Artificial polyenzyme (ArPoly) forms amphiphilic proteinpolymer conjugates that stabilize Pickering emulsions with tunable interfacial properties for biocatalysis. These emulsions enable efficient and robust single-step reactions, multi-step biocatalytic cascades, and whole-cell catalysis. Furthermore, ArPoly can be easily recycled via introducing a cationic polymer. These findings establish ArPoly as a versatile and robust platform for biocatalytic cascade synthesis.

Abstract

Artificial polyenzymes (ArPoly), combining universal protein scaffolds with polymers, have emerged as a transformative platform for biocatalysis. However, their application in stabilizing Pickering emulsions for biocatalytic cascades remains unexplored. Here, we introduce a robust and versatile ArPoly system based on bovine serum albumin as a noncatalytic protein scaffold, copolymerized with hydrophobic ligand monomers and charged styrene sulfonate monomers. The resulting ArPoly exhibits decent amphiphilicity, enabling stable Pickering emulsions with tunable interfacial properties. These emulsions demonstrate notable robustness, maintaining stability for over 24 h under optimized conditions. We highlight the versatility of ArPoly-enabled Pickering emulsions in biocatalytic cascade synthesis, successfully facilitating single-step reactions, multi-step cascades, and whole-cell catalysis. Furthermore, the tailored nature of ArPoly allows for efficient recycling through interaction with a cationic polymer, PolyN⁺, enabling recovery and reuse without compromising activity in a one-pot two-step cascade and a sequential three-step cascade reaction with Candida antarctica lipase B and transaminase for producing 1-phenylethanol and 1-phenylethyl butyrate. These findings establish ArPoly as a versatile and robust platform for biocatalytic cascade synthesis. Its recyclability, stability, and adaptability make it a promising alternative for complex chemical synthesis, offering a sustainable and efficient solution for industrial biotechnology.