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Exoglucanase (CBH) is the rate limiting enzyme in the process of cellulose degradation. The carbohydrate binding module (CBM) can improve the accessibility of cellulase to substrate, thereby promoting the enzymatic hydrolysis of cellulase. In this study, the influence of CBM on the properties of GH6 exoglucanase from Chaetomium thermophilum (CtCBH) is systematically explored from three perspectives: the fusion of exogenous CBM, the exogenous CBM replacement of its own CBM, and the deletion of its own CBM. The parental and reconstructed CtCBH presented the same optimum pH (6.0) and temperature (60 °C) for maximum activity. Fusion of exogenous CBM increased the binding capacity of CtCBH to Avicel by 8% and 9%, respectively, but it had no significant effect on its catalytic activity. The exogenous CBM replacement of its own CBM resulted in a 12% reduction in the binding ability of CtCBH to Avicel, and a 26% reduction in the catalytic activity of Avicel. The deletion of its own CBM significantly reduced the binding ability of CtCBH to Avicel by approximately 53%, but its catalytic activity was not obviously reduced. These observations suggest that binding ability of CBM is not necessary for the catalysis of CtCBH.

Rubichaetoglobin A (1), a new cytochalasan alkaloid, together with nine closely related known ones (2-10), were isolated from the ethyl acetate extracts of the endophytic fungus Chaetomium tectifimeti S104 harbored in the root of Rubia podantha Diels. Their structures were elucidated based on comprehensive spectroscopic analysis. All isolated compounds were tested for cytotoxic, antibacterial, and nitric oxide inhibitory activities. The results showed that 2, 4, and 5 possessed moderate cytotoxicity against MDA-MB-231 cells with the IC50 values of 19.14, 11.43, and 10.27 μM, respectively.

The herbicide butachlor has been used in huge quantities worldwide, affecting various environmental systems. Butachlor residues have been detected in soil, water, and organisms, and have been shown to be toxic to these non-target organisms. This paper briefly summarizes the toxic effects of butachlor on aquatic and terrestrial animals, including humans, and proposes the necessity of its removal from the environment. Due to long-term exposure, some animals, plants, and microorganisms have developed resistance toward butachlor, indicating that the toxicity of this herbicide can be reduced. Furthermore, we can consider removing butachlor residues from the environment by using such butachlor-resistant organisms. In particular, microbial degradation methods have attracted much attention, with about 30 kinds of butachlor-degrading microorganisms have been found, such as Fusarium solani, Novosphingobium chloroacetimidivorans, Chaetomium globosum, Pseudomonas putida, Sphingomonas chloroacetimidivorans, and Rhodococcus sp. The metabolites and degradation pathways of butachlor have been investigated. In addition, enzymes associated with butachlor degradation have been identified, including CndC1 (ferredoxin), Red1 (reductase), FdX1 (ferredoxin), FdX2 (ferredoxin), Dbo (debutoxylase), and catechol 1,2 dioxygenase. However, few reviews have focused on the microbial degradation and molecular mechanisms of butachlor. This review explores the biochemical pathways and molecular mechanisms of butachlor biodegradation in depth in order to provide new ideas for repairing butachlor-contaminated environments. KEY POINTS: • Biodegradation is a powerful tool for the removal of butachlor. • Dechlorination plays a key role in the degradation of butachlor. • Possible biochemical pathways of butachlor in the environment are described.

Acta crystallographica / Section D 77(6), 755 - 775 (2021). doi:10.1107/S2059798321003533, The FAD-dependent oxidoreductase from Chaetomium thermophilum (CtFDO) is a novel thermostable glycoprotein from the glucose–methanol–choline (GMC) oxidoreductase superfamily. However, CtFDO shows no activity toward the typical substrates of the family and high-throughput screening with around 1000 compounds did not yield any strongly reacting substrate. Therefore, protein crystallography, including crystallographic fragment screening, with 42 fragments and 37 other compounds was used to describe the ligand-binding sites of CtFDO and to characterize the nature of its substrate. The structure of CtFDO reveals an unusually wide-open solvent-accessible active-site pocket with a unique His–Ser amino-acid pair putatively involved in enzyme catalysis. A series of six crystal structures of CtFDO complexes revealed five different subsites for the binding of aryl moieties inside the active-site pocket and conformational flexibility of the interacting amino acids when adapting to a particular ligand. The protein is capable of binding complex polyaromatic substrates of molecular weight greater than 500 Da., Published by Wiley, Bognor Regis

Cytochalasins are a group of fungal secondary metabolites with diverse structures and bioactivities, including chaetoglobosin A production. Chaetoglobosin A is produced by Chaetomium globosum and has potential antifungal activity. Bioinformatics analysis of the chaetoglobosin A gene cluster (che) showed it that consists of nine open reading frames, including those encoding polyketide synthases (PKSs), PKS extender units, post-PKS modifications, and proposed regulators. Here, the role of the CgcheR regulator was investigated using gene disruption experiments. The CgcheR disruptant (ΔCgcheR) completely abolished the production of chaetoglobosin A, which was restored by the introduction of a copy of the wild-type CgcheR gene, suggesting that CgcheR is involved in chaetoglobosin A biosynthesis. A transcriptional analysis of the CgcheR disruptant indicated that CgCheR activates the transcription of chaetoglobosin biosynthetic genes in a pathway-specific manner. Furthermore, constitutive overexpression of CgcheR significantly improved the production of chaetoglobosin A from 52 to 260 mg/L. Surprisingly, CgcheR also played a critical role in sporulation; the CgcheR disruptant lost the ability to produce spores, suggesting that the regulator modulates cellular development. Our results not only shed light on the regulation of chaetoglobosin A biosynthesis, but also indicate a relationship between secondary metabolism and fungal morphogenesis.

Endophytic fungi always attract attention due to their generous production of bioactive and chemically novel compounds that have medical, agriculture, industrial applications. This review focused on Chaetomium as a model of endophytic fungi rich in therapeutic agents that have known medicinal and industrial application. Moreover, understanding the importance of this potent fungus encourage further studies to identify novel metabolites, and at the same time employing already known metabolites to evaluate their activity in order to be used in additional applications.

