Recalcitrant Substrates Research Articles

Parent material influences soil properties to shape bacterial community assembly processes, diversity, and enzyme-related functions

Soil parent material is the second most influential factor in pedogenesis, influencing soil properties and microbial communities. Different assembly processes shape diverse functional microbial communities. The question remains unresolved regarding how these ecological assembly processes affect microbial communities and soil functionality within soils on different parent materials. We collected soil samples developed from typical parent materials, including basalt, granite, metamorphic rock, and marine sediments across soil profiles at depths of 0–20, 20–40, 40–80, and 80–100 cm, within rubber plantations on Hainan Island, China. We determined bacterial community characteristics, community assembly processes, and soil enzyme-related functions using 16S rRNA high-throughput sequencing and enzyme activity analyses. We found homogeneous selection, dispersal limitation, and drift processes were the dominant drivers of bacterial community assembly across soils on different parent materials. In soils on basalt, lower pH and higher moisture triggered a homogeneous selection-dominated assembly process, leading to a less diverse community but otherwise higher carbon and nitrogen cycling enzyme activities. As deterministic process decreased, bacterial community diversity increased with stochastic process. In soils on marine sediments, lower water, carbon, and nutrient content limited the dispersal of bacterial communities, resulting in higher community diversity and an increased capacity to utilize relative recalcitrant substrates by releasing more oxidases. The r-strategy Bacteroidetes and genera Sphingomonas, Bacillus, Vibrionimonas, Ochrobactrum positively correlated with enzyme-related function, whereas k-strategy Acidobacteria, Verrucomicrobia and genera Acidothermus, Burkholderia-Caballeronia-Paraburkholderia, HSB OF53-F07 showed negative correlations. Our study suggests that parent material could influence bacterial community assembly processes, diversity, and soil enzyme-related functions via soil properties.

Engineering of glycoside hydrolase family 7 cellobiohydrolases directed by natural diversity screening

Protein engineering and screening of processive fungal cellobiohydrolases (CBHs) remain challenging due to limited expression hosts, synergy-dependency, and recalcitrant substrates. In particular, GH7 CBHs are a family critically important for the bioeconomy as well as a methodology for improving these and other difficult to engineer enzymes. Here, we target the discovery of highly active natural GH7 CBHs and engineering of variants with improved activity. Using experimentally assayed activities of genome mined CBHs, we applied sequence and structural alignments to top performers to identify key point mutations linked to improved activity. From ∼1500 known GH7 sequences, an evolutionarily diverse subset of 57 GH7 CBH genes was expressed in Trichoderma reesei and screened using a multiplexed activity screening assay. Ten catalytically enhanced natural variants were identified, produced, purified, and tested for efficacy using industrially-relevant conditions and substrates. Three key amino acids in CBHs with performance comparable or superior to Penicillium funiculosum Cel7A were identified and combinatorically engineered into P. funiculosum cel7a, expressed in T. reesei, and assayed on lignocellulosic biomass. The top performer generated using this combined approach of natural diversity genome mining, experimental assays, and computational modeling produced a 41% increase in conversion extent over native P. funiculosum Cel7A, a 55% increase over the current industrial standard T. reesei Cel7A, and 10% improvement over Aspergillus oryzae Cel7C, the best natural GH7 CBH previously identified in our laboratory.

Mono-Digestion of 5-Hydroxymethylfurfural Process-Wastewater in Continuously Operated Anaerobic Filters: A Cascade Utilization Approach

