Degradation of multiple PAHs and co-contaminants by microbial consortia and their toxicity assessment.

The anthropogenic activities toward meeting the energy requirements have resulted in an alarming rise in environmental pollution levels. Among pollutants, polycyclic aromatic hydrocarbons (PAHs) are the most predominant due to their persistent and toxic nature. Amidst the several pollutants depuration methods, bioremediation utilizing biodegradation is the most viable alternative. This study investigated the biodegradation efficacy using developed microbial consortium PBR-21 for 2-4 ringed PAHs named naphthalene (NAP), anthracene (ANT), fluorene (FLU), and pyrene (PYR). The removal efficiency was observed up to 100 ± 0.0%, 70.26 ± 4.2%, 64.23 ± 2.3%, and 61.50 ± 2.6%, respectively, for initial concentrations of 400 mg L-1 for NAP, ANT, FLU, and PYR respectively. Degradation followed first-order kinetics with rate constants of 0.39 d-1, 0.10 d-1, 0.08 d-1, and 0.07 d-1 and half-life t 1 / 2  of 1.8 h, 7.2 h, 8.5 h, and 10 h, respectively. The microbial consortia were found to be efficient towards the co-contaminants with 1 mM concentration. Toxicity examination indicated that microbial-treated PAHs resulted in lesser toxicity in aquatic crustaceans (Artemia salina) than untreated PAHs. Also, the study suggests that indigenous microbial consortia PBR-21 has the potential to be used in the bioremediation of PAH-contaminated environment.

Deciphering ligninolytic enzymes in the secretome of Pycnoporus sp. and their potential in degradation of 2-chlorophenol.

Chlorophenols and their derivatives are persistent environmental pollutants, posing a threat to terrestrial and aquatic life. The biological approach for eliminating toxic contaminants is an effective, sustainable, and environmental friendly method. In this study, the crude enzymes present in the secretome of white-rot fungus (Pycnoporus sp.) were explored for the degradation of 2-chlorophenol. The activity of ligninolytic enzymes in the secretome was analyzed and characterized for their kinetics and thermodynamic properties. Laccase and manganese peroxidase were prevalent ligninolytic enzymes and exhibited temperature stability in the range of 50-65 °C and pH 4-5, respectively. The kinetic parameters Michaelis constant (Km) and turnover number (Kcat) for Lac were 42.54 µM and 45 s-1 for 2,2'-azino-bis (3-ethylben- zothiazoline-6-sulfonic acid), and 93.56 µM and 48 s-1 towards 2,6-dimethoxyphenol whereas Km and Kcat for MnP were 2039 µM and 294 s-1 for guaiacol as substrate. Treatment with the crude enzymes laccase and manganese peroxidase results in the reduction of 2-chlorophenol concentration, confirmed by UV-visible absorption spectra and high-performance liquid chromatography analysis. The detoxification of 2-chlorophenol into less toxic forms was confirmed by the plate toxicity assay. This study demonstrated that crude enzymes produced by Pycnoporus sp. could potentially minimize the toxicity of phenolic compounds in a sustainable way.

Immobilized laccase: an effective biocatalyst for industrial dye degradation from wastewater.

Environmental concerns due to the release of industrial wastewater contaminated with dyes are becoming more and more intense with the increasing industrialization. Decolorization of industrial effluents has become the top priority due to the continuous demand for color-free discharge into the receiving water bodies. Different dye removal techniques have been developed, among which biodegradation by laccase enzyme is competitive. Laccase, as a green catalyst, has a high catalytic activity, generates less toxic by-products, and has been extensively researched in the field of remediation of dyes. However, laccase's significant catalytic activity could only be achieved after an effective immobilization step. Immobilization helps strengthen and stabilize the protein structure of laccase, thus enhancing its functional properties. Additionally, the reusability of immobilized laccase makes it an attractive alternative to traditional dye degradation technologies and in the realistic applications of water treatment, compared with free laccase. This review has elucidated different methods and the carriers used to immobilize laccase. Furthermore, the role of immobilized laccase in dye remediation and the prospects have been discussed.

