Quinone Redox Cycling Research Articles

Human NQO1 as a Selective Target for Anticancer Therapeutics and Tumor Imaging.

Human NAD(P)H-quinone oxidoreductase1 (HNQO1) is a two-electron reductase antioxidant enzyme whose expression is driven by the NRF2 transcription factor highly active in the prooxidant milieu found in human malignancies. The resulting abundance of NQO1 expression (up to 200-fold) in cancers and a barely detectable expression in body tissues makes it a selective marker of neoplasms. NQO1 can catalyze the repeated futile redox cycling of certain natural and synthetic quinones to their hydroxyquinones, consuming NADPH and generating rapid bursts of cytotoxic reactive oxygen species (ROS) and H2O2. A greater level of this quinone bioactivation due to elevated NQO1 content has been recognized as a tumor-specific therapeutic strategy, which, however, has not been clinically exploited. We review here the natural and new quinones activated by NQO1, the catalytic inhibitors, and the ensuing cell death mechanisms. Further, the cancer-selective expression of NQO1 has opened excellent opportunities for distinguishing cancer cells/tissues from their normal counterparts. Given this diagnostic, prognostic, and therapeutic importance, we and others have engineered a large number of specific NQO1 turn-on small molecule probes that remain latent but release intense fluorescence groups at near-infrared and other wavelengths, following enzymatic cleavage in cancer cells and tumor masses. This sensitive visualization/quantitation and powerful imaging technology based on NQO1 expression offers promise for guided cancer surgery, and the reagents suggest a theranostic potential for NQO1-targeted chemotherapy.

Induction of Extracellular Hydroxyl Radicals Production in the White-Rot Fungus Pleurotus eryngii for Dyes Degradation: An Advanced Bio-oxidation Process.

Among pollution remediation technologies, advanced oxidation processes (AOPs) are genuinely efficient since they are based on the production of strong, non-selective oxidants, mainly hydroxyl radicals (·OH), by a set of physicochemical methods. The biological counterparts of AOPs, which may be referred to as advanced bio-oxidation processes (ABOPs), have begun to be investigated since the mechanisms of induction of ·OH production in fungi are known. To contribute to the development of ABOPs, advanced oxidation of a wide number of dyes by the white-rot fungus Pleurotus eryngii, via a quinone redox cycling (QRC) process based on Fenton's reagent formation, has been described for the first time. The fungus was incubated with 2,6-dimethoxy-1,4-benzoquinone (DBQ) and Fe3+-oxalate, with and without Mn2+, leading to different ·OH production rates, around twice higher with Mn2+. Thanks to this process, the degradative capacity of the fungus increased, not only oxidising dyes it was not otherwise able to, but also increasing the decolorization rate of 20 dyes by more than 7 times in Mn2+ incubations. In terms of process efficacy, it is noteworthy that with Mn2+ the degradation of the dyes reached values of 90-100% in 2-4 h, which are like those described in some AOPs based on the Fenton reaction.

Immobilized bioreactor for enhanced ammonia, phosphorus, and phenol removal and effects of phenol on microbial communities, potential functions, and nitrogen metabolism

In the present study, an immobilized bioreactor was established to remove ammonia (NH4+-N), phosphate (PO43--P), and phenol using composite mycelium spheres (CMP) as the immobilization material in combination with Pseudomonas sp. Y1. Under optimal operating conditions, the bioreactor achieved 98.07, 91.71, and 92.57 % removal of NH4+-N, PO43--P, and phenol, respectively. The results showed that the bioreactor removed PO43--P by biomineralization and co-precipitation. Phenol removal relied on a Fenton-like reaction achieved by CMP-induced quinone redox cycling. High-throughput sequencing analysis and functional gene prediction indicated that Pseudomonas was the dominant genus and that the bioreactor had much potential for nitrogen removal, respectively. In addition, phenol affected the performance of functional genes and the associated enzymes, which influenced the nitrogen metabolism process in the bioreactor. This work serves as a guideline for the development of more stable and sustainable composite pollution removal technologies and fungal-bacterial symbiotic systems.