A novel thermostable xylanase gene from Chaetomium sp. CQ31 was cloned and codon–optimized (CsXynBop). The deduced protein sequence of the gene shared the highest similarity of 75% with the glycoside hydrolase (GH) family 10 xylanase from Achaetomium sp. Xz–8. CsXynBop was over–expressed in Pichia pastoris GS115 by high–cell density fermentation, with the highest xylanase yield of 10,017 U/mL. The recombinant xylanase (CsXynBop) was purified to homogeneity and biochemically characterized. CsXynBop was optimally active at pH 6.5 and 85 °C, respectively, and stable over a broad pH range of 5.0–9.5 and up to 60 °C. The enzyme exhibited strict substrate specificity towards oat–spelt xylan (2, 489 U/mg), beechwood xylan (1522 U/mg), birchwood xylan (1067 U/mg), and showed relatively high activity towards arabinoxylan (1208 U/mg), but exhibited no activity on other tested polysaccharides. CsXynBop hydrolyzed different xylans to yield mainly xylooligosaccharides (XOSs) with degree of polymerization (DP) 2–5. The application of CsXynBop (200 U/g malt) in malt mashing substantially decreased the filtration time and viscosity of malt by 42.3% and 8.6%, respectively. These excellent characteristics of CsXynBop may make it a good candidate in beer industry.

Chaetomium species are known as potential biocontrol agents against phytopathogens due to their multiple antagonistic mechanisms. Plant disease is controlled by Chaetomium exhibit complex interactions against plant pathogen under varied conditions. Previously, mycoparasitism and antibiosis have been reported as most effective mechanism exhibited by the C. globosum against Bipolaris sorokiniana . In the present study, the examination of major biosynthetic pathways underlying Chaetomium globosum biocontrol activity was elucidated. It was shown that the pathways related to biosynthesis of secondary metabolites, hydrolytic enzymes and other key regulator genes were involved in production of hydrolytic enzymes and antifungal metabolites. We identified various genes of biological function with significant log2 fold change such as phosphoribosyl aminoimidazole carboxylase (9.693), protease (8.18), cyanate hydratase (Cyanase) (6.7), Fe2OG dioxygenase domain-containing protein (5.9), superoxide dismutase (5.55), glycosidase (5.34), carboxylic ester hydrolase (5.27), alpha-1,2-Mannosidase (4.44), alpha-1,4 glucan phosphorylase (3.99), endochitinase (3.87), P53-like transcription factor (Fragment) (3.55), metalloprotease (3.4), polyketide synthase (3.35), Catalase-peroxidase (CP) (3.14), protein kinase domain-containing protein (3.18) and glutamate decarboxylase (2.1) which are involved in biosynthesis of secondary metabolites, polyketide synthase, antibiotic, hydrolytic enzymes and putative fungistatic metabolites. This data provides a good foundation for continued researches into C. globosum Cg2 biocontrol activity for facilitating widespread application under the field conditions.

Many representatives of soil saprotroph fungi can penetrate into plant tissues forming endophytic associations with a positive effect on both micro- and macroorganisms. The endophytic fungi may positive act on growth processes, immune status, and resistance of plants to stress factors. For some crops, including buckwheat, the ability to form endophytic associations with soil fungi remains unexplored. Thus the main objectives of our study were to establish the features of interaction between Chaetomium cochliodes3250 and buckwheat plants. Conventional biochemical (content of plant growth regulators, activity of succinate dehydrogenase, acid and alkaline phosphatase, exoglucanase, endoglucanase, β-glucosidase, polygalacturonase) and physicochemical (content of photosynthetic pigments) study methods were used during the work. We established the capability of C. cochliodes 3250 for growth-regulating substances synthesis. The fungus could produce indolyl 3-acetic acid, gibberellic acid, that stimulates growth and development of plants both with mediator molecules, synthesizes 2,4-epibrassinolide, cholesterol and ergosterol playing an important role in plant resistance against pathogens. We found out that C. cochliodes 3250 was capable for active synthesis of cellulase complex enzymes (exoglucanase, endoglucanase, β-glucosidase) and polygalacturonase, necessary for fungus penetration into the root tissues. Pre-sowing treatment of buckwheat seeds by fungus caused the main physiological responses of plants: an increase of total adsorption and active working surface of the roots, the length and weight of plants, the leaf area and the content of chlorophylls a and b. Thus, the ability of C. cochliodes3250 to form an efficient endophytic association with buckwheat plants been improved.

Condensin and cohesin, both members of the structural maintenance of chromosome (SMC) family, contribute to the regulation and structure of chromatin. Recent work has shown both condensin and cohesin extrude DNA loops and most likely work via a conserved mechanism. This review focuses on condensin complexes, highlighting recent in vitro work characterising DNA loop formation and protein structure. We discuss similarities between condensin and cohesin complexes to derive a possible mechanistic model, as well as discuss differences that exist between the different condensin isoforms found in higher eukaryotes.

Historic photographs are a precious cultural heritage. Despite this, there is a lack of studies on the microorganisms that are responsible for their biodeterioration and the underlying mechanisms. The aims of this study were twofold: (a) assess microorganism diversity on historic photographs using, for the first time, high-throughput sequencing on the Illumina platform; (b) assess the suitability of new methods for mass spectrometric analysis of metabolites on photographs, namely laser ablation-remote-electrospray ionization mass spectrometry imaging (LARESI MSI) in selected reaction monitoring mode (SRM), and surface-assisted laser desorption/ionization mass spectrometry (MS) and mass spectrometry imaging (MSI), on silver nanoparticle enhanced target (109AgNPET MS and MSI). Samples of 19th century Polish silver-gelatin photographs on baryta paper (paper substrate with layer of barium sulphate), obtained by the gelatin developing-out process, were studied. These photographs (after incubation in temperature: 25 ± 2 °C, relative humidity: 80%, time: 60 days) were contaminated with culturable bacteria at concentrations between 3.33 × 101 and 1.60 × 102 CFU/cm2 and fungi at concentrations between 1.57 × 105 and 9.93 × 105 CFU/cm2. Twenty-two phyla of bacteria and one of Archaea were identified on the photographs. Bacteria belonging to Proteobacteria predominated (relative abundance 75.25–84.94%); followed by Firmicutes (1.04–12.61%) and Actinobacteria. Fungi identified belonged mainly to Ascomycota. The dominant bacteria identified included: Mesorhizobium, Burkholderia, Delftia, Enhydrobacter, Paenibacillus, Saccharopolyspora and Olsenella. The dominant fungi belonged to the genera Aspergillus, Alternaria, Penicillium, Chaetomium, Talaromyces, Fusarium. These predominant microorganisms may be responsible for the biodeterioration of the photographs. Direct laser/desorption ionization mass spectrometry could be useful for the reconstruction of heavily damaged photographs. The LARESI MSI method was used to detect metabolites of microbial origin (organic acids and mycotoxins) on the surface of historic photographs, which are compatible with the analysis of fungal biodiversity. The biggest advantage of presented methods is that the tested photographs were not modified in any way before the tests which can be carried out in situ.

