SciCombinator

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Concept: Ligninase

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The objective of the present work was to develop a plant-bacterial synergistic system for efficient treatment of the textile effluents. Decolorization of the dye Scarlet RR and a dye mixture was studied under in vitro conditions using Glandularia pulchella (Sweet) Tronc., Pseudomonas monteilii ANK and their consortium. Four reactors viz. soil, bacteria, plant and consortium were developed that were subjected for treatment of textile effluents and dye mixture. Under in vitro conditions G. pulchella and P. monteilii showed decolorization of the dye Scarlet RR (SRR) by 97 and 84%, within 72 and 96 h respectively, while their consortium showed 100% decolorization of the dye within 48 h. In case of dye mixture G. pulchella, P. monteilii and consortium-PG showed an ADMI removal of 78, 67 and 92% respectively within 96 h. During decolorization of SRR G. pulchella showed induction in the activities of enzymes lignin peroxidase and DCIP reductase while P. monteilii showed induction of laccase, DCIP reductase and tyrosinase, indicating their involvement in the dye metabolism. High Performance Liquid Chromatography (HPLC), Fourier Transform Infra Red Spectroscopy (FTIR) and High Performance Thin Layer Chromatography (HPTLC) confirmed the biotransformation of SRR and dye mixture into different metabolites. Soil, bacteria, plant and consortium reactors performed an ADMI removal of 42, 46, 62 and 93% in the first decolorization cycle while it showed an average ADMI removal of 21, 27, 59 and 93% in the next three (second, third and fourth) decolorization cycles respectively for the dye mixture within 24 h. Consortium reactor showed an average ADMI removal of 95% within 48 and 60 h for textile effluents A and B respectively for three decolorization cycles, while it showed an average TOC, COD and BOD removal of 74, 70 and 70%, 66, 72 and 67%, and 70, 70 and 66% for three decolorization cycles of the dye mixture (second, third and fourth decolorization cycles), effluent A and effluent B respectively. Degradation of the textile effluents and dye mixture into different metabolites by the consortium reactor was confirmed using HPLC and FTIR. Phytotoxicity studies revealed the non-toxic nature of the metabolites of degradation of dye mixture, effluents A and B by consortium reactor. The developed consortial reactor system performed efficient treatment of the dye mixture and textile effluents, and can be used for treating large amounts of textile effluents when implemented as a constructed wetland by proper engineering approach.

Concepts: Chromatography, High performance liquid chromatography, Sewage treatment, Infrared spectroscopy, Thin layer chromatography, Paper chromatography, HPTLC, Ligninase

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The use of biomaterials or microorganisms in PAHs degradation had presented an eye-catching performance. Pleurotus eryngii is a white rot fungus, which is easily isolated from the decayed woods in the tropical rain forest, used to determine the capability to utilize naphthalene, a two-ring polycyclic aromatic hydrocarbon as source of carbon and energy. In the meantime, biotransformation of naphthalene to intermediates and other by-products during degradation was investigated in this study. Pleurotus eryngii had been incubated in liquid medium formulated with naphthalene for 14 days. The presence of metabolites of naphthalene suggests that Pleurotus eryngii begin the ring cleavage by dioxygenation on C1 and C4 position to give 1,4-naphthaquinone. 1,4-Naphthaquinone was further degraded to benzoic acid, where the proposed terepthalic acid is absent in the cultured extract. Further degradation of benzoic acid by Pleurotus eryngii shows the existence of catechol as a result of the combination of decarboxylation and hydroxylation process. Unfortunately, phthalic acid was not detected in this study. Several enzymes, including manganese peroxidase, lignin peroxidase, laccase, 1,2-dioxygenase and 2,3-dioxygenase are enzymes responsible for naphthalene degradation. Reduction of naphthalene and the presence of metabolites in liquid medium showed the ability of Pleurotus eryngii to utilize naphthalene as carbon source instead of a limited glucose amount.

Concepts: Metabolism, Enzyme, Carbon, Polycyclic aromatic hydrocarbon, Benzene, Aromaticity, Naphthalene, Ligninase

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Objective: To compare the decolorization efficiency of lignin peroxidase (LiP), manganese peroxidase (MnP) and laccase on eumelanin and pheomelanin, and to investigate the effect of topical administration of LiP solution on hyperpigmented guinea pigs skin induced by 308 nm excimer light. Methods: Pheomelanin-enriched specimens were prepared from human hair and cutaneous melanoma tissue using alkaline lysis method.Synthetic eumelanin was purchased from a commercial supplier.The same amount (0.02%) of melanin was incubated with the equal enzyme activity (0.2 U/ml) of ligninolytic enzymes for 3 h respectively.The absorbance at 475 nm (A(475)) in the enzyme-catalyzed solution was measured using ELISA microplate reader.The experimental hyperpigmentation model was established in the dorsal skin of brownish guinea pigs using 308 nm excimer light radiation.LiP and heat-inactivated LiP solution were topically applied at each site.Meanwhile, 3% hydroquinone and vehicle cream were used as control.The skin color (L value) was recorded using a CR-10 Minolta chromameter.Corneocytes were collected using adhesive taping method.The amount and distribution of melanin in the corneocytes and skin tissues was visualized by Fontana-Masson staining. Results: All three ligninolytic enzymes showed various degree of eumelanin and pheomelanin decolorization activity.The decolorization activity of LiP, MnP and laccase was 40%-70%, 22%-42% and 9%-21%, respectively.The similar lightening was shown in the skin treated with LiP solution and 3% hydroquinone.The amount of melanin granules in the corneocytes was 199±11 by LiP, which was less than that in untreated control (923±12) and heat-inactive control (989±13). The amount of melanin was decreased in the whole epidermis treated with hydroquinone, the epidermis thickness was increased as well. In contrast, melanin of LiP group was decreased only in the superficial epidermis, the epidermis thickness seemed to be normal. Conclusion: LiP exerts a potent decolorization activity for hair- or skin-derived pheomelanin as well as eumelanin.It remains to be further investigated whether LiP serves as a substitute for hydroquinone in skin lightening products.

