Molecular dynamics on laccase from Trametes versicolor to examine thermal stability induced by salt bridges

Herrera-Zúñiga, Leonardo David; Millán‐Pacheco, César; Viniegra‐González, Gustavo; Villegas, Elba; Arregui, Leticia; Rojo‐Domínguez, Arturo

doi:10.1016/j.chemphys.2018.10.019

Cited by 14 publications

(10 citation statements)

References 71 publications

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“…3a). However, the negatively charged E116 on the laccase domain D1 of the wildtype Lcc9 enzyme was H-bonded with the polar S501 from the laccase domain D3, suggesting that they may contribute to a pH-dependent 3-dimensional stability of the protein by the formation of interdomain bridges (Herrera-Zúñiga et al 2019). After the substitution to the positively charged K, the H-bond was abolished (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…In consequence, not many positions can be mutated without changing activities (Bloom and Arnold 2009). From structural simulation data, E116 may contribute to formation of interdomain bridges between D1 and D3 (Herrera-Zúñiga et al 2019). As a result, the identified serendipitous mutation E116K, although not directly involved in catalysis, may be involved in influencing the pH profile of the enzyme by influencing protein structure and stabilization (Maté et al 2010, 2013a; Scheiblbrandner et al 2017).…”

Section: Discussionmentioning

confidence: 99%

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The first fungal laccase with an alkaline pH optimum obtained by directed evolution and its application in indigo dye decolorization

et al. 2019

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Engineering of fungal laccases with optimum catalytic activity at alkaline pH has been a long-lasting challenge. In this study, a mutant library containing 3000 clones was obtained by error-prone PCR to adapt the optimum pH of a fungal laccase Lcc9 from the basidiomycete Coprinopsis cinerea. After three rounds of functional screening, a mutant with three amino acid changes (E116K, N229D, I393T) named PIE5 was selected. PIE5 showed an optimum pH of 8.5 and 8.0 against guaiacol and 2,6-DMP when expressed in Pichia pastoris, representing the first fungal laccase that possesses an optimum pH at an alkaline condition. Site directed mutagenesis disclosed that N229D contributed the most to the optimum pH increment. A single N229D mutation caused an increase in optimum pH by 1.5 units. When used in indigo dye decolorization, PIE5 efficiently decolorized 87.1 ± 1.1% and 90.9 ± 0.3% indigo dye at the optimum conditions of pH 7.0–7.5 and 60 °C, and with either methyl 3,5-dimethoxy-4-hydroxybenzoate or 2,2′-azino-bis(3-ethylbenzothazoline-6-sulfonate) as the mediator. In comparison, the commercially available fungal laccase TvLac from Trametes villosa decolorized 84.3 ± 1.8% of indigo dye under its optimum conditions (opt. pH 5.0 and 60 °C). The properties of an alkaline-dependent activity and the high indigo dye decolorization ability (1.3-fold better than the parental Lcc9) make the new fungal laccase PIE5 an alternative for specific industrial applications.

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Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

The first fungal laccase with an alkaline pH optimum obtained by directed evolution and its application in indigo dye decolorization

et al. 2019

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“…These approaches have successfully produced laccase mutants or derivatives with enhanced temperature or organic solvent stability; activities tailored to develop specificity to certain substrates; higher E ° in the T1 site, enhanced heterologous expression, the shift of pH-activity profiles, and tolerance to chemical inhibitors. In structural terms, these improvements were achieved by modifying the functional groups in the substrate binding site and T1 copper coordination [22], as well as introducing stabilizing mutants in the domain interface [163]. Nevertheless, the precise prediction of the effect of a specific mutation remains elusive [152, 155].…”

