Heterologous fusion gene expression and characterization of a novel carbohydrate binding module (Cbm36) to laccase (Lcc2)

Kurniati, Anita; Puspaningsih, Ni Nyoman Tri; Putri, Kartika Dwi Asni; Damayanti, Mamik; Purwani, Ni Nyoman; Rahmah, Sylvia Aulia; Purkan, Purkan; Fujiyama, Kazuhito; Sakka, Makiko; Sakka, Kazuo; Kimura, Takeshi; Rohman, Ali; Baktir, Afaf; Sanjaya, Rahmat Eko

doi:10.1016/j.bcab.2022.102377

Cited by 6 publications

(4 citation statements)

References 26 publications

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“…Laccases are multicopper oxidases capable of oxidizing a variety of phenolic and non-phenolic substrates. At the same time, CBMs are non-catalytic domains that can specifically bind to carbohydrates, facilitating the enzyme’s attachment to the complex structure of lignocellulosic biomass or synthetic polymers [ 32 , 33 , 34 , 35 ]. The potential benefit of fussing a CBM onto a laccase was also emphasized in this study, with wildtype Fom_lac catalyzed oxidation of lignin through long-distance electron transfer through an aqueous solution from lignin to mediator ( Figure 7 A).…”

Section: Discussionmentioning

confidence: 99%

An Engineered Laccase from Fomitiporia mediterranea Accelerates Lignocellulose Degradation

Pham,

Deng,

Choudhary

et al. 2024

Biomolecules

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Laccases from white-rot fungi catalyze lignin depolymerization, a critical first step to upgrading lignin to valuable biodiesel fuels and chemicals. In this study, a wildtype laccase from the basidiomycete Fomitiporia mediterranea (Fom_lac) and a variant engineered to have a carbohydrate-binding module (Fom_CBM) were studied for their ability to catalyze cleavage of β-O-4′ ether and C–C bonds in phenolic and non-phenolic lignin dimers using a nanostructure-initiator mass spectrometry-based assay. Fom_lac and Fom_CBM catalyze β-O-4′ ether and C–C bond breaking, with higher activity under acidic conditions (pH < 6). The potential of Fom_lac and Fom_CBM to enhance saccharification yields from untreated and ionic liquid pretreated pine was also investigated. Adding Fom_CBM to mixtures of cellulases and hemicellulases improved sugar yields by 140% on untreated pine and 32% on cholinium lysinate pretreated pine when compared to the inclusion of Fom_lac to the same mixtures. Adding either Fom_lac or Fom_CBM to mixtures of cellulases and hemicellulases effectively accelerates enzymatic hydrolysis, demonstrating its potential applications for lignocellulose valorization. We postulate that additional increases in sugar yields for the Fom_CBM enzyme mixtures were due to Fom_CBM being brought more proximal to lignin through binding to either cellulose or lignin itself.

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Section: Discussionmentioning

confidence: 99%

An Engineered Laccase from Fomitiporia mediterranea Accelerates Lignocellulose Degradation

Pham,

Deng,

Choudhary

et al. 2024

Biomolecules

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“…Meanwhile, the positive control had slower migration due to the increased molecular size in the presence of a 542 bp DNA insert. This is because large DNA fragments migrate slower than smaller DNA fragments (17)(18)(19).…”

Section: Discussionmentioning

confidence: 99%

Temperature and Time Optimization of pGEM-T Plasmid Transformation in Escherichia coli JM109

Merdekawati

2023

Gac Méd Caracas

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Introduction: The widely used method for transformation was the heat shock method, which was influenced by the time and temperature of heat shock treatment. However, there was a lack of research on the optimal temperature for pGEM-T plasmid transformation in Escherichia coli JM109. This study uses the heat shock method to determine the optimum temperature and time for pGEM-T transformation in Escherichia coli JM109.

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“…Like pH, the temperature was energy which played a vital role in the enzymatic reaction. The maximum laccase activity was achieved at 40 o C due to an increased speed in kinetic energy and also the enzyme interaction between protein molecules and substrates to a certain temperature range (Kurniati et al 2022). The reason for declined activity towards high temperature was due to protein denature and also loss of the three-dimensional structure.…”

Section: Dye Decolorization Box-behnken Methodsmentioning

confidence: 99%

Ameliorating Direct Blue Dye Degradation Using Trametes versicolor Derived Laccase Enzyme Optimized through Box–Behnken Design (BBD) via Submerged Fermentation

et al. 2022

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The major intend of this study was to elucidate the laccase production by Trametes versicolor under submerged fermentation using fruit waste peel as substrate. The textile dye was decolorized by the procured crude enzymatic extract using the response surface methodology. The submerged media with organic fruit peel waste extract (jackfruit, pineapple & kaffir) supplemented with gypsum, calcium carbonate, and nutrient broth were considered superior for laccase production. The produced laccase enzyme was used in dye decolorization at the optimum conditions using the Box-Behnken design. Subsequently, the experiment was designed with four variables (dye concentration, pH, temperature & time) with three factors to achieve the maximum direct blue dye decolorization. The highest laccase activity level was obtained from jackfruit peel extract with 3.86U/ml on 15th day at 25oC with pH 5.0 when compared to the other two extracts. The maximum laccase activity with guaiacol was obtained at optimum pH 4 and 40oC. The predicted value was experimentally validated by attaining 81.25% of dye color removal. From the result, the optimum conditions for direct blue color removal were: dye concentration 40ppm, pH 4.0, temperature 40oC at 24 hours. From the results of this study, it was concluded that the jack fruit peel was a more suitable substrate for laccase production. The dye decolorization results were recommended that Box-Behnken design for parameters optimization. The T. versicolor laccase was more proficient for textile dye decolorization. The opportunity was created by using the laccase enzyme for the biological treatment of textile dyeing effluent before discharging into the environment.

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Heterologous fusion gene expression and characterization of a novel carbohydrate binding module (Cbm36) to laccase (Lcc2)

Cited by 6 publications

References 26 publications

An Engineered Laccase from Fomitiporia mediterranea Accelerates Lignocellulose Degradation

An Engineered Laccase from Fomitiporia mediterranea Accelerates Lignocellulose Degradation

Temperature and Time Optimization of pGEM-T Plasmid Transformation in Escherichia coli JM109

Ameliorating Direct Blue Dye Degradation Using Trametes versicolor Derived Laccase Enzyme Optimized through Box–Behnken Design (BBD) via Submerged Fermentation

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