Cytoplasmic dynein is a eukaryotic motor protein complex that, along with its regulatory protein dynactin, is essential to the transport of organelles within cells. The interaction of dynein with dynactin is regulated by binding between the intermediate chain (IC) subunit of dynein and the p150Glued subunit of dynactin. Even though in the rat versions of these proteins this interaction primarily involves the single α-helix region at the N-terminus of the IC, in Drosophila and yeast ICs the removal of a nascent helix (H2) downstream of the single α-helix considerably diminishes IC-p150Glued complex stability. We find that for ICs from various species, there is a correlation between disorder in H2 and its contribution to binding affinity, and that sequence variations in H2 that do not change the level of disorder show similar binding behavior. Analysis of the structure and interactions of the IC from Chaetomium thermophilum demonstrates that the H2 region of C. thermophilum IC has a low helical propensity and establishes that H2 binds directly to the coiled-coil 1B (CC1B) domain of p150Glued, thus explaining why H2 is necessary for tight binding. Isothermal titration calorimetry, circular dichroism, and NMR studies of smaller CC1B constructs localize the region of CC1B most essential for a tight interaction with IC. These results suggest that it is the level of disorder in H2 of IC along with its charge, rather than sequence specificity, that underlie its importance in initiating tight IC-p150Glued complex formation. We speculate that the nascent H2 helix may provide conformational flexibility to initiate binding, whereas those species that have a fully folded H2 have co-opted an alternative mechanism for promoting p150Glued binding.

The essential process of iron-sulfur (Fe/S) cluster assembly (ISC) in mitochondria occurs in three major phases. First, [2Fe-2S] clusters are synthesized on the scaffold protein ISCU2; second, these clusters are transferred to the monothiol glutaredoxin GLRX5 by an Hsp70 system followed by insertion into [2Fe-2S] apoproteins; third, [4Fe-4S] clusters are formed involving the ISC proteins ISCA1–ISCA2–IBA57 followed by target-specific apoprotein insertion. The third phase is poorly characterized biochemically, because previous in vitro assembly reactions involved artificial reductants and lacked at least one of the in vivo-identified ISC components. Here, we reconstituted the maturation of mitochondrial [4Fe-4S] aconitase without artificial reductants and verified the [2Fe-2S]-containing GLRX5 as cluster donor. The process required all components known from in vivo studies (i.e., ISCA1–ISCA2–IBA57), yet surprisingly also depended on mitochondrial ferredoxin FDX2 and its NADPH-coupled reductase FDXR. Electrons from FDX2 catalyze the reductive [2Fe-2S] cluster fusion on ISCA1–ISCA2 in an IBA57-dependent fashion. This previously unidentified electron transfer was occluded during previous in vivo studies due to the earlier FDX2 requirement for [2Fe-2S] cluster synthesis on ISCU2. The FDX2 function is specific, because neither FDX1, a mitochondrial ferredoxin involved in steroid production, nor other cellular reducing systems, supported maturation. In contrast to ISC factor-assisted [4Fe-4S] protein assembly, [2Fe-2S] cluster transfer from GLRX5 to [2Fe-2S] apoproteins occurred spontaneously within seconds, clearly distinguishing the mechanisms of [2Fe-2S] and [4Fe-4S] protein maturation. Our study defines the physiologically relevant mechanistic action of late-acting ISC factors in mitochondrial [4Fe-4S] cluster synthesis, trafficking, and apoprotein insertion.

Box C/D RNA protein complexes (RNPs) catalyze site-specific 2′-O-methylation of RNA with specificity determined by guide RNAs. In eukaryotic C/D RNP, the paralogous Nop58 and Nop56 proteins specifically associate with terminal C/D and internal C'/D' motifs of guide RNAs, respectively. We have reconstituted active C/D RNPs with recombinant proteins of the thermophilic yeast Chaetomium thermophilum. Nop58 and Nop56 could not distinguish between the two C/D motifs in the reconstituted enzyme, suggesting that the assembly specificity is imposed by trans-acting factors in vivo. The two C/D motifs are functionally independent and halfmer C/D RNAs can also guide site-specific methylation. Extensive pairing between C/D RNA and substrate is inhibitory to modification for both yeast and archaeal C/D RNPs. N6-methylated adenine at box D/D' interferes with the function of the coupled guide. Our data show that all C/D RNPs share the same functional organization and mechanism of action and provide insight into the assembly specificity of eukaryotic C/D RNPs.

Genome regulation requires control of chromosome organization by SMC–kleisin complexes. The cohesin complex contains the Smc1 and Smc3 subunits that associate with the kleisin Scc1 to form a ring-shaped complex that can topologically engage chromatin to regulate chromatin structure. Release from chromatin involves opening of the ring at the Smc3–Scc1 interface in a reaction that is controlled by acetylation and engagement of the Smc ATPase head domains. To understand the underlying molecular mechanisms, we have determined the 3.2-A resolution cryo-electron microscopy structure of the ATPγS-bound, heterotrimeric cohesin ATPase head module and the 2.1-A resolution crystal structure of a nucleotide-free Smc1–Scc1 subcomplex from Saccharomyces cerevisiae and Chaetomium thermophilium. We found that ATP-binding and Smc1–Smc3 heterodimerization promote conformational changes within the ATPase that are transmitted to the Smc coiled-coil domains. Remodeling of the coiled-coil domain of Smc3 abrogates the binding surface for Scc1, thus leading to ring opening at the Smc3–Scc1 interface. Structural analysis reveals that ATP-binding and Smc1–Smc3 heterodimerization promotes conformational changes within the cohesin ATPase that are transmitted to the Smc coiled-coil domains, leading to ring opening at the Smc3–Scc1 interface.

