Manganese Peroxidase Production Research Articles

Enhancing Manganese Peroxidase: Innovations in Genetic Modification, Screening Processes, and Sustainable Agricultural Applications.

Manganese peroxidase (MnP), a vital extracellular enzyme for the degradation of lignin and other organic pollutants, has demonstrated immense potential for agricultural and environmental applications, including straw pretreatment, feed fermentation, mycotoxin degradation, and water treatment. However, current research remains in its exploratory phase, with naturally sourced MnP unable to meet industrial-scale demands and no mature commercial enzyme preparations available on the market. This comprehensive review innovatively constructs a framework for MnP research, probing into its molecular conformation and catalytic principles, while providing an overview of the advancements in high-throughput screening and In silco designing strategies. Specifically, this review focuses on the practical applications of MnP in sustainable agriculture, elaborating on its potential and challenges in straw resource utilization, efficient feed fermentation, mycotoxin control, and water quality improvement. Furthermore, this review summarizes the recent achievements in optimizing MnP activity through enzyme engineering techniques and discuss customized mutation strategies tailored to specific agricultural and environmental requirements, thereby laying a solid theoretical foundation and scientific basis for the industrial production and commercialization of MnP.

Evaluation of lignocellulatic activity of enzymes from microwave-irradiated Pleurotus sajor-caju cultivated with wheat straw

The production of lignocellulytic enzymes by microwave-radiated Pleurotus sajor-caju was assayed. Wheat straw was employed as substrate to P. sajor-caju for production of laccase, manganese peroxidase (MnPase), filter-paperase (FPase), carboxmethyl cellulase (CMCase), and cellulase (as evaluated using microcrystalline cellulose). P. sajor-caju exposed to 10 s of microwave radiation (MR) showed maximum growth with colony radius of 7.17 ± 0.45 cm, while with increasing the exposure time up to 50 s the growth decreased up to 2.67 ± 0.22 cm. Moreover, it failed to grow at 80 s of exposure time. Cellulase, MnPase, FPase, CMCase, and laccase activities were induced to 37 ± .0.54, 49 ± 2.36, 189 ± 2.12, 0.37 ± 0.06, and 1.58 ± 0.03 U/mL compared to that at control 31 ± 0.25, 46 ± 1.25, 177 ± 1.65, 0.28 ± 0.03, and 1.37 ± 0.12 U/mL, respectively as a result of P. sajor-caju exposure to 10 s of MR. As the exposure time increased, these enzymes activity decreased. Different levels of moisture with surfactant (polysorbate 80) were applied to optimize the enzymes activities at 10 s of exposure time. The optimum activities 3.15 ± 0.23, 0.62 ± 0.06, 269 ± 5.36, 65 ± 1.63, and 48 ± 0.98 U/mL were recorded for cellulase, MnPase, FPase, CMCase, and laccase, respectively at 70% of moisture and 0.15 mL/L of polysorbate 80.

Producción de oxidasas por Trametes versicolor crecido sobre residuos verdes y sobre espuma de poliuretano en fermentación en estado sólido: Un estudio comparativo

Green waste (GW) is generated by the maintenance of public or private green spaces. It is necessary to find ecological alternatives for GW utilization, aiming to avoid accumulation of this material at the environment. In this research, the production of laccase (Lac), lignin peroxidase (LiP), manganese peroxidase (MnP) and unspecific peroxygenase (UnP) produced by Trametes versicolor grown on GW as a substrate and on polyurethane foam (PUF) as an inert support in solid state fermentation was evaluated. T. versicolor showed higher values of Lac, MnP, UnP and LiP activities (34, 943, 1023 and 766 U/gS, respectively) when grown on GW than when grown on PUF (10.9, 588, 559 and 229 U/gS, respectively). These results suggest that T. versicolor produced Lac inducible and constitutively, while LiP, MnP and UnP were induced by GW at the beginning of fungal growth, however, these enzymes were constitutive and inducible during the rest of the fermentation. The production of oxidases and peroxidases was induced and increased by GW. It is suggested that LiP is involved (as a constitutive enzyme) at the beginning of the exponential phase, while MnP and UnP participate in fungal growth at the end of fermentation. To our knowledge, this is the first detailed study on the main lignincellulose-degrading fungal enzymes involved in GW degradation by fungi. In particular, the relevance of UnP was showed as peroxidase involved in lignocellulosic substrates biodegradation.

