Phase-Separated Multienzyme Condensates for Efficient Synthesis of Imines from Carboxylic Acids with Enhanced Dual-Cofactor Recycling DOI Open Access
Tao Guo,

Li-Fang Zeng,

Jiaxu Liu

et al.

International Journal of Molecular Sciences, Journal Year: 2025, Volume and Issue: 26(10), P. 4795 - 4795

Published: May 16, 2025

Enzyme catalysis represents a promising approach for sustainable chemical synthesis, yet its industrial applications face limitations due to the inefficient regeneration and high cost of essential cofactors, such as adenosine-5′-triphosphate (ATP) nicotinamide adenine dinucleotide phosphate (NADPH). While natural metabolic systems efficiently recycle cofactors through spatially organized enzymes, replicating this efficiency in vitro remains challenging. Here, we prepare five-enzyme condensate system using liquid–liquid phase separation (LLPS) mediated by intrinsically disordered proteins (IDPs). By colocalizing carboxylic acid reductase from Norcadia iowensis (NiCAR) with reductive aminase Aspergillus oryzae (AspRedAm) three cofactor-regenerating generated phase-separated catalytic that enhanced ATP NADPH recycling 4.7-fold 1.9-fold relative free respectively. Catalytic performance was correlated extent separation, confirmed fluorescence microscopy, which revealed clear enrichment within condensates. This proximity effect enabled efficient cofactor turnover one-step reaction, achieving substrate conversion above 90% 6 h enhancing space–time yield (STY) chiral imines 1.6-fold, only one-fifth standard load. creates scalable economic tool performing multienzyme cascade reactions are driven multiple cofactors.

Language: Английский

Access to Axially Chiral Biaryl Benzylamines via Ancestral Enzyme-Enabled Reductive Amination Desymmetrization DOI
W. Jim Zheng, Xinxin Zhu,

Zheng Zhu

et al.

ACS Catalysis, Journal Year: 2025, Volume and Issue: 15(3), P. 1522 - 1531

Published: Jan. 11, 2025

Axially chiral biaryl benzylamines are present in numerous natural products, pharmaceuticals, ligands, and catalysts. However, the direct catalytic synthesis of these functional molecules using a robust strategy remains formidable challenge. Reductive amination desymmetrization dialdehydes offers powerful approach for construction axially but suffers from extensive undesirable side reactions. Herein, we engineered ancestral imine reductases to enable reductive dialdehydes, allowing wide range with up 99% conversion enantiomeric excess (ee). The ratio product byproducts was 97:3 over 90:10 most cases. This work presents an alternative accessing will stimulate development associated bioactive catalysts/ligands.

Language: Английский

Citations

0
Phase-Separated Multienzyme Condensates for Efficient Synthesis of Imines from Carboxylic Acids with Enhanced Dual-Cofactor Recycling DOI Open Access
Tao Guo,

Li-Fang Zeng,

Jiaxu Liu

et al.

International Journal of Molecular Sciences, Journal Year: 2025, Volume and Issue: 26(10), P. 4795 - 4795

Published: May 16, 2025

Enzyme catalysis represents a promising approach for sustainable chemical synthesis, yet its industrial applications face limitations due to the inefficient regeneration and high cost of essential cofactors, such as adenosine-5′-triphosphate (ATP) nicotinamide adenine dinucleotide phosphate (NADPH). While natural metabolic systems efficiently recycle cofactors through spatially organized enzymes, replicating this efficiency in vitro remains challenging. Here, we prepare five-enzyme condensate system using liquid–liquid phase separation (LLPS) mediated by intrinsically disordered proteins (IDPs). By colocalizing carboxylic acid reductase from Norcadia iowensis (NiCAR) with reductive aminase Aspergillus oryzae (AspRedAm) three cofactor-regenerating generated phase-separated catalytic that enhanced ATP NADPH recycling 4.7-fold 1.9-fold relative free respectively. Catalytic performance was correlated extent separation, confirmed fluorescence microscopy, which revealed clear enrichment within condensates. This proximity effect enabled efficient cofactor turnover one-step reaction, achieving substrate conversion above 90% 6 h enhancing space–time yield (STY) chiral imines 1.6-fold, only one-fifth standard load. creates scalable economic tool performing multienzyme cascade reactions are driven multiple cofactors.

Language: Английский

Citations

0