Reaching New Biocatalytic Reactivity Using Continuous Flow Reactors

Cosgrove, Sebastian C.; Mattey, Ashley P.

doi:10.1002/chem.202103607

Cited by 23 publications

(19 citation statements)

References 83 publications

(89 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Besides screening of suitable enzymes for a particular transformation, investigation of its substrate scope, as well as reaction optimization and analysis of enzyme engineering approaches via directed evolution, will in particular greatly benefit from automated platforms to rapidly screen hundreds of different enzyme mutants. 73 The fact that our automatable chip-MS approach requires very little material is a strong advantage because genetically modified enzymes, in particular, are scarce and precious. To test the DMF-MS chip for such enzymatic transformations, we chose the biocatalytic bromination of aromatic compounds as a model reaction applying the vanadium-dependent haloperoxidase isolated from Acaryochloris marina (AmVHPO) recently established in the Gulder group.…”

Section: ■ Results and Discussionmentioning

confidence: 99%

On-the-Fly Mass Spectrometry in Digital Microfluidics Enabled by a Microspray Hole: Toward Multidimensional Reaction Monitoring in Automated Synthesis Platforms

et al. 2022

View full text Add to dashboard Cite

We report an approach for the online coupling of digital microfluidics (DMF) with mass spectrometry (MS) using a chip-integrated microspray hole (μSH). The technique uses an adapted electrostatic spray ionization (ESTASI) method to spray a portion of a sample droplet through a microhole in the cover plate, allowing its chemical content to be analyzed by MS. This eliminates the need for chip disassembly or the introduction of capillary emitters for MS analysis, as required by state-of-the-art. For the first time, this allows the essential advantage of a DMF devicefree droplet movementto be retained during MS analysis. The broad applicability of the developed seamless coupling of DMF and mass spectrometry was successfully applied to the study of various on-chip organic syntheses as well as protein and peptide analysis. In the case of a Hantzsch synthesis, we were able to show that the method is very well suited for monitoring even rapid chemical reactions that are completed in a few seconds. In addition, the strength of the low resource consumption in such on-chip microsyntheses was demonstrated by the example of enzymatic brominations, for which only a minute amount of a special haloperoxidase is required in the droplet. The unique selling point of this approach is that the analyzed droplet remains completely movable after the MS measurement and is available for subsequent on-DMF chip processes. This is illustrated here for the example of MS analysis of the starting materials in the corresponding droplets before they are combined to investigate the reaction progress by DMF-MS further. This technology enables the ongoing and almost unlimited tracking of multistep chemical processes in a DMF chip and offers exciting prospects for transforming digital microfluidics into automated synthesis platforms.

show abstract

Section: ■ Results and Discussionmentioning

confidence: 99%

On-the-Fly Mass Spectrometry in Digital Microfluidics Enabled by a Microspray Hole: Toward Multidimensional Reaction Monitoring in Automated Synthesis Platforms

et al. 2022

View full text Add to dashboard Cite

show abstract

“…Under these systems, reactions are conducted in a continuous stream and the different reaction steps can be fully segregated. Despite being a relatively new area of study, it has become very popular in recent years [72] . Examples on the application of continuous flow setups in multistep enzymatic cascades have led to higher productivities and easier downstream processes, and have demonstrated that previously incompatible cascades can now be run efficiently [73] .…”

Section: Discussionmentioning

confidence: 99%

“…Despite being a relatively new area of study, it has become very popular in recent years. [72] Examples on the application of continuous flow setups in multistep enzymatic cascades have led to higher productivities and easier downstream processes, and have demonstrated that previously incompatible cascades can now be run efficiently. [73] We clearly believe that the combination of continuous flow systems and chemoenzymatic cascades will lead to a greater uptake and use of these multistep processes at scale.…”

Section: Discussionmentioning

confidence: 99%

Construction of Chemoenzymatic Linear Cascades for the Synthesis of Chiral Compounds

