Laccase isozymes from Ganoderma lucidum MDU-7: Isolation, characterization, catalytic properties and differential role during oxidative stress

Kumar, Amit; Sharma, Krishna Kant; Kumar, Pramod; Ramchiary, Nirala

doi:10.1016/j.molcatb.2015.01.010

Cited by 48 publications

(30 citation statements)

References 49 publications

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“…For example, T. multicolor is reported to secrete five isozymes [50] whereas P. ostreatus secretes eight different laccase isozymes in response to Cu 2+ ions [51]. The number of laccase isozymes expressed/produced by different strains of the same species was also different [23–27, 37]. In this regard, we have summarized some physicochemical characteristics of several laccases isolated from a number of different strains of G. lucidum , Table 2.…”

Section: Discussionmentioning

confidence: 99%

Isolation and Physicochemical Characterization of Laccase fromGanoderma lucidum-CDBT1 Isolated from Its Native Habitat in Nepal

Shrestha

Joshi

et al. 2016

BioMed Research International

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At present, few organisms are known to and capable of naturally producing laccases and white rot fungi are one such group. In the present study, three fungal species, namely, Ganoderma lucidum-CDBT1, Ganoderma japonicum, and Lentinula edodes, isolated from their native habitat in Nepal were screened for laccase production, and G. lucidum-CDBT1 was found to express highest levels of enzyme (day 10 culture media showed 0.92 IU/mg total protein or 92 IU/mL laccase activity with ABTS as substrate). Lignin extracted from rice straw was used in Olga medium for laccase production and isolation from G. lucidum-CDBT1. Presence of lignin (5 g/L) and copper sulfate (30 μM) in the media increased the extracellular laccase content by 111% and 114%, respectively. The laccase enzyme produced by G. lucidum-CDBT1 was fractionated by ammonium sulfate and purified by DEAE Sepharose anion exchange chromatography. The purified enzyme was found to have a molecular mass of 43 kDa and exhibits optimal activity at pH 5.0 and 30°C. The isolated laccase was thermally stable for up to 70°C for 1 h and exhibited broad pH stability. The kinetic constants, K m, V max, and K cat, determined using 2,2′-azinobis-(-3-ethylbenzothiazoline-6-sulfonic acid) as substrate were found to be 110 μM, 36 μmol/min/mg, and 246 min−1, respectively. The isolated thermostable laccase will be used in future experiments for delignification process.

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Section: Discussionmentioning

confidence: 99%

Isolation and Physicochemical Characterization of Laccase fromGanoderma lucidum-CDBT1 Isolated from Its Native Habitat in Nepal

Shrestha

Joshi

et al. 2016

BioMed Research International

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“…I, CuSO 4 ; II, control (pH 4.0); III, tannic acid; IV, 3,5-dihydroxytoluene; V, 3,4-dihydroxybenzoic acid; VI, orcinol; VII, quinic acid; VIII, CuSO 4 ; IX, acetylsalicylic acid (1 mM); X, acetylsalicylic acid (5 mM); XI, catechol; XII, ethanol (3%); XIII, O -toluidine. (B) G. lucidum MDU-7 induced with 7 mM CuSO 4 (adopted from Kumar et al, 2015). (C) Ganoderma sp.…”

Section: Resultsmentioning

confidence: 99%

Gel-Based Purification and Biochemical Study of Laccase Isozymes from Ganoderma sp. and Its Role in Enhanced Cotton Callogenesis

et al. 2017

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Basidiomycetous fungi, Ganoderma lucidum MDU-7 and Ganoderma sp. kk-02 secreted multiple laccase isozymes under diverse growth condition. Aromatic compounds and metal salts were also found to regulate the differential expression of laccase isozymes from both the Ganoderma sp. Laccase isozymes induced in the presence of copper from G. lucidum MDU-7 were purified by gel-based (native-PAGE) purification method. The purity of laccase isozymes was checked by zymogram and SDS-PAGE. The SDS-PAGE of purified proteins confirmed the multimeric nature of laccase isozymes. The molecular mass of isozymes was found to be in the range of 40–66 kDa. Further, the purified laccase isozymes and their peptides were confirmed with the help of MALDI-TOF peptide fingerprinting. The biochemical characterization of laccase isozymes viz. Glac L2, Glac L3, Glac L4, and Glac L5 have shown the optimum temperature in the range of 30°–45°C and pH 3.0. The Km values of all the laccase isozymes determined for guaiacol were (96–281 μM), ABTS (15–83 μM) and O-tolidine (78–724 μM). Further, laccase isozymes from G. lucidum whole genome were studied using bioinformatics tools. The molecular modeling and docking of laccase isozymes with different substrates showed a significant binding affinity, which further validates our experimental results. Interestingly, copper induced laccase of 40 U/ml in culture medium was found to significantly induce cotton callogenesis. Interestingly, all the laccase isozymes were found to have an antioxidative role and therefore capable in free radicals scavenging during callogenesis. This is the first detailed study on the biochemical characterization of all the laccase isozymes purified by a gel-based novel method.

