Abstract

BackgroundPhysiological aggregation of a recombinant enzyme into enzymatically active inclusion bodies could be an excellent strategy to obtain immobilized enzymes for industrial biotransformation processes. However, it is not convenient to recycle “gelatinous masses” of protein inclusion bodies from one reaction cycle to another, as high centrifugation forces are needed in large volumes. The magnetization of inclusion bodies is a smart solution for large-scale applications, enabling an easier separation process using a magnetic field.ResultsMagnetically modified inclusion bodies of UDP–glucose pyrophosphorylase were recycled 50 times, in comparison, inclusion bodies of the same enzyme were inactivated during ten reaction cycles if they were recycled by centrifugation. Inclusion bodies of sialic acid aldolase also showed good performance and operational stability after the magnetization procedure.ConclusionsIt is demonstrated here that inclusion bodies can be easily magnetically modified by magnetic iron oxide particles prepared by microwave-assisted synthesis from ferrous sulphate. The magnetic particles stabilize the repetitive use of the inclusion bodies .

Highlights

  • Physiological aggregation of a recombinant enzyme into enzymatically active inclusion bodies could be an excellent strategy to obtain immobilized enzymes for industrial biotransformation processes

  • green fluorescent protein (GFP) (FPbase: TurboGFP; GenBank: ASW25889), sialic acid aldolase (SAA, UniProt: P0A6L4; NCBI-GeneID: 947742) and UDP–glucose pyrophosphorylase (GalU, UniProt: P0AEP3; NCBI-GeneID: 945730) genes were N-terminally fused with the cellulose-binding domain from Clostridium cellulovorans by cloning into plasmid pET-34b (Additional file 1)

  • The particles were supplied with Inclusion bodies (IBs) solution and the suspension was thoroughly mixed with a pipette

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Summary

Results

Modified inclusion bodies of UDP–glucose pyrophosphorylase were recycled 50 times, in com‐ parison, inclusion bodies of the same enzyme were inactivated during ten reaction cycles if they were recycled by centrifugation. Inclusion bodies of sialic acid aldolase showed good performance and operational stability after the magnetization procedure

Conclusions
Background
Methods
Results and discussion
Conclusion

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