Carbon as a Simple Support for Redox Biocatalysis in Continuous Flow

Poznansky, Barnabas; Thompson, Lisa A.; Warren, Sarah A.; Reeve, Holly A.; Vincent, Kylie A.

doi:10.1021/acs.oprd.9b00410

Cited by 14 publications

(9 citation statements)

References 41 publications

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“…Similar results were observed by Poznansky et al for the NADH-dependent bio catalytic conversion of pyruvate to lactate. In this case, the STY was 5.52 g L –1 d –1 for the batch process, while in the flow process, a STY of 269 g L –1 d –1 was reported, thus demonstrating that higher productivity can be achieved using the continuous-flow reactor …”

Section: Resultsmentioning

confidence: 73%

“…In this case, the STY was 5.52 g L −1 d −1 for the batch process, while in the flow process, a STY of 269 g L −1 d −1 was reported, thus demonstrating that higher productivity can be achieved using the continuous-flow reactor. 33 Comparative Evaluation of Alternative Immobilization Systems. For the purpose of benchmarking, different immobilization platforms, namely, CLEA (cross-linked enzyme aggregates) epoxy support, ECR8285, (Purolite, U.K.) were compared on the immobilization efficiency on apKRED-9 along with amino support LX1000HA and CEP platform.…”

Section: ■ Results and Discussionmentioning

confidence: 99%

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Highly Efficient Production of Optically Active (R)-Tetrahydrothiophene-3-ol in Batch and Continuous Flow by Using Immobilized Ketoreductase

Williams¹,

Cui²,

Zhao³

et al. 2022

Org. Process Res. Dev.

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In this work, an engineered ketoreductase, apKRED-9, derived from Acetobacter pasteurianus 386B was successfully immobilized on two platforms, namely, glutaraldehyde-activated amino polymer beads, LX1000 HA, and cofactor enriched poly(ethylenimine) (CEP) mediated coaggregation followed by glutaraldehyde cross-linking, respectively. The enzyme apKRED-9 immobilized on LX1000HA was evaluated in a packed bed reactor (PBR) for continuous-flow synthesis of (R)-tetrahydrothiophene-3-ol from 3-keto tetrahydrothiophene in an aqueous-isopropanol mixture, while the enzyme apKRED-9 immobilized on CEP was tested in batch mode until pilot scale for the same reaction. The long-term operational stability of the enzyme in both continuous-flow and batch modes was demonstrated, with high conversion of >99.0% and ee > 99.5% in both the cases. From the pilot-scale application of apKRED-9-CEP, (R)-tetrahydrothiophene-3-ol was obtained (118.0 g, GC purity 99.9%, chiral purity ee 99.9% and yield 76.3%). In the PBR flow reactor, the productivity in terms of space time yield (STY) 729 g L–1 d–1 was achieved with 64 h of continuous usage. Based on performance metrics, both platforms are scalable and reproducible, while CEP offers additional advantages on effective cost and adaptability to other enzymes.

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Section: Resultsmentioning

confidence: 73%

Section: ■ Results and Discussionmentioning

confidence: 99%

Highly Efficient Production of Optically Active (R)-Tetrahydrothiophene-3-ol in Batch and Continuous Flow by Using Immobilized Ketoreductase

Williams¹,

Cui²,

Zhao³

et al. 2022

Org. Process Res. Dev.

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“…A recent study demonstrated DoE as an effective method for optimizing multienzyme biocatalytic flow reactions, yet avoided optimization of the enzyme loadings despite a suggestion that this would offer further improvements. 37 Evidently, optimizing enzyme ratios is not a trivial problem when using immobilized enzymes in flow, even when applying DoE to reduce the necessary experimentation. Others attempt to tackle this challenge through the selection of the best ratio from a few arbitrary trials 38 or simply do not optimize enzyme ratios.…”

Section: ■ Introductionmentioning

confidence: 99%

Rapid Model-Based Optimization of a Two-Enzyme System for Continuous Reductive Amination in Flow

Finnigan

Citoler

Cosgrove

et al. 2020

Org. Process Res. Dev.

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Enzymes are increasingly combined into multienzyme systems for cost and productivity benefits. Further advantages can be gained through the use of immobilized enzymes, allowing continuous biotransformations in flow. However, the optimization of such multienzyme systems is challenging, particularly where immobilized enzymes are used. Here, we meet this challenge using both mechanistic and empirical modeling to optimize a reaction involving a reductive aminase and a glucose dehydrogenase for continuous biocatalytic reductive amination in flow. Crucially, the construction of the mechanistic model was achieved quickly, with only a few important parameters determined experimentally, and ensemble modeling used to facilitate the use of estimates or literature values. Upon reaching the limits of the mechanistic model's capabilities, we show that solution space can be further explored using a definitive screening design to generate an empirical model of the reaction, using the mechanistic model's prediction as a starting point. We demonstrate the use of this empirical model to design optimal processes for either high productivity or to minimize necessary cofactor and cosubstrate concentrations. Our results demonstrate that the synergistic use of both mechanistic and empirical modeling offers a route for rapid optimization of multienzyme systems of immobilized enzymes in flow with minimal experimental effort.

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“…First, the catalysts SH/C and SH + Hyd1/C were tested in H 2driven NADH-generating assays to determine the batch catalyst performance for the NADH generation step, Figure 3A. The enzymes were immobilized on carbon black (Black Pearls 2000, BP2000, ∼10 nm), which we have previously found to be a versatile enzyme support, according to the immobilization protocol described in General Enzyme Immobilization Procedure for Batch Studies (Poznansky et al, 2020). The enzyme-modified carbon particles were transferred into 800 μl of H 2 -saturated NAD + (0.1 mM) in Tris-HCl (100 mM, pH 8.0) in a cuvette, then the reaction was monitored in situ by UV-Vis.…”

Section: Heterogeneous Biocatalysts For H 2 -Driven Nadh Recycling In Batch Reactionsmentioning

confidence: 99%

Boosting the Productivity of H2-Driven Biocatalysis in a Commercial Hydrogenation Flow Reactor Using H2 From Water Electrolysis

et al. 2021

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Translation of redox biocatalysis into a commercial hydrogenation flow reactor, with in-built electrolytic H2 generation, was achieved using immobilized enzyme systems. Carbon-supported biocatalysts were first tested in batch mode, and were then transferred into continuous flow columns for H2-driven, NADH-dependent asymmetric ketone reductions. The biocatalysts were thus handled comparably to heterogeneous metal catalysts, but operated at room temperature and 1–50 bar H2, highlighting that biocatalytic strategies enable implementation of hydrogenation reactions under mild–moderate conditions. Continuous flow reactions were demonstrated as a strategy for process intensification; high conversions were achieved in short residence times, with a high biocatalyst turnover frequency and productivity. These results show the prospect of using enzymes in reactor infrastructure designed for conventional heterogeneous hydrogenations.

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Carbon as a Simple Support for Redox Biocatalysis in Continuous Flow

Cited by 14 publications

References 41 publications

Highly Efficient Production of Optically Active (R)-Tetrahydrothiophene-3-ol in Batch and Continuous Flow by Using Immobilized Ketoreductase

Highly Efficient Production of Optically Active (R)-Tetrahydrothiophene-3-ol in Batch and Continuous Flow by Using Immobilized Ketoreductase

Rapid Model-Based Optimization of a Two-Enzyme System for Continuous Reductive Amination in Flow

Boosting the Productivity of H2-Driven Biocatalysis in a Commercial Hydrogenation Flow Reactor Using H2 From Water Electrolysis

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