Novel Combi-lipase Systems for Fatty Acid Ethyl Esters Production

Toro, Esteban C.; proyectos, Serrano Rodríguez, Daniel, especialista de; Morales, Nelson; García, Lina; Godoy, César A.

doi:10.3390/catal9060546

Cited by 31 publications

(41 citation statements)

References 45 publications

(101 reference statements)

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“…No significant changes in the EPA/DHA specificity were observed among derivatives regardless of the pretreatment. The low ethyl esters yield values seen in the ethanolysis of palm olein with ethanol were also seen for BTL2 immobilized on glyoxyl supports under mild reaction conditions [43], which in that specific reaction context discards its use as part of more complex catalytic systems as Combi -lipases [44].…”

Section: Resultsmentioning

confidence: 99%

“…The sardine oil hydrolysis and the ethanolysis of palm olein with ethanol catalyzed by BTL2 and cc BTL2 CNBr-Sepharose ® derivatives with 6600 IU/g was carried out as described previously [43,44] with the following changes: ethanol 96% and temperature at 25°C. All the experiments were carried out by triplicate and the standard error was under 5% unless stated otherwise.…”

Section: Methodsmentioning

confidence: 99%

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Disulfide Engineered Lipase to Enhance the Catalytic Activity: A Structure-Based Approach on BTL2

Godoy

Klett

Geronimo

et al. 2019

IJMS

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Enhancement, control, and tuning of hydrolytic activity and specificity of lipases are major goals for the industry. Thermoalkaliphilic lipases from the I.5 family, with their native advantages such as high thermostability and tolerance to alkaline pHs, are a target for biotechnological applications. Although several strategies have been applied to increase lipases activity, the enhancement through protein engineering without compromising other capabilities is still elusive. Lipases from the I.5 family suffer a unique and delicate double lid restructuration to transition from a closed and inactive state to their open and enzymatically active conformation. In order to increase the activity of the wild type Geobacillus thermocatenulatus lipase 2 (BTL2) we rationally designed, based on its tridimensional structure, a mutant (ccBTL2) capable of forming a disulfide bond to lock the open state. ccBTL2 was generated replacing A191 and F206 to cysteine residues while both wild type C64 and C295 were mutated to serine. A covalently immobilized ccBTL2 showed a 3.5-fold increment in esterase activity with 0.1% Triton X-100 (2336 IU mg−1) and up to 6.0-fold higher with 0.01% CTAB (778 IU mg−1), both in the presence of oxidizing sulfhydryl agents, when compared to BTL2. The remarkable and industrially desired features of BTL2 such as optimal alkaliphilic pH and high thermal stability were not affected. The designed disulfide bond also conferred reversibility to the enhancement, as the increment on activity observed for ccBTL2 was controlled by redox pretreatments. MD simulations suggested that the most stable conformation for ccBTL2 (with the disulfide bond formed) was, as we predicted, similar to the open and active conformation of this lipase.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Disulfide Engineered Lipase to Enhance the Catalytic Activity: A Structure-Based Approach on BTL2

Godoy

Klett

Geronimo

et al. 2019

IJMS

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“…The transitions of MRM for atenolol were 267.20 > 116. 10 10. The optical purity of both substates and products as well as enantioselectivity of conducted enzyme-catalyzed biotransformation was calculated by using the equations based on peak areas from chromatogram of (R,S)-atenolol and its acetylated forms, which were described and employed in numerous studies [49,50].…”

Section: Chromatographic Conditionsmentioning

confidence: 99%

“…In addition, compound based catalyses are performed with a lot higher constancy, under mellow response conditions and are exceptionally effective as far as number of steps, giving them an edge over their synthetic partners. The one of a kind attributes of catalysts makes them exceptionally material for a number of synthetic change responses in pharmaceutical businesses, for example, particular acylation and deacylation, specific hydrolysis, deracemization, esterification, transesterification, and numerous others [10][11][12][13]. Therefore, these days, the utilization of enzymes in biotransformation has become a fascinating territory for researchers.…”

Section: Introductionmentioning

confidence: 99%

The Use of Ion Liquids as a Trojan Horse Strategy in Enzyme-Catalyzed Biotransformation of (R,S)-Atenolol

2020

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The enzymatic method was used for the direct biotransformation of racemic atenolol. The catalytic activities of commercially available lipases from Candida rugosa were tested for the kinetic resolution of (R,S)-atenolol by enantioselective acetylation in various two-phase reaction media containing ionic liquids. The composed catalytic system gave the possibility to easy separate substrates and products of the conducted enantioselective reaction and after specific procedure to reuse utilized enzymes in another catalytic cycle.

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“…That way, it has been shown that the combined use of different lipases, combining different enzyme specificities, selectivities or different sensibility to changes in the reaction conditions, produces a faster reaction and a higher conversion yield, mainly when using these heterofunctional substrates [43][44][45][46][47][48][49][50][51][52][53][54][55][56]. Even the use of the same lipase immobilized following different strategies may permit to get this positive effect [57]. The simultaneous use of several lipases has also proved to be useful in interesterification reactions [58].…”

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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Novel Combi-lipase Systems for Fatty Acid Ethyl Esters Production

Cited by 31 publications

References 45 publications

Disulfide Engineered Lipase to Enhance the Catalytic Activity: A Structure-Based Approach on BTL2

Disulfide Engineered Lipase to Enhance the Catalytic Activity: A Structure-Based Approach on BTL2

The Use of Ion Liquids as a Trojan Horse Strategy in Enzyme-Catalyzed Biotransformation of (R,S)-Atenolol

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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