Reduced graphene oxide promotes the biodegradation of sulfamethoxazole by white-rot fungus Phanerochaete chrysosporium under cadmium stress

The biodegradation of antibiotics in aquatic environment is consistently impeded by the widespread presence of heavy metals, necessitating urgent measures to mitigate or eliminate this environmental stress. This work investigated the degradation of sulfamethoxazole (SMX) by the white-rot fungus Phanerochaete chrysosporium (WRF) under heavy metal cadmium ion (Cd2+) stress, with a focus on the protective effects of reduced graphene oxide (RGO). The pseudo-first-order rate constant and removal efficiency of 5 mg/L SMX in 48 h by WRF decrease from 0.208 h−1 and 55.6% to 0.08 h−1 and 28.6% at 16 mg/L of Cd2+, while these values recover to 0.297 h−1 and 72.8% by supplementing RGO. The results demonstrate that RGO, possessing excellent biocompatibility, effectively safeguard the mycelial structure of WRF against Cd2+ stress and provide protection against oxidative damage to WRF. Simultaneously, the production of manganese peroxidase (MnP) by WRF decreases to 38.285 U/L in the presence of 24 mg/L Cd2+, whereas it recovers to 328.51 U/L upon the supplement of RGO. RGO can induce oxidative stress in WRF, thereby stimulating the secretion of laccase (Lac) and MnP to enhance the SMX degradation. The mechanism discovered in this study provides a new strategy to mitigate heavy metal stress encountered by WRF during antibiotic degradation.

Isolation and screening of potential lignocellulolytic microbes from Phra Nakhon Si Ayutthaya Province

Phra Nakhon Si Ayutthaya Province is an important area for rice cultivation in Thailand. However, large amounts of rice straw generated at harvest time are often burnt in the fields, causing environmental degradation and air pollution. To address this, rice straw composting by degrading microorganisms has been identified as an effective alternative for sustainable waste management, with potential applications in environmental biotechnology. For this study, 28 microorganism samples were collected from various sources in Phra Nakhon Si Ayutthaya Province, including a corrugated paper manufacturer, plant-cultivated soil, compost from dung, black soldier flies, and vermicompost. Lignocellulolytic microorganisms were screened for cellulase, xylanase, and manganese peroxidase production on agar plates and broth media using commercial and rice straw substrates. The lignocellulolytic activities were compared among the isolated and non-pathogenic microorganisms. A microbial consortium with high potential to degrade rice straw was identified, consisting of two thermophilic bacteria, Bacillus licheniformis BKT1 and BOT4, two mesophilic actinomycetes, Streptomyces ardesiacus AQ4 and An6, two thermophilic actinomycetes, Streptomyces thermoalcalitolerans APT3 and Streptomyces thermoviolaceus subsp. thermoviolaceus AQT2, and two fungi, Penicillium sp. Fh11 and Aspergillus sp. Fj6. This microbial consortium induced rapid composting of rice straw, thereby supporting sustainable agricultural waste management and reducing PM25 air pollution.

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.

Conversion of Corn Stover for Microbial Enzymes Production by Phanerochaete chrysosporium.

Phanerochaete chrysosporium, a white rot fungus, exhibits remarkable capabilities in producing various extracellular enzymes. These microbial enzymes find extensive applications in disrupting the intricate structure of plant cell walls, decolorizing synthetic dyes, and facilitating pulp extraction, among other functions. The process of solid-state fermentation stands out as an economical and sustainable approach, ideal for achieving high yields in enzyme production using lignocellulosic biomass as a substrate. In this research paper, both untreated and alkali pretreated corn stover materials served as substrates for enzyme production, leveraging the fungal strain's capacity to generate enzymes like cellulases and manganese peroxidase. The maximum production of endoglucanase was notably observed, reaching 121.21 ± 0.90 U/gds on the 9th day for untreated biomass and 79.75 ± 0.57 U/gds on the 6th day for treated biomass. Similarly, the peak exoglucanase production was recorded at 2.46 ± 0.008 FPU/ml on the 3rd day for untreated biomass and 0.92 ± 0.002 FPU/ml on the 6th day for treated biomass. Furthermore, the highest production of manganese peroxidase was achieved, with values of 5076.81 U/l on the 6th day for untreated biomass and 1127.58 ± 0.23 U/l on the 3rd day for treated biomass. These results collectively emphasize the potential of corn stover as a renewable and promising substrate for the production of essential enzymes.