The enzyme Tpt1 removes an internal RNA 2′-PO4 via a two-step reaction in which: (i) the 2′-PO4 attacks NAD+ to form an RNA-2′-phospho-(ADP-ribose) intermediate and nicotinamide; and (ii) transesterification of the ADP-ribose O2″ to the RNA 2′-phosphodiester yields 2′-OH RNA and ADP-ribose-1″,2″-cyclic phosphate. Because step 2 is much faster than step 1, the ADP-ribosylated RNA intermediate is virtually undetectable under normal circumstances. Here, by testing chemically modified nucleic acid substrates for activity with bacterial Tpt1 enzymes, we find that replacement of the ribose-2′-PO4 nucleotide with arabinose-2′-PO4 selectively slows step 2 of the reaction pathway and results in the transient accumulation of high levels of the reaction intermediate. We report that replacing the NMN ribose of NAD+ with 2′-fluoroarabinose (thereby eliminating the ribose O2″ nucleophile) results in durable trapping of RNA-2′-phospho-(ADP-fluoroarabinose) as a “dead-end” product of step 1. Tpt1 enzymes from diverse taxa differ in their capacity to use ara-2″F-NAD+ as a substrate.

The superfamily 2 helicase XPB is an integral part of the general transcription factor TFIIH and assumes essential catalytic functions in transcription initiation and nucleotide excision repair. In both processes the ATPase activity of XPB is essential. We investigated the interaction network that regulates XPB via the p52 and p8 subunits with functional mutagenesis based on a crystal structure of the full p52/p8 complex and current cryo-EM structures. Importantly, we show that XPB’s ATPase can be activated either by DNA or by the interaction with the p52/p8 proteins. Intriguingly, we observe that the ATPase activation by p52/p8 is significantly weaker than the activation by DNA and when both p52/p8 and DNA are present, p52/p8 dominates the maximum activation. We therefore define p52/p8 as the master regulator of XPB that acts as an activator and speed limiter at the same time. A correlative analysis of the ATPase and translocase activities of XPB shows that XPB only acts as a translocase within the context of complete core TFIIH and that XPA increases the processivity of the translocase complex without altering XPB’s ATPase activity. Our data unravel an intricate network that tightly controls the activity of XPB during transcription and nucleotide excision repair.

The incidence of infections caused by uncommon Chaetomiaceae (Chaetomium and related species) in humans has increased in the recent years. The in vitro activity of eight antifungal drugs (amphotericin B, five azoles, two echinocandins) against 42 morphologically identified Chaetomium strains was determined according to the Clinical and Laboratory Standards Institute (CLSI) guideline. The strains were subsequently identified based on sequences of the internal transcribed spacer 1 and 2 including the intervening 5.8S nrDNA region (ITS) and the partial β tubulin gene (tub2). Chaetomium globosum (n = 24), was the most frequently isolated species, followed by Amesia atrobrunnea (syn. Chaetomium atrobrunnea, n = 6), Dichotomopilus dolichotrichus (syn. Chaetomium dolichotrichum, n = 2) and Acrophialophora jodhpurensis, Chaetomium coarctatum, C. elatum, C. gracile, C. subaffine, C. tarraconense, C. unguicola, Dichotomopilus sp., Dichotomopilus variostiolatus, Ovatospora brasiliensis (all represented by a single strain). The geometric means of the minimum inhibitory concentrations/minimum effective concentrations (MICs/MECs) of the antifungals across all strains were (in increasing order): micafungin 0.12 µg/ml, itraconazole and posaconazole 0.21 µg/ml, amphotericin B 0.25 µg/ml, voriconazole 0.45 µg/ml, isavuconazole 0.54 µg/ml, caspofungin 2.57 µg/ml, and fluconazole 45.25 µg/ml. Micafungin had the lowest geometric mean followed by amphotericin B which had the largest range against tested isolates. All examined C. globosum strains had similar antifungal susceptibility patterns. Fluconazole and caspofungin could not be considered as an option for treatment of infections caused by Chaetomium and chaetomium-like species. Lay summary Infections caused by uncommon fungi such as Chaetomium have increased in the recent years. Chaetomium globosum has been reported from onychomycosis and phaeohyphomycosis. This species often induces superficial infections in immunocompetent patients. The taxonomy of Chaetomium spp. has changed dramatically in the last years. Antifungal treatment is a crucial step for managing these kinds of infections. Therefore, the in vitro activity of eight antifungal drugs against Chaetomium strains was determined and β-tubulin (tub2) sequencing was applied to identify the strains. Chaetomium globosum was the most frequent species in our dataset. Based on the results of susceptibility testing, micafungin had the lowest geometric mean followed by amphotericin B. Fluconazole and caspofungin cannot be considered a proper treatment option for infections caused by Chaetomium and chaetomium-like species.

Chemical investigation on the deep-sea-derived fungus Chaetomium globosum led to the isolation of nine compounds. By extensive analyses of the1D and 2D NMR as well as HRESIMS spectra, their structures were elucidated as xylariol A ( 1 ), 1,3-dihydro-4,5,6-trihydroxy-7-methyl-isobenzofuran ( 2 ), epicoccone B ( 3 ) epicoccolide B ( 4 ), chaetoglobosin G ( 5 ), chaetoglobosin Fex ( 6 ), cochliodone A ( 7 ), cochliodone B ( 8 ), and chaetoviridin A ( 9 ), assorting as four phenolics ( 1 - 4 ), two cytochalosans ( 5 - 6 ), and three azaplilones ( 7 - 9 ). Compounds 1 - 3 were firstly reported from C. globosum . Under the concentrations of 20 μg/mL, 1 , 2 , and 3 exhibited potent in vitro anti-HIV activity with the inhibition rates of 70%, 75%, and 88%, respectively.

Chaetomium species for plant disease control are reported to be antagonize many plant pathogens. It is a new broad spectrum biological fungicide from Chaetomium species which firstly discovered and patented No. 6266, International Code: AO 1 N 25/12, and registered as Ketomium® mycofungicide for plant disease control in Thailand, Laos, Vietnam, Cambodia and China. Chaetoimum biofungicide and biostimulants are applied to implement integrated plant disease control. It showed protective and curative effects in controlling plant disease and promoting plant growth. It has been successfully applied to the infested soils with integrated cultural control for the long-term protection against rice blast (Magnaporte oryzae), durian and black Pepper rot (Piper nigram L.) (Phytophthora palmivora), citrus rot (Phytophthora parasitica) and strawberry rot (Fragaria spp.) caused by Phytophthora cactorum, wilt of tomato (Fusarium oxysporum f. sp. lycopersici), basal rot of corn (Sclerotium rolfsii) and anthracnose (Colletotrichum spp.) etc. Further research is reported on the other bioactive compounds from active strains of Chaetomium spp. We have discovered various new compounds from Ch. globosum, Ch. cupreum, Ch. elatum, Ch. cochliodes, Ch. brasiliense, Ch. lucknowense, Ch. longirostre and Ch. siamense. These new compounds are not only inhibiting human pathogens (anti-malaria, anti-tuberculosis, anti-cancer cell lines and anti-C. albicans etc) but also plant pathogens as well. These active natural products from different strains of Chaetomium spp. are further developed to be biodegradable nanoparticles from active metabolites as a new discovery of scientific investigation which used to induce plant immunity, namely microbial degradable nano-elicitors for inducing immunity through phytoalexin production in plants e.g. inducing tomato to produce alpha-tomaline against Fusarium wilt of tomato, capsidiol against chili anthracnose, sakuranitin and oryzalexin B against rice blast, scopletin and anthrocyaidin against Phytophthora or Pythium rot Durian and scoparone against Phytophthora or Pythium rot of citrus. Chaetomium biofungicide can be applied instead of toxic chemical fungicides to control plant diseases.

