Novel method of enzymes stabilization on crosslinked thermosensitive carriers

Hamerska-Dudra, Agnieszka; Bryjak, Jolanta; Trochimczuk, Andrzej W.

doi:10.1016/j.enzmictec.2005.08.019

Cited by 22 publications

(12 citation statements)

References 15 publications

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“…These results indicate that a high concentration of enzyme crosslinker influence the enzyme activity negatively by restricting the enzyme mobility and causing changes in the enzyme conformation, often into a less favorable one, which can result in the deactivation of the enzyme [25]. Furthermore, some undesirable side reactions can occur and connect two copolymers into one and make those copolymers inactive for covalent attachment of enzyme [26].…”

Section: Resinmentioning

confidence: 99%

Surface modification of macroporous poly(glycidyl methacrylate-co-ethylene glycol dimethacrylate) resins for improved Candida antarctica lipase B immobilization

Miletić

Rohandi

Vuković

et al. 2009

Reactive and Functional Polymers

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Crosslinked macroporous hydrophilic poly(glycidyl methacrylate-co-ethylene glycol dimethacrylate) [abbreviated poly(GMA-co-EGDMA)] with identical chemical structure (60% of glycidyl methacrylate) but with varied average pore sizes (from 30 to 560 nm), specific surface areas (from 13.2 to 106.0 m 2 / g), specific volumes (from 0.755 to 1.191 cm 3 /g) and particle sizes (<100 lm-630 lm) were synthesized via suspension polymerization. Modifications of poly(GMA-co-EGDMA) with various diamines (1,2diaminoethane, 1,4-diaminobutane, 1,6-diaminohexane and 1,8-diaminooctane), 2-fluoroethylamine, glutaraldehyde and cyanuric chloride were carried out. The influence of the interaction between Candida antarctica lipase B (Cal-B) and various carriers during immobilization on the loading and hydrolytic activity (hydrolysis of para-nitrophenyl acetate) of the immobilized Cal-B were studied. Immobilization of Cal-B was performed at different temperatures and pH values. Cal-B immobilized at 30°C and pH 6.8 was leading to increased activities. Purely physical adsorption between enzyme and copolymer was observed on carriers in which amine or fluorine groups were introduced into the carrier structure by modification with various diamines or 2-fluoroethylamine. As a consequence enzyme loading and activity decreases. In contrary, modification of the poly(GMA-co-EGDMA) with glutaraldehyde and cyanuric chloride results in a covalent connection between enzyme and carrier. The obtained results show a significant increase in Cal-B activity. The influence of the amount of glutaraldehyde and cyanuric chloride used for modification was screened. Increasing the amount of glutaraldehyde or cyanuric chloride used for modification resulted in an increase of the enzyme loading. Consequently, higher amount of glutaraldehyde used led to a higher fraction of the enzyme molecules that are covalently connected on to the carrier. As the amount of glutaraldehyde or cyanuric chloride used for modifications increases, activity of immobilized C. antarctica lipase B primarily increases, showing the highest value for 0.66% and 0.050% w/w, respectively, and subsequently decreases. We could show that Cal-B immobilized on epoxy-containing copolymer modified with glutaraldehyde and cyanuric chloride performs higher activity than free enzyme powder.

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Section: Resinmentioning

confidence: 99%

Surface modification of macroporous poly(glycidyl methacrylate-co-ethylene glycol dimethacrylate) resins for improved Candida antarctica lipase B immobilization

Miletić

Rohandi

Vuković

et al. 2009

Reactive and Functional Polymers

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“…In addition, various materials used in enzyme immobilization have been reported in the literature. Synthetic environmentally sensitive carriers such as thermosensitive carriers 28 , 29 , pH sensitive hydrogel 30 , magnetic response carriers (including shell/core structure 31 , core/shell structure 32 and heterofunctional structure 33 ) and multi-responsive carriers 34 , can respond to environmental stimulation factors. These factors include temperature, light, magnetic field, electric field, pH value of buffer, solvent, reactant, ion or stress.…”

Section: Introductionmentioning

confidence: 99%

Burkholderia cepacia lipase immobilized on heterofunctional magnetic nanoparticles and its application in biodiesel synthesis

