Purification and characterization of a fungal laccase from the ascomycete Thielavia sp. and its role in the decolorization of a recalcitrant dye

Mtibaà, Rim; Barriuso, Jorge; Eugenio, Laura de; Aranda, Elisabet; Belbahri, Lassaâd; Nasri, Monçef; Martı́nez, Marı́a Jesús; Mechichi, Tahar

doi:10.1016/j.ijbiomac.2018.09.175

Cited by 52 publications

(40 citation statements)

References 63 publications

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“…This is extremely important because only a few cold-adapted laccases have been described so far. Moreover, G3 laccase is the first enzyme of its kind for which activity was also observed at 0 and 4 • C. Most of the known multicopper oxidases are thermophilic enzymes characterized by high optimal operating temperatures, and the fungal laccases from the literature usually have T opt of 50-60 • C [29,35,45,[71][72][73][74]. Homologues with lower T opt were also described, e.g., laccases from A. pullulans [51,52], Monilinia fructicola [39], Aureobasidium melanogenum [75], V. volvacea V14 [31] and Cerrena sp.…”

Section: Properties Of the Preliminary Purified K Bupleuri Laccase G3mentioning

confidence: 99%

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Screening of Novel Laccase Producers—Isolation and Characterization of Cold-Adapted Laccase from Kabatiella bupleuri G3 Capable of Synthetic Dye Decolorization

2021

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Psychrophilic laccases catalyzing the bond formation in mild, environmentally friendly conditions are one of the biocatalysts at the focus of green chemistry. Screening of 41 cold-adapted strains of yeast and yeast-like fungi revealed a new laccase-producing strain, which was identified as Kabatiella bupleuri G3 IBMiP according to the morphological characteristics and analysis of sequences of the D1/D2 regions of 26S rDNA domain and the ITS1–5,8S–ITS2 region. The extracellular activity of laccase in reaction with 2,2’-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) at the optimal pH 3.5 was 215 U/L after 15 days of growth in a medium with waste material and 126 U/L after 25 days of cultivation in a defined medium. Copper (II) ions (0.4 mM), Tween 80 (1.0 mM) and ascorbic acid (5.0 mM) increased the production of laccase. The optimum temperature for enzyme operation is in the range of 30–40 °C and retains over 60% of the maximum activity at 10 °C. New laccase shows high thermolability—half-life at 40 °C was only 60 min. Enzyme degradation of synthetic dyes was the highest for crystal violet, i.e., 48.6% after 1-h reaction with ABTS as a mediator. Outcomes of this study present the K. bupleuri laccase as a potential psychrozyme for environmental and industrial applications.

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Section: Properties Of the Preliminary Purified K Bupleuri Laccase G3mentioning

confidence: 99%

“…20% of the maximum activity [28]. However, most of the reported laccases remain stable for at least 1 h at 50 °C [52,71,73,74,77,78]. The influence of pH on the activity of G3 laccase produced by K. bupleuri was determined.…”

Section: Properties Of the Preliminary Purified K Bupleuri Laccase G3mentioning

confidence: 99%

Screening of Novel Laccase Producers—Isolation and Characterization of Cold-Adapted Laccase from Kabatiella bupleuri G3 Capable of Synthetic Dye Decolorization

2021

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“…If one compares the kcat and kcat/Km for all three unpurified samples, it can be noticed that both values are the highest for 4A9 mutant (not shown in the table; unpurified 4A9 exhibited kcat of 48 s −1 and catalytic efficiency of 0.2 µM −1 s −1 ). The Km of many fungal laccases is in the range of 20-30 µM [36][37][38] , which is 10 times lower than for 4A9 mutant. The laccase from Thielavia sp.…”