A proper remedy for the overexploitation of biomass and biobased materials in the bioeconomy is the valorization of biorefineries’ side streams into meaningful products. Hence, in pursuit of a cascade utilization of renewables, a unique biorefinery byproduct was investigated for its biogas potential, specifically methane, in continuously operated anaerobic filters. For this purpose, 5-Hydroxymethylfurfural process-wastewater, after supplementation of necessary nutrients, was diluted down to 10, 20, 30, 40, and 50 gCOD/L concentrations and thereafter tested individually at 43 °C and 55 °C. Maximum methane conversion efficiency at either temperature was observed for test substrates with 10 gCOD/L and 20 gCOD/L concentrations. At 43 °C, the anaerobic filters exhibited their highest biogas yields when supplied with the 30 gCOD/L feedstock. Further exposure of the mesophilic and thermophilic consortia to the ensuing 5-Hydroxymethylfurfural process-wastewater dilutions compromised the stability of the anaerobic process due to the soaring concentrations of short-chained volatile fatty acids. The supplementation of necessary nutrients to unlock the methane potential of the given recalcitrant substrate appears insufficient. Techniques like micro aeration, photolysis, or the use of activated carbon in the fixed bed might have the ability to enhance the biochemical methane conversion of such feedstock; otherwise, the introduction of trace elements alone may be adequate if aiming for platforms (volatile fatty acids) via anaerobic technologies.

(Invited) Commercializing Single Walled Carbon Nanotube-Based Sensors - Enzyme Activity and Beyond

The ECS carbon nanostructures community is well informed on the potential of single walled carbon nanotube (SWCNT)-based sensors and likewise the challenges associated with realizing practical applications outside of an academic laboratory. This talk will discuss our latest efforts in facile separation and sensor preparation methods. It will discuss potential use cases in improving high throughput screening of custom enzymes, with a current focus on recalcitrant substrates such as conventional polymers. This talk will also focus on latest efforts of reducing to practice optical fluorimeters (single, multi-channel, spatial, and temperature controlled) for field use applications. Finally, this talk will detail the on-going adventure of commercialization at Zymosense Inc. which we hope will serve as a practical guide to other groups considering transferring their technology from the lab.

Conversion of steppe to cropland increases spatial heterogeneity of soil functional genes.

The microbiome function responses to land use change are important for the long-term prediction and management of soil ecological functions under human influence. However, it has remains uncertain how the biogeographic patterns of soil functional composition change when transitioning from natural steppe soils (NS) to agricultural soils (AS). We collected soil samples from adjacent pairs of AS and NS across 900 km of Mollisol areas in northeast China, and the soil functional composition was characterized using shotgun sequencing. AS had higher functional alpha-diversity indices with respect to KO trait richness and a higher Shannon index than NS. The distance-decay slopes of functional gene composition were steeper in AS than in NS along both spatial and environmental gradients. Land-use conversion from steppe to farmland diversified functional gene profiles both locally and spatially; it increased the abundances of functional genes related to labile carbon, but decreased those related to recalcitrant substrate mobilization (e.g., lignin), P cycling, and S cycling. The composition of gene functional traits was strongly driven by stochastic processes, while the degree of stochasticity was higher in NS than in AS, as revealed by the neutral community model and normalized stochasticity ratio analysis. Alpha-diversity of core functional genes was strongly related to multi-nutrient cycling in AS, suggesting a key relationship to soil fertility. The results of this study challenge the paradigm that the conversion of natural to agricultural habitat will homogenize soil properties and biology while reducing local and regional gene functional diversity.

Endophyte inoculation enhanced microbial metabolic function in the rhizosphere benefiting cadmium phytoremediation by Phytolaccaacinosa

Microbial metabolic activities in rhizosphere soil play a critical role in plant nutrient utilization and metal availability. However, its specific characteristics and influence on endophyte assisted phytoremediation remains unclear. In this study, an endophyte strain Bacillus paramycoides (B. paramycoides) was inoculated in the rhizosphere of Phytolacca acinosa (P. acinosa), and microbial metabolic characteristics of rhizosphere soils were analyzed using Biolog system to investigate how they influence phytoremediation performance of different types of cadmium contaminated soil. The results indicated that endophyte B. paramycoides inoculation enhanced bioavailable Cd percentage by 9–32%, resulting in the increased Cd uptake (32–40%) by P. acinosa. With endophyte inoculation, the utilization of carbon sources was significantly promoted by 4–43% and the microbial metabolic functional diversity increased by 0.4–36.8%. Especially, B. paramycoides enhanced the utilization of recalcitrant substrates carboxyl acids, phenolic compounds and polymers by 48.3–225.6%, 42.4–65.8% and 15.6–25.1%, respectively. Further, the microbial metabolic activities were significant correlated with rhizosphere soil microecology properties and impact phytoremediation performance. This study provided new insight into the microbial processes during endophyte assisted phytoremediation.

