Influence of phosphate anions on the stability of immobilized enzymes. Effect of enzyme nature, immobilization protocol and inactivation conditions

Kornecki, Jakub F.; Carballares, Diego; Morellon‐Sterling, Roberto; Siar, El Hocine; Kashefi, Saeid; Mazri, Chafiaa; Arana-Peña, Sara; Rios, Nathália Saraiva; Gonçalves, Luciana Rocha Barros; Fernández-Lafuente, Roberto

doi:10.1016/j.procbio.2020.02.025

Cited by 36 publications

(24 citation statements)

References 77 publications

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“…The structure of HA is composed of phosphates, hydroxyls, and calcium groups, which are useful for ionic interactions with proteins. Our results thus suggested that the main acting force was ionic bonds [ 10 , 27 ].…”

Section: Resultsmentioning

confidence: 88%

Immobilization of Dextranase on Nano-Hydroxyapatite as a Recyclable Catalyst

Ding¹,

Zhang²,

Wang³

et al. 2020

Materials

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The immobilization technology provides a potential pathway for enzyme recycling. Here, we evaluated the potential of using dextranase immobilized onto hydroxyapatite nanoparticles as a promising inorganic material. The optimal immobilization temperature, reaction time, and pH were determined to be 25 °C, 120 min, and pH 5, respectively. Dextranase could be loaded at 359.7 U/g. The immobilized dextranase was characterized by field emission gun-scanning electron microscope (FEG-SEM), X-ray diffraction (XRD), and Fourier-transformed infrared spectroscopy (FT-IR). The hydrolysis capacity of the immobilized enzyme was maintained at 71% at the 30th time of use. According to the constant temperature acceleration experiment, it was estimated that the immobilized dextranase could be stored for 99 days at 20 °C, indicating that the immobilized enzyme had good storage properties. Sodium chloride and sodium acetic did not desorb the immobilized dextranase. In contrast, dextranase was desorbed by sodium fluoride and sodium citrate. The hydrolysates were 79% oligosaccharides. The immobilized dextranase could significantly and thoroughly remove the dental plaque biofilm. Thus, immobilized dextranase has broad potential application in diverse fields in the future.

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Section: Resultsmentioning

confidence: 88%

Immobilization of Dextranase on Nano-Hydroxyapatite as a Recyclable Catalyst

Ding¹,

Zhang²,

Wang³

et al. 2020

Materials

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“…A standard colorimetric assay was developed on the basis of previously reported methods with slight modifications ( Liu and Zhu, 2015 ; Kornecki et al, 2020 ; Wang et al, 2020 ). In brief, 25 μL appropriately diluted fermentation broth (containing cells) was added to 75 μL of 5 mM p NPG.…”

Section: Methodsmentioning

confidence: 99%

Development of a New High-Cell Density Fermentation Strategy for Enhanced Production of a Fungus β-Glucosidase in Pichia pastoris

et al. 2020

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“…Biocatalysts were incubated in 50 mM Tris HCL at pH 7.0 and 60 • C. The pH was adjusted at 25 • C, thus some changes in the real pH value may be expected at 60 • C, although this should not be relevant for our purposes. Tris HCL was used as buffer to avoid the deleterious effects of phosphate buffer on lipase stability [125,126]. Periodically, samples were withdrawn and their residual activities were measured using the p-NPB assay described above.…”

Section: Lipase Biocatalysts Thermal Inactivationsmentioning

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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Influence of phosphate anions on the stability of immobilized enzymes. Effect of enzyme nature, immobilization protocol and inactivation conditions

Cited by 36 publications

References 77 publications

Immobilization of Dextranase on Nano-Hydroxyapatite as a Recyclable Catalyst

Immobilization of Dextranase on Nano-Hydroxyapatite as a Recyclable Catalyst

Development of a New High-Cell Density Fermentation Strategy for Enhanced Production of a Fungus β-Glucosidase in Pichia pastoris

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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