Purification and characterization of novel manganese peroxidase from Trichoderma parestonica and its bio-conversion study of toxic arylamine

Trichoderma parestonica is a novel source of manganese peroxidase (MnP), [E.C.1.11.1.13] enzyme. The following was the sequence in which T. parestonica induced the production of MnP in the liquid culture medium: rice husk, corncob, sawdust, coir-dust and dried luffa on days 3, 4, 5, 2 and 3, respectively. The molecular weight of the enzyme was found to be 45kDa, determined from SDS-PAGE analysis which was further confirmed by mass analysis and was found to be 51.9kDa. Enzyme characterization parameters like optimum pH, temperature, Km, Kcat and catalytic efficiency are 5, 20ᵒC, 0.014mM (for Mn2+) and 0.028mM (for H2O2), 23.9 S-1 and 1.7 μM-1 S-1 respectively. Effect of various metal ions, such as bivalent Copper(Cu), Cobalt(Co), Mercury(Hg), Nickel(Ni), Cadmium(Cd), trivalent Chromium(Cr), Iron(Fe), Tetra-valent cerium(Ce), Penta-valent vanadium(V) and hexavalent molybdenum(Mo) the enzyme-catalyzed steady-state velocity was studied. The inhibition constant and nature of inhibition by these metal ions were determined. In the current investigation, MnP catalytic activities demonstrated that it could effectively catalyze the oxidation of phenols, aromatic amines and wastewater treatment. This communication suggested potential pathways for aromatic amine bioconversion mediated by MnP

Elucidating the role of media nitrogen in augmenting the production of lignin-depolymerizing enzymes by white-rot fungi.

Indigenous white-rot fungal isolates Schizophyllum commune, Phanerochaete chrysosporium, Ganoderma racenaceum, and Lentinus squarrosulus, demonstrating the ability to depolymerize lignin of the crop residues, were studied for their potential to produce ligninolytic enzymes using modified production media under conditions of limiting and excess nitrogen for higher enzymatic expressions. Secretome-rich media on the investigation confirmed the successful production of lignin-depolymerizing enzymes, viz. laccase, lignin peroxidase, manganese peroxidase, and versatile peroxidase. Production of laccases and peroxidases was statistically significant in nitrogen-limiting media with and without the substrate, across all white-rot fungal cultures at 95% confidence interval. Nitrogen-limiting media with the substrate on analysis extracellularly expressed 99.27 U of laccase and 68.48 U of manganese peroxidase in Schizophyllum commune, while 195.14 U of lignin peroxidase was produced by Phanerochaete chrysosporium. Lentinus squarrosulus expressed 455.34 U of laccase and 357.13 U of versatile peroxidase with 250.09 U of laccase and 206.95 U of manganese peroxidase produced by Ganoderma racenaceum for every milliliter of the media used. Nitrogen-limiting media triggered the production of laccase during the initial stages of growth while the expression of peroxidases was predominant at a later stage. Also, this media evinced increased enzymatic yields with low biomass content compared to nitrogen-excess conditions. The extant study confirmed the positive influence of nitrogen-limiting media in the efficient production of ligninolytic enzymes and their suggestive degradation potential for environmental pollutants, making these enzymes a safe, clean alternative to the use of chemicals and the media to be effective for large-scale production of ligninolytic enzymes. IMPORTANCE Lignin on account of its high abundance, complex polymeric structure, and biochemical properties is identified as a promising candidate in renewable energy and bioproduct manufacturing. However, depolymerization of lignin remains a major challenge in lignin utilization, entailing the employment of harsh treatments representing not only an environmental concern but also a waste of economic potential. Developing an alternative green technology to minimize this impact is imperative. Methods using enzymes to depolymerize lignin are the focus of recent studies. Current research work emphasized the efficient expression of the major lignin-depolymerizing enzymes: laccases, lignin peroxidases, manganese peroxidases, and versatile peroxidases from native isolates of white-rot fungus for several biotechnological applications as well as treatment of crop residues for use as ruminant feed in improving productivity. The importance of nitrogen in augmenting the expression of lignin-depolymerizing enzymes and providing a media recipe for the cost-effective production of ligninolytic enzymes is highlighted.

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.