Controlling Catalyst Behavior in Lewis Acid-Catalyzed Carbonyl-Olefin Metathesis.

Lewis acid-catalyzed carbonyl-olefin metathesis has introduced a new means for revealing the behavior of Lewis acids. In particular, this reaction has led to the observation of new solution behaviors for FeCl3 that may qualitatively change how we think of Lewis acid activation. For example, catalytic metathesis reactions operate in the presence of superstoichiometric amounts of carbonyl, resulting in the formation of highly ligated (octahedral) iron geometries. These structures display reduced activity, decreasing catalyst turnover. As a result, it is necessary to steer the Fe-center away from inhibiting pathways to improve the reaction efficiency and augment yields for recalcitrant substrates. Herein, we examine the impact of the addition of TMSCl to FeCl3-catalyzed carbonyl-olefin metathesis, specifically for substrates that are prone to byproduct inhibition. Through kinetic, spectroscopic, and colligative experiments, significant deviations from the baseline metathesis reactivity are observed, including mitigation of byproduct inhibition as well as an increase in the reaction rate. Quantum chemical simulations are used to explain how TMSCl induces a change in catalyst structure that leads to these kinetic differences. Collectively, these data are consistent with the formation of a silylium catalyst, which induces the reaction through carbonyl binding. The FeCl3 activation of Si-Cl bonds to give the silylium active species is expected to have significant utility in enacting carbonyl-based transformations.

Nitrogen application influences the effect of bacteria on the belowground allocation of photosynthesized carbon under elevated CO2

Elevated carbon dioxide (eCO2) affects the translocation of photosynthesized carbon (C) from plants to soil, which is predominantly executed by soil microorganisms. However, the response strategy and mediation of the bacterial community on photosynthesized C allocation belowground remains elusive under eCO2 coupled with nitrogen (N) availability. Here, the spring wheat (Triticum aestivum L.) was continuously labeled with 13CO2 under ambient (aCO2, 350 ppm) or elevated (eCO2, 600 ppm) at three N application levels, and the incorporation of 13C into soil organic C was measured to indicate photosynthesized C accrual in soil. High-throughput MiSeq sequencing of 16S rRNA gene amplicons was used to explore soil bacterial response strategy. Compared with aCO2, eCO2 increased the relative abundance of Chloroflexi and Nitrospirae at the lowest N application level, while it increased that of Bacteroidetes at the highest N level. This shift of the response phyla from oligotrophic to copiotrophic species was closely associated with the increase in root biomass and its decomposability under eCO2 coupled with increasing N availability. The keystone taxa in the bacterial network might have strong ability to regulate such response of bacterial phyla to C and N availability. Specifically, the lower decomposability of the root-derived C caused by the intensive N deficiency under eCO2 stimulated oligotrophic bacteria, Saccharibacteria, Chloroflexi and Nitrospirae as keystone phyla, which could denote that the decomposition ability on recalcitrant substrate of bacterial phyla was enhanced before the activation of N transformation-related bacteria. Comparatively, sufficient N application under eCO2 facilitated copiotrophic bacteria as keystones for utilizing labile C derived from wheat roots, and thus the translocation efficiency of photosynthesized C belowground decreased. Our study sheds new light on the functional traits and mechanisms of the bacterial community in regulating C and N stoichiometry under eCO2.

Decomposition rate and biochemical fate of carbon from natural polymers and microplastics in boreal lakes.