Enhanced ammonia nitrogen and phenol removal by immobilized bacteria through composite mycelium pellet-driven quinone redox cycle

The composite mycelium pellet (CMP) was coupled with Pseudomonas sp. Y1 (CMP-Y1) to remove phenol and ammonia nitrogen (NH4+-N). The CMP was formed by the self-assembly of fungal mycelium with sponge iron (SIO), gallic acid (GA), and oxalic acid. The results showed that CMP with abundant pore size and successful internal loading of sponge iron containing iron nanoparticles. CMP could induce GA redox cycle to form Fenton-like reaction and thus achieve efficient phenol removal (93.32%, 24 h). Meanwhile, the removal efficiencies of phenol, NH4+-N, and chemical oxygen demand (COD) using CMP-Y1 at 12 h were 93.71, 92.40, and 89.00%, respectively. The increase in the electron transfer activity of strain Y1 by the addition of CMP could facilitate the nitrogen removal processes. In addition, high-throughput sequencing results indicated the abundance of antioxidant and repair genes was increased, which might be a strategy of strain Y1 to cope with oxidative stress. This strategy provided the possibility for the practical application of the combination of advanced oxidation and biological treatment, and offered new insights into the symbiotic system of fungi and bacteria.

Iron reduction in composting environment synergized with quinone redox cycling drives humification and free radical production from humic substances

The aim of this paper was to investigate the influence of Fe (III) on humification and free radicals evolution. The experimental data showed that the experimental group (CT) with Fe2(SO4)3 had a better degree of humification than the control group (CK). The humic substances (HS) content was 10% higher in CT (23.94 mg·g−1) than in CK (21.54 mg·g−1) in the final. Fe (III) contributed significantly to the formation of free radicals in HS. The amount of H2O2 in CT increased to 74.8 mmol·kg−1, while CK was only 46.5 mmol·kg−1. The content of semiquinone free radical was 10.32 × 1011 spins/mm3 in CT, 5.11 × 1011 spins/mm3 in CK in the end. Several iron-reducing bacteria were detected in composting, among which Paenibacillus was dominant. The above findings suggested that the application of Fe2(SO4)3 enhanced the iron reduction synergistic quinone redox cycling and promoted the generation of free radicals during the humification of composting.

Redox-Cycling "Mitocans" as Effective New Developments in Anticancer Therapy.

Our study proposes a pharmacological strategy to target cancerous mitochondria via redox-cycling "mitocans" such as quinone/ascorbate (Q/A) redox-pairs, which makes cancer cells fragile and sensitive without adverse effects on normal cells and tissues. Eleven Q/A redox-pairs were tested on cultured cells and cancer-bearing mice. The following parameters were analyzed: cell proliferation/viability, mitochondrial superoxide, steady-state ATP, tissue redox-state, tumor-associated NADH oxidase (tNOX) expression, tumor growth, and survival. Q/A redox-pairs containing unprenylated quinones exhibited strong dose-dependent antiproliferative and cytotoxic effects on cancer cells, accompanied by overproduction of mitochondrial superoxide and accelerated ATP depletion. In normal cells, the same redox-pairs did not significantly affect the viability and energy homeostasis, but induced mild mitochondrial oxidative stress, which is well tolerated. Benzoquinone/ascorbate redox-pairs were more effective than naphthoquinone/ascorbate, with coenzyme Q0/ascorbate exhibiting the most pronounced anticancer effects in vitro and in vivo. Targeted anticancer effects of Q/A redox-pairs and their tolerance to normal cells and tissues are attributed to: (i) downregulation of quinone prenylation in cancer, leading to increased mitochondrial production of semiquinone and, consequently, superoxide; (ii) specific and accelerated redox-cycling of unprenylated quinones and ascorbate mainly in the impaired cancerous mitochondria due to their redox imbalance; and (iii) downregulation of tNOX.

