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Expression and purification of horseradish peroxidase in insect larvae

Paper ID Volume ID Publish Year Pages File Format Full-Text
35922 45114 2008 5 PDF Available
Title
Expression and purification of horseradish peroxidase in insect larvae
Abstract

A strategy for obtaining a high-level expression of horseradish peroxidase isozyme C (HRPC) in insect larvae is herein described. The HRPC-6xHis coding sequence was inserted into the AcNPV genome via the pAcGP67HRPC-6xHis transfer vector to originate an AcHRPC-6xHis recombinant baculovirus of phenotype occ−. Rachiplusia nu larvae were injected with a viral stock derived from culture supernatants of Sf9 cells infected with AcHRPC-6xHis. Enzyme concentration at day 3 post-infection was 230 ± 10 mg/kg haemolymph and 100 ± 14 mg/kg larvae, an expression level never attained by other expression systems. HRPC-6xHis was purified from the crude larval extract or haemolymph by immobilised metal ion affinity chromatography with yields of 88.8% and 89.0% and purification factors of 18.9 and 14.0, respectively, in a single step. The purity of the final product was 90%.

Keywords
Horseradish peroxidase; Expression; Purification; Insect larvae; Rachiplusia nu; Haemolymph
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Expression and purification of horseradish peroxidase in insect larvae
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Publisher
Database: Elsevier - ScienceDirect
Journal: Process Biochemistry - Volume 43, Issue 1, January 2008, Pages 103–107
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
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