Microbial mineralization of organic compounds is essential for carbon recycling in food webs. Microbes can decompose terrestrial recalcitrant and semi-recalcitrant polymers such as lignin and cellulose, which are precursors for humus formation. In addition to naturally occurring recalcitrant substrates, microplastics have been found in various aquatic environments. However, microbial utilization of lignin, hemicellulose, and microplastics as carbon sources in freshwaters and their biochemical fate and mineralization rate in freshwaters is poorly understood. To fill this knowledge gap, we investigated the biochemical fate and mineralization rates of several natural and synthetic polymer-derived carbon in clear and humic lake waters. We used stable isotope analysis to unravel the decomposition processes of different 13C-labeled substrates [polyethylene, polypropylene, polystyrene, lignin/hemicellulose, and leaves (Fagus sylvatica)]. We also used compound-specific isotope analysis and molecular biology to identify microbes associated with used substrates. Leaves and hemicellulose were rapidly decomposed compared to microplastics which were degraded slowly or below detection level. Furthermore, aromatic polystyrene was decomposed faster than aliphatic polyethylene and polypropylene. The major biochemical fate of decomposed substrate carbon was in microbial biomass. Bacteria were the main decomposers of all studied substrates, whereas fungal contribution was poor. Bacteria from the family Burkholderiaceae were identified as potential leaf and polystyrene decomposers, whereas polypropylene and polyethylene were not decomposed.

Synergistic Hydrolysis of Cellulose by a Blend of Cellulase-Mimicking Polymeric Nanoparticle Catalysts.

Enzyme-like catalysts by design have been a long sought-after goal of chemists but difficult to realize due to the challenges in the construction of multifunctionalized active sites with accurately positioned catalytic groups for complex substrates. Hydrolysis of cellulose is a key step in biomass utilization and requires multiple enzymes to work in concert to overcome the difficulty associated with hydrolyzing the recalcitrant substrate. We here report methods to construct synthetic versions of these enzymes through covalent molecular imprinting and strategic postmodification of the imprinted sites. The synthetic catalysts cleave a cellulose chain endolytically at multiple positions or exolytically from the nonreducing end by one or three glucose units at a time, all using the dicarboxylic acid motif found in natural cellulases. By mimicking the endocellulase, exocellulase, and β-glucosidase, the synthetic catalysts hydrolyze cellulose in a synergistic manner, with an activity at 90 °C in pH 6.5 buffer more than doubled that of Aspergillus niger cellulase at pH 5 and 37 °C and 44% of that of a commercial cellulase blend (from Novozyme). As robust cross-linked polymeric nanoparticles, the synthetic catalysts showed little changes in activity after preheating at 90 °C for 3 days and could be reused, maintaining 76% of activity after 10 reaction cycles.

A Comprehensive Review on Pretreatment Methods for Enhanced Biogas Production from Sewage Sludge

The treatment of municipal wastewater is considered a cornerstone for the protection of public health and environment. However, a major issue derived from this process is the large quantities of produced sewage sludge. Although anaerobic digestion is a widely applied method in Wastewater Treatment Plants (WWTPs) aiming to stabilize the sludge and to recover energy in the form of methane, it is usually limited due to the reduced decomposition efficiency and slow biodegradation rate of this recalcitrant substrate. For this reason, various pretreatment methods have been proposed aiming to modify the sludge structure, solubilize the organic matter, and decrease the crystallinity of sludge so as to accelerate hydrolysis and consequently enhance methane production. The current research is a comprehensive collection of recent advances in pretreatment technologies that can be potentially applied in wastewater treatment facilities. The critical review analysis presented herein reveals the several advantages and drawbacks, as well as the technical opportunities of the pretreatment methods and provides an assessment of their feasibility/applicability from an energetic, environmental, and economic point of view.

Enhanced in situ H2O2 production explains synergy between an LPMO with a cellulose-binding domain and a single-domain LPMO