Redox cycling of quinones reduced by ascorbic acid

In aqueous solutions containing ascorbic acid and O2, many quinones undergo reduction to the semiquinone followed by reoxidation. This redox cycling mediates the oxidation of ascorbic acid and the reduction of O2 to superoxide and ultimately hydrogen peroxide. For that reason, redox cycling has attracted attention as a source of reactive oxygen species. This redox cycling is paradoxical, however, because the one-equivalent reduction potentials of the reactants are unfavorable, so the concentrations of the products, monodehydroascorbate and superoxide, must be kept extremely low. Disproportionation is not fast enough to eliminate these products. We have investigated the mechanism of ascorbate-driven redox cycling by monitoring the redox status of the quinone and the rate of redox cycling in parallel. Evidence is presented for a mechanism in which monodehydroascorbate is oxidized by the semiquinone. The result is that cycling of the semiquinone and hydroquinone mediates a rapid disproportionation of monodehydroascorbate. This mechanism accounts for the dependence of the redox cycling rate on quinone and ascorbate concentrations as well as on the reduction potential of the quinone. Therefore, it predicts how fast ascorbate-driven redox cycling will generate hydrogen peroxide under a variety of conditions and with different quinones.

Quinone redox cycling drives lignocellulose depolymerization and degradation in composting environments based on metagenomics analysis

In this study, the effect of Fe3+ on the quinone redox cycling driving lignocellulosic degradation in composting systems was investigated. The results showed that the degradation rates of cellulose, hemicellulose, and lignin were higher in the experimental group (CT) with Fe2(SO4)3 addition than in the blank group (CK) (CT, 52.55 %, 45.14 %, 56.98 %; CK, 49.63 %, 37.34 %, 52.3 %). Changes in the abundance of key enzymes for quinone reduction (AA3_1, AA3_2, AA6) and the structural succession of microbial communities were analyzed by metagenomic analysis. Among them, Fe2(SO4)3 had the most significant effect on AA3_2, with an approximately 8-fold increase in abundance compared to the beginning of composting. The dominant phylum in the composting process was Actinobacteria. In conclusion, the addition of Fe2(SO4)3 contributed to the quinone redox cycling and effectively improved the degradation rate of lignocellulose in composting.

Development of signal multiplication system for quinone linked immunosorbent assay (Multi-QuLISA) by using poly-l-lysine dendrigraft and 1,2-naphthoquinone-4-sulfonate as enzyme-free tag

A sensitive and stable signal multiplied quinone-linked immunosorbent assay (Multi-QuLISA) was developed. In Multi-QuLISA, an oligomerized quinone linked to biotin, namely biotin-8mer-naphthoquinone (Bio8mer-NQ), is used as a signal-generating label. Bio8mer-NQ is formed from a dendrigraft poly-l-lysine generation 1 (DPLL G1), a controlled branched oligomer composed of eight lysine moieties with nine free amino groups as a backbone. One of the nine amino groups of DPLL G1 is attached to biotin moiety, while the other eight are attached to 1,2-naphthoquinone-4-sulfonate (NQS). Bio8mer-NQ labels a biotinylated detection antibody using avidin as a co-binder. Then, multi-quinones in Bio8mer-NQ undergo a redox cycle with dithiothreitol and luminol, generating strong chemiluminescence. Standard ELISA uses a label enzyme that suffers from vulnerability in different conditions and poor stability. Bio8mer-NQ showed better stability than the enzyme (biotin-HRP) under different drastic pH and temperature conditions, hydrolytic enzymes, etc. Furthermore, Bio8mer-NQ was used as both chemiluminescence and colorimetric label based on the redox cycle of quinone, and it had LODs of 1.5 and 6.5 nM, respectively. The method could detect biotinylated immunocomplex in an in-house designed immunoassay down to 0.2 nM, which is about 25 times more sensitive than biotin HRP. Eventually, Bio8mer-NQ was applied successfully in Multi-QuLISA for detecting β-casein with a sensitivity of 3.2 ng/mL, while the conventional ELISA had an LOD of 35 ng/mL. Overall, Bio8mer-NQ is a stable compound that could be used as an excellent replacement for the enzyme in immunoassay and can be used in both colorimetric and chemiluminescence assays with good sensitivity.

