Toward modular construction of cell-free multienzyme systems

Zhang, Yinchen; Nie, Ning; Zhang, Yifei

doi:10.1016/s1872-2067(21)64002-0

“…In contrast, multi-enzyme catalysis can involve relatively complex chemical reaction sequences where each step is mediated by specific enzymes, and the activity of each enzyme is regulated by the presence or absence of a specific cofactor. 14 An in vitro CFS may comprise an assembly of multiple enzymes and/or cofactors in the absence of external membrane barriers. 4 Finally, whole-cell fermentation involves the in vivo transformation of a substrate into different products through interconnected metabolic pathways constituting a complex metabolic system.…”

Section: Cell-free Systems: An In Vitro Approach To Bioproductsmentioning

confidence: 99%

“…[9][10][11][12] Several other inadequacies include feedback inhibition, the degeneration and exhaustion of cofactors, and the loss of reaction intermediates when multi-enzyme catalysed reactions are operated. 5,13,14 This necessitates the development of advanced and stable CFSs for the large-scale production of different materials and their industrial applications.…”

Section: Introductionmentioning

confidence: 99%

Cell-free systems for biosynthesis: towards a sustainable and economical approach

Manan

¹

,

Ul-Islam

²

,

Khattak³

et al. 2023

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Modularization of Immobilized Multienzyme Cascades for Continuous‐Flow Enantioselective C−H Amination

Kong,

Huang,

Zhou

et al. 2024

Angewandte Chemie

0

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Multienzyme cascades (MECs) have gained much attention in synthetic chemistry but remain far from being a reliable synthetic tool. Here we report a four‐enzyme cascade comprising a cofactor‐independent and a cofactor self‐sustaining bienzymatic module for the enantioselective benzylic C‐H amination of arylalkanes, a challenging transformation from bulk chemicals to high value‐added chiral amines. The two modules were subsequently optimized by enzyme co‐immobilization with microenvironmental tuning, and finally integrated in a gas‐liquid segmented flow system, resulting in simultaneous improvements in enzyme performance, mass transfer, system compatibility, and productivity. The flow system enabled continuous C‐H amination of arylalkanes (up to 100 mM) utilizing the sole cofactor NADH (0.5 mM) in >90% conversion, achieving a high space‐time yield (STY) of 3.6 g·L ‐1 ·h ‐1 , which is a 90‐fold increase over previously reported values.

show abstract

Modularization of Immobilized Multienzyme Cascades for Continuous‐Flow Enantioselective C−H Amination

Kong,

Huang,

Zhou

et al. 2024

Angew Chem Int Ed

0

View full text Add to dashboard Cite

Multienzyme cascades (MECs) have gained much attention in synthetic chemistry but remain far from being a reliable synthetic tool. Here we report a four‐enzyme cascade comprising a cofactor‐independent and a cofactor self‐sustaining bienzymatic module for the enantioselective benzylic C‐H amination of arylalkanes, a challenging transformation from bulk chemicals to high value‐added chiral amines. The two modules were subsequently optimized by enzyme co‐immobilization with microenvironmental tuning, and finally integrated in a gas‐liquid segmented flow system, resulting in simultaneous improvements in enzyme performance, mass transfer, system compatibility, and productivity. The flow system enabled continuous C‐H amination of arylalkanes (up to 100 mM) utilizing the sole cofactor NADH (0.5 mM) in >90% conversion, achieving a high space‐time yield (STY) of 3.6 g·L ‐1 ·h ‐1 , which is a 90‐fold increase over previously reported values.

show abstract

Toward modular construction of cell-free multienzyme systems

Cited by 5 publications

References 95 publications

Cell-free systems for biosynthesis: towards a sustainable and economical approach

Cell-free systems for biosynthesis: towards a sustainable and economical approach

Modularization of Immobilized Multienzyme Cascades for Continuous‐Flow Enantioselective C−H Amination

Modularization of Immobilized Multienzyme Cascades for Continuous‐Flow Enantioselective C−H Amination

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