Evaluation of a Microreactor for Flow Biocatalysis by Combined Theory and Experiment

Burgahn, Teresa; Pietrek, Philip; Dittmeyer, Roland; Rabe, Kersten S.; Niemeyer, Christof M.

doi:10.1002/cctc.202000145

Cited by 19 publications

(17 citation statements)

References 41 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…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 or simply do not optimize enzyme ratios. − With enzymes increasingly combined into multienzyme cascades, the optimization of enzyme ratios in flow reactors is likely to become a progressively necessary challenge to overcome, requiring an efficient solution to maximize space-time yield (STY) and biocatalyst productivity. , …”

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.

View full text Add to dashboard Cite

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.

show abstract

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.

View full text Add to dashboard Cite

show abstract

“…Table S1: Comparison of spectrophotometric data for Gre2p. Gre2p is HOB-tagged in the work by Burgahn et al 4 Data from this work is identical to data in Table 1 in the main text. Vmax was converted to turnover numbers (kcat), assuming a molecular weight for Gre2p of 39040 Da (eq.…”

Section: Figure S1: Gre2p Catalyzed Conversion Of Nohk To the Diol Gre2p Requires The Nadph Cofactor For Catalysis And The Conversion Of mentioning

confidence: 85%

Towards Reproducible Enzyme Modeling with Isothermal Titration Calorimetry

Ott¹,

Rabe²,

Niemeyer³

et al. 2021

Preprint

Self Cite

View full text Add to dashboard Cite

<div> <p>An experimental workflow to provide detailed information of the molecular mechanisms of enzymes is described. This workflow will help in the application of enzymes in technical processes by providing crucial parameters needed to plan, model and implement biocatalytic processes more efficiently. These parameters are homogeneity of the enzyme sample (HES), kinetic and thermodynamic parameters of enzyme kinetics and binding of reactants to enzymes. The techniques used to measure these properties are dynamic light scattering (DLS), UV-Vis spectrophotometry and isothermal titration calorimetry (ITC) respectively. The workflow is standardized by the use of SOPs and python-scripted data analysis. </p> <p>We have used the NADPH-dependent alcohol dehydrogenase Gre2p as a challenging enzyme to demonstrate the power of this workflow. Our work highlights the utility for combined binding and kinetic studies for such complex multi-substrate reactions and the importance of sample quality control during experiments.</p> </div>

show abstract

“…Flow chemistry has been performed in organic chemistry laboratories in academia and industry for the development, optimization, and intensification of chemical processes for several decades [ 23 ]. More recently, the first biocatalytic reactions performed in microfluidics were reported [ 24 , 25 ]; however, flow biocatalysis today remains in its infancy compared to flow chemistry. This is surprising considering the many advantages microfluidics offer, such as the possibility of continuously operated reactions, much higher surface-to-volume ratios and hence short diffusion paths, and tremendously increased mass and heat transfer rates [ 26 , 27 ].…”

Section: Biocatalysis At the Microscalementioning

confidence: 99%

Two-Phase Biocatalysis in Microfluidic Droplets

et al. 2021

View full text Add to dashboard Cite

This Perspective discusses the literature related to two-phase biocatalysis in microfluidic droplets. Enzymes used as catalysts in biocatalysis are generally less stable in organic media than in their native aqueous environments; however, chemical and pharmaceutical compounds are often insoluble in water. The use of aqueous/organic two-phase media provides a solution to this problem and has therefore become standard practice for multiple biotransformations. In batch, two-phase biocatalysis is limited by mass transport, a limitation that can be overcome with the use of microfluidic systems. Although, two-phase biocatalysis in laminar flow systems has been extensively studied, microfluidic droplets have been primarily used for enzyme screening. In this Perspective, we summarize the limited published work on two-phase biocatalysis in microfluidic droplets and discuss the limitations, challenges, and future perspectives of this technology.

show abstract

Evaluation of a Microreactor for Flow Biocatalysis by Combined Theory and Experiment

Cited by 19 publications

References 41 publications

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

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

Towards Reproducible Enzyme Modeling with Isothermal Titration Calorimetry

Two-Phase Biocatalysis in Microfluidic Droplets

Contact Info

Product

Resources

About