Characterization of a Deep-Sea Actinobacterium Strain Uncovers Its Prominent Capability of Utilizing Taurine and Polyvinyl Alcohol.

Actinobacteria represent a large group of important prokaryotes with great application potentials and widely distribute in diverse natural environments including the ocean. However, compared to their terrestrial cultured members, there are much less available marine Actinobacteria, especially deep-sea counterparts. Here, we cultured a bacterial strain of deep-sea actinobacterium, Marmoricola sp. TYQ2, by using a basal medium supplemented with taurine. Consistently, the growth of strain TYQ2 was significantly promoted by the supplement of taurine. Transcriptomic analysis showed that the expressions of genes encoding proteins associated with taurine metabolization and utilization as well as energy generation were evidently up-regulated when taurine was added. Moreover, strain TYQ2 was demonstrated to degrade polyvinyl alcohol (PVA) with the involvement of the redox cycle of extracellular quinol and quinone and the reduction of iron to ferrous, and strain TYQ2 could utilize the degradation products for energy production, thereby supporting bacterial growth. Overall, our experimental results demonstrate the prominent degradation capabilities of Marmoricola sp. TYQ2 toward the organics taurine and PVA.

Effects of Acidity on Reactive Oxygen Species Formation from Secondary Organic Aerosols.

Reactive oxygen species (ROS) play a critical role in the chemical transformation of atmospheric secondary organic aerosols (SOA) and aerosol health effects by causing oxidative stress in vivo. Acidity is an important physicochemical property of atmospheric aerosols, but its effects on the ROS formation from SOA have been poorly characterized. By applying the electron paramagnetic resonance spin-trapping technique and the Diogenes chemiluminescence assay, we find highly distinct radical yields and composition at different pH values in the range of 1–7.4 from SOA generated by oxidation of isoprene, α-terpineol, α-pinene, β-pinene, toluene, and naphthalene. We observe that isoprene SOA has substantial hydroxyl radical (•OH) and organic radical yields at neutral pH, which are 1.5–2 times higher compared to acidic conditions in total radical yields. Superoxide (O2•–) is found to be the dominant species generated by all types of SOAs at lower pH. At neutral pH, α-terpineol SOA exhibits a substantial yield of carbon-centered organic radicals, while no radical formation is observed by aromatic SOA. Further experiments with model compounds show that the decomposition of organic peroxide leading to radical formation may be suppressed at lower pH due to acid-catalyzed rearrangement of peroxides. We also observe 1.5–3 times higher molar yields of hydrogen peroxide (H2O2) in acidic conditions compared to neutral pH by biogenic and aromatic SOA, likely due to enhanced decomposition of α-hydroxyhydroperoxides and quinone redox cycling, respectively. These findings are critical to bridge the gap in understanding ROS formation mechanisms and kinetics in atmospheric and physiological environments.

Functionalized Dioxonaphthoimidazoliums: A Redox Cycling Chemotype with Potent Bactericidal Activities against Mycobacterium tuberculosis.

