Manganese Peroxidase Activity Research Articles

Harnessing the potential of a novel lignin-degrading Streptomyces sp. MS-S2 from wood-feeding termite for malachite green decolorization and detoxification

Lignin and synthetic dyes share complex structural similarities and relevant management difficulties, which should be effectively addressed towards environmental sustainability. Under this scope, the quest for oxidative enzyme-producing microorganisms with a view to efficiently valorizing organic pollutants is of utmost importance. The intestinal bacteria of wood-feeding termites represent a unique source of valuable enzymes. The decolorization efficiency of five different dyes, including malachite green (MG), methyl violet, anihiline blue, reactive red, and reactive blue, after 12 h of degradation by the MS-S2 bacterial strain was recorded to be 98.2, 17.5, 8.7, 4.6, and 3.28%, respectively. Several physico-chemical parameters were then optimized to accelerate MG degradation, including pH, temperature, glucose, and yeast extract concentration. Streptomyces sp. strain MS-S2 completely decolorized MG (50 mg/L) within 2 h under optimized conditions when the cultivation medium was amended with 5 g/L glucose and 0.08 g/L yeast extract at pH 8 and incubated at 28 °C. After MG degradation, the enzyme activities of manganese peroxidase (4.605 U/L) and laccase (45.185 U/L) were estimated. To comprehend the degradation process of MG by the MS-S2 strain, UV-Vis spectroscopy, Fourier transform infrared (FTIR), and gas chromatography-mass spectrometry (GC-MS) analyses were carried out to ascertain the possible degradation mechanism. The metabolites identified, such as 2,6-Bis (tert-butyl) phenol and [4-(dimethylamino) phenyl] phenyl, suggest that MG was broken down into less toxic compounds for further degradation. MS-S2 exhibited outstanding characteristics that could make it suitable for the bioremediation of MG dye in an industrial setting.

Expression of cellobiose dehydrogenase gene in Aspergillus niger C112 and its effect on lignocellulose degrading enzymes.

Cellobiose dehydrogenase (CDH) is one of the cellulase auxiliary proteins, which is widely used in the field of biomass degradation. However, how to efficiently and cheaply apply it in industrial production still needs further research. Aspergillus niger C112 is a significant producer of cellulase and has a relatively complete lignocellulose degradation system, but its CDH activity was only 3.92 U. To obtain a recombinant strain of A. niger C112 with high cellulases activity, the CDH from the readily available white-rot fungus Grifola frondose had been heterologously expressed in A. niger C112, under the control of the gpdA promoter. After cultivation in the medium with alkali-pretreated poplar fiber as substrate, the enzyme activity of recombinant CDH reached 36.63 U/L. Compared with the original A. niger C112, the recombinant A. niger transformed with Grifola frondosa CDH showed stronger lignocellulase activity, the activities of cellulases, β-1, 4-glucosidase and manganese peroxidase increased by 28.57, 35.07 and 121.69%, respectively. The result showed that the expression of the gcdh gene in A. niger C112 could improve the activity of some lignocellulose degrading enzymes. This work provides a theoretical basis for the further application of gcdh gene in improving biomass conversion efficiency.

Caatinga, Amazon and Atlantic Forest as natural sources for microbial lignocellulolytic enzymes.