Screening of Conditions for Manganese Peroxidase Production from White Rot Fungi (Pleurotus djamor)

The need for alternative and environmentally friendly methods for waste clean-up has led to the use of enzymes in bioremediation. In this study, white rot fungi were isolated from decaying plant parts using standard microbiological and biochemical techniques. The isolated fungi were identified and screened using standard substrates (2, 6-DMP) to determine their capability to produce manganese peroxidase. Pure manganese peroxidase was achieved after four distinct purification phases. Crude extract of the homogenate proteins obtained through optimized submerged fermentation system were precipitated using ammonium sulfate. Ammonium sulfate used at 60% concentration at pH 4.5 precipitated proteins with highest Manganese peroxidase activity (322 U/mg). The precipitated proteins were desalted through dialysis for twelve hours with buffer exchange at interval of six hours and activity of 343.91 U/mg was recorded afterwards. DEAE-cellulose was used for the ion exchange purification of the dialyzed protein, NaCl gradients of 0.3-0.6 M was found to be best in washing off bound protein from the exchange resin and activity of 434.18 U/mg was recorded from the pooled fraction tubes. Sephadex G-100 was used for separation of the proteins into molecular sizes and weights. 2.8 purification folds of the enzyme were achieved after ion exchange (DEAE-cellulose) and gel filtration (sephadex G-100) with percentage yield of 2.20%. Specific activity of the enzyme increased to 602.00% after gel filtration. The partial purified enzyme was further characterized for determination of optimal pH (4.5), temperature (40 OC) and kinetic properties Km of 3.4 mM and Vmax of 250µmol/min.

The reverse function of lignin-degrading enzymes: The polymerization ability to promote the formation of humic substances in domesticated composting

This study aimed to confirm the ability of lignin peroxidase (LiP) and manganese peroxidase (MnP) in promoting the formation of humic substances (HS) during domesticated composting. Three raw materials with different lignin types were used for composting, including rice straw, tree branches, and pine needles. Results suggested that LiP and MnP activity increased during domesticated composting. But HS formation was only promoted by LiP. The effect of MnP was insignificant, which might be caused by the lack of enzyme cofactors like Mn2+. Meanwhile, bacteria highly associated with LiP and MnP production were identified as core bacteria. Function prediction of 16S-PICRUSt2 showed that the function of core bacteria was consistent with total bacterial functions which mainly promoted compost humification. Therefore, it speculated that LiP and MnP had the ability to promote HS formation during composting. Accordingly, it is a new understanding of the role of biological enzymes in composting.

Screening of conditions for manganese peroxidase production from white rot fungi (Pleurotus djamor)

Screening of conditions for manganese peroxidase production from white rot fungi (Pleurotus djamor)

Studies on the Rapid and Simple DNA Extraction Method, Antibacterial Activity and Enzyme Activity Involved in Plant Biomass Conversion by Cookeina sulcipes and C. tricholoma (Cup Fungi)

Cookeina sulcipes and C. tricholoma are a cup fungi (Ascomycota) collected in Saraburi, Thailand. The fungi have been isolated, cultured and confirmed as respective species. For morphology, both Cookeina sp. are white mycelium and the growth rate on potato dextrose agar (PDA) result present C. sulcipes is faster than C. tricholoma. The molecular characterization from a rapid and simple DNA extraction method that is modified based on thermolysis method, The DNA extraction is finish in thirty minutes and efficiency to continuous with polymerase chain reaction (PCR) amplification to fungi species level identification. The DNA sequence from internal transcribed spacer (ITS) gene regions by universal primer pairs ITS5/ITS4 is effective to confirm Cookeina species level that C. sulcipes has 616 bp and C. tricholoma has 570 bp. Including, DNA sequence of large subunit (LSU) gene regions by universal primer pairs LROR/LR5 is generate that C. sulcipes has 912 bp and C. tricholoma has 906 bp. The cultures are screened for antibacterial activity by agar plug diffusion method and found that both isolates have been no activity against test strains (Bacillus subtilis, Escherichia coli, Kocuria rhizophila (Micrococcus luteus), Pseudomonas aeruginosa, Staphylococcus aureus and S. epidermidis). In a preliminary screening test of enzymes involved in plant biomass breakdown by agar plate method, both Cookeina sp. show cellulolytic and hemicellulolytic enzymatic activity, and manganese peroxidase (MnP) productivity. In contrast, only C. sulcipes had additional laccase activity. Neither isolate generate pectinolytic and lignin peroxidase (LiP) activities. Thus, Cookeina spp. proved the potentiality to break down lignocelluloses.