Correction: Removal of Petroleum Contaminants Through Bioremediation with Integrated Concepts of Resource Recovery: A Review.

[This corrects the article DOI: 10.1007/s12088-021-00928-4.].

Nano-bioremediation: an eco-friendly and effective step towards petroleum hydrocarbon removal from environment.

Global concern about petroleum hydrocarbon pollution has intensified and gained scientific interest due to its noxious nature, high persistence in environmental matrices, and low degradability. One way to address this is by combining remediation techniques that could overcome the constraints of traditional physio-chemical and biological remediation strategies. The upgraded concept of bioremediation to nano-bioremediation in this direction offers an efficient, economical, and eco-friendly approach to mitigate petroleum contaminants. Here, we review the unique attributes of different types of nanoparticles and their synthesis procedures in remediating various petroleum pollutants. This review also highlights the microbial interaction with different metallic nanoparticles and their consequential alteration in microbial as well as enzymatic activity which expedites the remediating process. Besides, the latter part of the review explores the application of petroleum hydrocarbon degradation and the application of nano supports as immobilizing agents for microbes and enzymes. Further, the challenges and the future prospects of nano-bioremediation have also been discussed.

Trends in fisheries waste utilization: a valuable resource of nutrients and valorized products for the food industry.

The rise in fisheries production worldwide has caused a remarkable increase in associated anthropogenic waste. This poses significant concerns due to adverse environmental impacts and economic losses. Owing to its renewability, high abundance, and potential as a rich source of many nutrients and bioactive compounds, strategies have been developed to convert fish waste into different value-added products. Conventional and improved methods have been used for the extraction of biomolecules from fish waste. The extracted fish waste-derived value-added products such as enzymes, peptides, fish oil, etc. have been used to fortify different food products. This review aims to provide an overview of the nature and composition of fish waste, strategies for extracting biomolecules from fish waste, and the potential application of fish waste as a source of calcium and other nutrients in food fortification and animal feed has been discussed. In context to fishery waste mitigation, valorization, and circular bioeconomy approach are gaining momentum, aiming to eliminate waste while producing high-quality value-added food and feed products from fishery discards.

Deciphering the Antimicrobial Potential of Taxus wallichiana Zucc: Identification and Characterization Using Bioassay-Guided Fractionation.

Taxus wallichiana Zucc. is a high valued medicinal plant and has been mainly studied for its anti-cancer properties. However, research on its other important biological activities, such as its antimicrobial potential, still needs attention. The focus of the present study is to investigate the antimicrobial activity of secondary metabolites of T. wallichiana needles against 3 different groups of microorganisms, i. e., bacteria, actinobacteria, and fungi. Bioactive compounds from T. wallichiana needles were separated through column chromatography, and, TLC-bioautography. Mobile phases were optimized using Snyder's selectivity triangle. Antimicrobial spots were fractionated and compounds were identified by gas chromatography-mass spectroscopy (GC/MS) and liquid chromatography-mass spectrometry (LC/MS). Functional groups were characterized using Fourier transform infrared spectrometry (FTIR) and nuclear magnetic resonance (NMR) was used to identify the molecular structures. GC/MS and LC/MS data analysis confirm the presence of fatty acids (arachidic acid, behenic acid, palmitic acid, and stearic acid), vitamins (nicotinamide), and alkaloids (cinchonine, timolol), aminobenzamides (procainamide), carbocyclic sugar (myoinositol), and alkane hydrocarbon (hexadecane), having antimicrobial activity in the needles of T. wallichiana. To the best of our knowledge, this is the first report on the isolation and characterization of antimicrobial compounds from the needles of Taxus wallichiana (Himalayan yew). The data obtained from the present study will be supportive to the new drug discoveries in modern medicine with various combinations of medicinal plant's active constituents that can be used for curing many diseases.

