Purification and characterisation of new laccase from Trametes polyzona WRF03

Ezike, Tobechukwu Christian; Ezugwu, Arinze Linus; Udeh, Jerry Okwudili; Eze, Sabinus Oscar Onyebuchi; Chilaka, Ferdinand Chiemeka

doi:10.1016/j.btre.2020.e00566

Cited by 34 publications

(39 citation statements)

References 69 publications

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“…On the other hand, the laccase activity of Trametes villosa (1179 U/mg) is similar to that previously reported in isolates from tropical environments [53,54], demonstrating the efficiency and production in the short term, compared with the activity of isolated fungi in temperate environments [35,55,56]. Additionally, this laccase activity can be improved by adding Cu 2+ to the culture medium as an inducer [57,58] and/or changing the carbon source [59,60].…”

Section: Discussionsupporting

confidence: 81%

Enzymatic Bioprospecting of Fungi Isolated from a Tropical Rainforest in Mexico

Peraza-Jiménez

Rosa-García

Huijara-Vasconcelos

et al. 2021

JoF

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The humid tropical environment provides an ideal place for developing a high diversity of plants; this is why it is an interesting site for the enzymatic bioprospecting of fungi that are responsible for the recycling of organic matter in an efficient and accelerated way and whose enzymes could have multiple biotechnological applications. For this study, 1250 isolates of macroscopic and microscopic fungal morphotypes were collected from soil, leaf litter, and wood. One hundred and fifty strains (50 from each source) were selected for the enzymatic screening. From the first phase, 51 strains with positive activity for laccase, protease, amylase, xylanase, and lipase enzymes were evaluated, of which 20 were isolated from leaf litter, 18 from the soil, and 13 from wood. The 10 best strains were selected for the enzymatic quantification, considering the potency index and the production of at least two enzymes. High laccase activity was detected for Trametes villosa FE35 and Marasmius sp. CE25 (1179 and 710.66 U/mg, respectively), while Daedalea flavida PE47 showed laccase (521.85 U/mg) and protease activities (80.66 U/mg). Fusarium spp. PH79 and FS400 strains had amylase (14.0 U/mg, 49.23 U/mg) and xylanase activities (40.05 U/mg, 36.03 U/mg) respectively. These results confirm the enzymatic potential of fungi that inhabit little-explored tropical rainforests with applications in industry.

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Section: Discussionsupporting

confidence: 81%

Enzymatic Bioprospecting of Fungi Isolated from a Tropical Rainforest in Mexico

Peraza-Jiménez

Rosa-García

Huijara-Vasconcelos

et al. 2021

JoF

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“…Laccase was produced in 250 ml Erlenmeyer flasks containing 20 ml of fermentation medium composed of (NH 4 ) 2 SO 4 (0.22 g/L), KH 2 PO 4 (0.2 g/L), MgSO 4 .7H 2 O (0.05 g/L), thiamine (0.001 g/L), CuSO 4 (0.08 g/L), CaCl 2 (0.01 g/L), NaMO 4 (0.05 g/L), FeSO 4 (0.03 g/L), MnSO 4 (0.07 g/L), ZnSO 4 (0.043 g/L), glucose (10 g/L) and solidified with wheat bran (g/L) [ 28 ]. Three PDA agar plugs cut from the growing edge of pure Trametes polyzona WRF03 cultures were inoculated into the media.…”

Section: Methodsmentioning

confidence: 99%

“…In line with the modified method of Sahay et al. [ 31 ], crude enzyme extract from Trametes polyzona WRF03 was purified by three protein purification steps – ammonium sulfate precipitation (90 % ammonium sulfate saturation)/dialysis, anion exchange chromatography (DEAE-cellulose column; 2 × 30 cm) and gel filtration (Sephadex G-100 column; (2 × 76 cm) as previously reported [ 28 ].…”