Concepts: Enzyme, Skin, Melanin, Epidermis, Human skin color, Melanocyte, Ligninase, Skin whitening

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Studies have determined that the white-rot basidiomycete Phanerochaete chrysosporium is capable of biodegrading the atrazine herbicide with its broad-specificity enzymes, but the particular role of biocatalysts is still unclear. In the case of lignin peroxidase, a ligand access channel connected to the active heme cofactor provides access to the active site for potential small-sized substrates. Experimental results show that lignin peroxidase is unable to degrade atrazine, therefore, the primary goal was to determine whether there is any connection between the structural and dynamical properties of the enzyme and its incapability to degrade atrazine. The results of protein-ligand docking and molecular dynamics study correlate with relevant, published NMR and molecular dynamics data, and give the answer to the lack of atrazine degradation by lignin peroxidase which has already been established by numerous authors using experimental methods. Atrazine has no access to heme edge due to the electric charges of the delocalized s-triazine ring. The detected phenomenon suggests that the small size of the ligands only is not a sufficient condition to access the active site. Their physicochemical properties influence the structural behaviour of the channel.

Concepts: Enzyme, Catalysis, Enzyme inhibitor, Heme, Enzymes, Herbicide, Atrazine, Ligninase

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The combination of superheated steam (SHS) with ligninolytic enzyme laccase pretreatment together with size reduction was conducted in order to enhance the enzymatic hydrolysis of oil palm biomass into glucose. The oil palm empty fruit bunch (OPEFB) and oil palm mesocarp fiber (OPMF) were pretreated with SHS and ground using a hammer mill to sizes of 2, 1, 0.5 and 0.25 mm before pretreatment using laccase to remove lignin. This study showed that reduction of size from raw to 0.25 mm plays important role in lignin degradation by laccase that removed 38.7% and 39.6% of the lignin from OPEFB and OPMF, respectively. The subsequent saccharification process of these pretreated OPEFB and OPMF generates glucose yields of 71.5% and 63.0%, which represent a 4.6 and 4.8-fold increase, respectively, as compared to untreated samples. This study showed that the combination of SHS with laccase pretreatment together with size reduction could enhance the glucose yield.

Concepts: Protein, Adenosine triphosphate, Enzyme, Starch, Lignin, Hydrolysis, Fructose, Ligninase

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A mycoremedial study was undertaken for decolourization of synthetic dyes using wood rot fungal culture Lenzites elegans WDP2. The culture was isolated from decaying wood as fruiting body, and identified on the basis of 5.8S ITS rRNA gene sequence analysis. Qualitative plate screening of culture showed extracellular laccase and lignin peroxidase production, while only laccase enzyme was produced in higher amount (156.793 Uml-1) in minimal salt broth medium containing glucose and veratryl alcohol. Laccase activity was increased up to 189.25 Uml-1 after optimization of laccase production by optimization of one variable at a time approach. Molecular characterization of laccase enzyme was done using SDS PAGE and Native PAGE based isozyme analyses. The culture was able to decolorize three synthetic dying compounds (congo red, Malachite green and brilliant green) in broth media, while showed very less decolourization in plate assay. The fungal culture varied in their dye decolourizing potential in broth culture, showing 92.77%, 21.27% and 98.8% maximum decolourization of brilliant green, malachite green and congo red respectively. The congo red dye was completely bio-absorbed by fungal culture within one month. The fungal decolourized broth also revealed the extracellular laccase activity; varied from 10 Uml-1 to 68.5 Uml-1 in all the three cases, supports the involvement of laccase enzyme in decolorization. Phase contrast microscopy clearly revealed bio-sorption of the dyes by fungal culture into the mycelium/spores in the photomicrographs.