Ascaso-Alegre

MANGAS

2022

Eur J Org Chem

View full text Add to dashboard Cite

Juan Mangas Sanchez was nominated to be part of this collection by EurJOC Board Member Silvia Osuna OliverasInspired by nature, synthetic chemists try to mimic the efficient metabolic networks in living organisms to build complex molecules by combining different types of catalysts in the same reaction vessel. These multistep cascade processes provide many advantages to synthetic procedures, resulting in higher productivities with lower waste generation and cost. However, combining different chemo-and biocatalysts can be challenging as reaction conditions might differ greatly. As a highly multidisciplinary field that benefits from advances in chemical catalysis, molecular biology and reaction engineering, this area of study is rapidly progressing. In this Review, we highlight recent trends and advances in the construction of multistep chemoenzymatic one-pot cascades to access chiral compounds as well as the different strategies to solve current challenges in the field.

show abstract

“…Flow chemistry is an important tool for process intensification and has been reported to reduce reaction times concomitantly improving enantioselectivities by suppressing undesired side reactions. Additionally, it enhances safety and among others reduces reaction volumes [25][26][27][28][29][30][31][32]. At the same time, it requires the enzymes to be immobilised, which often improves enzyme stability.…”

Section: Introductionmentioning

confidence: 99%

Batch and Flow Nitroaldol Synthesis Catalysed by Granulicella tundricola Hydroxynitrile Lyase Immobilised on Celite R-633

et al. 2022

View full text Add to dashboard Cite

Granulicella tundricola hydroxynitrile lyase (GtHNL) catalyses the synthesis of chiral (R)-cyanohydrins and (R)-β-nitro alcohols. The triple variant GtHNL-A40H/V42T/Q110H (GtHNL-3V) was immobilised on Celite R-633 and used in monophasic MTBE saturated with 100 mM KPi buffer pH 7 for the synthesis of (R)-2-nitro-1-phenylethanol (NPE) in batch and continuous flow systems. Nitromethane was used as a nucleophile. A total of 82% of (R)-NPE and excellent enantioselectivity (>99%) were achieved in the batch system after 24 hours of reaction time. GtHNL-3V on Celite R-633 was successfully recycled five times. During more recycling steps a significant decrease in yield was observed while the enantioselectivity remained excellent over eight cycles. The use of a flow system enabled the continuous synthesis of (R)-NPE. A total of 15% formation of (R)-NPE was reached using a flow rate of 0.1 mL min−1; unfortunately, the enzyme was not stable, and the yield decreased to 4% after 4 hours on stream. A similar yield was observed during 15 hours at a rate of 0.01 mL min−1. Surprisingly the use of a continuous flow system did not facilitate the process intensification. In fact, the batch system displayed a space-time-yield (STY/mgenzyme) of 0.10 g L−1 h−1 mgenzyme−1 whereas the flow system displayed 0.02 and 0.003 g L−1 h−1 mgenzyme−1 at 0.1 and 0.01 mL min−1, respectively. In general, the addition of 1 M nitromethane potentially changed the polarity of the reaction mixture affecting the stability of Celite-GtHNL-3V. The nature of the batch system maintained the reaction conditions better than the flow system. The higher yield and productivity observed for the batch system show that it is a superior system for the synthesis of (R)-NPE compared with the flow approach.

show abstract

Reaching New Biocatalytic Reactivity Using Continuous Flow Reactors

Cited by 23 publications

References 83 publications

On-the-Fly Mass Spectrometry in Digital Microfluidics Enabled by a Microspray Hole: Toward Multidimensional Reaction Monitoring in Automated Synthesis Platforms

On-the-Fly Mass Spectrometry in Digital Microfluidics Enabled by a Microspray Hole: Toward Multidimensional Reaction Monitoring in Automated Synthesis Platforms

Construction of Chemoenzymatic Linear Cascades for the Synthesis of Chiral Compounds

Batch and Flow Nitroaldol Synthesis Catalysed by Granulicella tundricola Hydroxynitrile Lyase Immobilised on Celite R-633

Contact Info

Product

Resources

About