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
Summary
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
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Disclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.