Brazilian biomes are important sources for environmental microorganisms, including efficient metabolic machineries, like actinomycetes. These bacteria are known for their abilities to produce many bioactive compounds, including enzymes with multiple industrial applications. The present work aimed to evaluate lignocellulolytic abilities of actinomycetes isolated from soil and rhizosphere samples collected at Caatinga, Atlantic and Amazon Forest. Laccase (Lac), lignin peroxidase (LiP), manganese peroxidase (MnP) and cellulase were evaluated for their efficiency. These enzymes have an essential role in lignin decomposition, through oxidation of phenolic and non-phenolic compounds, as well as enzymatic hydrolysis of vegetal biomass. In this sense, a total of 173 actinomycetes were investigated. Eleven (11) of them were selected by their enzymatic performance. The actinomycete AC166 displayed some activity in all analysed scenarios in terms of Lac, MnP and LiP activity, while AC171 was selected as the most promising strain, showing the following activities: 29.7 U.L-1 for Lac; 2.5 U.L-1 for LiP and 23 U.L-1 for MnP. Cellulolytic activities were evaluated at two pH conditions, 4.8 and 7.4, obtaining the following results: 25 U.L-1 and 71 U.L-1, respectively. Thermostability (4, 30 and 60 o C) and salinity concentrations (0 to 4M) and pH variation (2.0 to 9.0) stabilities of the obtained LiP and Lac enzymatic extracts were also verified. The actinomycete strain AC171 displayed an adaptable response in distinct pH and salt profiles, indicating that bacterial LiP was some halophilic type. Additionally, the strain AC149 produced an alkali and extreme halophilic lignin peroxidase, which are promising profiles for their future application under lignocellulosic biomass at bioethanol biorefineries.

Use of sawdust for production of ligninolytic enzymes by white-rot fungi and pharmaceutical removal.

Use of white-rot fungi for enzyme-based bioremediation of wastewater is of high interest. These fungi produce considerable amounts of extracellular ligninolytic enzymes during solid-state fermentation on lignocellulosic materials such as straw and sawdust. We used pure sawdust colonized by Pleurotus ostreatus, Trametes versicolor, and Ganoderma lucidum for extraction of ligninolytic enzymes in aqueous suspension. Crude enzyme suspensions of the three fungi, with laccase activity range 12-43 U/L and manganese peroxidase activity range 5-55 U/L, were evaluated for degradation of 11 selected pharmaceuticals spiked at environmentally relevant concentrations. Sulfamethoxazole was removed significantly in all treatments. The crude enzyme suspension from P. ostreatus achieved degradation of wider range of pharmaceuticals when the enzyme activity was increased. Brief homogenization of the colonized sawdust was also observed to be favorable, resulting in significant reductions after a short exposure of 5 min. The highest reduction was observed for sulfamethoxazole which was reduced by 84% compared to an autoclaved control without enzyme activity and for trimethoprim which was reduced by 60%. The compounds metoprolol, lidocaine, and venlafaxine were reduced by approximately 30% compared to the control. Overall, this study confirmed the potential of low-cost lignocellulosic material as a substrate for production of enzymes from white-rot fungi. However, monitoring over time in bioreactors revealed a rapid decrease in enzymatic ligninolytic activity.

Characterization and mechanism of simultaneous degradation of aflatoxin B1 and zearalenone by an edible fungus of Agrocybe cylindracea GC-Ac2.

Contamination with multiple mycotoxins is a major issue for global food safety and trade. This study focused on the degradation of aflatoxin B1 (AFB1) and zearalenone (ZEN) by 8 types of edible fungi belonging to 6 species, inclulding Agaricus bisporus, Agrocybe cylindracea, Cyclocybe cylindracea, Cyclocybe aegerita, Hypsizygus marmoreus and Lentinula edodes. Among these fungi, Agrocybe cylindracea strain GC-Ac2 was shown to be the most efficient in the degradation of AFB1 and ZEN. Under optimal degradation conditions (pH 6.0 and 37.4°C for 37.9 h), the degradation rate of both AFB1 and ZEN reached over 96%. Through the analysis of functional detoxification components, it was found that the removal of AFB1 and ZEN was primarily degraded by the culture supernatant of the fungus. The culture supernatant exhibited a maximum manganese peroxidase (MnP) activity of 2.37 U/mL. Interestingly, Agrocybe cylindracea strain GC-Ac2 also showed the capability to degrade other mycotoxins in laboratory-scale mushroom substrates, including 15A-deoxynivalenol, fumonisin B1, B2, B3, T-2 toxin, ochratoxin A, and sterigmatocystin. The mechanism of degradation of these mycotoxins was speculated to be catalyzed by a complex enzyme system, which include MnP and other ligninolytic enzymes. It is worth noting that Agrocybe cylindracea can degrade multiple mycotoxins and produce MnP, which is a novel and significant discovery. These results suggest that this candidate strain and its enzyme system are expected to become valuable biomaterials for the simultaneous degradation of multiple mycotoxins.

