Effect of solid-phase chemical modification on the features of the lipase from Thermomyces lanuginosus

Galvis, Magaly; Barbosa, Oveimar; Torres, Rodrigo; Ortíz, Claudia; Fernández‐Lafuente, Roberto

doi:10.1016/j.procbio.2011.12.001

Cited by 34 publications

(21 citation statements)

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“…The enzyme preparations were filtered and washed with 25 mM sodium phosphate at pH 7 and distilled water. The recovered activities after this chemical modification for the standard pNPB measurement was 105% for A-CALB, 103% for A-RML and 117% for A-TLL, qualitatively fitting with previous reports [55,64].…”

Section: Chemical Modification Of Lipases Immobilized On Octyl Agarosesupporting

confidence: 89%

“…Modification started with the addition of EDAC up to a final concentration of 10 mM for modification of 100% of the superficial carboxylic acids [55,64]. After 90 min of gentle stirring at 25 • C, the aminated derivatives were filtered and washed 5 times with distilled water and stored at 4 • C. Then, the immobilized and chemically modified preparations were incubated for 24 h in 1 M hydroxylamine at pH 8 to recover the DEC-modified tyrosines [69].…”

Section: Chemical Modification Of Lipases Immobilized On Octyl Agarosementioning

confidence: 99%

“…Chemical modification of lipases adsorbed on octyl agarose have been described as a simple way of modifying these proteins, and the enzyme is easily desorbed after chemical modification by using detergents and later immobilized on glyoxyl supports [52][53][54]. Moreover, it has been shown that the chemical amination of the lipase surface may, in some instances, improve some enzyme properties (e.g., lipase from Thermomyces lanuginosus immobilized on octyl agarose improved activity and stability after modification) [55], but now, our objective is just to enrich the enzyme in reactive groups and in that way simplify the reaction between the enzyme and the glyoxyl groups in the support [45]. An additional advantage of this strategy is the lower pK of the new amino groups [56] that open the possibility of reacting the enzymes against glyoxyl groups at lower pH values [53].…”

Section: Introductionmentioning

confidence: 99%

“…Thus, the use of octylglyoxyl with these 3 enzymes may be greatly benefited from the use of aminated enzymes. CALB [52,64], TLL [55] and RML [65] have been already aminated with good results in terms of activity and stability recovery.…”

Section: Introductionmentioning

confidence: 99%

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

et al. 2016

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Section: Chemical Modification Of Lipases Immobilized On Octyl Agarosesupporting

confidence: 89%

Section: Chemical Modification Of Lipases Immobilized On Octyl Agarosementioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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

et al. 2016

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“…Dez gramas de Lecitase imobilizada, Lecitase-TNBS ou Lecitase-GLU foram adicionadas a 100 mL de 1M, a pH 4,75 EDA, sob agitação contínua, como descrito no trabalho de Galvis et al (2012). A modificação começou a partir da adição de DEC sólido (para uma concentração final de 10 mM).…”

Section: Aminação Em Fase Sólida De Lecitaseunclassified

MODIFICAÇÃO QUÍMICA DE LECITASE ULTRA EM FASE SÓLIDA: EFEITO DO PROTOCOLO DE IMOBILIZAÇÃO (um espaço) (um espaço)

Gonçalves¹,

Santos²,

Garcia‐Galan³

et al. 2015

Anais Do XX Congresso Brasileiro De Engenharia Química

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em octilagarose. Os derivados foram submetidos a diferentes modificações químicas (aminação, glutaraldeído ou ácido 2,4,6-trinitrobenzensulfonic (TNBS)), a fim de verificar o efeito nas propriedades catalíticas das enzimas imobilizadas. A modificação com glutaraldeído da enzima imobilizada ou modificada e aminada permitiu melhorar a estabilidade da enzima imobilizada, tanto em pH7 e 9 (em torno de 10 vezes), mas apenas a enzima aminada melhorou a estabilidade em pH5. A aminação melhorou a estabilidade de octilLecitase, ao mesmo tempo em que reduziu a estabilidade da imobilizada por covalência. E a modificação com TNBS apenas melhora a estabilidade da enzima imobilizada covalentemente, por um fator de 10 vezes, em pH9.(um espaço) INTRODUÇÃOAs fosfolipases A1 hidrolisam o grupo 1-acilo de um fosfolipídio a um lisofosfolipido e a um ácido graxo livre. Uma enzima com atividade de fosfolipase A1, nomeadamente Lecitase Ultra ® , foi patenteada e disponibilizada comercialmente, esta enzima é uma preparação obtida a partir da fusão dos genes da lipase de Thermomyces lanuginosus e a fosfolipase de Fusarium oxysporum. Essa nova enzima apresenta estabilidade da lipase de T. lanuginosus e a atividade da enzima de F. oxysporum, de acordo com Garcia-Galan et al. (2014) e De Maria et al. (2007.Neste trabalho foram estudados os efeitos de diferentes modificações químicas sobre as diversas características catalíticas de Lecitase. Esta enzima, como as lipases, tem um mecanismo catalítico chamado ativação interfacial, que seria uma tampa que cobre o centro ativo no meio homogêneo (forma fechada) em equilíbrio com uma forma em que o centro ativo é exposto ao meio (forma aberta), de acordo com estudos de Garcia-Galan et al. (2014).A modificação química foi realizada em Lecitase imobilizada, isto simplifica o processo, evita a precipitação da proteína, durante ou depois da modificação, e pode até mesmo permitir a pré-estabilização da enzima, como temos referencia no trabalho de Rodrigues et al. (2011).

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The effects of the chemical modification on immobilized lipase features are affected by the enzyme crowding in the support

Abellanas‐Perez,

Carballares,

Rocha‐Martin

et al. 2023

Biotechnology Progress

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In this article, we have analyzed the interactions between enzyme crowding on a given support and its chemical modification (ethylenediamine modification via the carbodiimide route and picryl sulfonic (TNBS) modification of the primary amino groups) on the enzyme activity and stability. Lipase from Thermomyces lanuginosus (TLL) and lipase B from Candida antarctica (CALB) were immobilized on octyl‐agarose beads at two very different enzyme loadings, one of them exceeding the capacity of the support, one well under this capacity. Chemical modifications of the highly loaded and lowly loaded biocatalysts gave very different results in terms of activity and stability, which could increase or decrease enzyme activity depending on the enzyme support loading. For example, both lowly loaded biocatalysts increased their activity after modification while the effect was the opposite for the highly loaded biocatalysts. Additionally, the modification with TNBS of highly loaded CALB biocatalyst increased its stability while decrease the activity.

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Effect of solid-phase chemical modification on the features of the lipase from Thermomyces lanuginosus

Cited by 34 publications

References 50 publications

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

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

MODIFICAÇÃO QUÍMICA DE LECITASE ULTRA EM FASE SÓLIDA: EFEITO DO PROTOCOLO DE IMOBILIZAÇÃO (um espaço) (um espaço)

The effects of the chemical modification on immobilized lipase features are affected by the enzyme crowding in the support

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