Proteomic response of Pseudomonas aeruginosa IIPIS-8 during rapid and efficient degradation of naphthalene.

Polycyclic aromatic hydrocarbons (PAHs) are widely distributed in the ecosystem and are of significant concern due to their toxicity and mutagenicity. Bioremediation of PAHs is a popular and benign approach that ameliorates the environment. This study investigated the biodegradation and proteome response of Pseudomonas aeruginosa IIPIS-8 for two-ringed PAH: naphthalene (NAP) to understand proteome alteration during its bioremediation. Rapid biodegradation was observed up to 98 ± 1.26% and 84 ± 1.03%, respectively, for initial concentrations of 100 mg L-1 and 500 mg L-1 of NAP. Degradation followed first-order kinetics with rate constants of 0.12 h-1 and 0.06 h-1 and half-life (t1/2) of 5.7 h and 11.3 h, respectively. Additionally, the occurrence of key ring cleavage and linear chain intermediates, 2,3,4,5,6, -pentamethyl acetophenone, 1-octanol 2-butyl, and hexadecanoic acid supported complete NAP degradation. Proteomics study of IIPIS-8 throws light on the impact of protein expression, in which 415 proteins were quantified in sequential windowed acquisition of all theoretical fragment ion mass spectra (SWATH-MS) analysis, of which 97 were found to be significantly up-regulated and 75 were significantly down-regulated by ≥ 2-fold change (p values ≤ 0.05), during the NAP degradation. The study also listed the up-regulation of several enzymes, including oxido-reductases, hydrolases, and catalases, potentially involved in NAP degradation. Overall, differential protein expression, through proteomics study, demonstrated IIPIS-8's capability to efficiently assimilate NAP in their metabolic pathways even in a high concentration of NAP.

Innovative strategies in algal biomass pretreatment for biohydrogen production.

Biohydrogen is one of the cleanest renewable energies with a high calorific value. Algal biomass can be utilized as a sustainable feedstock for biohydrogen production via dark fermentation. However, the recovery of fermentable sugar from algal biomass is challenging because of the diversity and complex cell wall composition and therefore, requires an additional pretreatment step. However, most of the conventional pretreatment strategies suffer from limited technological feasibility and poor economic viability. In this context, this review aims to present the structural complexities of the cell wall of algae and highlight the innovative approaches such as the use of hybrid technologies, biosurfactants, nanoparticles, and genetic engineering approaches for the hydrolysis of algal biomass and improved biohydrogen production. Additionally, a comprehensive discussion of the comparative evaluation of various pretreatment methods, and the techno-economic and life cycle assessment of algal biohydrogen production is also presented in this review.

Pyrene remediation by Trametes maxima: an insight into secretome response and degradation pathway.

The rapid pace of economic development has resulted in the release of several polycyclic aromatic hydrocarbons (PAHs) into the environment. Microbial degradation using white-rot fungi is a promising method for the removal of PAHs from the environment. In the present study, biodegradation of recalcitrant PAH by a white-rot fungus, Trametes maxima IIPLC-32, was investigated using pyrene. The pyrene concentration decreased by 79.80%, 65.37%, and 56.37% within 16 days from the initial levels of 10 mg L-1, 25 mg L-1, and 50 mg L-1, respectively. Gas chromatographic-mass spectrometric identification of prominent metabolites 1-hydroxypyrene, 2-methyl-1-naphthyl acetic acid, di-n-butyl phthalate, and diethyl phthalate helped in determining the pyrene degradation pathway. The presence of 81 extracellular proteins was revealed by secretome analysis. The identified proteins up-regulated in response to pyrene degradation were classified into detoxification proteins (6.12%), redox proteins (6.12%), stress proteins (4.08%), metabolic-related proteins (26.53%), translation and transcriptional proteins (49%), catalytic proteins (49%), and other proteins (8.16%). Knowledge of secretome analysis in pyrene degradation helped to understand the degradation mechanism of pyrene. Also, the study suggests that T. maxima IIPLC-32 has the potential to be used in the bioremediation of PAH contaminated aquatic environment.