Section: Methodsmentioning

confidence: 99%

Substrate specificity of a new laccase from Trametes polyzona WRF03

Ezike

Udeh

Joshua

et al. 2021

Heliyon

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Various aromatic compounds that are structurally analogous to lignin were tested as possible/preferred substrates for purified laccase from newly isolated white rote fungi, Trametes polyzona WRF03. The pH optima were tested using different substrates and kinetic studies were conducted at these pH optima. The pH optima in the presence of ABTS, α-naphthol, o-dianisidine, and catechol were 4.5 but 5.0 and 5.5 in the presence of guaiacol and pyrogallol, respectively. The initial velocities obtained from the kinetic study were analyzed using Graph Pad Prism 7 and Lineweaver-Burk plot to obtain kinetic constants ( k m and Vmax ) which were used to calculate substrate specificity. Amongst all the substrates tested, ABTS had the highest specificity-constant (181.51 M −1 s −1 ), and therefore, the most preferred substrate was followed by α-naphthol, o -dianisidine, guaiacol, pyrogallol, and catechol. Resorcinol, orcinol, and veratryl alcohol did not display any considerable chemical shift in the presence of Trametes polyzona WRF03 laccase. Also, oxidation of phenolic substrates appeared to be dependent on the nature of the substituent groups and their relative position on the aromatic nucleus. Since most of these substrates are structural analogs of lignin and many recalcitrant environmental pollutants, the enzyme may find application in delignification, treatment of wastewater containing dyes, and polycyclic aromatic hydrocarbons (PAHs).

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“…Acetone had a positive effect on the enzyme, causing a 28% and 45% increase in activity at a concentration of 20% and 40% (v/v), respectively [58]. On the other hand, for TpL laccase from Trametes polyzonia WRF03, a slight negative effect of polar solvents at a concentration of 10% (v/v) was found, similarly to KbLcc1 laccase, while 50% (v/v) solvent addition resulted in a loss of almost 85% of activity in the case of ethanol, 55% for methanol, and 50% for acetone after 1 h of incubation [59]. Yang et al [49] obtained opposite results for the laccase-like enzyme Lac1326 from a marine metagenomic library after 12-h incubation.…”

Section: Influence Of Organic Solvents On Laccase Kblcc1 Activitymentioning

confidence: 95%

Novel Cold-Adapted Recombinant Laccase KbLcc1 from Kabatiella bupleuri G3 IBMiP as a Green Catalyst in Biotransformation

Wiśniewska

Twarda-Cłapa

Białkowska

2021

IJMS

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Cold-adapted enzymes are useful tools in the organic syntheses conducted in mixed aqueous-organic or non-aqueous solvents due to their molecular flexibility that stabilizes the proteins in low water activity environments. A novel psychrophilic laccase gene from Kabatiella bupleuri, G3 IBMiP, was spliced by Overlap‑Extension PCR (OE-PCR) and expressed in Pichia pastoris. Purified recombinant KbLcc1 laccase has an optimal temperature of 30 °C and pH of 3.5, 5.5, 6.0, and 7.0 in the reaction with 2,2′‑azino‑bis(3‑ethylbenzothiazoline‑6‑sulfonic acid) (ABTS), guaiacol, sinapic acid, and syringaldazine, respectively. Moreover, laccase KbLcc1 is highly thermolabile, as it loses 40% of activity after 30 min at 40 °C and is inactivated at 50 °C after the same period of incubation. The new enzyme remained active with 1 mM of Ni2+, Cu2+, Mn2+, and Zn2+ and with 2 mM of Co2+, Ca2+, and Mg2+, but Fe2+ greatly inhibited the laccase activity. Moreover, 1% ethanol had no impact on KbLcc1, although acetone and ethyl acetate decreased the laccase activity. The presence of hexane (40%, v/v) caused a 58% increase in activity. Laccase KbLcc1 could be applied in the decolorization of synthetic dyes and in the biotransformation of ferulic acid to vanillin. After 5 days of reaction at 20 °C, pH 3.5, with 1 mM ABTS as a mediator, the vanillin concentration was 21.9 mg/L and the molar yield of transformation reached 14.39%.

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Purification and characterisation of new laccase from Trametes polyzona WRF03

Abstract: Graphical abstract

Cited by 34 publications

References 69 publications

Enzymatic Bioprospecting of Fungi Isolated from a Tropical Rainforest in Mexico

Enzymatic Bioprospecting of Fungi Isolated from a Tropical Rainforest in Mexico

Substrate specificity of a new laccase from Trametes polyzona WRF03

Novel Cold-Adapted Recombinant Laccase KbLcc1 from Kabatiella bupleuri G3 IBMiP as a Green Catalyst in Biotransformation

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