Endre søk
RefereraExporteraLink to record
Permanent link

Direct link
Referera
Referensformat
  • apa
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • Annet format
Fler format
Språk
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Annet språk
Fler språk
Utmatningsformat
  • html
  • text
  • asciidoc
  • rtf
One-step enzyme extraction and immobilization for biocatalysis applications
KTH, Skolan för bioteknologi (BIO), Biokemi.
KTH, Skolan för bioteknologi (BIO), Biokemi.
Vise andre og tillknytning
2011 (engelsk)Inngår i: Biotechnology Journal, ISSN 1860-6768, E-ISSN 1860-7314, Vol. 6, nr 4, s. 463-469Artikkel i tidsskrift (Fagfellevurdert) 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.

sted, utgiver, år, opplag, sider
2011. Vol. 6, nr 4, s. 463-469
Emneord [en]
Condition promiscuity, Enzyme in organic solvent, Enzymatic synthesis, HisSi Immobilization, His(6)-tag
HSV kategori
Identifikatorer
URN: urn:nbn:se:kth:diva-39187DOI: 10.1002/biot.201000357ISI: 000289214500011PubMedID: 21381205Scopus ID: 2-s2.0-79953685174OAI: oai:DiVA.org:kth-39187DiVA, id: diva2:439510
Tilgjengelig fra: 2011-09-08 Laget: 2011-09-08 Sist oppdatert: 2020-03-09bibliografisk kontrollert
Inngår i avhandling
1. Tools in biocatalysis: enzyme immobilisation on silica and synthesis of enantiopure amines
Åpne denne publikasjonen i ny fane eller vindu >>Tools in biocatalysis: enzyme immobilisation on silica and synthesis of enantiopure amines
2010 (engelsk)Licentiatavhandling, med artikler (Annet vitenskapelig)
Abstract [en]

This thesis presents two techniques in the field of biocatalysis:

An enzyme immobilisation method based on the His6-tag for attachment on modified silica oxide beads, and it’s employment in aqueous and organic medium for synthesis applications. The method functions as a one step extraction and immobilisation protocol.

An equilibrium displacement system which enables complete conversion in reactions with ω-transaminases where isopropylamine is the donor, a route for synthesis of pharmaceutically interesting enantiopure amines.

Biocatalysis is predicted to be a paramount technology for an environmentally sustainable chemical industry, to which every newly developed method represents a small but important step. The work done here is aimed to be a part of this development.

 

Abstract [sv]

I denna avhandling presenteras två tekniker inom ämnet biokatalys:

En metod för immobilisering av His6-enzym på modifierad kiseloxid, och användning av detta konstrukt för kemiska synteser i vatten och organiska lösningsmedel. Detta system fungerar även som en snabb extraherings- och immobiliseringsmetod.

Ett jämviksförskjutningssystem som möjliggör fullständig omsätt-ning i reaktioner med ω-transaminaser där isopropylamin är amino-donator, en syntesväg för tillverkning av farmakologiskt intressanta kirala aminer.

Biokatalys förutspås att bli en ovärderlig teknologi i en miljömässigt hållbar kemisk industri, i vilken varje ny metod är en liten men dock viktig del. Detta arbete är menat som en del i denna utveckling.

Publisher
s. 27
Serie
Trita-BIO-Report, ISSN 1654-2312 ; 2010:11
HSV kategori
Identifikatorer
urn:nbn:se:kth:diva-12936 (URN)
Presentation
2010-05-28, 15:00 (engelsk)
Opponent
Veileder
Merknad
QC 20100519Tilgjengelig fra: 2010-05-19 Laget: 2010-05-19 Sist oppdatert: 2011-11-29bibliografisk kontrollert

Open Access i DiVA

Fulltekst mangler i DiVA

Andre lenker

Forlagets fulltekstPubMedScopus

Personposter BETA

Berglund, Per

Søk i DiVA

Av forfatter/redaktør
Cassimjee, Karim EngelmarkLarsen, Marianne WittrupThanh, Nguyen HongBerglund, Per
Av organisasjonen
I samme tidsskrift
Biotechnology Journal

Søk utenfor DiVA

GoogleGoogle Scholar

doi
pubmed
urn-nbn

Altmetric

doi
pubmed
urn-nbn
Totalt: 487 treff
RefereraExporteraLink to record
Permanent link

Direct link
Referera
Referensformat
  • apa
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • Annet format
Fler format
Språk
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Annet språk
Fler språk
Utmatningsformat
  • html
  • text
  • asciidoc
  • rtf