Lytic polysaccharide monooxygenases (LPMOs) are mono-copper enzymes that catalyze oxidative depolymerization of recalcitrant substrates such as chitin or cellulose. Recent work has shown that LPMOs catalyze fast peroxygenase reactions and that, under commonly used reaction set-ups, access to in situ generated H2O2 likely limits catalysis. Based on a hypothesis that the impact of a cellulose-binding module (CBM) on LPMO activity could relate to changes in in situ H2O2 production, we have assessed the interplay between CBM-containing ScLPMO10C and its truncated form comprising the catalytic domain only (ScLPMO10CTR). The results show that truncation of the linker and CBM leads to elevated H2O2 production and decreased enzyme stability. Most interestingly, combining the two enzyme forms yields strong synergistic effects, which are due to the combination of high H2O2 generation by ScLPMO10CTR and efficient productive use of H2O2 by the full-length enzyme. Thus, cellulose degradation becomes faster, while enzyme inactivation due to off-pathway reactions with excess H2O2 is reduced. These results underpin the complexity of ascorbic acid-driven LPMO reactions and reveal a potential mechanism for how LPMOs may interact synergistically during cellulose degradation.

Impact of substrate digestibility on microbial community stability in methanogenic digestors: The mechanism and solution

This study investigated the temporal dynamics of digestion efficiency and community stability in digesters fed with waste activated sludge (WAS), straw (STR-AD), food waste (FW-AD) and mixture of straw-and-food waste (STR-FW-AD). Results showed that carbon removals of recalcitrant substrates (i.e., 48.2 ± 3.9% in WAS-AD and 57.8 ± 4.9% in STR-AD) were lower than that of labile substrates (i.e., 70.7 ± 4.0% in FW-AD). Nonetheless, carbon removal of recalcitrant substrates was largely improved through co-digestion (70.3 ± 3.2% in STR-FW-AD). In contrast to monopoly communities (e.g., the highly enriched Paludibacter) fed with the labile substrates, recalcitrant substrates supported highly diverse communities. Accordingly, the medians of negative/positive cohesions of communities in WAS-AD, STR-AD, STR-FW-AD and FW-AD decreased from 0.86 to 0.63, suggesting their decreasing community stability. Microbial source tracking analyses showed the major contribution of the STR-AD community to the co-digestion community. This study provided unprecedented mechanistic insight into stability improvement of substrate co-digestion on the methanogenic digestion microbiome.

One-pot conversion of industrial hemp residue into fermentable feedstocks using green catalyst and enzyme cocktails generated by solid-state fermentation

This study isolated fascinating novel fungal consortium from pomelo peel (PP) to discover its potential for on-site production of lignocellulose degrading enzymes through solid-state fermentation (SSF). SSF strategy was developed to use low and high recalcitrant substrates including PP and industrial hemp residue (IHR), respectively. The result revealed that SSF using a mixture of these substrates was significantly enhanced as compared with the corresponding individual substrate. IHR was pretreated with autohydrolysis, thermal hydrolysis, and thermochemical hydrolysis. A green and sustainable catalyst, oxalic acid, was employed to further optimize the thermochemical pretreatment of IHR. Pretreatment with 3% (w/v) oxalic acid resulted the highest sugar which is 8.3 and 3.24 folds compared to autohydrolysis and thermal hydrolysis, respectively. Finally, a novel approach dubbed one-pot pretreatment and saccharification was implemented using IHR treated with 2% oxalic acid and enzymes generated by SSF of mixed substrates. This strategy resulted the highest sugar production of 39.49 g/L. The current study can be considered as a proof-of-concept for sustainable and economic production of bio-based products from waste biomass resources.

An in-depth study of substrate effect on the performance of BOD sensor type microbial fuel cell

Nowadays, instant assessment of the organic content in wastewater is an urgent requirement to reduce water pollution. Microbial fuel cells (MFCs) can be used as effective biosensors for rapidly measuring BOD concentration of wastewater. However, wastewaters from different sources may consist of diverse chemical components, which may affect the BOD-measuring performance of MFC-type biosensors. Therefore, in this research, we tested different input substrates for the BOD sensor type MFC (MFC_BOD) to investigate their effects on the performance of the MFC. The substrates belonging to diverse groups such as carbohydrates, organic acids, amino acids and some chlorinated compounds (xenobiotics) were tested at different concentrations equivalent to BOD levels from 10 to 200 mg L-1. Concurrently, we also analyzed the alteration of the bacterial community in the anode of the MFC when tested with those different substrates by using PCR-DGGE. Our results showed that the MFC_BOD could have linear current-to-BOD responses (with the respective R2 values >0.9) to more metabolizable substrates such as carbohydrates, organic acid and glycerol; while it responded less sensitively at different degrees to some amino acids (serine, threonine and methionine) and did not respond to chloroform and chlorobenzene (chlorinated compounds). PCA and bacterial community analysis results surprisingly imply that such different responses may be solely due to different bio(electro)chemical processes associated with the substrates but not due to changes in the composition of the bacterial community. The results suggest that, to enable the MFC_BOD to accurately sense the BODs of the wastewaters containing recalcitrant or toxic substrates, special procedures are required to enrich in the anode the bacterial communities acclimated to the substrates right from the beginning

