Immobilization of lipase from Pseudomonas fluorescens on glyoxyl-octyl-agarose beads: Improved stability and reusability

Rios, Nathália Saraiva; Mendez-Sanchez, Carmen; Arana-Peña, Sara; Rueda, Nazzoly; Ortíz, Claudia; Gonçalves, Luciana Rocha Barros; Fernández-Lafuente, Roberto

doi:10.1016/j.bbapap.2019.06.005

Cited by 42 publications

(23 citation statements)

References 59 publications

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“…That way, at 70 • C the activity remained almost unaltered after 5 h using the highly loaded preparations. This effect of enzymeenzyme interactions on enzyme stability had been previously described, but it is clearly visible using this enzyme (Fernandez-Lopez et al, 2017;Zaak et al, 2017b;Rios et al, 2019c).…”

Section: Activity With Different Substrates Of Different Pfl Preparatsupporting

confidence: 60%

“…The presence of CaCl 2 during the immobilization produced a much more stable biocatalyst than the enzyme immobilized in the absence of this additive (Table 3). This occurred even though calcium has not effects when added in the inactivation solution of immobilized PFL (Rios et al, 2019c). In fact, this biocatalyst maintained over 80% of the activity after 24 h, while the biocatalysts prepared in absence of this compound presented a half-life of only 1.5 h. Using lowly loaded preparations, this positive effect was almost negligible (see Table 1).…”

Section: Effect Of the Pfl Immobilization Conditions On The Biocatalymentioning

confidence: 96%

“…Although the immobilization using hydrophobic supports is based on weak physical interactions, it involves many groups of the enzyme, that is, the adsorption is caused by many weak enzyme-surface interactions and becomes quite strong . The most serious problem of this immobilization protocol is the risk of enzyme release under drastic conditions or in the presence of detergent-like substrates or products Hirata et al, 2016;Rios et al, 2019c;Rodrigues et al, 2019). This may be solved using different strategies, e.g., intermolecular crosslinking (physical or chemical), use of heterofunctional supports Suescun et al, 2015;Rios et al, 2019c), trapping of the enzyme in some solid matrix (Wiemann et al, 2009;Bhattacharya et al, 2012;Nieguth et al, 2017), etc.…”

Section: Introductionmentioning

confidence: 99%

“…The most serious problem of this immobilization protocol is the risk of enzyme release under drastic conditions or in the presence of detergent-like substrates or products Hirata et al, 2016;Rios et al, 2019c;Rodrigues et al, 2019). This may be solved using different strategies, e.g., intermolecular crosslinking (physical or chemical), use of heterofunctional supports Suescun et al, 2015;Rios et al, 2019c), trapping of the enzyme in some solid matrix (Wiemann et al, 2009;Bhattacharya et al, 2012;Nieguth et al, 2017), etc. In fact, the immobilization of lipases on hydrophobic supports via interfacial activation is used for the preparations of the most popular commercial immobilized lipase biocatalysts: Novozym 435 (Ortiz et al, 2019).…”

Section: Introductionmentioning

confidence: 99%

“…On the other hand, phosphate anions have been described to have a very significant destabilizing effect on lipases immobilized on these supports (Zaak et al, 2017a;Arana-Peña et al, 2018), including PFL (Rios et al, 2019c). Comparing two enzyme loads, it may be possible to determine if the enzyme-enzyme interactions can alter the effect of the immobilization conditions on the biocatalysts performance.…”

Section: Introductionmentioning

confidence: 99%

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Effects of Enzyme Loading and Immobilization Conditions on the Catalytic Features of Lipase From Pseudomonas fluorescens Immobilized on Octyl-Agarose Beads

Arana-Peña

Rios

Carballares

et al. 2020

Front. Bioeng. Biotechnol.