Pseudomonas-associated bacteria play a key role in obtaining nutrition from bamboo for the giant panda (Ailuropoda melanoleuca).

Although giant pandas only feed on bamboo, the mechanism of lignin digestion in pandas is unclear. Here, the metabolic pathways for lignin degradation in wild pandas were explored by comparing gut metagenomic from species with different feeding habits. Results showed that lignin degradation central pathways, including beta-ketoadipate and homogentisate pathway, were enriched in the gut of wild bamboo-eating pandas. Genes from pathways involved in degrading ferulate and p-coumarate via beta-ketoadipate pathway were also enriched in bamboo-eating pandas. The final products of the above process, such as acetyl-CoA, can potentially provide the raw materials for metabolism in pandas. Specifically, Pseudomonas, as the most dominant gut bacteria genus, mainly provides genes involved in lignin degradation. Herein, Pseudomonas-associated strains isolated from the feces of pandas could degrade extracellular lignin. These findings suggest that gut microbiome of pandas is crucial in obtaining nutrition from lignin via Pseudomonas, as the main lignin-degrading bacteria.

SCREENING OF LACCASE AND MANGANESE PEROXIDASE ACTIVITIES PRODUCED BY ESCHERICHIA COLI AND PENICILLIUM ITALICUM ISOLATED FROM PLASTIC WASTE SITE IN AKURE, NIGERIA

Objective: To determine the microbial growth of Echerichia coli and Penicillium Italicum on polyethylene (PE) and screen for the activities of Manganese Peroxidase (MnP) and Laccase produced by the two microbial strain (Echerichia coli and Penicillium Italicum). Methods: Polyethylene (PE) used were obtained from Elizade university dumpsite Ilara-mokin, Ondo State. The polyethylene (PE) were cut into tiny pieces, rinsed with distilled water and then used as the sole carbon source for the growth of microorganisms in an orbital shaker flask. Laccase and manganese (Mnp) peroxidase activity were assayed in Escherichia coli and Penicillium italicum spectrophometrically as they utilize polyethylene (PE) as a carbon source. Results: Escherichia coli growth was at 0.002 at 0 h, rose to the exponential phase at 96 h, and declined to the death phase at 144 h. Penicillium italicum growth was at 0.004 at 0 h, rose to the exponential phase at 72 h, and declined to the death phase at 144 h. Laccase activity was 9.2 (U/ml) in Echerichia coli and manganese peroxidase (MnP)was 5.25 (U/ml) in Echerichia coli. Manganese peroxidase (MnP) was 10.643(U/ml) in Penicillium italicum while laccase activity was 9.5(U/ml) in Penicillium italicum. Conclusion: Echerichia coli and penicillium italicum showed Manganese peroxidase and Lacasse activities as they utilized polyethylene (PE) as carbon source. Hence, they should be explored for biodegradation of polyethylene (PE).

Activity of Lignin-Modifying Enzyme of Selected Medicinal Mushrooms in Submerged Fermentation of Lignocellulosic Materials

In the present study, wide diversity in the set and activity of lignin-modifying enzymes (LME) was revealed during submerged fermentation of mandarin peel with 15 strains of white rot Basidiomycetes. Among them, <i>Trametes pubescens</i> BCC153 was distinguished by the simultaneous production of laccase, manganese peroxidase (MnP), and lignin peroxidase (LiP). Supplementation of CuSO<sub>4</sub> at a concentration of 1 mM in the media for the cultivation of four <i>Trametes</i> species manifold increased the production of laccase. The diverse effects of chemically different lignocellulosic growth substrates and nitrogen sources on the production of individual LME have been established. The maximum laccase activity of <i>T. pubescens</i> was observed when the fungus was cultivated on media containing mandarin peel and wheat bran, whereas the highest MnP and LiP activities were detected in the submerged fermentation of tobacco residue. Peptone and casein hydrolysate appeared to be the best sources of nitrogen to produce laccase and both peroxidases by <i>T. pubescens</i> BCC153 whereas KNO<sub>3</sub> was the worst nitrogen-containing compound for the production of all enzymes.