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“…In terms of intestinal uptake, apart from their particle size, nanoparticle nature and charge properties seem to influence the uptake by intestinal epithelia [15]. NPs based on hydrophilic polymers, negatively charged, showed a strong increase in bioadhesive properties and were absorbed by both M cells and absorptive enterocytes [18].…”

Section: Nanoparticles Characterizationmentioning

confidence: 99%

“…They exhibited a maximum at pH 3.0 but decreased on varying the pH from 3.0 to 6.0. Increase in pH might have caused a conformational change in laccase structure and specially on its catalytic site, thus inhibiting internal electron transfer and differing reaction product [18]. This suggests that enzyme encapsulation neither affect protein conformation nor enzyme function, keeping this laccase the characteristic behavior of the blue ones [18][19][20].…”

Section: Nanoparticles Characterizationmentioning

confidence: 99%

“…Increase in pH might have caused a conformational change in laccase structure and specially on its catalytic site, thus inhibiting internal electron transfer and differing reaction product [18]. This suggests that enzyme encapsulation neither affect protein conformation nor enzyme function, keeping this laccase the characteristic behavior of the blue ones [18][19][20]. The inhibition of T2 Cu site might be explained by the presence of OHions, which prevent oxygen reduction to water via the reaction O2 − + 2H + → H2O [17].…”

Section: Nanoparticles Characterizationmentioning

confidence: 99%

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Double-emulsion technique applied to laccase immobilization on polymeric nanoparticles

Chong-Cerda¹,

Levin²,

Castro‐Ríos³

et al. 2020

Preprint

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One primary drawback of enzyme catalysis at industrial scale is the short term service life of the enzymes. Enzymes lose their activity due to oxidation or other processes which results in less stability and a shorter lifetime thereby rendering them less efficient. An effective way to increase the stability, longevity and reusability of the enzymes is to attach them to nanoparticles by applying the double-emulsion technique. In this work the polymer Eudragit® L 100-55 sensitive to pH was used to prepare laccase polymeric nanoparticles by the double-emulsion solvent evaporation approach. The size and morphology of the nanoparticles obtained was evaluated by Scanning Electron Microscope and particle size distribution was assessed by Photon Correlation Spectroscopy. Encapsulation efficiency and zeta potential were calculated. The effect of pH on laccase activity and stability was compared between free laccase and the immobilized one. Their stability was also determined in a sequential assay involving acidic pHs up to alkaline ones. The nanoparticles had a spherical shape with a mean size of 289 nm, zeta potential of -22.7 mV at pH 7.0, and load efficiency of 87%. The optimum pH of both free and immobilized laccases was 3.0, being the nanoparticles more stable at acidic pHs (2.0-4.0). Howevwe, this last kept 80% of enzyme activity at pH 2.0 approx., after 24 h. The polymer Eudragit® L 100-55 also conferred them resistance towards the pHs usually found at the gastrointestinal tract. These results suggest the potential use of nanoparticles as adjuvants in animal feed, serving as carriers for oral laccase delivery.

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Laccase isozymes from Ganoderma lucidum MDU-7: Isolation, characterization, catalytic properties and differential role during oxidative stress

Cited by 48 publications

References 49 publications

Isolation and Physicochemical Characterization of Laccase fromGanoderma lucidum-CDBT1 Isolated from Its Native Habitat in Nepal

Isolation and Physicochemical Characterization of Laccase fromGanoderma lucidum-CDBT1 Isolated from Its Native Habitat in Nepal

Gel-Based Purification and Biochemical Study of Laccase Isozymes from Ganoderma sp. and Its Role in Enhanced Cotton Callogenesis

Double-emulsion technique applied to laccase immobilization on polymeric nanoparticles

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