l -asparaginase is a chemotherapeutic drug used in the treatment of acute lymphoblastic leukemia, a malignant disorder in children. l -asparaginase helps in removing acrylamide found in fried and baked foods which is carcinogenic in nature. The search for new therapeutic enzymes is of great interest in both medical and food applications. The present work aims to isolate the intracellular l -asparaginase from endophytic fungi Chaetomium sp. The intracellular enzyme was partially purified by chromatographic techniques. Molecular weight of enzyme was found to be ~66 kDa by SDS PAGE analysis. The enzyme is highly specific for l -asparagine and did not show glutaminase and urease activity. Maximum enzyme activity was found to be 58 ± 5 U/mL at 40 °C, pH 7.0 with 2 μg of protein. Intracellular l -asparaginase from Chaetomium sp. exhibited anticancer activity on human blood cancer (MOLT-4) cells.

l-Asparaginase catalyzes the hydrolysis of asparagine into aspartic acid and ammonia. The present work elaborates the isolation and identification of a novel endophytic fungal isolate producing l-glutaminase and urease-free l-asparaginase. Cell growth and enzyme production were investigated for large production. The isolated endophytic fungi were identified at molecular levels and a phylogenetic tree was constructed. The enzyme synthesis was evaluated by cultivating the isolated microorganisms in potato dextrose agar medium. Out of 27 isolated endophytes, nine were producing "l-glutaminase and urease-free l-asparaginase." l-Asparaginase from Chaetomium sp. exhibited superior enzyme activity than from the other isolates. Observed optimal conditions for l-asparaginase activity were 25 min of incubation time, 0.5 mg of enzyme source, 40°C of temperature, and pH 7.0. l-Asparaginase from Chaetomium sp. exhibited anticancer activity on human blood cancer (MOLT-4) cells. The current study has demonstrated the production of contaminant-free l-asparaginase enzyme from endophytic fungal species. The results showed that: (a) maximum enzyme activity was observed for l-asparaginase from Chaetomium sp., (b) concentration of glucose in the medium as a carbon source suppressed the enzyme production. Chaetomium sp. is a novel source for "l-glutaminase and urease-free l-asparaginase," which may play a major role in pharmacotherapy.

This study was aimed to explore molecular characters of fungal isolates and analyze the phylogenic relationship between isolates. Soil samples collected from different habitats of Jaipur, were analyzed for the prevalence of keratinophilic fungi using the hair baiting technique. The physiological conditions of soil samples were analyzed to study the effect of environmental factors on the growth and occurrence of fungi. Keratinophilic fungi were isolated and then identified on the basis of their macro and micro morphological characteristics. The identification of fungi was confirmed by DNA sequencing by PCR amplification using ITS4 and ITS5 gene primers. The fungal species were identified by the nBLAST using fungal DNA sequences against the data available in NCBI/GenBank. The isolated keratinophilic fungal species were identified as Chaetomium seminis-citrulli and Penicillium guttulosum. These two fungi were reported first time from Rajasthan, India using hair baiting technique. Phylogenic trees were constructed using MEGA7 software with the help of keratinophilic fungi reference data retrieved from NCBI GenBank data, to study the phylogenic relationship between the isolated fungi.

High-resolution structural analysis of flexible proteins is frequently challenging and requires the synergistic application of different experimental techniques. For these proteins, small-angle X-ray scattering (SAXS) allows for a quantitative assessment and modeling of potentially flexible and heterogeneous structural states. Here, we report SAXS characterization of the condensin HEAT-repeat subunit Ycg1(Cnd3) in solution, complementing currently available high-resolution crystallographic models. We show that the free Ycg1 subunit is flexible in solution but becomes considerably more rigid when bound to its kleisin-binding partner protein Brn1(Cnd2). The analysis of SAXS and dynamic and static multiangle light scattering data furthermore reveals that Ycg1 tends to oligomerize with increasing concentrations in the absence of Brn1. Based on these data, we present a model of the free Ycg1 protein constructed by normal mode analysis, as well as tentative models of Ycg1 dimers and tetramers. These models enable visualization of the conformational transitions that Ycg1 has to undergo to adopt a closed rigid shape and thereby create a DNA-binding surface in the condensin complex.

Ribosome assembly is a complex process reliant on the coordination of trans-acting enzymes to produce functional ribosomal subunits and secure the translational capacity of cells. The endoribonuclease (RNase) Las1 and the poly-nucleotide kinase (PNK) Grc3 assemble into a multienzyme complex, herein designated RNase PNK, to orchestrate processing of precursor ribosomal RNA. RNase PNK belongs to the functionally-diverse HEPN nuclease superfamily, whose members rely on distinct cues for nuclease activation. To establish how RNase PNK coordinates its dual enzymatic activities, we solved a series of cryo-electron microscopy structures of Chaetomium thermophilum RNase PNK in multiple conformational states. The structures reveal that RNase PNK adopts a butterfly-like architecture harboring a composite HEPN nuclease active site flanked by discrete RNA kinase sites. We identify two molecular switches that coordinate nuclease and kinase function. Together our structures and corresponding functional studies establish a new mechanism of HEPN nuclease activation essential for ribosome production.