Section: Expression and Engineering Of Laccasementioning

confidence: 88%

Evolved Fusarium oxysporum laccase expressed in Saccharomyces cerevisiae

Kwiatos

Jędrzejczak-Krzepkowska

Krzemińska

et al. 2020

Sci Rep

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Fusarium oxysporum laccase was functionally expressed in Saccharomyces cerevisiae and engineered towards higher expression levels and higher reactivity towards 2,6-dimethoxyphenol, that could be used as a mediator for lignin modification. A combination of classical culture optimization and protein engineering led to around 30 times higher activity in the culture supernatant. The winner mutant exhibited three times lower Km, four times higher kcat and ten times higher catalytic efficiency than the parental enzyme. The strategy for laccase engineering was composed of a combination of random methods with a rational approach based on QM/MM MD studies of the enzyme complex with 2,6-dimethoxyphenol. Laccase mediator system with 2,6-dimethoxyphenol caused fulvic acids release from biosolubilized coal. Laccases are oxidoreductases that catalyze the 4-electron reduction of O 2 to water with simultaneous oxidation of organic substrates. Laccases are able to catalyze oxidation of a substrate of interest directly or indirectly by the formation of a radical, which then may take part in a non-enzymatic event that effects in the oxidation of a substrate of interest. Laccases oxidize phenolic substrates but are also able to oxidize non-phenolic or bigger substrates by Laccase Mediator System (LMS), where a small phenolic compound acts as a mediator 1. Laccases contain 4 copper atoms buried in their 3D structure, which are located in two separate Cu centers. T1 copper ion is a mononuclear center, whereas T2 copper ion and two T3 copper ions are positioned in T2/T3 copper center. The substrate is oxidized closed to T1 copper ion and then the electrons are transferred to the tri-nuclear center where O 2 is reduced to water. The coppers are coordinated by nearby located residues: the T1 copper ion by one Cys and two His residues, the T2 copper by two His residues and a solvent molecule and T3 copper to three His residues 2,3. Biosolubilization of brown coal is a clean coal technology that aims at the conversion of the lignite to its cleaner form or to change its structure to gain new features 4,5. Such solubilized material could be used as a source of value-added products 4. The liquid form of coal-microorganisms, such as Fusarium oxysporum reported before as an excellent coal solubilizer 6 , uses their metabolites, alkaline substances, biosurfactants and enzymes to turn the solid polymer to black liquid. Laccases, from bacteria and fungi, for example from species Pleurotus 7 or Streptomyces 8 , are known to take part in the degradation of lignin and lignite. These two polymers are similar in structure, thus, the mechanism of their degradation is expected to be the same. Lignin comprises only 10-20% of phenolic subunits. In theory, laccase could act on those subunits present on the lignin surface and modify the polymer. However, the possibility of lignin subunits entering the laccase active site is very limited. Moreover, it is not known to what extent lignin could be modified in this way 9. According to literature data, the treatme...

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“…Laccase is a copper-containing oxidase mainly involved in the catalysis of lignin degradation, pigment synthesis, and fungal pathogenicity (Mtibaa et al, 2018). The phenotype of lighter conidia color and attenuated pathogenicity led us to analyze the laccase activity.…”

Section: Discussionmentioning

confidence: 99%

“…Laccase activity was quantified as described previously (Mtibaa et al, 2018). Based on the oxidation of ABTS, the activated strains of wild type, AaPdel, AaPdeh, and complementary mutants were inoculated on the ABTS medium at 28 • C. The morphology of these colonies was observed and photographed at 6 dpi.…”

Section: Laccase Activity Assaymentioning

confidence: 99%

Pdel, Encoding a Low-Affinity cAMP Phosphodiesterase, Regulates Conidiation and Pathogenesis in Alternaria alternata Tangerine Pathotype

Kong

Zhou

et al. 2020

Front. Microbiol.

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Based on intracellular second messenger cAMP, the cyclic AMP-protein kinase A (cAMP-PKA) pathway transforms extracellular stimuli to activate effectors and downstream signaling components, mediating physiological processes in filamentous fungi. The concentration of intracellular cAMP was regulated by adenylate cyclase biosynthesis and cAMP phosphodiesterase (PDEs) hydrolysis, which mediate signal transduction and termination. In this study, we used a gene deletion and complementary strategy to characterize the functions of AaPdel and AaPdeh genes, which encoded low-affinity PDEs (Pdel) and high-affinity PDEs (Pdeh), respectively, in Alternaria alternata. AaPdel, but not AaPdeh, was found to be a key regulator in conidiation and pathogenesis in A. alternata. ΔAaPdel showed defects in conidiation, producing approximately 65% reduced conidiation and forming lowly pigmented aberrant structures. In response to osmotic stress, ΔAaPdel was more sensitive to non-ionic osmotic stress than ionic osmotic stress. Moreover, AaPdel deletion mutants had defects in vegetative growth and hyphal growth. Further analyses showed that the high chitin content of ΔAaPdel might account for the sensitivity to Congo red. Based on the attenuated pathogenicity and lowly pigmented aberrant structures, the laccase activity analysis found that both AaPdel and AaPdeh were involved in laccase activity regulation. Our data further support the PKA-mediated cAMP signaling pathway, as we have found that AaPdel was involved in intracellular cAMP levels in A. alternata.

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Purification and characterization of a fungal laccase from the ascomycete Thielavia sp. and its role in the decolorization of a recalcitrant dye

Abstract: Purification and characterization of a fungal laccase from the ascomycete Thielavia sp. and its role in the decolourization of a recalcitrant dye

Cited by 52 publications

References 63 publications

Screening of Novel Laccase Producers—Isolation and Characterization of Cold-Adapted Laccase from Kabatiella bupleuri G3 Capable of Synthetic Dye Decolorization

Screening of Novel Laccase Producers—Isolation and Characterization of Cold-Adapted Laccase from Kabatiella bupleuri G3 Capable of Synthetic Dye Decolorization

Evolved Fusarium oxysporum laccase expressed in Saccharomyces cerevisiae

Pdel, Encoding a Low-Affinity cAMP Phosphodiesterase, Regulates Conidiation and Pathogenesis in Alternaria alternata Tangerine Pathotype

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