Hydrated lime promoted the polysaccharide content and affected the transcriptomes of Lentinula edodes during brown film formation.

Brown film formation, a unique developmental stage in the life cycle of Lentinula edodes, is essential for the subsequent development of fruiting bodies in L. edodes cultivation. The pH of mushroom growth substrates are usually adjusted with hydrated lime, yet the effects of hydrated lime on cultivating L. edodes and the molecular mechanisms associated with the effects have not been studied systemically. We cultivated L. edodes on substrates supplemented with 0% (CK), 1% (T1), 3% (T2), and 5% (T3) hydrated lime (Ca (OH)2), and applied transcriptomics and qRT-PCR to study gene expression on the brown film formation stage. Hydrated lime increased polysaccharide contents in L. edodes, especially in T2, where the 5.3% polysaccharide content was approximately 1.5 times higher than in the CK. The addition of hydrated lime in the substrate promoted laccase, lignin peroxidase and manganese peroxidase activities, implying that hydrated lime improved the ability of L. edodes to decompose lignin and provide nutrition for its growth and development. Among the annotated 9,913 genes, compared to the control, 47 genes were up-regulated and 52 genes down-regulated in T1; 73 genes were up-regulated and 44 were down-regulated in T2; and 125 genes were up-regulated and 65 genes were down-regulated in T3. Differentially expressed genes (DEGs) were enriched in the amino acid metabolism, lipid metabolism and carbohydrate metabolism related pathways. The carbohydrate-active enzyme genes up-regulated in the hydrated lime treatments were mostly glycosyl hydrolase genes. The results will facilitate future optimization of L. edodes cultivation techniques and possibly shortening the production cycle.

Application of Miscanthus substrates in the cultivation of Ganoderma lingzhi.

Ganoderma lingzhi is a traditional Chinese medicine that has been used to improve health and longevity for thousands of years. It is usually cultivated on hardwood log- or sawdust-based formulations. Conversely, in this study, we used Miscanthus sacchariflorus (MSF), M. floridulus, and M. sinensis (MSS), fast-growing perennial grasses widely distributed in China, for G. lingzhi cultivation. Mycelial growth rate, activities of lignin-degrading enzymes on colonized mushroom substrates, and expression levels of CAZymes and laccase genes based on different substrates were analyzed. Total triterpenoids, sterols, and polysaccharides content of fruiting bodies obtained from different substrates were investigated. The activities of laccase and manganese peroxidase in mycelia increased in the MSF- and MSS-based formulations compared with that in the sawdust-based formulation. The results of mycelial growth- and cultivation-related experiments showed that the Miscanthus substrates could be used as the substrates for cultivating G. lingzhi. The content of activeingredients, namely triterpenoids, sterols, and polysaccharides, in fruiting bodies cultivated on the Miscanthus substrates did not decrease compared with those in substrate obtained from the sawdust-based formulation. Therefore, the present study provides alternative substrates for the cultivation of G. lingzhi, and a reference for better utilization of inexpensive substrate in future.

Optimisation of Manganese Peroxidase (MnP) Activity of Enterobacter wuhouensis Using Response Surface Method and Evaluation of Its Maillard Reaction Products Along with Lignin Degradation Ability.

Optimisation of Manganese Peroxidase (MnP) Activity of Enterobacter wuhouensis Using Response Surface Method and Evaluation of Its Maillard Reaction Products Along with Lignin Degradation Ability.