Fenton-Mediated Chlorophenol Degradation by Iron-Reducing Compounds Isolated from Endophytic Fungi in Atacama Puna Plateau Lecanicillium ATA01

Low-molecular-mass iron-reducing compounds (IRCs) were produced by entomopathogenic endophytic fungi Lecanicillium sp. ATA01 in liquid cultures. The extracellular hydrophilic extract contained three IRCs formed by peptides, iron and phenolate structures with molecular masses of 1207, 567 and 550 Da. These compounds were able to chelate and mediate the reduction of Fe+3 to Fe+2 and oxidized recalcitrant lignin-model substrates such as veratryl alcohol (VA), 2,6-dimethoxyphenol (DMP), and 2,2′-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid (ABTS) with or without hydrogen peroxide. Besides, IRCs can promote the degradation of chlorophenols. The maximal degradation of p-chlorophenol, 2,4-dichlorophenol, 2,4,6-trichlorophenol, and pentachlorophenol was conducted at optimal degradation conditions for IRCs (pH 3.5, iron 100 mM, and H2O2 10 mM). Furthermore, Fenton-like reactions using the synthetic iron chelates DTPA and EDTA and free Fe+2 and Fe+3 were also carried out in order to compare with the reaction mediated by IRCs. The ferric IRCs displayed the ability to enhance the hydroxylation of chlorophenols as a part of a degradation mechanism of the IRC-assisted Fenton reaction. The complexed iron was more efficient than free iron in the Fenton-like reaction, and between them, the fungal chelates were more efficient than the synthetic mill chelates.

The Novel Strain Acidomyces acidophilum Isolated from Acidophilic Biofilms (Snottites) Located in the Sheki-Heh Cave (North Caucasus).

A novel acidophilic fungal strain isolated from snottites in the active sulfuric acid speleogenesis (SAS) Sheki-Heh Cave (North Caucasus, Chechen Republic) was identified and characterized. The Sheki-Heh Cave is one of three cavities of the joint SAS speleosystem; to date, it remains the only of such cave explored in Russia. Highly acidic biofilms termed snottites are found sporadically on the cave roof in sulfurous water degassing zones. Only dark-colored micromycete colonies were isolated from these microbial biofilms using direct inoculation onto Czapek agar. The dominant fungal isolate was selected for further characterization. This work aimed to identify the micromycete strain isolated from cave snottites and explore its growth characteristics. Based on the phylogenetic analysis of the rDNA ITS region (540bp), the novel fungal strain was identified as Acidomyces acidophilum with a similarity level of 99.26%. The physiological properties of the strain were examined; the optimal pH and temperature for its growth were pH 3 and 20-28°C, respectively. Strain IB-G85 is able to grow under NaCl concentrations up to 3%. Although IB-G85 was isolated from an oligotrophic environment and was growing under nutrient deficiency, it could utilize some sugars and proteins as well as recalcitrant substrates, such as chitin and tannin. Compared to base Czapek-Dox Agar, lactic acid and colloidal chitin as the sole carbon sources enhanced fungal growth by 100 and 59%, respectively. The occurrence of A. acidophilum and closely related fungal species within acidophilic microbial communities inhabiting sulfur-containing ecosystems is discussed in view of their contribution to snottite structure formation in SAS caves.