ATP-dependent chromatin remodeling factors of SWI/SNF2 family including ISWI, SNF2, CHD1 and INO80 subfamilies share a conserved but functionally non-interchangeable ATPase domain. Here we report cryo-electron microscopy (cryo-EM) structures of the nucleosome bound to an ISWI fragment with deletion of the AutoN and HSS regions in nucleotide-free conditions and the free nucleosome at ∼ 4 Å resolution. In the bound conformation, the ATPase domain interacts with the super helical location 2 (SHL 2) of the nucleosomal DNA, with the N-terminal tail of H4 and with the α1 helix of H3. Density for other regions of ISWI is not observed, presumably due to disorder. Comparison with the structure of the free nucleosome reveals that although the histone core remains largely unchanged, remodeler binding causes perturbations in the nucleosomal DNA resulting in a bulge near the SHL2 site. Overall, the structure of the nucleotide-free ISWI-nucleosome complex is similar to the corresponding regions of the recently reported ADP bound ISWI-nucleosome structures, which are significantly different from that observed for the ADP-BeFx bound structure. Our findings are relevant to the initial step of ISWI binding to the nucleosome and provide additional insights into the nucleosome remodeling process driven by ISWI.

D-glucuronic acid (GlcUA) is an important intermediate with numerous applications in the food, cosmetics, and pharmaceutical industries. Its biological production routes which employ myo-inositol oxygenase (MIOX) as the key enzyme are attractive. In this study, five diverse MIOX-encoding genes, from Cryptococcus neoformans, Chaetomium thermophilum, Arabidopsis thaliana, Thermothelomyces thermophila, and Mus musculus were overexpressed in Escherichia coli, respectively. A novel MIOX from Thermothelomyces thermophila (TtMIOX) exhibited high specific activity, and efficiently converted myo-inositol to GlcUA. Meanwhile, the degradation of GlcUA was inhibited by inactivation of uxaC from the Escherichia coli genome. Finally, the BWΔuxaC whole-cell biocatalyst harboring TtMIOX resulted in the production of 106 g/L GlcUA within 12 h in a 1-L bioreactor, corresponding to a conversion of 91% and productivity of 8.83 g/L/h. This study provides a feasible method for the industrial production of GlcUA.

Histone H3 lysine 36 methylation (H3K36me) is a conserved histone modification deposited by the Set2 methyltransferases. Recent findings show that over-expression or mutation of Set2 enzymes promotes cancer progression, however, mechanisms of H3K36me are poorly understood. Set2 enzymes show spurious activity on histones and histone tails, and it is unknown how they obtain specificity to methylate H3K36 on the nucleosome. In this study, we present 3.8 Å cryo-EM structure of Set2 bound to the mimic of H2B ubiquitinated nucleosome. Our structure shows that Set2 makes extensive interactions with the H3 αN, the H3 tail, the H2A C-terminal tail and stabilizes DNA in the unwrapped conformation, which positions Set2 to specifically methylate H3K36. Moreover, we show that ubiquitin contributes to Set2 positioning on the nucleosome and stimulates the methyltransferase activity. Notably, our structure uncovers interfaces that can be targeted by small molecules for development of future cancer therapies., Set2 methyltransferase catalyzes the lysine 36 methylation of histone H3 (H3K36me) and the enzyme is mutated in many cancers. The authors provide mechanistic insights into how Set2 methylates nucleosomes by determining the 3.8 Å cryo-EM structure of Set2 bound to a H2B ubiquitinated nucleosome core particle.

Endophytic fungus, as a new kind of microbial resources and separated from plants, has attracted increasing attention due to its ability to synthesize the same or similar bioactive secondary metabolites as the host plants. Nevertheless, the effects of the symbiotic relationship between microorganisms and elicitors existed in host plant on metabolite production are not adequately understood. In the present work, the impacts of elicitors (ginseng saponin and puerarin) and symbiotic microorganisms on endophytic fungus Chaetomium globosum CGMCC 6882 synthesizing polysaccharide were evaluated. Results show that the polysaccharide titers increased from 2.36 to 3.88 g/L and 3.67 g/L with the addition of 16 μg/L ginseng saponin and puerarin, respectively. Moreover, the maximum polysaccharide titer reached 4.55 g/L when C. globosum CGMCC 6882 was co-cultured with UV-irradiated G. pentaphyllum. This work brings a significant contribution to the research and interpretation of the relationship between endophytic fungus and its host plant.

Balanced fusion and fission are key for the proper function and physiology of mitochondria1,2. Remodelling of the mitochondrial inner membrane is mediated by the dynamin-like protein mitochondrial genome maintenance 1 (Mgm1) in fungi or the related protein optic atrophy 1 (OPA1) in animals3–5. Mgm1 is required for the preservation of mitochondrial DNA in yeast6, whereas mutations in the OPA1 gene in humans are a common cause of autosomal dominant optic atrophy—a genetic disorder that affects the optic nerve7,8. Mgm1 and OPA1 are present in mitochondria as a membrane-integral long form and a short form that is soluble in the intermembrane space. Yeast strains that express temperature-sensitive mutants of Mgm19,10 or mammalian cells that lack OPA1 display fragmented mitochondria11,12, which suggests that Mgm1 and OPA1 have an important role in inner-membrane fusion. Consistently, only the mitochondrial outer membrane—not the inner membrane—fuses in the absence of functional Mgm113. Mgm1 and OPA1 have also been shown to maintain proper cristae architecture10,14; for example, OPA1 prevents the release of pro-apoptotic factors by tightening crista junctions15. Finally, the short form of OPA1 localizes to mitochondrial constriction sites, where it presumably promotes mitochondrial fission16. How Mgm1 and OPA1 perform their diverse functions in membrane fusion, scission and cristae organization is at present unknown. Here we present crystal and electron cryo-tomography structures of Mgm1 from Chaetomium thermophilum. Mgm1 consists of a GTPase (G) domain, a bundle signalling element domain, a stalk, and a paddle domain that contains a membrane-binding site. Biochemical and cell-based experiments demonstrate that the Mgm1 stalk mediates the assembly of bent tetramers into helical filaments. Electron cryo-tomography studies of Mgm1-decorated lipid tubes and fluorescence microscopy experiments on reconstituted membrane tubes indicate how the tetramers assemble on positively or negatively curved membranes. Our findings convey how Mgm1 and OPA1 filaments dynamically remodel the mitochondrial inner membrane. Crystal and electron cryo-tomography structures of Mgm1 from Chaetomium thermophilum reveal that Mgm1 forms bent tetramers, which further assemble into helical filaments on both positively and negatively curved membranes.