Disruption of redox homeostasis in mycobacteria causes irreversible stress induction and cell death. Here, we report the dioxonaphthoimidazolium scaffold as a novel redox cycling antituberculosis chemotype with potent bactericidal activity against growing and nutrient-starved phenotypically drug-resistant nongrowing bacteria. Maximal potency was dependent on the activation of the redox cycling quinone by the positively charged scaffold and accessibility to the mycobacterial cell membrane as directed by the lipophilicity and conformational characteristics of the N-substituted side chains. Evidence from microbiological, biochemical, and genetic investigations implicates a redox-driven mode of action that is reliant on the reduction of the quinone by type II NADH dehydrogenase (NDH2) for the generation of bactericidal levels of the reactive oxygen species (ROS). The bactericidal profile of a potent water-soluble analogue 32 revealed good activity against nutrient-starved organisms in the Loebel model of dormancy, low spontaneous resistance mutation frequency, and synergy with isoniazid in the checkerboard assay.

Lytic polysaccharide monooxygenases promote oxidative cleavage of lignin and lignin-carbohydrate complexes during fungal degradation of lignocellulose.

Overcoming lignocellulosic biomass recalcitrance, especially the cleavage of cross-linkages in lignin-carbohydrate complexes (LCCs) and lignin, is essential for both the carbon cycle and industrial biorefinery. Lytic polysaccharide monooxygenases (LPMOs) are copper-containing enzymes that play a key role in fungal polysaccharide oxidative degradation. Nevertheless, comprehensive analysis showed that LPMOs from a white-rot fungus, Pleurotus ostreatus, correlated well with the Fenton reaction and were involved in the degradation of recalcitrant nonpolysaccharide fractions in this research. Thus, LPMOs participated in the extracellular Fenton reaction by enhancing iron reduction in quinone redox cycling. A Fenton reaction system consisting of LPMOs, hydroquinone, and ferric iron can efficiently produce hydroxy radicals and then cleave LCCs or lignin linkages. This finding indicates that LPMOs are underestimated auxiliary enzymes in eliminating biomass recalcitrance.

Enhanced Fenton Reaction for Xenobiotic Compounds and Lignin Degradation Fueled by Quinone Redox Cycling by Lytic Polysaccharide Monooxygenases.

The Fenton reaction is considered to be of great significance in the initial attack of lignocellulose in wood-decaying fungi. Quinone redox cycling is the main way to induce the Fenton reaction in fungi. We show that lytic polysaccharide monooxygenases (LPMOs), through LPMO-catalyzed oxidation of hydroquinone, can efficiently cooperate with glucose dehydrogenase (GDH) to achieve quinone redox cycling. The LPMO/GDH system can enhance Fe3+-reducing activity, H2O2 production, and hydroxyl radical generation, resulting in a fueled Fenton reaction. The system-generated hydroxyl radicals exhibited a strong capacity to decolorize different synthetic dyes and degrade lignin. Our results reveal a potentially critical connection between LPMOs and the Fenton reaction, suggesting that LPMOs could be involved in xenobiotic compound and lignin degradation in fungi. This new role of LPMOs may be exploited for application in biorefineries.

Hydrogen-dependent current generation and energy conservation by Shewanella oneidensis MR-1 in bioelectrochemical systems

Bioelectrochemical systems (BESs) are engineered systems that utilize electrochemical interactions between electrochemically active bacteria (EAB) and electrodes. BESs have attracted considerable attention for their utility in biotechnological processes. In a BES, hydrogen is generated by the reduction of water on low-potential cathode electrodes. However, limited information is available on the effect of hydrogen on the metabolism and growth of EAB and current generation in BESs. Here, we investigated the effect of hydrogen on current generation by a model EAB, Shewanella oneidensis MR-1. We found that this strain utilizes hydrogen as an electron donor for electrode respiration, thereby facilitating current generation and cell growth in the presence of organic substrates. Inner membrane (IM) quinones (i.e., ubiquinone and menaquinone), IM quinone-reactive hydrogenase Hya, and IM-bound quinone reductase CymA are involved in hydrogen-dependent current generation, suggesting that the redox cycling of IM quinones catalyzed by Hya and CymA contributes to the generation of the proton motive force and the synthesis of ATP via F0F1-ATPase. These findings indicate that the evolution of hydrogen on the cathode facilitates energy metabolism and growth of hydrogen-utilizing EAB associated with anodes. The results also suggest that hydrogen cycling between cathodes and anodes can hinder quantitative evaluation of organic substrate-dependent current generation in BESs.