Study of the conditions for determining some chloromethoxybenzoic acid derivatives

Chloromethoxybenzoic acid (CBA) Derivatives are active ingredients, are an integral part of many pesticides, important among which 3-chlorine-5-methoxybenzoic acid have, which is used as isopropylamine salts, as well as 2-chloro-4-methoxybenzoic acid used in the form of butylocutyl esther as herbicidal preparations. The residual amounts of these and other pesticides have a detrimental effect on the environment. An important role in the detoxification of pesticides have the soil microorganisms, but, unfortunately, the CBA derivatives are a recalcitrant substrate and are poorly decomposed by these microorganisms, since the CBA prevents the proliferation of such microorganisms. Therefore, the control of residues of CBA derivatives in environmental objects is important, as well as the development of new highly sensitive methods for determining CBA derivatives in such objects. To determine derivatives, CBA is most often used the Revers Phases HPLC method.

Lignocellulose degradation for the bioeconomy: The potential of enzyme synergies between xylanases, ferulic acid esterase and laccase for the production of arabinoxylo-oligosaccharides

The success of establishing bioeconomies replacing current economies based on fossil resources largely depends on our ability to degrade recalcitrant lignocellulosic biomass. This study explores the potential of employing various enzymes acting synergistically on previously pretreated agricultural side streams (corn bran, oat hull, soluble and insoluble oat bran). Degrees of synergy (oligosaccharide yield obtained with the enzyme combination divided by the sum of yields obtained with individual enzymes) of up to 88 were obtained. Combinations of a ferulic acid esterase and xylanases resulted in synergy on all substrates, while a laccase and xylanases only acted synergistically on the more recalcitrant substrates. Synergy between different xylanases (glycoside hydrolase (GH) families 5 and 11) was observed particularly on oat hulls, producing a yield of 57%. The synergistic ability of the enzymes was found to be partly due to the increased enzyme stability when in combination with the substrates.

Enzyme-assisted bioremediation approach for synthetic dyes and polycyclic aromatic hydrocarbons degradation.

Environmental protection from emerging pollutants has become a significant challenge for mankind as an increasing number of contaminants, including synthetic dyes and polycyclic aromatic hydrocarbons (PAHs), represent a serious risk to ecological and environmental balance. Most synthetic dyes have complex aromatic structures and are resistant to degrade by classical approaches, such as physical and chemical processes, including adsorption, chemical coagulation, flocculation, ion exchange, membrane separation, froth flotation, and reverse osmosis. Enzymes-assisted catalytic transformation of pollutants has become a potential alternative to classical methods because of their ability to react with complex compounds, a quick degradation rate, and producing less harmful by-products. Plant peroxidases, and microbial laccase and lignin-degrading peroxidases (manganese and lignin peroxidase) have gained significant attention for treating aromatic waste due to their capability of oxidizing and detoxifying a wide range of recalcitrant xenobiotics, including PAHs and synthetic dyes. Peroxidases being efficient biocatalysts detoxify an array of toxic compounds by simple free-radical mechanism resulting in the formation of oxidized and depolymerized products of significantly reduced toxicity. Moreover, it is an ecofriendly and economically favorable approach towards the biodegradation of recalcitrant and toxic industrial waste. Among microbial and plant peroxidases, bacterial enzymes have broad substrate specificity and can transform a wide range of recalcitrant substrates. Ligninolytic enzymes oxidize the aromatic ring into quinones and acids by producing free hydroxyl radicals instead of dihydrodiols and mineralize aromatic hydrocarbon in combination with cytochrome P450, monooxygenases, and epoxide hydrolases. In the review, an attempt has been made to provide detailed knowledge about the availability of inexpensive peroxidases sources, their mechanism of action, and degradation potential. The present review summarizes the exploitation of peroxidases from plants, bacteria, and fungus (manganese peroxidase, lignin peroxidase, and laccases) for detoxification and degradation of textile dyes as well as PAHs. Conclusively, peroxidases have great potential to react with almost all classes of synthetic dyes and most PAHs due to broad substrate specificity and transformed them into less harmful metabolites.