Development of manganese peroxidase based voltammetric biosensor for detection of textile Azo dyes RR 195 & RB 221

Azo dyes are extensively applied in textile and leather industries. Monitoring the toxicity and rate of exhaustion of such pollutants besides their mineralization are extremely important for environmental safety and sustainability. Detection of dyes based on enzyme inhibition mechanism and exploration of effect of various dyes on enzymes such as manganese peroxidase (MnP) for such purpose are not studied previously. The goal of the study is to develop MnP based biosensor and to determine whether all dyes impart inhibitory effect on enzyme or not. Thereby, two different dyes i.e. Reactive Red 195 (RR195) and Reactive Blue 221 (RB221) were used for the study and purified MnP was co-immobilized on carbon felt electrode for quantitative measurement of these dyes using electrochemical signals. Voltammetric analysis was carried out to determine the effects of different dye concentrations on the enzyme activity in the form electric signal (I) at an applied voltage of −1 to +1. It was revealed that increase in concentrations(10 μg/L −50 mg/L) of RR195 and RB221 transfuse inhibitory and non-inhibitory effect on MnP activity resulting in current (I) output reduction and enhancement respectively. To validate the study, another study was also conducted, which investigated the effect of these dyes on fungal growth and manganese peroxidase enzyme activity in-vitro. The results obtained authenticated the biosensing results obtained by CV as RR195 inhibit MnP activity (highly toxic) and RB221 impart no effect on MnP activity (less toxic). The fabricated biosensors demonstrated reproducibility, durability, and low threshold of detection with little interference from other substances. Even 100 days later, the MnP-based biosensors showed minor fluctuation in its current (I) outputs with 96–36 % and 86-65 % stability in case of RR195 and RB221 respectively. The study demonstrated the application of enzyme-based biosensors for both detection and toxicological monitoring of dyes.

Removal of perfluorooctanoic acid (PFOA) in the liquid culture of Phanerochaete chrysosporium

Perfluorooctanoic acid (PFOA) is becoming a concern due to its persistence, bioaccumulation, and potential harmful effects on humans and the environment. In this study, the fungus Phanerochaete chrysosporium (P. chrysosporium) was used to remove the PFOA in liquid culture system. The results showed that the average activities of laccase (Lac), lignin peroxidase (LiP), and manganese peroxidase (MnP) enzymes secreted by P. chrysosporium were 0.0003 U/mL, 0.013 U/mL, and 0.0059 U/mL, respectively, during the incubation times of 0–75 days. The pH of 3 and incubation time of 45–55 days were the optimum parameters for the three enzymes activities. The enzyme activities in P. chrysosporium incubation system were firstly inhibited by adding PFOA and then they were enhanced after 14 days. The maximum removal efficiency of PFOA (69.23%) was achieved after 35 days in P. chrysosporium incubation system with an initial PFOA concentration of 0.002 mM and no veratryl alcohol (VA). Adsorption was not a main pathway for PFOA removal and the PFOA adsorbed in fungi mycelial mat accounted for merely 1.91%. The possible products of PFOA contained partially fluorinated aldehyde, alcohol, and aromatic ring. These partially fluorinated compounds might result from PFOA degradation via a combination of cross-coupling and rearrangement of free radicals.

Production of high-value oxidative enzymes by Cyathus bulleri on agricultural and agri-food wastes for application in the textile sector.