Jute sticks biomass delignification through laccase-mediator system for enhanced saccharification and sustainable release of fermentable sugar.

Jute sticks obtained after the extraction of jute fiber are an excellent biomass feedstock with a significant amount of carbohydrates that makes it an attractive resource for sustainable energy generation. However, the high lignin content in the jute stick hinders the cellulosic component of the cell wall from enzymatic hydrolysis.This work demonstrates the lignin degradation of jute stick biomass by Trametes maxima laccase in the presence of mediator Hydroxybenzotriazole and improvement in its subsequent saccharification. Lignin component in jute stick is reduced by 21.8% in a single reaction treatment with laccase-mediator compared to the untreated jute stick sample used as control. The yield of fermentable sugar is increased by 19.5% that verifies enhanced saccharification after lignin removal. Delignification of jute stick was corroborated through different analytical techniques. The Pyrolysis gas chromatography/mass spectrometry results further confirms abundance of S lignin unit in the jute stick compared to the H and G unit and modification in lignin polymer as a change in the syringyl-to-guaiacyl ratio. Hence, this work demonstrates that jute stick can be effectively delignified using biocatalyst-mediator system and utilized as biomass source, thus contributing in circular bio-economy through waste valorization.

Removal of Petroleum Contaminants Through Bioremediation with Integrated Concepts of Resource Recovery: A Review.

There is an upsurge in industrial production to meet the rising demands of the rapidly growing population globally. The enormous energy demand of the growing economies still depends upon petroleum. It has also resulted in environmental pollution due to the release of petroleum origin pollutants. Soil and aquifers, especially in the direct impact zones of petroleum refineries, are the worst hit. The integrated concept of bioremediation and resource recovery offers a sustainable solution to mitigate environmental pollution. It involves biodegradation, a benign utilization of toxic wastes, and the recycling of natural resources. Bioremediation is considered an integral contributor to the emerging concepts of bio-economy and sustainable development goals. This review article aims to provide an updated overview of bioremediation involving petroleum-based contaminants. Microbial degradation is discussed as a promising strategy for petroleum refinery effluent and sludge treatment. The review also provides an insight into resource reuse and recovery as a holistic approach towards sustainable refinery waste treatment. Furthermore, the integrated technologies that deserve in-depth exploration for future study in the refinery sector are highlighted in the present study.

Laccase-mediated delignification and detoxification of lignocellulosic biomass: removing obstacles in energy generation.

The rising global population and worldwide industrialization have led to unprecedented energy demand that is causing fast depletion of fossil reserves. This has led to search for alternative energy sources that are renewable and environment friendly. Use of lignocellulosic biomass for energy generation is considered a promising approach as it does not compete with food supply. However, the lignin component of the biomass acts as a natural barrier that prevents its efficient utilization. In order to remove the lignin and increase the amount of fermentable sugars, the lignocellulosic biomass is pretreated using physical and chemical methods which are costly and hazardous for environment. Moreover, during the traditional pretreatment process, numerous inhibitory compounds are generated that adversely affect the growth of fermentative microbes. Alternatively, biological methods that use microbes and their enzymes disrupt lignin polymers and increase the accessibility of the carbohydrates for the sugar generation. Microbial laccases have been considered as an efficient biocatalyst for delignification and detoxification offering a green initiative for energy generation process. The present review aims to bring together recent studies in bioenergy generation using laccase biocatalyst in the pretreatment processes. The work provides an overview of the sustainable and eco-friendly approach of biological delignification and detoxification through whole-cell and enzymatic methods, use of laccase-mediator system, and immobilized laccases for this purpose. It also summarizes the advantages, associated challenges, and potential prospects to overcome the limitations.

