Chemical amination of lipases improves their immobilization on octyl-glyoxyl agarose beads

Rueda, Nazzoly; Santos, José Cleiton Sousa dos; Ortíz, Claudia; Barbosa, Oveimar; Fernández-Lafuente, Roberto; Torres, Rodrigo

doi:10.1016/j.cattod.2015.05.027

Cited by 67 publications

(39 citation statements)

References 74 publications

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“…The constituents of the enzymatic extract possibly help the conservation of the enzyme structure during the immobilization process. In fact, when the PMMA/PMMA support was employed, CalB exhibited a hyper‐activation (147% of recovered activity), which had not been described before for this enzyme using other substrates and octyl agarose …”

Section: Resultssupporting

confidence: 59%

Pilot‐scale development of core–shell polymer supports for the immobilization of recombinant lipase B from Candida antarctica and their application in the production of ethyl esters from residual fatty acids

Cipolatti

Pinto

Robert

et al. 2018

J of Applied Polymer Sci

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A recombinant lipase B from Candida antarctica (LipB) in Pichia pastoris was synthesized through submerged fermentation using crude glycerin as substrate. The immobilization of this enzyme on the core–shell polymeric supports is an effective alternative for its application. The supports with distinct levels of hydrophobicity were produced through combined suspension and emulsion polymerization in pilot scale. Particles with distinct compositions were synthesized (PMMA/PMMA; PMMA‐co‐DVB/PMMA‐co‐DVB; and PS‐co‐DVB/PS‐co‐DVB) and employed on the immobilization of the produced lipase (LipB) and the commercial enzyme (CalB). The morphological properties (specific area, average pore diameter, specific volume of pores, and hydrophobicity level) and the influence of the polymerization conditions on the morphology of the supports were studied. The thermal stability of such biocatalysts was also investigated in the presence of calcium cation (Ca+2), maintained 100% of the activity after 3 h at 50°C when the PMMA‐co‐DVB/PMMA‐co‐DVB was employed. The synthesized enzyme and supports manufactured in pilot scale were employed successfully for production of esters using residual fatty acids as substrates, adding value to these raw materials and increasing the ranges of possible applications.

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Section: Resultssupporting

confidence: 59%

Pilot‐scale development of core–shell polymer supports for the immobilization of recombinant lipase B from Candida antarctica and their application in the production of ethyl esters from residual fatty acids

Cipolatti

Pinto

Robert

et al. 2018

J of Applied Polymer Sci

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“…These configurations were checked by polarimetry, by which the value of specific optical rotation of ( R )‐indanol was [α] D = −30,1 (c 1,0; CHCl 3 ) e.e. = 97%, while the value reported in the literature was [ α ] D = −30.6 (c 1.0; CHCl 3 ) e.e. > 99% .…”

Section: Resultsmentioning

confidence: 69%

“…SDS‐polyacrylamide gel electrophoresis was performed according to the procedure described by Laemmli using a Miniprotean tetra‐cell (Biorad), 12% running gel. One hundred milligrams of the immobilized enzyme samples was resuspended in 1 mL of rupture buffer, which was prepared according to the methodology described by Rueda et al The samples were boiled for 10 min and 12 µL of supernatants were used in the experiment. This treatment released all enzyme just interfacially activated on the support but not the covalently attached .…”

Section: Methodsmentioning

confidence: 99%

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Kinetic resolution of drug intermediates catalyzed by lipase B from Candida antarctica immobilized on immobead‐350

Pinheiro

Rios

Fonseca

et al. 2018

Biotechnology Progress

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108

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Novozyme 435, which is a commercial immobilized lipase B from Candida antarctica (CALB), has been proven to be inadequate for the kinetic resolution of rac-indanyl acetate. As it has been previously described that different immobilization protocols may greatly alter lipase features, in this work, CALB was covalently immobilized on epoxy Immobead-350 (IB-350) and on glyoxyl-agarose to ascertain if better kinetic resolution would result. Afterwards, all CALB biocatalysts were utilized in the hydrolytic resolution of rac-indanyl acetate and rac-(chloromethyl)-2-(o-methoxyphenoxy) ethyl acetate. After optimization of the immobilization protocol on IB-350, its loading capacity was 150 mg protein/g dried support. Furthermore, the CALB-IB-350 thermal and solvent stabilities were higher than that of the soluble enzyme (e.g., by a 14-fold factor at pH 5-70°C and by a 11-fold factor in dioxane 30%-65°C) and that of the glyoxyl-agarose-CALB (e.g., by a 12-fold factor at pH 10-50°C and by a 21-fold factor in dioxane 30%-65°C). The CALB-IB-350 preparation (with 98% immobilization yield and activity versus p-nitrophenyl butyrate of 6.26 ± 0.2 U/g) was used in the hydrolysis of rac-indanyl acetate using a biocatalyst/substrate ratio of 2:1 and a pH value of 7.0 at 30°C for 24 h. The conversion obtained was 48% and the enantiomeric excess of the product (e.e. ) was 97%. These values were much higher than the ones obtained with Novozyme 435, 13% and 26% of conversion and e.e.p, respectively. © 2018 American Institute of Chemical Engineers Biotechnol. Prog., 34:878-889, 2018.

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“…Although the stability may be improved using some additives,13 the non‐absolute covalent immobilization of the adsorbed enzymes reduced interest in this strategy. Amination of the enzyme after interfacial activation on octyl glyoxyl permitted both problems to be solved (the lipases may be covalently attached at pH 9), and also improved the stabilization obtained 42. This was achieved even faster at pH 9 than using the non‐aminated enzyme at pH 10.…”

Section: Modification Of Proteins To Improve Enzyme Immobilizationmentioning

confidence: 99%

Chemical Modification in the Design of Immobilized Enzyme Biocatalysts: Drawbacks and Opportunities

Rueda

Santos

Ortíz

et al. 2016

The Chemical Record

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192

106

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Chemical modification of enzymes and immobilization used to be considered as separate ways to improve enzyme properties. This review shows how the coupled use of both tools may greatly improve the final biocatalyst performance. Chemical modification of a previously immobilized enzyme is far simpler and easier to control than the modification of the free enzyme. Moreover, if protein modification is performed to improve its immobilization (enriching the enzyme in reactive groups), the final features of the immobilized enzyme may be greatly improved. Chemical modification may be directed to improve enzyme stability, but also to improve selectivity, specificity, activity, and even cell penetrability. Coupling of immobilization and chemical modification with site-directed mutagenesis is a powerful instrument to obtain fully controlled modification. Some new ideas such as photoreceptive enzyme modifiers that change their physical properties under UV exposition are discussed.

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Chemical amination of lipases improves their immobilization on octyl-glyoxyl agarose beads

Cited by 67 publications

References 74 publications

Pilot‐scale development of core–shell polymer supports for the immobilization of recombinant lipase B from Candida antarctica and their application in the production of ethyl esters from residual fatty acids

Pilot‐scale development of core–shell polymer supports for the immobilization of recombinant lipase B from Candida antarctica and their application in the production of ethyl esters from residual fatty acids

Kinetic resolution of drug intermediates catalyzed by lipase B from Candida antarctica immobilized on immobead‐350

Chemical Modification in the Design of Immobilized Enzyme Biocatalysts: Drawbacks and Opportunities

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