Agro-industrial wastes revalorization as feedstock: production of lignin-modifying enzymes extracts by solid-state fermentation using white rot fungi

The purpose of the study was to evaluate the production of lignin-modifying enzyme extracts and delignified biomass from agro-industrial wastes using white rot fungi (Inonotus sp. Sp2, Stereum hirsutum Ru-104, Bjerkandera sp. BOS55, Pleurotus eryngii IJFM 169 and Phanerochaete chrysosporium BKM-F-1767). These were screened based on their adaptability and colonization ability on different substrates, as well as by the Laccase, Manganese peroxidase, and Lignin peroxidase enzymatic production. Native strains (Inonotus sp. Sp2 and S. hirsutum Ru-104) showed the highest growth kinetics under the solid-substrate fermentation conditions and the growth rate parameters of the kinetic logistic model for the different substrates were between 0.39–0.81 (1/d) and 0.42–0.83 (1/d), respectively; the determination coefficients were ≥0.99. Inonotus sp. Sp2 was subsequently cultured in static flasks to produce crude enzyme extracts, obtaining manganese peroxidase activity levels of 18.5 and 31.3 (U/g) when growing in corn cob husk and spent tea leaves, respectively. Besides, it was to establish that the best conditions for lignin-modifying enzymes production using corn cob husk are 70% of initial moisture and 2.12 mm of particle size; reaching after 30 incubation days a manganese peroxidase activity of 21 ± 6 (U/g) under these conditions; enzyme that showed a suitable thermostability.

High Potential Decolourisation of Textile Dyes from Wastewater by Manganese Peroxidase Production of Newly Immobilised Trametes hirsuta PW17-41 and FTIR Analysis.

Coloured wastewater from the textile industry is a very serious global problem. Among 16 different white-rot fungal isolates, Trametes hirsuta PW17-41 revealed high potential for decolourisation of mixed textile dyes (Navy EC-R, Ruby S3B and Super Black G) from real industrial wastewater samples. The efficiency of dye decolourisation was evaluated using the American Dye Manufacturers’ Institute (ADMI) standard methodology. The suitable support for fungal mycelium immobilisation was nylon sponges. The optimal dye decolourisation (95.39%) was achieved by using palm sugar and ammonium nitrate as carbon and nitrogen sources, respectively. The initial pH was 5 and the agitation speed was 100 rpm at 30 °C. The ADMI values of textile dyes decreased from 2475 to 114 within two days, reducing the treatment time from seven days before optimisation. The major mechanism of dye decolourisation was biodegradation, which was confirmed by UV–visible and FTIR spectra. Manganese peroxidase (MnP) (4942 U L−1) was found to be the main enzyme during the decolourisation process at an initial dye concentration of 21,200 ADMI. The results indicated the strong potential of immobilised fungal cells to remove high concentrations of textile dyes from industrial wastewater and their potential ability to produce high MnP and laccase activities that can be used in further application.

Modelling and Environmental Profile Associated with the Valorization of Wheat Straw as Carbon Source in the Biotechnological Production of Manganese Peroxidase

Interest in the development of biorefineries and biotechnological processes based on renewable resources has multiplied in recent years. This driving force is the result of the availability of lignocellulosic biomass and the range of applications that arise from its use and valorization. The approach of second-generation sugars from lignocellulosic biomass opens up the possibility of producing biotechnological products such as enzymes as a feasible alternative in the framework of biorefineries. It is in this context that this manuscript is framed, focusing on the modelling of a large-scale fermentative biotechnological process to produce the enzyme manganese peroxidase (MnP) by the fungus Irpex lacteus using wheat straw as a carbon source. The production scheme is based on the sequence of four stages: pretreatment of wheat straw, seed fermenters, enzyme production and downstream processes. For its environmental assessment, the Life Cycle Assessment methodology, which allows the identification and quantification of environmental impacts associated with the process, was utilized. As the main finding, the stages of the process with the highest environmental burdens are those of pretreatment and fermentation, mainly due to energy requirements. With the aim of proposing improvement scenarios, sensitivity analyses were developed around the identified hotspots. An improvement in the efficiency of steam consumption leads to a reduction of environmental damage of up to 30%.