Ligninolytic and other oxidative enzymes have emerged as promising biocatalysts in several industries. Since their production at a low cost is necessary for any large-scale application, we demonstrate the use of rice bran (RB), an agricultural waste and agri-food wastes such as potato peelings (PP), banana peelings (BP), and green pea peelings (GPP) for their production. High activity of laccase (12 U/ml), manganese peroxidase (16.11 ± 1.43 U/ml), and aryl alcohol oxidase (1.25 U/ml) was obtained on the PP on the 12th day of growth and ~ 6 U/ml of lytic polysaccharide monooxygenase was obtained on the 14th day of growth demonstrating PP to be a good substrate for their production. RB served as the next best substrate for the production of these enzymes. While the GPP was effective for the production of laccase (9.2 U/ml), this and the BP were not good substrates for the production of other enzymes. Efficient (48-82%) decolorization of several azo-, triarylmethane- dyes, and real textile effluent, without the addition of any mediator, demonstrated the high oxidative ability of the crude culture filtrate produced on the PP (CF-PP), which was a significant improvement compared to the treatment given by the previously reported culture filtrate obtained on wheat bran (CF-WB). An extensive breakdown of Reactive Orange (RO) 16 was demonstrated using CF-PP resulting in the formation of a new product at m/z of 294.05 (6-acetamido-3,4-dioxo-3,4-dihydronapthalene-2-sulfonate), previously reported to be produced on ozonation/advanced oxidation of RO16. The predominant laccase and manganese peroxidase isoforms produced on the PP were also identified.

Biodegradation of Benzo[a]pyrene by a White-Rot Fungus Phlebia acerina: Surfactant-Enhanced Degradation and Possible Genes Involved.

Polycyclic aromatic hydrocarbons (PAHs) are persistent environmental pollutants that pose a threat to human health. Among these PAHs, benzo[a]pyrene (BaP), a five-ring compound, exhibits high resistance to biodegradation. White-rot fungus Phlebia acerina S-LWZ20190614-6 has demonstrated higher BaP degradation capabilities compared with Phanerochaete chrysosporium and P. sordida YK-624, achieving a degradation rate of 57.7% after 32 days of incubation under a ligninolytic condition. To further enhance the biodegradation rate, three nonionic surfactants were used, and the addition of 1 or 2 g·L-1 of polyethylene glycol monododecyl ether (Brij 30) resulted in nearly complete BaP biodegradation by P. acerina S-LWZ20190614-6. Interestingly, Brij 30 did not significantly affect the activity of manganese peroxidase and lignin peroxidase, but it did decrease laccase activity. Furthermore, the impact of cytochrome P450 on BaP degradation by P. acerina S-LWZ20190614-6 was found to be relatively mild. Transcriptomic analysis provided insights into the degradation mechanism of BaP, revealing the involvement of genes related to energy production and the synthesis of active enzymes crucial for BaP degradation. The addition of Brij 30 significantly upregulated various transferase and binding protein genes in P. acerina S-LWZ20190614-6. Hence, the bioremediation potential of BaP by the white-rot fungus P. acerina S-LWZ20190614-6 holds promise and warrants further exploration.

Exploring the cellulolytic and hemicellulolytic activities of manganese peroxidase for lignocellulose deconstruction

