Cell-free enzyme cascades — application and transition from development to industrial implementation

Teshima, Mariko; Willers, Vivian Pascal; Sieber, Volker

doi:10.1016/j.copbio.2022.102868

Cited by 18 publications

(8 citation statements)

References 53 publications

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“…86 The use of cell-free biocatalysis in multistep enzymatic synthesis of metabolites and natural products is not only of interest for the early development phase, but also for reaction engineering and the translation to sustainable manufacturing processes. 87,88…”

Section: Reaction Chemistry and Engineering Reviewmentioning

confidence: 99%

Route selection and reaction engineering for sustainable metabolite synthesis

Wohlgemuth

2023

React. Chem. Eng.

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Section: Reaction Chemistry and Engineering Reviewmentioning

confidence: 99%

Route selection and reaction engineering for sustainable metabolite synthesis

Wohlgemuth

2023

React. Chem. Eng.

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“…The use of immobilized enzymatic cascades in packed-bed reactors is of great interest due to its potential for enhanced space-time yields, high specificity, and sustainable production (less waste generation since less solvent and chemicals are required) of a variety of chemicals [5][6][7][8][9][10][11][12]. These multi-enzymatic pro-cesses, where several enzymes perform sequential reactions without the isolation and purification of intermediates, have experienced rapid growth in scientific and industrial applications [13][14][15][16][17][18][19][20][21][22][23][24][25][26].…”

Section: Introductionmentioning

confidence: 99%

Modeling Enzymatic Cascade Reactions Immobilized in Plug‐Flow Reactors for Flow Biocatalysis

Paschalidis,

Arana‐Peña,

Sieber

et al. 2024

Chemie Ingenieur Technik

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Flow biocatalysis has emerged as a promising technology for the sustainable production of chemicals. Multiple immobilized enzymes are often used as biocatalysts to perform enzymatic cascade reactions in flow biocatalytic systems. When designing packed‐bed reactors with multiple immobilized enzymes, the enzymes are distributed in porous particles, and the porous particles are distributed in catalyst zones. A recently developed methodology is extended for selecting spatial immobilization distributions in batch reactors to packed‐bed reactors. Mechanistic models are used to compare the three most basic designs for a packed‐bed reactor, i.e., mixed individually immobilized enzymes, separated individually immobilized enzymes, and mixed co‐immobilized enzymes. For first‐order microkinetics, the design with co‐immobilized enzymes was found to always outperform the other two. Separating individually immobilized enzymes in two zones was proven to outperform mixing them in one zone for any positive order reaction kinetic.

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“…The idea of an organic synthesis, realized by conversion of expedient substrates in several chemical steps without the isolation of intermediates, has strong appeal (Kara & Rudroff, 2021; Rosenthal & Lütz, 2018; Ruales‐Salcedo et al, 2019; Schrittwieser et al, 2018; Sheldon & Brady, 2019). The view of biochemical engineering on enzyme cascades is however mixed (Burek et al, 2022; Kuschmierz et al, 2022; Nazor et al, 2021; Teshima et al, 2023). Advantage of economy in the processing steps used must be balanced against the drawback of expanse in process system complexity.…”

Section: Introductionmentioning

confidence: 99%

Pushing the boundaries of phosphorylase cascade reaction for cellobiose production I: Kinetic model development

Sigg,

Klimacek,

Nidetzky

2023

Biotech & Bioengineering

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One‐pot cascade reactions of coupled disaccharide phosphorylases enable an efficient transglycosylation via intermediary α‐d‐glucose 1‐phosphate (G1P). Such transformations have promising applications in the production of carbohydrate commodities, including the disaccharide cellobiose for food and feed use. Several studies have shown sucrose and cellobiose phosphorylase for cellobiose synthesis from sucrose, but the boundaries on transformation efficiency that result from kinetic and thermodynamic characteristics of the individual enzyme reactions are not known. Here, we assessed in a step‐by‐step systematic fashion the practical requirements of a kinetic model to describe cellobiose production at industrially relevant substrate concentrations of up to 600 mM sucrose and glucose each. Mechanistic initial‐rate models of the two‐substrate reactions of sucrose phosphorylase (sucrose + phosphate → G1P + fructose) and cellobiose phosphorylase (G1P + glucose → cellobiose + phosphate) were needed and additionally required expansion by terms of glucose inhibition, in particular a distinctive two‐site glucose substrate inhibition of the cellobiose phosphorylase (fromCellulumonas uda). Combined with mass action terms accounting for the approach to equilibrium, the kinetic model gave an excellent fit and a robust prediction of the full reaction time courses for a wide range of enzyme activities as well as substrate concentrations, including the variable substoichiometric concentration of phosphate. The model thus provides the essential engineering tool to disentangle the highly interrelated factors of conversion efficiency in the coupled enzyme reaction; and it establishes the necessary basis of window of operation calculations for targeted optimizations toward different process tasks.

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Cell-free enzyme cascades — application and transition from development to industrial implementation

Cited by 18 publications

References 53 publications

Route selection and reaction engineering for sustainable metabolite synthesis

Route selection and reaction engineering for sustainable metabolite synthesis

Modeling Enzymatic Cascade Reactions Immobilized in Plug‐Flow Reactors for Flow Biocatalysis

Pushing the boundaries of phosphorylase cascade reaction for cellobiose production I: Kinetic model development

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