Concepts: Molecular biology, Enzyme, RNA, Ribosomal RNA, SDS-PAGE, Dye, Triarylmethane dyes, Ligninase

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The biological pretreatment of lignocellulosic biomass is a low-cost and eco-friendly method for facilitating enzymatic hydrolysis. In this study, strains with lignin depletion capability were screened using a high-throughput screening method. Sixty-three strains were screened out and Myrothecium verrucaria secreted three lignin-degrading enzymes simultaneously during the bio-pretreatment process. The activity levels of laccase, lignin peroxidase and manganese peroxidase were 6.61, 0.78 and 1.31 U g-1dry biomass. The content of lignin in corn stover decreased by 42.30% after bio-pretreatment, and the conversion rate increased by 123.84% during the subsequent saccharification process in comparison with the untreated corn stover. Furthermore, the effects of bio-pretreatment on the structure of corn stover were presented using a scanning electron microscope (SEM), Brunauer-Emmet-Teller (BET), X-ray diffractometer (XRD) and Fourier transform infrared spectroscopy (FTIR). The results showed that M.V. is a promising lignin-degrading fungus. This research demonstrated an efficient pretreatment approach for enhancing the enzymatic saccharification of corn stover.

Concepts: Spectroscopy, Enzyme, Fungus, Starch, Scientific techniques, Scanning electron microscope, Fourier transform spectroscopy, Ligninase

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Lignocellulose is considered a major source for the production of valuable chemicals. Efficient degradation of lignin as the natural carrier of the lignocellulosic biomass represents a key limiting factor in biomass digestibility. Recently, biological delignification methods have been promoted as sustainable and environmentally friendly alternatives to the traditional technologies. In this study, porous nanocomposite of Fe3O4@SiO2@KIT-6 with magnetic properties was synthesized. The immobilized laccase supported on nanocomposite with enhanced stability in a hydrophobic ionic liquid has been developed for both olive pomace delignification and degradation of phenolic extracts from the pomace. After 6 h incubation, the degradation rate of lignin and phenol by the immobilized laccase was estimated to be 77.3% and 76.5%, respectively. The immobilized laccase retained 70% of its initial degradation ability after 11 successive batch treatments of olive pomace. Furthermore, the immobilized laccase retained >70% of its initial activity after 21 days of storage at room temperature. The obtained results indicated that the immobilized laccase on magnetic mesoporous support together with (1-Butyl-3-methylimidazolium hexafluorophosphate) ([Bmim][PF6]) could potentially provide a promising procedure for an improved enzymatic degradation of lignin and phenol in the related industries.

Concepts: Magnetism, Ionic liquid, Immobilized enzyme, Vanillin, 1-Butyl-3-methylimidazolium hexafluorophosphate, Hexafluorophosphate, Ionic liquids, Ligninase

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Biomimetic mineralization has emerged as a novel tool for generating excellent supports for enzyme stabilization. In this work, protamine was used to induce titanium (IV) bis(ammonium lactato) dihydroxide (Ti-BALDH) into titania nanoparticles. This biomimetic titanification process was adopted for laccase immobilization. Laccase-biotitania biocatalyst was prepared and the effect of different parameters (buffer solution, titania precursor concentration, protamine concentration, and enzyme loading) on the encapsulation efficiency and recovery of laccase were evaluated. Compared with free laccase, the thermal and pH stability of immobilized laccase were improved significantly. In addition, laccase loaded on titania was effective at enhancing its storage stability. After seven consecutive cycles, the immobilized laccase still retained 51% of its original activity. Finally, laccase-biotitania biocatalysts showed good performance on decolorization of malachite green (MG), which can be attributed to an adsorption and degradation effect. The intermediates of the MG degradation were identified by gas chromatography-mass spectrometry (GC-MS) analysis, and the most probable degradation pathway was proposed. This study provides deeper understanding of the laccase-biotitania particles as a fast biocatalyst for MG decolorization.

Concepts: Enzyme, Catalysis, Chemical equilibrium, PH, Buffer solution, Immobilized enzyme, Ligninase

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This study deals the biodegradation of crystal violet dye by a ligninolytic enzyme producing bacterium isolated from textile wastewater that was characterized and identified as Aeromonas hydrophila based on the 16 S rRNA gene sequence analysis. The degradation of crystal violet dye was studied under different environmental and nutritional conditions, and results showed that the isolated bacterium was effective to decolourize 99% crystal violet dye at pH 7 and temperature 35 °C in presence of sucrose and yeast extract as C and N source, respectively. This bacterium also produced lignin peroxidase and laccase enzyme, which were characterized by the SDS-PAGE analysis and found to have the molecular weight of ~ 40 and ~ 60 kDa, respectively. Further, the GC-MS analysis showed that CV dye was biotransformed into phenol, 2, 6-bis (1,1-dimethylethyl), 2',6'-dihydroxyacetophenone and benzene by the isolated bacterium and the toxicity of CV dye was reduced upto a significant level as it showed 60%, 56.67% and 46.67% inhibition in seed germination. But, after the bacterial degradation/decolourization, it showed only 43.33%, 36.67% and 16.67% inhibition in seed germination after 24, 48 and 72 h, respectively. Thus, this study concluded that the isolated bacterium has high potential for the degradation/decolourization of CV dye as well to reduce its toxicity upto a significant level.

Concepts: DNA, Bacteria, Molecular biology, Metabolism, Enzyme, RNA, Brewing, Ligninase