BackgroundA cost-effective pretreatment and saccharification process is a necessary prerequisite for utilizing lignocellulosic biomass (LCB) in biofuel and biomaterials production. Utilizing a multifunctional enzyme with both pretreatment and saccharification functions in a single step for simultaneous biological pretreatment and saccharification process (SPS) will be a green method of low cost and high efficiency. Manganese peroxidase (MnP, EC 1.11.1.13), a well-known lignin-degrading peroxidase, is generally preferred for the biological pretreatment of biomass. However, exploring the role and performance of MnP in LCB conversion will promote the application of MnP for lignocellulose-based biorefineries.ResultsIn this study, we explored the ability of an MnP from Moniliophthora roreri, MrMnP, in LCB degradation. With Mn2+ and H2O2, MrMnP decomposed 5.0 g/L carboxymethyl cellulose to 0.14 mM of reducing sugar with a conversion yield of 5.0 mg/g, including 40 μM cellobiose, 70 μM cellotriose, 20 μM cellotetraose, and 10 μM cellohexaose, and degraded 1.0 g/L mannohexaose to 0.33 μM mannose, 4.08 μM mannotriose, and 4.35 μM mannopentaose. Meanwhile, MrMnP decomposed 5.0 g/L lichenan to 0.85 mM of reducing sugar with a conversion yield of 30.6 mg/g, including 10 μM cellotriose, 20 μM cellotetraose, and 80 μM cellohexose independently of Mn2+ and H2O2. Moreover, the versatility of MrMnP in LCB deconstruction was further verified by decomposing locust bean gum and wheat bran into reducing sugars with a conversion yield of 54.4 mg/g and 29.5 mg/g, respectively, including oligosaccharides such as di- and tri-saccharides. The catalytic mechanism underlying MrMnP degraded lignocellulose was proposed as that with H2O2, MrMnP oxidizes Mn2+ to Mn3+. Subsequently, it forms a complex with malonate, facilitating the degradation of CMC and mannohexaose into reducing sugars. Without H2O2, MrMnP directly oxidizes malonate to hydroperoxyl acetic acid radical to form compound I, which then attacks the glucosidic bond of lichenan.ConclusionThis study identified a new function of MrMnP in the hydrolysis of cellulose and hemicellulose, suggesting that MrMnP exhibits its versatility in the pretreatment and saccharification of LCB. The results will lead to an in-depth understanding of biocatalytic saccharification and contribute to forming new enzymatic systems for using lignocellulose resources to produce sustainable and economically viable products and the long-term development of biorefinery, thereby increasing the productivity of LCB as a green resource.

Establishing Experimental Conditions to Produce Lignin-Degrading Enzymes on Wheat Bran by Trametes versicolor CM13 Using Solid State Fermentation

Valorisation of wheat bran can be achieved by solid state fermentation (SSF), through application of this material as a growth substrate for a natural white rot fungal isolate, Trametes versicolor CM13, to produce lignin-degrading enzymes. One of the main challenges in optimising and upscaling (SSF) processes is the accurate adjustment and maintenance of moisture conditions. This factor was assessed in the scale up of microcosms and was evaluated over 28 days, under two slightly different moisture contents, reflecting minor differences in experimental conditions during set up and operation of the SSF process. In addition, the microcosms were processed differently from the initial trial using homogenisation of whole microcosms to create a homogeneous mixture prior to sampling. This appeared to result in less variation among the collected samples from the microcosms. Variation of measured parameters as a percentage of actual values measured ranged from 1.33% to 144% in the unmixed microcosms and from 0.77% to 36.0% in the pre-mixed microcosms. Decomposition in the more saturated microcosms progressed more quickly as hemicellulose content decreased and reached a steady state after 14 days, whereas hemicellulose content continued to decrease until 21 days in the less saturated microcosms. Lignin-degrading enzyme activities were not significantly different between either sets of experiments except for laccase on day 7. Laccase and manganese peroxidase activities were highest on day 21 and were similar in both sets of experiments. Enzyme activities on day 21 in the microcosms at moisture content of 42.9% and at 54.6% for laccase activities were 750 ± 30.5 and 820 ± 30.8 units, and for manganese peroxidase, activities were 23.3 ± 6.45 and 21.4 ± 21.4 units, respectively. These results revealed different decomposition rates during the early stage of solid-state fermentation as a function of the initial moisture content, whereas final enzyme activities and fibre content during the later stage were similar in microcosms having different moisture contents at the start.

Screening of lignin-degrading fungi and bioaugmentation on the directional humification of garden waste composting

