One-step enzyme extraction and immobilization for biocatalysis applications.

Karim Engelmark Cassimjee, Robert Kourist, Diana Lindberg, Marianne Wittrup Larsen, Nguyen Hong Thanh, Mikael Widersten, Uwe T Bornscheuer, Per Berglund

Research output: Contribution to journalArticleResearchpeer-review

Abstract

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.
Original languageEnglish
Pages (from-to)463-469
Number of pages7
JournalMolecular biotechnology
Volume6
DOIs
Publication statusPublished - 2011

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Biocatalysis
Immobilization
Lipases
Enzymes
Lipase
His-His-His-His-His-His
Bacilli
Esterases
Organic solvents
Bacillus subtilis
Epoxide Hydrolases
Hydrolases
Candida
Enzyme activity
Cyclohexane
Acetonitrile
Silicon Dioxide
Oxides
Metal ions
Solanum tuberosum

Keywords

  • condition promiscuity enzymatic synthesis enzyme in organic solvent his 6 -tag hissi immobilization

Cite this

Cassimjee, K. E., Kourist, R., Lindberg, D., Wittrup Larsen, M., Thanh, N. H., Widersten, M., ... Berglund, P. (2011). One-step enzyme extraction and immobilization for biocatalysis applications. Molecular biotechnology, 6, 463-469. https://doi.org/10.1002/biot.201000357

One-step enzyme extraction and immobilization for biocatalysis applications. / Cassimjee, Karim Engelmark; Kourist, Robert; Lindberg, Diana; Wittrup Larsen, Marianne; Thanh, Nguyen Hong; Widersten, Mikael; Bornscheuer, Uwe T; Berglund, Per.

In: Molecular biotechnology, Vol. 6, 2011, p. 463-469.

Research output: Contribution to journalArticleResearchpeer-review

Cassimjee, KE, Kourist, R, Lindberg, D, Wittrup Larsen, M, Thanh, NH, Widersten, M, Bornscheuer, UT & Berglund, P 2011, 'One-step enzyme extraction and immobilization for biocatalysis applications.' Molecular biotechnology, vol. 6, pp. 463-469. https://doi.org/10.1002/biot.201000357
Cassimjee, Karim Engelmark ; Kourist, Robert ; Lindberg, Diana ; Wittrup Larsen, Marianne ; Thanh, Nguyen Hong ; Widersten, Mikael ; Bornscheuer, Uwe T ; Berglund, Per. / One-step enzyme extraction and immobilization for biocatalysis applications. In: Molecular biotechnology. 2011 ; Vol. 6. pp. 463-469.
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