Evaluation of the performance of differently immobilized recombinant lipase B from Candida antarctica preparations for the synthesis of pharmacological derivatives in organic media

Manoel, Evelin Andrade; Robert, Julia de Macedo; Pinto, Maria do Carmo F. R.; Machado, Antônio C.O.; Besteti, Marina D.; Coelho, Maria Alice Zarur; Simas, Alessandro B. C.; Fernández-Lafuente, Roberto; Pinto, José Carlos; Freire, Denise Maria Guimarães

doi:10.1039/c5ra22508f

Cited by 28 publications

(38 citation statements)

References 64 publications

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“…The objective of this article was to optimize the immobilization of CALB on IB‐350 via multipoint covalent attachment and to compare the performance of the new biocatalyst with Novozyme 435 and gyoxyl‐agarose‐CALB. There are many reports stating that the properties of CALB may be strongly modulated via immobilization, and we expected that the new immobilized CALB biocatalyst may have fully different properties when compared to Novozyme 435, perhaps exhibiting better properties in the resolution of the target compounds.…”

Section: Introductionmentioning

confidence: 99%

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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Section: Introductionmentioning

confidence: 99%

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

Pinheiro

Rios

Fonseca

et al. 2018

Biotechnology Progress

Self Cite

108

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Section: Introductionmentioning

confidence: 99%

“…This polymerization technique can be very advantageous since it allows the production of micrometric porous particles and permits functionalization of the core and the shell of the polymer carriers obtained, without additional reaction steps . The use of these porous core–shell supports has been reported in some studies for the immobilization of the recombinant lipase B from Candida antarctica (CALB) . Such biocatalysts were used in some standard esterification reactions, using oleic acid and ethanol as substrates, for example .…”

Section: Introductionmentioning

confidence: 99%

“…Lipases (EC 3.1.1.3) are an important group of biocatalysts used in many biotechnological applications, such as in the synthesis of biopolymers, biodiesel, biolubricants, pharmaceuticals, agrochemicals, compounds used in the cosmetics industry, among others. [1][2][3][4][5][6][7][8][9][10] These enzymes are able to catalyze distinct reactions, such as hydrolysis, esterification and transesterification, presenting enormous versatility in different types of organic synthesis, exhibiting high specificity and selectivity. [11][12][13][14] In esterification reactions, for example, depending on the alcohol or carboxylic acid chain length, ester molecules exhibiting distinct properties are produced and can be employed for different purposes, such as emulsifiers, lubricants, flavor or cosmetic compounds.…”

Section: Introductionmentioning

confidence: 99%

“…33,38,39 The use of these porous core-shell supports has been reported in some studies for the immobilization of the recombinant lipase B from Candida antarctica (CALB). 6,36,40 Such biocatalysts were used in some standard esterification reactions, using oleic acid and ethanol as substrates, for example. 6,36 However, no study has been found that used such home-made biocatalysts for the synthesis of the ester isopropyl palmitate, a product that has direct application in the cosmetics field.…”

Section: Introductionmentioning

confidence: 99%

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Production and optimization of isopropyl palmitate via biocatalytic route using home‐made enzymatic catalysts

Barsé

Graebin

Cipolatti

et al. 2018

J of Chemical Tech & Biotech

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BACKGROUND This study investigated the use of core–shell biocatalysts, synthesized using recombinant lipase B from Candida antarctica (CALB), for the production of isopropyl palmitate and the optimization of this biocatalytic route. RESULTS Initially, three distinct core–shell biocatalysts, were evaluated for the production of isopropyl palmitate and the CALB‐P(S‐co‐DVB)/P(S‐co‐DVB) biocatalyst (biocatalyst made of polystyrene‐co‐divinylbenzene core–shell particles) was the most suitable biocatalyst for this purpose. The effect of organic solvents – n‐hexane, isooctane, n‐heptane and cyclohexane – in the ester conversion was evaluated, and the most suitable solvent was the isooctane. The optimal reaction conditions were pursued conducting an optimization statistical analysis. The most suitable reaction conditions were identified: reaction temperature, 55°C; substrate molar ratio, 1:1.42 (palmitic acid: isopropyl alcohol); biocatalyst content, 24% w/w. Finally, the reuse of the biocatalyst was evaluated and the relative enzymatic activity remained up to 50% after the fourth cycle. CONCLUSION Results were compared with commercial biocatalyst and the home‐made enzymatic biocatalyst (CALB‐P(S‐co‐DVB/PS‐co‐DVB) showed promise for application in the synthesis of isopropyl palmitate. The optimization study improved reaction conversion from 30% to 78%. The home‐made biocatalysts exhibited better performance for the synthesis of isopropyl palmitate compared with the commercial Novozym 435, using the same enzymatic activity. © 2018 Society of Chemical Industry

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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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Evaluation of the performance of differently immobilized recombinant lipase B from Candida antarctica preparations for the synthesis of pharmacological derivatives in organic media

Abstract: This paper shows the production of lipase B from Candida antarctica (LIPB) after cloning the gene that encoded it in Pichia pastoris using PGK as a constitutive promoter. The lipase was immobilized on different home-made supports for distinct reactions.

Cited by 28 publications

References 64 publications

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

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

Production and optimization of isopropyl palmitate via biocatalytic route using home‐made enzymatic catalysts

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

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