Phanerochaete chrysosporium-driven quinone redox cycling promotes degradation of imidacloprid

This study systematically investigated the advanced oxidation processes driven by the white-rot fungus (WRF) Phanerochaete chrysosporium in the removal of the neonicotinoid pesticide imidacloprid (IMI). The quinone redox cycling driven by P. chrysosporium promotes the degradation of IMI in presence of Mn2+ and Fe (III)-oxalate. Results showed that the degradation efficiency varied with the type of quinine mediator, and gallic acid (GA) was a more effective mediator than 1,4-benzoquinone (BQ). The removal rate of IMI was increased by 1.8-fold in the presence of 100 μM of Mn2+ compared with that of the system without Mn2+. The addition of oxalic acid and EDTA promoted the degradation of IMI, and the effect of oxalic acid was more pronounced at the later stage (day 5 and day 6); the optimal Fe3+/oxalate ratio was 1:3. Time-course experiments revealed the maximum removal rates of IMI were 97.37%, 82.0% and 73.52% at the initial IMI concentration of 10, 20, and 30 mg/L, respectively, in the presence of 100 μM Mn2+, 100 μM Fe3+-300 μM of oxalate and 500 μM gallic acid. Under quinone redox cycling conditions, the good correlation between ·OH production rates and the enzymes activity was concluded, the quinines redox cycles promote enzyme activity, as well as the rate of ·OH production. Results from this study indicated the GA mediator-based system using P. chrysosporium offers a new method to advance the bio-oxidation process owing to its simple culture system, low-cost precursors, unique fungi, and degradation stability.

Quinone-based antibody labeling reagent for enzyme-free chemiluminescent immunoassays. Application to avidin and biotinylated anti-rabbit IgG labeling

Chemiluminescence-enzyme immunoassays make it possible to measure trace components with high sensitivity and selectivity due to the high specificity of the antigen-antibody reaction and the high sensitivity of chemiluminescence assays. However, using an enzyme-labeled antibody suffers from many problems such as low reproducibility due to the instability of the enzyme and inhibition of antigen-antibody reaction due to its steric effect. Therefore, herein we report an innovative non-enzymatic chemiluminescence immunoassays labeling reagent through using quinone as a signal-generating tag coupled with biotin as a binder, to overcome enzymatic labeling problems. Biotinylated-1,4-naphthoquinone (biotin-NQ) was synthesized and characterized and it could produce long-lasting chemiluminescence upon mixing with dithiothreitol and luminol based on the redox cycle of quinone. Biotin-NQ showed exceptional stability towards different stress factors that may be encountered during performing the immunoassay such as high temperatures, highly acidic and alkaline conditions, and repeated freeze-thaw cycles. On the other hand, all these conditions lead to decreased labeling enzyme reactivity due to possible denaturation of its protein structure. Finally, the measurement of the biotin-labeled antibody was successfully performed using biotin-NQ and avidin. As a result, the antibody could be detected down to 25.7 nM which is 2.5 times sensitive than biotin-HRP chemiluminescence-enzyme immunoassays. Moreover, our method was applied successfully for determination of avidin using immobilized biotinylated antibody and biotin-NQ, which simulates immunoassays. Avidin could be detected down to 23.4 nM with excellent linearity (r = 0.996). Accordingly, our developed reagent, biotin-NQ, could be used as a universal highly stable, cost-effective, and steric free non-enzymatic label for immunoassays.

Enhanced Photooxidation of Hydroquinone by Acetylacetone, a Novel Photosensitizer and Electron Shuttle.