Application of laccase immobilized rice straw biochar for anthracene degradation.

The present study explores the immobilization of ligninolytic enzyme-laccase on the surface of rice straw biochar and evaluates its application for anthracene biodegradation. The rice straw biochar was acid-treated to generate carboxyl functionality on its surface, followed by detailed morphological and chemical characterization. The surface area of functionalized biochar displayed a two-fold increase compared to the untreated biochar. Laccase was immobilized on functionalized biochar, and an immobilization yield of 66% was obtained. The immobilized enzyme demonstrated operational stability up to six cycles while retaining 40% of the initial activity. Laccase immobilization was further investigated by performing adsorption and kinetic studies, which revealed the highest immobilization concentration of 500 U g-1 at 25 °C. The adsorption followed the Langmuir isotherm model at equilibrium, and the kinetic study confirmed pseudo-second-order kinetics. The equilibrium rate constant (K2) at 25 °C and 4 °C were 3.6 × 10-3 g U-1 min-1 and 4 × 10-3 g U-1 min-1 respectively for 100 U g-1 of enzyme loading. This immobilized system was applied for anthracene degradation in the aqueous batch mode, which resulted in complete degradation of 50 mg L-1 anthracene within 24 h of interaction exposure.

Ethanol fermentation from molasses at high temperature by thermotolerant yeast Kluyveromyces sp. IIPE453 and energy assessment for recovery.

High temperature ethanol fermentation from sugarcane molasses B using thermophilic Crabtree-positive yeast Kluyveromyces sp. IIPE453 was carried out in batch bioreactor system. Strain was found to have a maximum specific ethanol productivity of 0.688 g/g/h with 92 % theoretical ethanol yield. Aeration and initial sugar concentration were tuning parameters to regulate metabolic pathways of the strain for either cell mass or higher ethanol production during growth with an optimum sugar to cell ratio 33:1 requisite for fermentation. An assessment of ethanol recovery from fermentation broth via simulation study illustrated that distillation-based conventional recovery was significantly better in terms of energy efficiency and overall mass recovery in comparison to coupled solvent extraction-azeotropic distillation technique for the same.

Design and optimization of ethanol production from bagasse pith hydrolysate by a thermotolerant yeast Kluyveromyces sp. IIPE453 using response surface methodology.

Ethanol production from sugarcane bagasse pith hydrolysate by thermotolerant yeast Kluyveromyces sp. IIPE453 was analyzed using response surface methodology. Variables such as Substrate Concentration, pH, fermentation time and Na2HPO4 concentration were found to influence ethanol production significantly. In a batch fermentation, optimization of key process variables resulted in maximum ethanol concentration of 17.44 g/L which was 88% of the theoretical with specific productivity of 0.36 g/L/h.

Co-expression of recombinant human CYP2C9 with human cytochrome P450 reductase in protease deficient S. cerevisiae strain at a higher scale yields an enzyme of higher specific activity.

Cytochrome P450 (CYP450) isozymes play an important role in the study of drug metabolism and drug discovery. A number of reports are available that describe recombinant expression of CYP450 isozymes. In this paper, human CYP2C9 and human cytochrome P450 reductase cDNAs were cloned and expressed in Premas proprietary yeast episomal and integrative vectors respectively under the influence of GAL1 promoter. Yeast cells were grown and induced at optimal parameters to make microsomal membranes. Isolated microsomal membranes were analyzed for CYP2C9 and cytochrome P450 reductase activity, CYP2C9 content and inhibition properties. We report heterologous expression of human CYP2C9 along with human cytochrome P450 reductase in protease deficient S. cerevisiae at a 5 litre scale resulting in high yields (8-10 nmols/litre) of enzyme with higher specific activity (2-3 fold higher). This yields a superior enzyme and makes it amenable to miniaturization of screening assays with concomitant lowering of costs.