Lecitase ultra: A phospholipase with great potential in biocatalysis

Virgen-Ortíz, José J.; Santos, José Cleiton Sousa dos; Ortíz, Claudia; Berenguer-Murcia, Ángel; Barbosa, Oveimar; Rodrigues, Rafael C.; Fernández-Lafuente, Roberto

doi:10.1016/j.mcat.2019.110405

Cited by 50 publications

(39 citation statements)

References 225 publications

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“…This coimmobilization strategy has been reported using the lipase B from Candida antarctica (CALB) (a very stable enzyme) [48] and Lecitase Ultra (LU) (a phospholipase) [49] and the lipase from Rhizomucor miehei (RML) [50][51][52]. As explained above, one requirement of the strategy is that the enzyme covalently attached to the support must be the most stable one under the operational conditions.…”

Section: Introductionmentioning

confidence: 99%

Reuse of Lipase from Pseudomonas fluorescens via Its Step-by-Step Coimmobilization on Glyoxyl-Octyl Agarose Beads with Least Stable Lipases

et al. 2019

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Coimmobilization of lipases may be interesting in many uses, but this means that the stability of the least stable enzyme determines the stability of the full combilipase. Here, we propose a strategy that permits the reuse the most stable enzyme. Lecitase Ultra (LU) (a phospholipase) and the lipases from Rhizomucor miehei (RML) and from Pseudomonas fluorescens (PFL) were immobilized on octyl agarose, and their stabilities were studied under a broad range of conditions. Immobilized PFL was found to be the most stable enzyme under all condition ranges studied. Furthermore, in many cases it maintained full activity, while the other enzymes lost more than 50% of their initial activity. To coimmobilize these enzymes without discarding fully active PFL when LU or RML had been inactivated, PFL was covalently immobilized on glyoxyl-agarose beads. After biocatalysts reduction, the other enzyme was coimmobilized just by interfacial activation. After checking that glyoxyl-octyl-PFL was stable in 4% Triton X-100, the biocatalysts of PFL coimmobilized with LU or RML were submitted to inactivation under different conditions. Then, the inactivated least stable coimmobilized enzyme was desorbed (using 4% detergent) and a new enzyme reloading (using in some instances RML and in some others employing LU) was performed. The initial activity of immobilized PFL was maintained intact for several of these cycles. This shows the great potential of this lipase coimmobilization strategy.

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Section: Introductionmentioning

confidence: 99%

Reuse of Lipase from Pseudomonas fluorescens via Its Step-by-Step Coimmobilization on Glyoxyl-Octyl Agarose Beads with Least Stable Lipases

et al. 2019

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“…In other cases, enzyme activity may be improved, in some instances by fixing more active enzyme conformations (e.g., the case of lipases interfacially activated versus hydrophobic supports), in others by making more resilient enzymes under the measurement conditions. These effects of immobilization on enzyme activity have been reviewed [34][35][36][37][38][39][40][41]. Immobilization may also improve enzyme selectivity or/and selectivity in the industrially-targeted process, reduce enzyme inhibition, increase enzyme purity, etc.…”

Section: Biocatalysis and Biocatalysts Designmentioning

confidence: 99%

Genipin as An Emergent Tool in the Design of Biocatalysts: Mechanism of Reaction and Applications

et al. 2019

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Genipin is a reagent isolated from the Gardenia jasminoides fruit extract, and whose low toxicity and good crosslinking properties have converted it into a reactive whose popularity is increasing by the day. These properties have made it widely used in many medical applications, mainly in the production of chitosan materials (crosslinked by this reactive), biological scaffolds for tissue engineering, and nanoparticles of chitosan and nanogels of proteins for controlled drug delivery, the genipin crosslinking being a key point to strengthen the stability of these materials. This review is focused on the mechanism of reaction of this reagent and its use in the design of biocatalysts, where genipin plays a double role, as a support activating agent and as inter- or intramolecular crosslinker. Its low toxicity makes this compound an ideal alterative to glutaraldehyde in these processes. Moreover, in some cases the features of the biocatalysts prepared using genipin surpassed those of the biocatalysts prepared using other standard crosslinkers, even disregarding toxicity. In this way, genipin is a very promising reagent in the design of biocatalysts.

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“…Moreover, this should reinforce the PEI ionic adsorption on the biocatalysts (to reduce PEI release during the incubation in high salt concentration utilized to release the least stable and inactivated enzyme from the biocatalyst) [95]. As enzymes, we have selected some of the most used ones, such as lipases A and B from Candida antarctica (CALA and CALB) [96][97][98][99], Thermomyces lanuginosus (TLL) [100] or Rhizomucor miehei (RML) [101,102], and the commercial and artificial phospholipase Lecitase ultra (LEU) [103].…”

Section: Introductionmentioning

confidence: 99%

Multi-Combilipases: Co-Immobilizing Lipases with Very Different Stabilities Combining Immobilization via Interfacial Activation and Ion Exchange. The Reuse of the Most Stable Co-Immobilized Enzymes after Inactivation of the Least Stable Ones

et al. 2020

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The lipases A and B from Candida antarctica (CALA and CALB), Thermomyces lanuginosus (TLL) or Rhizomucor miehei (RML), and the commercial and artificial phospholipase Lecitase ultra (LEU) may be co-immobilized on octyl agarose beads. However, LEU and RML became almost fully inactivated under conditions where CALA, CALB and TLL retained full activity. This means that, to have a five components co-immobilized combi-lipase, we should discard 3 fully active and immobilized enzymes when the other two enzymes are inactivated. To solve this situation, CALA, CALB and TLL have been co-immobilized on octyl-vinyl sulfone agarose beads, coated with polyethylenimine (PEI) and the least stable enzymes, RML and LEU have been co-immobilized over these immobilized enzymes. The coating with PEI is even favorable for the activity of the immobilized enzymes. It was checked that RML and LEU could be released from the enzyme-PEI coated biocatalyst, although this also produced some release of the PEI. That way, a protocol was developed to co-immobilize the five enzymes, in a way that the most stable could be reused after the inactivation of the least stable ones. After RML and LEU inactivation, the combi-biocatalysts were incubated in 0.5 M of ammonium sulfate to release the inactivated enzymes, incubated again with PEI and a new RML and LEU batch could be immobilized, maintaining the activity of the three most stable enzymes for at least five cycles of incubation at pH 7.0 and 60 °C for 3 h, incubation on ammonium sulfate, incubation in PEI and co-immobilization of new enzymes. The effect of the order of co-immobilization of the different enzymes on the co-immobilized biocatalyst activity was also investigated using different substrates, finding that when the most active enzyme versus one substrate was immobilized first (nearer to the surface of the particle), the activity was higher than when this enzyme was co-immobilized last (nearer to the particle core).

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Lecitase ultra: A phospholipase with great potential in biocatalysis

Cited by 50 publications

References 225 publications

Reuse of Lipase from Pseudomonas fluorescens via Its Step-by-Step Coimmobilization on Glyoxyl-Octyl Agarose Beads with Least Stable Lipases

Reuse of Lipase from Pseudomonas fluorescens via Its Step-by-Step Coimmobilization on Glyoxyl-Octyl Agarose Beads with Least Stable Lipases

Genipin as An Emergent Tool in the Design of Biocatalysts: Mechanism of Reaction and Applications

Multi-Combilipases: Co-Immobilizing Lipases with Very Different Stabilities Combining Immobilization via Interfacial Activation and Ion Exchange. The Reuse of the Most Stable Co-Immobilized Enzymes after Inactivation of the Least Stable Ones

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