Section: Structure Of Laccases and Comparative Structure Analysesmentioning

confidence: 99%

Laccases: structure, function, and potential application in water bioremediation

et al. 2019

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The global rise in urbanization and industrial activity has led to the production and incorporation of foreign contaminant molecules into ecosystems, distorting them and impacting human and animal health. Physical, chemical, and biological strategies have been adopted to eliminate these contaminants from water bodies under anthropogenic stress. Biotechnological processes involving microorganisms and enzymes have been used for this purpose; specifically, laccases, which are broad spectrum biocatalysts, have been used to degrade several compounds, such as those that can be found in the effluents from industries and hospitals. Laccases have shown high potential in the biotransformation of diverse pollutants using crude enzyme extracts or free enzymes. However, their application in bioremediation and water treatment at a large scale is limited by the complex composition and high salt concentration and pH values of contaminated media that affect protein stability, recovery and recycling. These issues are also associated with operational problems and the necessity of large-scale production of laccase. Hence, more knowledge on the molecular characteristics of water bodies is required to identify and develop new laccases that can be used under complex conditions and to develop novel strategies and processes to achieve their efficient application in treating contaminated water. Recently, stability, efficiency, separation and reuse issues have been overcome by the immobilization of enzymes and development of novel biocatalytic materials. This review provides recent information on laccases from different sources, their structures and biochemical properties, mechanisms of action, and application in the bioremediation and biotransformation of contaminant molecules in water. Moreover, we discuss a series of improvements that have been attempted for better organic solvent tolerance, thermo-tolerance, and operational stability of laccases, as per process requirements.

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“…White rot fungi are known to be active lignin degraders as they harness a plethora of ligninolytic enzymes and the depolymerization rate by fungi ranges from 20% to nearly 100%, depending on different lignin sources 39 . S.commune has been used as a model organism for the production of ligninolytic enzymes 40 and there has been an increasing interest in exploiting the potential of S.commune in various biotechnological processes due to the presence of protein-coding genes like laccases, glycoside hydrolases, lytic polysaccharide monooxygenases (LPMOs) and expansins—like proteins 41 , production of ligninolytic enzymes 14 , 42 , 43 , use in mixed cultures 44 . Various types of ligninolytic enzymes are produced by WRF in nature based on the substrate upon which it acts.…”

Section: Resultsmentioning

confidence: 99%

Biological depolymerization of lignin using laccase harvested from the autochthonous fungus Schizophyllum commune employing various production methods and its efficacy in augmenting in vitro digestibility in ruminants

Kumar

Sridhar

Rao

2022

Sci Rep

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A laccase-producing hyper performer, Schizophyllum commune, a white-rot fungus, was evaluated for its ability to selectively degrade lignin of diverse crop residues in vitro. Relative analysis of crop residue treatment using laccase obtained from immobilized cells demonstrated degradation of 30–40% in finger millet straw and sorghum stover, 27–32% in paddy straw, 21% in wheat straw, and 26% in maize straw, while 20% lignin degradation was observed when purified and recombinant laccase was used. Further investigations into in vitro dry matter digestibility studies gave promising results recording digestibility of 54–59% in finger millet straw 33–36% in paddy straw and wheat straw, 16% in maize straw for laccase obtained from cell immobilization method, whereas 14% digestibility was observed when purified and recombinant laccase was used. Sorghum stover recorded digestibility of 13–15% across all straws treated with laccase. The results obtained elucidated the positive influence of laccase treatment on lignin degradation and in vitro dry matter digestibility. The present research gave encouraging figures confirming the production of laccase using the cell immobilization method to be an efficient production method commensurate with purified and recombinant laccase under conditions of submerged cultivation, proclaiming a cost-effective, environmentally safe green technology for effectual lignin depolymerization.

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Molecular dynamics on laccase from Trametes versicolor to examine thermal stability induced by salt bridges

Cited by 14 publications

References 71 publications

The first fungal laccase with an alkaline pH optimum obtained by directed evolution and its application in indigo dye decolorization

The first fungal laccase with an alkaline pH optimum obtained by directed evolution and its application in indigo dye decolorization

Laccases: structure, function, and potential application in water bioremediation

Biological depolymerization of lignin using laccase harvested from the autochthonous fungus Schizophyllum commune employing various production methods and its efficacy in augmenting in vitro digestibility in ruminants

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