Preparation of activated supports containing low pK amino groups. A new tool for protein immobilization via the carboxyl coupling method

Fernández‐Lafuente, Roberto; Rosell, Cristina M.; Rodríguez, Verónica; Santana, César Costapinto; Soler, Gloria; Bastida, Ágatha; Guisán, José M.

doi:10.1016/0141-0229(93)90016-u

Cited by 247 publications

(149 citation statements)

References 4 publications

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“…41 Inicialmente, 40 mL de solução de etilenodiamina (EDA) 2 M a pH 10,0 foram adicionados a 10 g de glioxil-suportes ativados com glicidol (glioxil--bagaço de cana e glioxil-PHB) e mantido sob agitação por 2 h. Em seguida, 0,57 g de boro-hidreto de sódio (NaBH 4 ) foi adicionado ao sistema e mantido por mais 2 h sob agitação branda em frasco aberto à temperatura ambiente. Foram adicionados aos suportes aminados (glioxil-amino), previamente lavados com água destilada e filtrados sob vácuo, solução de glutaraldeído a 25% (16,8 mL) diluída em tampão fosfato de sódio 200 mM pH 7,0 (11,2 mL) e mantidos sob agitação a 25 °C por 18 h. Os suportes glioxil-amino-glutaraldeído preparados foram lavados exaustivamente com água destilada, filtrados a vácuo e estocados sob refrigeração.…”

Section: Preparação De Suportes Glioxil-amino-glutaraldeídounclassified

Triagem de suportes orgânicos e protocolos de ativação na imobilização e estabilização de lipase de Thermomyces lanuginosus

Mendes¹,

Castro²,

Giordano³

2013

Quím. Nova

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Thermomyces lanuginosus. Lipase from Thermomyces lanuginosus was covalently immobilized on activated poly-hydroxybutyrate, sugarcane bagasse and the chemically modified hybrid hydrogel chitosan-alginate prepared by different strategies. Among the tested supports, chitosan-alginate chemically modified with 2,4,6-trinitrobenzenesulfonic acid rendered derivatives with the highest hydrolytic activity and thermal-stability, 45-fold more stable than soluble lipase and was then selected for further studies. The pH of maximum activity was similar for both immobilized and free lipase (pH 8.0) while optimum temperature was 5 -10 ºC higher for the immobilized lipase. Higher yields in the butyl butyrate synthesis were found for the derivatives prepared by activation with glycidol and epichlorohydrin.

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Section: Preparação De Suportes Glioxil-amino-glutaraldeídounclassified

Triagem de suportes orgânicos e protocolos de ativação na imobilização e estabilização de lipase de Thermomyces lanuginosus

Mendes¹,

Castro²,

Giordano³

2013

Quím. Nova

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“…Considering that the pK a of these groups is around 7 for the primary and around 10 for the secondary amine, a titration with 10 mM NaOH from pH 4.8 to 8.8 will determine the concentration of the first group. It is recommended that 1.5 M of KCl be added to the medium 41 . (v) The remaining epoxy groups can be blocked with mercaptoethanol by incubation with the blocking solution 2 for 14-18 h to avoid reaction of the enzyme with the epoxy groups.…”

Section: Likely Applicationsmentioning

confidence: 99%

Immobilization of enzymes on heterofunctional epoxy supports

et al. 2007

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“…Monoaminoethyl-N-aminoethyl (MANAE)-agarose, was prepared as described elsewhere [18] and the glutaraldehyde-agarose support was prepared from MANAE agarose [19,20]. Briefly respectively.…”

Section: Preparation Of Supportmentioning

confidence: 99%

Co-immobilization and stabilization of xylanase, β-xylosidase and α-l-arabinofuranosidase from Penicillium janczewskii for arabinoxylan hydrolysis

Terrasan

Trobo-Maseda

Moreno-Pérez

et al. 2016

Process Biochemistry

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Differently activated agarose-based supports were evaluated for co-immobilization of a crude extract from Penicillium janczewskii containing xylanase, β-xylosidase and α-Larabinofuranosidase activities. Adequately selecting support and immobilization conditions (8 h, using agarose with 10% crosslinking) increased enzyme levels substantially, mainly in relation to the xylanase (2-fold). A coating with dextran aldehyde MW 6,000 Da, partially oxidized, covalently attached the enzymes to the support. Optimum activity was verified in the pH range 2-4, and at 50, 65 and 80 °C for the xylanase, α-L-arabinofuranosidase and β-xylosidase, 2 respectively. The xylanase was highly thermostable retaining more than 70% of activity even after 24 h incubation at 60 and 70 °C; and at 80 °C its half-life was 1.7 h. The half-lives of the β-xylosidase and α-L-arabinofuranosidase at 50 °C were 2.3 and 3.8 h, respectively. The coimmobilization of the enzymes on a single support give raise to a functional multi-enzymatic biocatalyst acting in the complete hydrolysis of different and complex substrates such as oat spelt and wheat arabinoxylans, with xylose yield higher than 40%. The xylanase and the α-Larabinofuranosidase presented high stability retaining 86.6 and 88.0% of activity after 10 reuse cycles.

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Preparation of activated supports containing low pK amino groups. A new tool for protein immobilization via the carboxyl coupling method

Cited by 247 publications

References 4 publications

Triagem de suportes orgânicos e protocolos de ativação na imobilização e estabilização de lipase de Thermomyces lanuginosus

Triagem de suportes orgânicos e protocolos de ativação na imobilização e estabilização de lipase de Thermomyces lanuginosus

Immobilization of enzymes on heterofunctional epoxy supports

Co-immobilization and stabilization of xylanase, β-xylosidase and α-l-arabinofuranosidase from Penicillium janczewskii for arabinoxylan hydrolysis

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