Effective biodeterioration of a common endocrine disruptor 17β-estradiol using mixed microbial cultures isolated from waste water

Twelve bacteria were isolated from wastewater for the degradation of 17β-estradiol (strain K1, K2, K3, K4, L5, L6, L7, L8, M9, M10, M11 and M12) by enrichment method. Initial screening revealed maximum degradation (78 ± 1.1%, and 86 ± 4.2%) of 17β-estradiol by strains L7, and M10, respectively. The production of manganese peroxidase by bacteria gets involved in the degradation of various contaminants. Almost all strains isolated from the enriched medium produced manganese peroxidase. Based on initial screening on 17β-estradiol degradation and manganese peroxidase production, two bacteria (Flavobacterium longum L7 and Pseudomonas aeruginosa M10) were selected to analyze 17β-estradiol degradation in co-culture system. The co-cultured strains could rapidly remove 17β-estradiol (91.3%) from the culture medium at 10 mg/L 17β-estradiol initial concentration. Moreover, degradation of 17β-estradiol was only 82% and 75% when the strains L7 and M10 were treated individually. Initial 17β-estradiol concentration was 10 mg/L and 0.3% inoculum were optimum and achieved >90% degradation. Alkaline pH was optimum (pH 7.5) and reached 98.4 ± 7.5% degradation in co-culture. Glucose and trehalose improved the growth and degradation of 17β-estradiol. The organic nitrogen sources such as, yeast extract and peptone improved the removal of 17β-estradiol. The selected bacterial strains were resistant against penicillin, erythromycin, oxytetracycline and tetracycline. The present findings demonstrated the application of F. longum L7 and P. aeruginosa M10 co-culture for the removal of steroid hormones in the wastewater.

Efficient biodegradation of Congo red dye using fungal consortium incorporated with Penicillium oxalicum and Aspergillus tubingensis.

A novel approach had been carried out to develop fungal consortium, namely, RH-2, containing two marine procured fungal isolates in order to evaluate biodegradation of recalcitrant diazo dye Congo red. The fungi were isolated from the seacoast of Diu, India. According to the ITS sequencing, the strains were identified as Penicillium oxalicum (DS-2) and Aspergillus tubingensis (DS-4). Discs of 12mm were cut out from the edge of both the fungal isolates (DS-2 and DS-4) and inoculated in flasks consisting of potato dextrose broth with 100mg/L Congo red for the development of fungal consortium RH-2. The degradation by the fungal consortium RH-2 was more effective than the fungal monocultures DS-2 and DS-4 with the respective degradation reaching 97.15 ± 0.15%, 68.96 ± 0.09%, and 29.96 ± 0.21% in addition of yeast extract (1% w/v) within 12h. The influence of dextrose (1% w/v), yeast extract (1% w/v), pH 5, and salt concentration (1% w/v) enhanced the degradation potential of fungal consortium RH-2. The maximal degradation was correlated with the production of laccase (12.498 ± 0.21 U/mL) and manganese peroxidase (10.314 ± 0.25 U/mL). The catabolism of Congo red was confirmed by UV-Visible spectroscopic analysis (Congo red λ-max = 499nm) and ATR-FTIR spectroscopic analysis. The filtrates obtained after Congo red degradation were also evaluated for microbial toxicity against bacteria (Bacillus haynesii) and phytotoxicity analysis on plant seed (Trigonella foenum) which revealed that the filtrate acquired after the treatment of Congo red by fungal consortium RH-2 was less toxic than the original dye in nature. A novel aspect is determined by the evidence of mutualistic interaction between two different fungi for the rapid decolorization and degradation of dye providing a prospective of utilizing the developed consortium RH-2 as a cost-effective approach in textile wastewater treatment for cleaner environment.

Process performance improvement for the simultaneous production of ligninolytic enzymes in solid culture using agricultural wastes through the Taguchi method

Despite a large amount of published research on the production of ligninolytic enzymes, the latter are not yet being applied to combat environmental pollution. No cost-effective process has been developed to date. This study describes an improvement of the solid-state fermentation procedure for the production of ligninolytic enzymes via Phanerochaete chrysosporium ATX by applying the Taguchi method and using an agro-industrial waste as substrate. The production of lignin peroxidase (LiP), manganese peroxidase (MnP), and laccase (Lac) were simultaneously increased within a packed-bed column. The factors and levels studied were humidity (A: 60, 70, 80%), inoculum concentration (B: 7.5, 10.0, 12.5 × 105 spores/mL), packed density (C: 0.14, 0.16, 0.18 g/mL), and time (D: 6, 8, 10 days). The results showed that humidity was the factor with a higher effect upon LiP and Lac's production, while time was for MnP. Humidity exerted the greatest influence on the global desirability of the process. Improved conditions (A, 60%; B, 1.0 × 106 spores/mL; C, 0.17 g/mL; D, 8 days) were further validated: the results revealed an overall desirability increase of 237% over the unoptimized process. Process performance was likewise maintained at a higher scale (1:10). The results contribute to establishing a cost-effective bioprocess to produce ligninolytic enzymes by reducing the cost associated with raw materials and purification steps.