The effective lignin-degrading strains mesophilic Sordaria tomentoalba (MSDA1) and thermophilic Thermomyces lanuginosus (HDGA2) were screened and used for the directional humification of garden waste (GW) composting. The strains MSDA1 and HDGA2 demonstrated significant Laccase (Lac), Lignin peroxidase (LiP), and Manganese peroxidase (MnP) activities, of which MnP activity was the highest at 746.31 U/L and 901.48 U/L, respectively. The composting BA-1 group was inoculated with the fungi MSDA1, HDGA2, M1 (mesophilic cellulose-degrading strain Aspergillus oryzae), and ZJ3 (thermophilic cellulose-degrading strain Thermoascus aurantiacus) on day 0, while BA-2 group was inoculated with the fungi MSDA1 and M1 on day 0 and the fungi HDGA2 and ZJ3 on day 3, respectively. The attained humic acid (HA) was 122.2 g/kg and 131.81 g/kg in BA-1 and BA-2 groups, respectively, significantly higher than the control group (72.97 g/kg). The organic matter contents of BA-2 (51.28%) were higher than that of CK (30.55%) and BA-1 (22.04%), and HA/fulvic acid (FA) were 1.26 (CK), 2.04 (BA-1) and 3.10 (BA-2), respectively. The results revealed that bioaugmentation could promote the directional humification by providing more FA precursors with the addition of lignocellulosic degradation fungi. Ammonium nitrogen (NH4+−N) could be fixed by FA during the high-temperature period, promoting NH4+−N being assimilated into amino acids and then converted to HA. Bioaugmentation effectively increased the HA and N content of the compost. The sequential addition of mesophilic and thermophilic fungi improved the quality of the directional humification compost.

Comparison of Fungal Thermophilic and Mesophilic Catalase-Peroxidases for Their Antioxidative Properties.

Catalase-peroxidases (KatGs) are unique bifunctional oxidoreductases that contain heme in their active centers allowing both the peroxidatic and catalatic reaction modes. These originally bacterial enzymes are broadly distributed among various fungi allowing them to cope with reactive oxygen species present in the environment or inside the cells. We used various biophysical, biochemical, and bioinformatics methods to investigate differences between catalase-peroxidases originating in thermophilic and mesophilic fungi from different habitats. Our results indicate that the architecture of the active center with a specific post-translational modification is highly similar in mesophilic and thermophilic KatG and also the peroxidatic acitivity with ABTS, guaiacol, and L-DOPA. However, only the thermophilic variant CthedisKatG reveals increased manganese peroxidase activity at elevated temperatures. The catalatic activity releasing molecular oxygen is comparable between CthedisKatG and mesophilic MagKatG1 over a broad temperature range. Two constructed point mutations in the active center were performed selectively blocking the formation of described post-translational modification in the active center. They exhibited a total loss of catalatic activity and changes in the peroxidatic activity. Our results indicate the capacity of bifunctional heme enzymes in the variable reactivity for potential biotech applications.

Screening and optimization of Manganese peroxidase (MnP) production by Pseudoduganella violacea (SMB4), a bacterial isolate from Similipal Biosphere Reserve, Odisha and evaluation of Maillard reaction products degradation

The Maillard reaction products (Melanoidin) are major environmental pollutants of the distillery and fermentation industries. Manganese peroxidase (MnP) was found to be an efficient ligninolytic enzyme capable of degrading Maillard products. MnP can convert Mn2 ions in wood and soils to the more reactive Mn2+ ions. The current study aims to isolate MnP producing bacteria from Similipal Biosphere Reserve (SBR), Odisha, and determine its biodegradation ability towards millards products. In total 10 ligninolytic bacterial colonies were isolated from the soil sample of SBR and their MnP activity was screened using phenol red dye as an indicator. The isolate, SMB4 was found to be the most efficient MnP-producing bacterium showing the highest MnP activity of 3.89 U/ml/min in un-optimized conditions. The bacterium was identified as Pseudoduganella violacea based on morphological, biochemical and molecular characterization. MnP activity of SMB4 was optimized using response surface methodology. Under optimized conditions SMB4 showed MnP activity of 6.12 U/ml/min which is 1.57 times from un-optimized condition. The isolate SMB4 was found to show significant decolorization of maillards products up to 83.68 % at temperature 37 °C within 192 h of incubation. The degraded product was analyzed using UV–vis and FTIR analyses. The current investigation suggests that MnP produced by Pseudoduganella violacea may be successfully exploited for bioremediation purposes to degrade Maillard products.