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Non-covalent binding tags for batch and flow biocatalysis.
Rocha, Raquel A; Esquirol, Lygie; Rolland, Vivien; Hands, Philip; Speight, Robert E; Scott, Colin.
Afiliación
  • Rocha RA; School of Biology and Environmental Science, Faculty of Science, Queensland University of Technology (QUT), Brisbane, Qld 4000, Australia; CSIRO Environment, Black Mountain Science and Innovation Park, Canberra, ACT 2601, Australia.
  • Esquirol L; CSIRO Environment, Black Mountain Science and Innovation Park, Canberra, ACT 2601, Australia.
  • Rolland V; CSIRO Agriculture and Food, Black Mountain Science and Innovation Park, Canberra, ACT 2601, Australia.
  • Hands P; CSIRO Agriculture and Food, Black Mountain Science and Innovation Park, Canberra, ACT 2601, Australia.
  • Speight RE; School of Biology and Environmental Science, Faculty of Science, Queensland University of Technology (QUT), Brisbane, Qld 4000, Australia; ARC Centre of Excellence in Synthetic Biology, Queensland University of Technology (QUT), Brisbane, Qld 4000, Australia.
  • Scott C; CSIRO Environment, Black Mountain Science and Innovation Park, Canberra, ACT 2601, Australia. Electronic address: colin.scott@csiro.au.
Enzyme Microb Technol ; 169: 110268, 2023 Sep.
Article en En | MEDLINE | ID: mdl-37300919
Enzyme immobilization offers considerable advantage for biocatalysis in batch and continuous flow reactions. However, many currently available immobilization methods require that the surface of the carrier is chemically modified to allow site specific interactions with their cognate enzymes, which requires specific processing steps and incurs associated costs. Two carriers (cellulose and silica) were investigated here, initially using fluorescent proteins as models to study binding, followed by assessment of industrially relevant enzyme performance (transaminases and an imine reductase/glucose oxidoreductase fusion). Two previously described binding tags, the 17 amino acid long silica-binding peptide from the Bacillus cereus CotB protein and the cellulose binding domain from the Clostridium thermocellum, were fused to a range of proteins without impairing their heterologous expression. When fused to a fluorescent protein both tags conferred high avidity specific binding with their respective carriers (low nanomolar Kd values). The CotB peptide (CotB1p) induced protein aggregation in the transaminase and imine reductase/glucose oxidoreductase fusions when incubated with the silica carrier. The Clostridium thermocellum cellulose binding domain (CBDclos) allowed immobilization of all the proteins tested, but immobilization led to loss of enzymatic activity in the transaminases (< 2-fold) and imine reductase/glucose oxidoreductase fusion (> 80%). A transaminase-CBDclos fusion was then successfully used to demonstrate the application of the binding tag in repetitive batch and a continuous-flow reactor.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Celulosa / Enzimas Inmovilizadas Idioma: En Revista: Enzyme Microb Technol Año: 2023 Tipo del documento: Article País de afiliación: Australia Pais de publicación: Estados Unidos
Texto completo
Añadir a Mi BVS
Imprimir
XML
PubMed Links
Buscar en Google

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Celulosa / Enzimas Inmovilizadas Idioma: En Revista: Enzyme Microb Technol Año: 2023 Tipo del documento: Article País de afiliación: Australia Pais de publicación: Estados Unidos