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One-step enzyme extraction and immobilization for biocatalysis applications

Cassimjee, Karim Engelmark (author)
KTH,Biokemi
Kourist, Robert (author)
Lindberg, Diana (author)
Uppsala universitet,Institutionen för biokemi och organisk kemi
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Larsen, Marianne Wittrup (author)
KTH,Biokemi
Thanh, Nguyen Hong (author)
KTH,Biokemi
Widersten, Mikael (author)
Uppsala universitet,Biokemi
Bornscheuer, Uwe T (author)
Berglund, Per (author)
KTH,Biokemi
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 (creator_code:org_t)
2011-03-07
2011
English.
In: Biotechnology Journal. - : Wiley. - 1860-6768 .- 1860-7314. ; 6:4, s. 463-469
Related links:
https://urn.kb.se/re...
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  • Journal article (peer-reviewed)
Abstract Subject headings
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  • 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.

Subject headings

NATURVETENSKAP  -- Biologi -- Biokemi och molekylärbiologi (hsv//swe)
NATURAL SCIENCES  -- Biological Sciences -- Biochemistry and Molecular Biology (hsv//eng)

Keyword

Condition promiscuity
Enzyme in organic solvent
Enzymatic synthesis
HisSi Immobilization
His(6)-tag
Biochemistry
Biokemi

Publication and Content Type

ref (subject category)
art (subject category)

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