{"database":"biostudies-literature","file_versions":[],"scores":null,"additional":{"submitter":["Humer D"],"funding":["Austrian Science Fund FWF"],"pagination":["E4625"],"full_dataset_link":["https://www.ebi.ac.uk/biostudies/studies/S-EPMC7369975"],"repository":["biostudies-literature"],"omics_type":["Unknown"],"volume":["21(13)"],"pubmed_abstract":["Horseradish peroxidase (HRP), an enzyme omnipresent in biotechnology, is still produced from hairy root cultures, although this procedure is time-consuming and only gives low yields. In addition, the plant-derived enzyme preparation consists of a variable mixture of isoenzymes with high batch-to-batch variation preventing its use in therapeutic applications. In this study, we present a novel and scalable recombinant HRP production process in Escherichia coli that yields a highly pure, active and homogeneous single isoenzyme. We successfully developed a multi-step inclusion body process giving a final yield of 960 mg active HRP/L culture medium with a purity of ≥99% determined by size-exclusion high-performance liquid chromatography (SEC-HPLC). The Reinheitszahl, as well as the activity with 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) and 3,3',5,5'-tetramethylbenzidine (TMB) as reducing substrates, are comparable to commercially available plant HRP. Thus, our preparation of recombinant, unglycosylated HRP from E. coli is a viable alternative to the enzyme from plant and highly interesting for therapeutic applications."],"journal":["International journal of molecular sciences"],"pubmed_title":["Scalable High-Performance Production of Recombinant Horseradish Peroxidase from E. coli Inclusion Bodies."],"pmcid":["PMC7369975"],"funding_grant_id":["P30872-B26"],"pubmed_authors":["Spadiut O","Humer D","Ebner J"],"additional_accession":[]},"is_claimable":false,"name":"Scalable High-Performance Production of Recombinant Horseradish Peroxidase from E. coli Inclusion Bodies.","description":"Horseradish peroxidase (HRP), an enzyme omnipresent in biotechnology, is still produced from hairy root cultures, although this procedure is time-consuming and only gives low yields. In addition, the plant-derived enzyme preparation consists of a variable mixture of isoenzymes with high batch-to-batch variation preventing its use in therapeutic applications. In this study, we present a novel and scalable recombinant HRP production process in Escherichia coli that yields a highly pure, active and homogeneous single isoenzyme. We successfully developed a multi-step inclusion body process giving a final yield of 960 mg active HRP/L culture medium with a purity of ≥99% determined by size-exclusion high-performance liquid chromatography (SEC-HPLC). The Reinheitszahl, as well as the activity with 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) and 3,3',5,5'-tetramethylbenzidine (TMB) as reducing substrates, are comparable to commercially available plant HRP. Thus, our preparation of recombinant, unglycosylated HRP from E. coli is a viable alternative to the enzyme from plant and highly interesting for therapeutic applications.","dates":{"release":"2020-01-01T00:00:00Z","publication":"2020 Jun","modification":"2025-04-22T19:31:29.273Z","creation":"2025-04-06T02:48:47.81Z"},"accession":"S-EPMC7369975","cross_references":{"pubmed":["32610584"],"doi":["10.3390/ijms21134625"]}}