Purpose To assess in vitro activities of six antifungal agents (amphotericin B, itraconazole, voriconazole, posaconazole, caspofungin and terbinafine) and the combined effects of eight pairs of them (caspofungin or terbinafine with amphotericin B, itraconazole, voriconazole or posaconazole) against 22 isolates of Chaetomium spp. Methodology The broth microdilution method drafted by the Clinical and Laboratory Standards Institute and the checkerboard method were used in this study to evaluate in vitro activities of antifungal drugs both alone and in combination against Chaetomium spp. Results Amphotericin B and triazoles exhibited lower geometric mean, MIC50 and MIC90 than caspofungin and terbinafine. Besides, all the paired drugs displayed varying degrees of synergism, with the interactions between caspofungin and itraconazole ranking first (86.36 %). Conclusion Our study illustrated varying degrees of synergism between caspofungin or terbinafine and itraconazole, voriconazole, posaconazole or amphotericin B towards Chaetomium spp., which could be a reference for the clinical treatment of Chaetomium spp. infections.

The aim of this study was to examine the impact of microbiocides on the reduction of the microbial population on paper in order to protect it against biodeterioration. The cellulose products were modified with the cationic gemini surfactant hexamethylene-1,6-bis-(N,N-dimethyl-N-dodecylammonium bromide) C6 and its monomeric analog (didecyldimethylammonium chloride) DDAC. The microbiocides were introduced into the paper by coating and spraing. In the coating method the microbiocides were mixed with starch solution and applied to paper surface as a wet film with a thickness of 24 and 50 µm. In the spraying method the surfactants were applied at 3% concentration in water to get 0.005 ml/cm2 of paper. Experiments were performed with moulds of the genera Aspergillus, Penicilllium, Chaetomium and Trichoderma and bacteria represented by the genera Bacilllus and Pseudomonas. The antimicrobial properties of the paper modified with surfactants were determined by qualitative and quantitative methods. The zones of inhibition were measured using the parallel streak method. Macroscopic assessment of mould growth on the surface of the paper samples was performed over 21 days of incubation. In quantitative analysis, the survival rate of microorganisms on the modified paper samples was determined over 24 h. Aspergillus brasiliensis was the least sensitive mould, with no observable inhibition zones. No growth of this mould was observed either on or underneath the sample. Both of the surfactants applied as coatings or sprays protected the paper effectively against the growth of bacteria and mould. However, the spraying method is simpler to use. Over 24 h, the number of spores and bacteria in all the samples containing surfactants was reduced to below 1 log/10 cm2 (reduction factor up to 99.9%). The compounds may therefore be applied as antimicrobial agents for the protection of paper.

Soil microbial fuel cells (MFCs) provide an inexhaustible electron acceptor for the removal of metolachlor and in situ biocurrent stimulation for fungal activity was investigated. The metolachlor degradation rates enhanced by 33%–36% upon the introduction of electrodes after 23 d. In closed MFCs, the abundance of Mortierella as the most dominant genus increased to 43%–54% from 17% in the original soil, whereas those of Aphanoascus and Penicillium decreased to 0.24%–0.39% and 0.38–0.72% from 14% to 11%, respectively. Additionally, a 10-fold amplification of unique OTUs was observed, mainly from increase on the electrode surface. The different treatments were clustered, especially samples near the cathode. The linear discriminant analysis showed that Aphanoascus fulvescens acted as a biomarker between the original and treated soils. The co-occurrence networks demonstrated that Mortierella universally competed for growth with coexisting species while Cladosporium exhibited the most affiliations with species from the 36 other genera present. The correlation analysis indicated that the species associated with degradation belonged to Mortierella, Kernia, Chaetomium and Trichosporon, while the species associated with electrogenesis were Debaryomyces hansenii and Mortierella polycephala. Importantly, this study is the first to reveal fungal community structure in soil MFCs with degrading pollutants and producing electricity.

The fungus Chaetomium sp. is a causative agent of infections in humans and is ubiquitous in the natural environment. The secondary metabolites of this genus exhibit many biological activities, including antifungal activity and toxicity in mitochondria. In this study, we isolated cristazine from the fungus C. cristatum, which has the potential to inhibit the growth of human epidermoid carcinoma (A431) cells in a dose- and time-dependent manner. Its inhibitory activity was examined using a cell viability assay and cell death was elucidated by western blot analysis. Cristazine triggered apoptotic cell death via the Type I death receptor pathway including the activation of caspases and other target proteins. However, cristazine did not have any effect on mitochondrial apoptotic proteins such as Bid, cytochrome c, and apoptosis-inducing factor. Cristazine inhibited the cell cycle progression by arresting the G1 /S phase and up-regulating the inhibitory proteins of cyclin-dependent kinases. Thus, cristazine has great potential for inducing apoptosis in A431 cells via both cell cycle arrest and the inhibition of cell growth.

The best support reported for dextransucrase (DS) and dextranase (DN) immobilization was Eupergit C, but this carrier was discontinued and is no longer available in the market. Thus, epoxy-agarose support is suggested as an alternative to co-immobilize DS from Leuconostoc mesenteroides B-512F and DN from Chaetomium erraticum. The co-immobilization approach improved DS performance, compared to immobilized DS, due to the removal of the dextran layer accumulated on DS by DN, and enhanced both enzymes catalytic activities under a wide range of pH and temperature. Since DS and DN have the same optimum pH and temperature, the co-immobilization was done in a single step. The EPA-DS-DN0.5 (co-immobilized DS (26.16 UI) and DN (2.76 UI) in epoxy-agarose) showed the highest recovered activity (59.54%). The biocatalyst was stable at 4 °C, retaining above 70% of activity for 60 days and was efficient producing oligosaccharides, yielding 12.68 ± 0.09 g/L of oligosaccharides with a degree of polymerization up to 5.

