiments indicated that genes responsible for the degradation of Endosulfan are present on the chromosome and not on the plasmid, as growth of Ps. aeruginosa was observed on modified non-sulfur medium with Endosulfan after the plasmid was cured with ethidium bromide. The results of PCR indicated that there is no amplified product of ~1350 bp expected for esd gene, in Ps. aeruginosa, although there were some non-specific bands. Enzymatic degradation studies indicated that the enzymes involved in the degradation of Endosulfan are intracellular. With this investigation, it was indicated that immobilized cells of Ps. aeruginosa have the potential to be used in the bioremediation of water contaminated with Endosulfan.

Dataset Information

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

Publications

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

Similar Datasets

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