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One-step enzyme extraction and immobilization for biocatalysis applications
Uppsala University, Disciplinary Domain of Science and Technology, Chemistry, Department of Biochemistry and Organic Chemistry.
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2011 (English)In: Biotechnology Journal, ISSN 1860-6768, Vol. 6, no 4, 463-469 p.Article in journal (Refereed) Published
Abstract [en]

An extraction/immobilization method for HIs(6) -tagged enzymes for use in synthesis applications is presented. By modifying silica oxide beads to be able to accommodate metal ions, the enzyme was tethered to the beads after adsorption of Co(II). The beads were successfully used for direct extraction of C. antarctica lipase B (CalB) from a periplasmic preparation with a minimum of 58% activity yield, creating a quick one-step extraction-immobilization protocol. This method, named HisSi Immobilization, was evaluated with five different enzymes [Candida antarctica lipase B (CalB), Bacillus subtilis lipase A (BslA), Bacillus subtilis esterase (BS2), Pseudomonas fluorescence esterase (PFE), and Solanum tuberosum epoxide hydrolase 1 (StEH1)]. Immobilized CalB was effectively employed in organic solvent (cyclohexane and acetonitrile) in a transacylation reaction and in aqueous buffer for ester hydrolysis. For the remaining enzymes some activity in organic solvent could be shown, whereas the non-immobilized enzymes were found inactive. The protocol presented in this work provides a facile immobilization method by utilization of the common His(6) -tag, offering specific and defined means of binding a protein in a specific location, which is applicable for a wide range of enzymes.

Place, publisher, year, edition, pages
2011. Vol. 6, no 4, 463-469 p.
Keyword [en]
Condition promiscuity, Enzyme in organic solvent, Enzymatic synthesis, HisSi Immobilization, His(6)-tag
National Category
Biochemistry and Molecular Biology
Identifiers
URN: urn:nbn:se:uu:diva-149282DOI: 10.1002/biot.201000357ISI: 000289214500011PubMedID: 21381205OAI: oai:DiVA.org:uu-149282DiVA: diva2:404387
Available from: 2011-03-16 Created: 2011-03-16 Last updated: 2011-05-02Bibliographically approved

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Widersten, Mikael

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