Quinones are important electron shuttles as well as micropollutants in the nature. Acetylacetone (AA) is a newly recognized electron shuttle in aqueous media exposed to UV irradiation. Herein, we studied the interactions between AA and hydroquinone (QH2) under steady-state and transient photochemical conditions to clarify the possible reactions and consequences if QH2 and AA coexist in a solution. Steady-state experimental results demonstrate that the interactions between AA and QH2 were strongly affected by dissolved oxygen. In O2-rich solutions, the phototransformation of QH2 was AA-independent. Both QH2 and AA utilize O2 as the electron acceptor, but in O2-insufficient solutions, AA became an important electron acceptor for the oxidation of QH2. In all cases, the coexistence of AA increased the phototransformation of QH2, whereas the decomposition of AA in O2-saturated and oversaturated solutions was inhibited by the presence of QH2. The underlying mechanisms were investigated by a combination of laser flash photolysis (LFP) and reduction potential analysis. The LFP results show that the excited AA serves as a better electron shuttle than QH2. As a consequence, AA might regulate the redox cycling of quinones, leading to significant effects on many processes, ranging from photosynthesis and respiration to photodegradation.

Development of ultrafast colorimetric microplate assay method for ubiquinone utilizing the redox cycle of the quinone

A simple, rapid, and sensitive microplate assay method for determination of ubiquinone in cosmetic products has been developed and validated. This method is based on measuring reactive oxygen species (ROS) generated from the redox cycle of quinone using redox colorimetric reagent, 2-(4-Iodophenyl)-3-(4-nitrophenyl)-5-phenyl-2H-tetrazolium chloride (INT). The reaction with ROS converts INT to a formazan dye having a strong absorbance at 510 nm that is increased with increasing concentration of ubiquinone. On the basis of this reaction, we developed a microplate assay method that allows high-throughput measurement of ubiquinone. This method uses microplate reader for the measurement of absorbance and it can determine many samples at one time. Under the optimized conditions, the proposed method was linear in the range of 0.02–4 μM and the limit of detection (blank +3SD) was 14.8 nM. We applied the proposed method to determine ubiquinone contents in cosmetic products, and the measurement values were in good agreement with the indicated value. The proposed colorimetric method should be useful as a high-throughput analysis for quality management of ubiquinone containing products.

The secretome of two representative lignocellulose-decay basidiomycetes growing on sugarcane bagasse solid-state cultures

Secretome evaluations of lignocellulose-decay basidiomycetes can reveal new enzymes in selected fungal species that degrade specific substrates. Proteins discovered in such studies can support biorefinery development. Brown-rot (Gloeophyllum trabeum) and white-rot (Pleurotus ostreatus) fungi growing in sugarcane bagasse solid-state cultures produced 119 and 63 different extracellular proteins, respectively. Several of the identified enzymes are suitable for in vitro biomass conversion, including a range of cellulases (endoglucanases, cellobiohydrolases and β-glucosidases), hemicellulases (endoxylanases, α-arabinofuranosidases, α-glucuronidases and acetylxylan esterases) and carbohydrate-active auxiliary proteins, such as AA9 lytic polysaccharide monooxygenase, AA1 laccase and AA2 versatile peroxidase. Extracellular oxalate decarboxylase was also detected in both fungal species, exclusively in media containing sugarcane bagasse. Interestingly, intracellular AA6 quinone oxidoreductases were also exclusively produced under sugarcane bagasse induction in both fungi. These enzymes promote quinone redox cycling, which is used to produce Fenton’s reagents by lignocellulose-decay fungi. Hitherto undiscovered hypothetical proteins that are predicted in lignocellulose-decay fungi genomes appeared in high relative abundance in the cultures containing sugarcane bagasse, which suggests undisclosed, new biochemical mechanisms that are used by lignocellulose-decay fungi to degrade sugarcane biomass. In general, lignocellulose-decay fungi produce a number of canonical hydrolases, as well as some newly observed enzymes, that are suitable for in vitro biomass digestion in a biorefinery context.