Strategies of covalent immobilization of a recombinant Candida antarctica lipase B on pore-expanded SBA-15 and its application in the kinetic resolution of ( R , S )-Phenylethyl acetate

Rios, Nathália Saraiva; Pinheiro, Maísa Pessoa; Santos, José Cleiton Sousa dos; Fonseca, Thiago de Sousa; Lima, Lara Dias; Mattos, Marcos Carlos de; Freire, Denise Maria Guimarães; Silva, Ivanildo J. da; Rodríguez‐Aguado, Elena; Gonçalves, Luciana Rocha Barros

doi:10.1016/j.molcatb.2016.08.009

Cited by 74 publications

(42 citation statements)

References 58 publications

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“…The activities of soluble and immobilized CALB (CALB‐IB‐350 and CALB‐GLX) were determined by hydrolysis of 0.5 mM p NPB. The reaction was followed by spectrophotometry at 400 nm, recording the increase in absorbance caused by the release of the reaction product, p ‐nitrophenol, ( ɛ under these conditions is 10.052 mol −1 cm −1 ). One activity unit (U) was defined as the amount of CALB capable of hydrolyzing 1 µmol of p NPB per min under these conditions .…”

Section: Methodsmentioning

confidence: 99%

“…The reaction was followed by spectrophotometry at 400 nm, recording the increase in absorbance caused by the release of the reaction product, p ‐nitrophenol, ( ɛ under these conditions is 10.052 mol −1 cm −1 ). One activity unit (U) was defined as the amount of CALB capable of hydrolyzing 1 µmol of p NPB per min under these conditions . Protein concentration was determined using the methodology described by Bradford and bovine serum albumin was used as the reference protein.…”

Section: Methodsmentioning

confidence: 99%

“…One activity unit (U) was defined as the amount of CALB capable of hydrolyzing 1 mmol of pNPB per min under these conditions. 61 Protein concentration was determined using the methodology described by Bradford 62 and bovine serum albumin was used as the reference protein.…”

Section: Immobilization Of Calb On Glyoxyl-agarose Supportmentioning

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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Section: Methodsmentioning

confidence: 99%

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

Self Cite

108

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“…Numerous methods and support materials have been described for the immobilization of a wide variety of enzymes [55]. Depending on the support material used, immobilization can improve the stability of an enzyme towards harsher operating conditions, such as elevated temperatures or the presence of organic co-solvents [56][57][58], since bonds are formed between the enzyme and the support. However, in some cases, these bonds, particularly strong covalent bonds, may prevent the enzyme from adopting a more stable conformation or one that is required to catalyze the desired reaction and so, immobilization of an enzyme can also negatively affect its stability or activity [59,60].…”

Section: Immobilizationmentioning

confidence: 99%

Reactor Selection for Effective Continuous Biocatalytic Production of Pharmaceuticals

Lindeque

Woodley

2019

Catalysts

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Enzyme catalyzed reactions are rapidly becoming an invaluable tool for the synthesis of many active pharmaceutical ingredients. These reactions are commonly performed in batch, but continuous biocatalysis is gaining interest in industry because it would allow seamless integration of chemical and enzymatic reaction steps. However, because this is an emerging field, little attention has been paid towards the suitability of different reactor types for continuous biocatalytic reactions. Two types of continuous flow reactor are possible: continuous stirred tank and continuous plug-flow. These reactor types differ in a number of ways, but in this contribution, we focus on residence time distribution and how enzyme kinetics are affected by the unique mass balance of each reactor. For the first time, we present a tool to facilitate reactor selection for continuous biocatalytic production of pharmaceuticals. From this analysis, it was found that plug-flow reactors should generally be the system of choice. However, there are particular cases where they may need to be coupled with a continuous stirred tank reactor or replaced entirely by a series of continuous stirred tank reactors, which can approximate plug-flow behavior. This systematic approach should accelerate the implementation of biocatalysis for continuous pharmaceutical production.Catalysts 2019, 9, 262 2 of 17 counterparts) and thereby the use of flow technology is hard to justify. In reality, the arguments for flow technology are perhaps a little different for enzymes and, in this brief review, we will discuss the issue of reactor selection, in order to capitalize upon the benefits of both flow technology and biocatalysis. Downstream unit operations are not included in this discussion. Figure 1 shows schematic diagrams of the three ideal reactor types, namely the batch stirred tank reactor (BSTR), the continuous stirred tank reactor (CSTR) and the continuous plug-flow reactor (CPFR). The characteristics of these reactors have been described in extensive detail elsewhere [18] and will therefore be only briefly summarized here. Reactor TypesCatalysts 2019, 9, x FOR PEER REVIEW 2 of 17 (compared to their chemical counterparts) and thereby the use of flow technology is hard to justify. In reality, the arguments for flow technology are perhaps a little different for enzymes and, in this brief review, we will discuss the issue of reactor selection, in order to capitalize upon the benefits of both flow technology and biocatalysis. Downstream unit operations are not included in this discussion. Reactor Types

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“…Various immobilization techniques have been developed, including adsorption, covalent binding, entrapment, encapsulation and cross-linking [17][18][19][20][21][22][23]. Selection of materials for immobilization has an effect on the properties of immobilized enzymes [24][25][26]. Immobilization of the biocatalysts onto solid supports has become a robust and widely accepted industrial technique.…”

Section: Introductionmentioning

confidence: 99%

Bisepoxide-activated Hollow Silica Microspheres for Covalent Immobilization of Lipase from Burkholderia cepacia

Nagy

Szabó

Bugovics

et al. 2019

Period. Polytech. Chem. Eng.

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An efficient and easy-to-perform method was developed for covalent immobilization of lipase from Burkholderia cepacia (Lipase PS) on hollow silica microspheres (M540) by bisepoxide activation. For immobilization, various bisepoxides of different length, rigidity and hydrophobicity in their linkers were applied to activate the amino groups on the M540 support. Effect of the individual bisepoxides on the catalytic performance of the immobilized Lipase PS was studied by using lipase-catalyzed kinetic resolution (KR) of racemic 1-phenylethanol (rac-1) with vinyl acetate in batch mode. Catalytic activity, enantiomer selectivity, recyclability and thermal stability of the new immobilized Lipase PS biocatalysts were investigated. The optimal enzyme / support ratio with the support activated by the most efficient bisepoxide, i.e. poly(ethylene glycol) diglycidyl ether (PDE), was 1:5. The most efficient Lipase PS on PDE activated M540 showed an almost five fold higher biocatalytic activity value (rbatch = 42.8 U/g) with enhanced selectivity (ee(R)-2 = 99.1 %) to the free form of Lipase PS (rbatch = 9.0 U/g; ee(R)-2 = 98.9 %). The Lipase PS on PDE-M540 was compared to a commercially available immobilized Lipase PS biocatalyst (Lipobond Lipase PS) and also applied in a packed-bed enzyme reactor operated in continuous-flow mode, where the optimal temperature of M540-PDE-PS reached the 70 °C, while the optimum for Lipobond Lipase PS was 50 °C.

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Strategies of covalent immobilization of a recombinant Candida antarctica lipase B on pore-expanded SBA-15 and its application in the kinetic resolution of ( R , S )-Phenylethyl acetate

Cited by 74 publications

References 58 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

Reactor Selection for Effective Continuous Biocatalytic Production of Pharmaceuticals

Bisepoxide-activated Hollow Silica Microspheres for Covalent Immobilization of Lipase from Burkholderia cepacia

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