Fan

et al. 2017

Sci Rep

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Biodiesel production using immobilized lipase as a biocatalyst is a promising process. The performance of immobilized lipase is mainly determined by supporting materials and immobilization method. To avoid the shortcomings of adsorption and covalent bonding methods, in this study, we developed a novel heterofunctional carrier of being strengthened anion exchange and weakened covalent binding to avoid activity loss and improve operational stability of the immobilized lipase. 2,3-epoxypropyltrimethylammonium chloride with epoxy and quaternary ammonium group and glutaraldehyde were grafted onto aminated magnetic nanoparticles (AMNPs) to generate a new matrix, named GEAMNP. Then Burkholderia cepacia lipase (BCL) was immobilized on GEAMNP via anion exchange and covalent bonding. The transesterification between soybean oil and methanol was used to test the activities. Activity recovery of the immobilized BCL was up to 147.4% and the corresponding transesterification activity was 1.5-fold than that of BCL powder. The immobilized lipase was further used for biodiesel production to confirm its feasibility. The fatty acid methyl esters conversion yield could reach 96.8% in the first 12 h. Furthermore, the immobilized lipase, BCL-GEAMNP showed markedly improved operational stability, better reusability and higher esters than BCL-GAMNP, where MNPs were only modified with (3-aminopropyl) triethoxysilane and glutaraldehyde.

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“…Enzyme immobilization has attracted interest in biotechnological processes due to its high operational stability and durability, easy separation of products, and low cost of industrial applications [5]. A wide variety of immobilization techniques can be used, including adsorption on solid supports [6,7], covalent attachment [8,9], or entrapment in silica/polymer matrices [2]. Immobilization techniques are usually very easy to apply, but the bonding of the biocatalyst to the surface of the matrix is relatively weak.…”

Section: Introductionmentioning

confidence: 99%

Gold and silver nanoparticles for biomolecule immobilization and enzymatic catalysis

et al. 2012

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In this work, a simple method for alcohol synthesis with high enantiomeric purity was proposed. For this, colloidal gold and silver surface modifications with 3-mercaptopropanoic acid and cysteamine were used to generate carboxyl and amine functionalized gold and silver nanoparticles of 15 and 45 nm, respectively. Alcohol dehydrogenase from Thermoanaerobium brockii (TbADH) and its cofactor (NADPH) were physical and covalent (through direct adsorption and using cross-linker) immobilized on nanoparticles' surface. In contrast to the physical and covalent immobilizations that led to a loss of 90% of the initial enzyme activity and 98% immobilization, the use of a cross-linker in immobilization process promoted a loss to 30% of the initial enzyme activity and >92% immobilization. The yield of NADPH immobilization was about 80%. The best results in terms of activity were obtained with Ag-citr nanoparticle functionalized with carboxyl groups (Ag-COOH), Au-COOH(CTAB), and Au-citr functionalized with amine groups and stabilized with CTAB (Au-NH2(CTAB)) nanoparticles treated with 0.7% and 1.0% glutaraldehyde. Enzyme conformation upon immobilization was studied using fluorescence and circular dichroism spectroscopies. Shift in ellipticity at 222 nm with about 4 to 7 nm and significant decreasing in fluorescence emission for all bioconjugates were observed by binding of TbADH to silver/gold nanoparticles. Emission redshifting of 5 nm only for Ag-COOH-TbADH bioconjugate demonstrated change in the microenvironment of TbADH. Enzyme immobilization on glutaraldehyde-treated Au-NH2(CTAB) nanoparticles promotes an additional stabilization preserving about 50% of enzyme activity after 15 days storage. Nanoparticles attached-TbADH-NADPH systems were used for enantioselective (ee > 99%) synthesis of (S)-7-hydroxy-2-tetralol.

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Novel method of enzymes stabilization on crosslinked thermosensitive carriers

Cited by 22 publications

References 15 publications

Surface modification of macroporous poly(glycidyl methacrylate-co-ethylene glycol dimethacrylate) resins for improved Candida antarctica lipase B immobilization

Surface modification of macroporous poly(glycidyl methacrylate-co-ethylene glycol dimethacrylate) resins for improved Candida antarctica lipase B immobilization

Burkholderia cepacia lipase immobilized on heterofunctional magnetic nanoparticles and its application in biodiesel synthesis

Gold and silver nanoparticles for biomolecule immobilization and enzymatic catalysis

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