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The lipase from Pseudomonas fluorescens (PFL) has been immobilized on octyl-agarose beads under 16 different conditions (varying pH, ionic strength, buffer, adding some additives) at two different loadings, 1 and 60 mg of enzyme/g of support with the objective of check if this can alter the biocatalyst features. The activity of the biocatalysts versus p-nitrophenyl butyrate and triacetin and their thermal stability were studied. The different immobilization conditions produced biocatalysts with very different features. Considering the extreme cases, using 1 mg/g preparations, PFL stability changed more than fourfolds, while their activities versus pNPB or triacetin varied a 50-60%. Curiously, PFL specific activity versus triacetin was higher using highly enzyme loaded biocatalysts than using lowly loaded biocatalysts (even by a twofold factor). Moreover, stability of the highly loaded preparations was higher than that of the lowly loaded preparations, in many instances even when using 5 • C higher temperatures (e.g., immobilized in the presence of calcium, the highly loaded biocatalysts maintained after 24 h at 75 • c a 85% of the initial activity, while the lowly loaded preparation maintained only 27% at 70 • C). Using the highly loaded preparations, activity of the different biocatalysts versus pNPB varied almost 1.7-folds and versus triacetin 1.9-folds. In this instance, the changes in stability caused by the immobilization conditions were much more significant, some preparations were almost fully inactivated under conditions where the most stable one maintained more than 80% of the initial activity. Results suggested that immobilization conditions greatly affected the properties of the immobilized PFL, partially by individual molecule different conformation (observed using lowly loaded preparations) but much more relevantly using highly loaded preparations, very likely by altering some enzymeenzyme intermolecular interactions. There is not an optimal biocatalyst considering all parameters. That way, preparation of biocatalysts using this support may be a powerful tool to tune enzyme features, if carefully controlled.

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Section: Activity With Different Substrates Of Different Pfl Preparatsupporting

confidence: 60%

Section: Effect Of the Pfl Immobilization Conditions On The Biocatalymentioning

confidence: 96%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Effects of Enzyme Loading and Immobilization Conditions on the Catalytic Features of Lipase From Pseudomonas fluorescens Immobilized on Octyl-Agarose Beads

Arana-Peña

Rios

Carballares

et al. 2020

Front. Bioeng. Biotechnol.

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Application of lipase immobilized on a hydrophobic support for the synthesis of aromatic esters

Santos

Gama

Tavares

et al. 2020

Biotech and App Biochem

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The present study aimed at preparing three biocatalysts via physical adsorption of lipases from Candida rugosa (CRL), Mucor javanicus, and Candida sp. on a hydrophobic and mesoporous support (Diaion HP-20). These biocatalysts were later applied to the synthesis of aromatic esters of apple peel and citrus (hexyl butyrate), apple and rose (geranyl butyrate), and apricot and pineapple (propyl butyrate). Scanning electron microscopy and gel electrophoresis confirmed a selective adsorption of lipases on Diaion, thus endorsing simultaneous immobilization and purification. Gibbs free energy (G) evinced the spontaneity of the process (-17.9 kJ/mol ≤ G ≤ -5.1 kJ/mol). Maximum immobilized protein concentration of 30 mg/g support by CRL. This biocatalyst was the most active in olive oil hydrolysis (hydrolytic activity of 126.0 ± 2.0 U/g) and in the synthesis of aromatic esters. Maximum conversion yield of 89.1% was attained after 150 Min for the synthesis of hexyl butyrate, followed by the synthesis of geranyl butyrate (87.3% after 240 Min) and propyl butyrate (80.0% after 150 Min). CRL immobilized on Diaion retained around 93% of its original activity after six consecutive cycles of 150 Min for the synthesis of hexyl butyrate.

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Immobilization of Amano lipase AK from Pseudomonas fluorescens on different types of chitosan-containing supports: use in the kinetic resolution of rac-indanol

Fonseca

Oliveira

et al. 2021

Bioprocess Biosyst Eng

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Immobilization of lipase from Pseudomonas fluorescens on glyoxyl-octyl-agarose beads: Improved stability and reusability

Cited by 42 publications

References 59 publications

Effects of Enzyme Loading and Immobilization Conditions on the Catalytic Features of Lipase From Pseudomonas fluorescens Immobilized on Octyl-Agarose Beads

Effects of Enzyme Loading and Immobilization Conditions on the Catalytic Features of Lipase From Pseudomonas fluorescens Immobilized on Octyl-Agarose Beads

Application of lipase immobilized on a hydrophobic support for the synthesis of aromatic esters

Immobilization of Amano lipase AK from Pseudomonas fluorescens on different types of chitosan-containing supports: use in the kinetic resolution of rac-indanol

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