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This study is the first report on purification, characterization, and application of laccase derived from the white-rot fungus, Pleurotus ostreatus HK35 (Hungary strain), in Congo Red decolorization. The purification process involved ammonium sulfate precipitation, dialysis, anion exchange chromatography, and ultrafiltration, yielding a specific laccase activity of 15.26 U/mg and a 30.21% recovery rate. The purified enzyme, with a molecular weight of approximately 34 kilodaltons, displayed optimal activity at a temperature of 60 °C and pH 4.0 when using 2,2'-azino-bis (3-ethylbenzthiazoline-6-sulphonic acid) (ABTS) as a substrate. The enzyme maintained over 82.02 ± 1.01% of its activity at temperatures up to 50 °C after 180 min but displayed less than 5% of its activity at 60 and 70 °C. Notably, the enzyme's activity was significantly enhanced by Pb(NO3)2, whereas ß-mercaptoethanol completely inhibited the activity. Utilizing the Box-Behnken design, we optimized Congo Red decolorization efficiency to 91.05 ± 0.82% at 100 mg/L Congo Red, 1.5 mM mediator concentration, and 1.6 U/mL laccase activity. Analysis of Variance (ANOVA) suggested the model was significant, and all variables significantly influenced decolorization efficiency. Supplementary Information: The online version contains supplementary material available at 10.1007/s13205-024-03926-7.

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Vegetable waste, including mixed cabbage residue (MCR), is considered a promising raw material for bioenergy production because of its high lignocellulosic component. In this study, the pretreatment of MCR by ionic liquid (IL) 1-ethyl-3-methylimidazolium acetate ([Emim][OAc]) was optimized based on response surface methodology. The optimal condition for MCR pretreatment was determined at 55.8 °C, with a reaction of 2.65 h and liquid-solid ratio of 4.60:1 v/w. Hydrolysis of pretreated MCR from optimal pretreatment conditions generated a maximum glucose yield of 156.65 ± 7.66 mg/g MCR. Untreated and pretreated MCRs were successfully characterized by scanning electron microscopy, X-ray diffraction, and Fourier transform infrared spectroscopy. The pretreated MCR exhibited increased clear pores and incomplete structure. Moreover, compared with untreated biomass, decreased lignin, decreased hemicellulose, increased surface area, and cellulose crystallinity were observed. Thus, [Emim][OAc] pretreatment is a promising alternative approach for higher glucose production from MCR.

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Laccase is an effective biocatalyst in bioremediation process; however, the application of the enzyme is limited due to its cost, recovery, and stability. In this study, we developed, characterized and evaluated the efficiency of immobilized laccase on zinc oxide nanostructure to catalyze biodegradation of TBA in comparison to the suspended enzyme. The results showed that both immobilized and suspended laccase were capable of catalyzing TBA biodegradation; however, the efficiency of the immobilized laccase on TBA removal was higher than that of the suspended enzyme. The repeatability testing revealed the potential of the immobilized laccase for repeatedly catalyzing TBA biodegradation with storage capacity. While the Vmax of immobilized enzyme was higher than suspended laccase (2.25 ± 0.542 mg TBA/h∙U vs. 1.47 ± 0.185 mg TBA/h∙U), the km of the immobilized enzyme was higher than the suspended laccase (67.9 ± 20.5 mg TBA/L vs. 33.5 ± 7.10 mg TBA/L). This suggests that the immobilized laccase is better in TBA removal, but has lower affinity with TBA than the suspended enzyme. Thus, immobilization of the enzyme can be applied to increase the efficiency and minimize the use of laccase for TBA remediation.

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Here, we report the high-quality draft genome sequences of Opitutaceae sp. strains TAV3 and TAV4, which were isolated from the hindgut of the wood-feeding termite Reticulitermes flavipes Using a combination of Illumina and PacBio sequencing, we constructed nearly complete assemblies totaling 5.84 and 5.91 Mbp in length for strains TAV3 and TAV4, respectively. In addition, we report an in silico analysis of potential lignocellulose-digesting enzymes present in these strains.

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The Opitutaceae bacterium strain TAV5, a member of the phylum Verrucomicrobia, was isolated from the wood-feeding termite hindgut. We report here its complete genome sequence, which contains a chromosome and a plasmid of 7,317,842 bp and 99,831 bp, respectively. The genomic analysis reveals genes for methylotrophy, lignocellulose degradation, and ammonia and sulfate assimilation.

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Termite hindguts are populated by a dense and diverse community of microbial symbionts working in concert to transform lignocellulosic plant material and derived residues into acetate, to recycle and fix nitrogen, and to remove oxygen. Although much has been learned about the breadth of microbial diversity in the hindgut, the ecophysiological roles of its members is less understood. In this study, we present new information about the ecophysiology of microorganism Diplosphaera colotermitum strain TAV2, an autochthonous member of the Reticulitermes flavipes gut community. An integrated high-throughput approach was used to determine the transcriptomic and proteomic profiles of cells grown under hypoxia (2% O2) or atmospheric (20% O2) concentrations of oxygen. Our results revealed that genes and proteins associated with energy production and utilization, carbohydrate transport and metabolism, nitrogen fixation, and replication and recombination were upregulated under 2% O2. The metabolic map developed for TAV2 indicates that this microorganism may be involved in biological nitrogen fixation, amino-acid production, hemicellulose degradation and consumption of O2 in the termite hindgut. Variation of O2 concentration explained 55.9% of the variance in proteomic profiles, suggesting an adaptive evolution of TAV2 to the hypoxic periphery of the hindgut. Our findings advance the current understanding of microaerophilic microorganisms in the termite gut and expand our understanding of the ecological roles for members of the phylum Verrucomicrobia.

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Microbial communities in the termite hindgut are essential for degrading plant material. We present the high-quality draft genome sequence of the Opitutaceae bacterium strain TAV1, the first member of the phylum Verrucomicrobia to be isolated from wood-feeding termites. The genomic analysis reveals genes coding for lignocellulosic degradation and nitrogen fixation.

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