The progress of research on silver nanoparticles (AgNPs) has led to their inclusion in many consumer products (chemicals, cosmetics, clothing, water filters, and medical devices) as a biocide. Despite the widespread use of AgNPs, their biocidal activity is not yet fully understood and is usually associated with various factors (size, composition, surface, red-ox potential, and concentration) and, obviously, specific features of microorganisms. There are merely a few studies concerning the interaction of molds with AgNPs. Therefore, the determination of the minimal AgNPs concentration required for effective growth suppression of five fungal species (Paecilomyces variotii, Penicillium pinophilum, Chaetomium globosum, Trichoderma virens, and Aspergillus brasiliensis), involved in the deterioration of construction materials, was particularly important. Inhibition of bacteria (Pseudomonas aeruginosa, Staphylococcus aureus, and Escherichia coli) and yeasts (Candida albicans and Yarrowia lipolytica) was also assessed as the control of AgNPs effectiveness. AgNPs at the concentrations of 9-10.7 ppm displayed high inhibitory activity against moulds, yeast, and bacteria. The TEM images revealed that 20 nm AgNPs migrated into bacterial, yeast, and fungal cells but aggregated in larger particles (50-100 nm) exclusively inside eukaryotic cells. The aggregation of 20 nm AgNPs and particularly their accumulation in the cell wall, observed for A. brasiliensis cells, are described here for the first time. The progress of research on silver nanoparticles (AgNPs) has led to their inclusion in many consumer products (chemicals, cosmetics, clothing, water filters, and medical devices) as a biocide. Despite the widespread use of AgNPs, their biocidal activity is not yet fully understood and is usually associated with various factors (size, composition, surface, red-ox potential, and concentration) and, obviously, specific features of microorganisms. There are merely a few studies concerning the interaction of molds with AgNPs. Therefore, the determination of the minimal AgNPs concentration required for effective growth suppression of five fungal species (Paecilomyces variotii, Penicillium pinophilum, Chaetomium globosum, Trichoderma virens, and Aspergillus brasiliensis), involved in the deterioration of construction materials, was particularly important. Inhibition of bacteria (Pseudomonas aeruginosa, Staphylococcus aureus, and Escherichia coli) and yeasts (Candida albicans and Yarrowia lipolytica) was also assessed as the control of AgNPs effectiveness. AgNPs at the concentrations of 9–10.7 ppm displayed high inhibitory activity against moulds, yeast, and bacteria. The TEM images revealed that 20 nm AgNPs migrated into bacterial, yeast, and fungal cells but aggregated in larger particles (50–100 nm) exclusively inside eukaryotic cells. The aggregation of 20 nm AgNPs and particularly their accumulation in the cell wall, observed for A. brasiliensis cells, are described here for the first time.

Rationale Microbial natural products are often biosynthesized as classes of structurally related compounds that have similar tandem mass spectrometry (MS/MS) fragmentation patterns. Mining MS/MS datasets for precursor ions that share diagnostic or common features enables entire chemical classes to be identified, including novel derivatives that have previously been unreported. Analytical data analysis tools that can facilitate a class-targeted approach to rapidly dereplicate known compounds and identify structural variants within complex matrices would be useful for the discovery of new natural products. Methods A diagnostic fragmentation filtering (DFF) module was developed for MZmine to enable the efficient screening of MS/MS datasets for class-specific product ions(s) and/or neutral loss(es). This approach was applied to series of the structurally related chaetoglobosin and cytochalasin classes of compounds. These were identified from the culture filtrates of three fungal genera: Chaetomium globosum, a putative new species of Penicillium (called here P. cf. discolor: closely related to P. discolor), and Xylaria sp. Extracts were subjected to LC/MS/MS analysis under positive electrospray ionization and operating in a data-dependent acquisition mode, performed using a Thermo Q-Exactive mass spectrometer. All MS/MS datasets were processed using the DFF module and screened for diagnostic product ions at m/z 130.0648 and 185.0704 for chaetoglobosins, and m/z 120.0808 and 146.0598 for cytochalasins. Results Extracts of C. globosum and P. cf. discolor strains revealed different mixtures of chaetoglobosins, whereas the Xylaria sp. produced only cytochalasins; none of the strains studied produced both classes of compounds. The dominant chaetoglobosins produced by both C. globosum and P. cf. discolor were chaetoglobosins A, C, and F. Tetrahydrochaetoglobosin A was identified from P. cf. discolor extracts and is reported here for the first time as a natural product. The major cytochalasins produced by the Xylaria sp. were cytochalasin D and epoxy cytochalasin D. A larger unknown "cytochalasin-like" molecule with the molecular formula C38 H47 NO10 was detected from Xylaria sp. culture filtrate extracts and is a current target for isolation and structural characterization. Conclusions DFF is an effective LC/MS data analysis approach for rapidly identifying entire classes of compounds from complex mixtures. DFF has proved useful in the identification of new natural products and allowing for their partial characterization without the need for isolation.

By feeding L-tyrosine into the culture medium, nine undescribed compounds, termed as armochaetoglasins A–I, together with three known analogues, namely armochaetoglobin E, chaetoglobosin V, and chaetoglobosin J, were isolated and identified from the medicinal terrestrial arthropod-derived fungus Chaetomium globosum TW1-1. Their structures were elucidated by means of NMR spectroscopy, single-crystal X-ray crystallography, and comparison of their electronic circular dichroism (ECD) spectra. Structurally, armochaetoglasin A represented the first tyrosine-derived cytochalasan alkaloid characterized by a 13-membered carbocyclic ring system; armochaetoglasins B and C possessed a rare 19,20-seco-chaetoglobosin skeleton. Armochaetoglasin B, chaetoglobosin V, and chaetoglobosin J showed weak cytotoxic activity with IC50 values ranging from 19.5 to 34.72 μM.

In food industries UV-C irradiation is used to achieve decontamination of some packaging devices, such as plastic caps or laminated foils, and of those smooth surfaces that can be directly irradiated. Since its effectiveness can be checked by microbial validation tests, some ascospore-forming molds (Aspergillus hiratsukae, Talaromyces bacillisporus, Aspergillus montevidensis, and Chaetomium globosum) were compared with one of the target microorganisms actually used in industrial bio-validations (Aspergillus brasiliensis ATCC 16404) to find the species most resistant to UV-C. Tests were carried out with an UV-C lamp (irradiance = 127 μW/cm2; emission peak = 253.7 nm) by inoculating HDPE caps with one or more layers of spores. Inactivation kinetics of each strain were studied and both the corresponding 1D-values and the number of Logarithmic Count Reductions (LCR) achieved were calculated. Our results showed the important role played by the type of inoculum (one or more layers) and by the differences in cell structure (thickness, presence of protective solutes, pigmentation, etc.) of the strains tested. With a single-layer inoculum, Chaetomium globosum showed the highest resistance to UV-C irradiation (1D-value = 100 s). With a multi-layer inoculum, Aspergillus brasiliensis ATCC 16404 was the most resistant fungus (1D-value = 188 s), even if it reached a number of logarithmic reductions that was higher than those of some ascospore-forming mycetes (Aspergillus montevidensis, Talaromyces bacillisporus) tested.

β-1,3-1,4-Glucanases are a type of hydrolytic enzymes capable of catalyzing the strict cleavage of β-1,4 glycosidic bonds adjacent to β-1,3 linkages in β-D-glucans and have exhibited great potential in food and feed industrials. In this study, a novel glycoside hydrolase (GH) family 12 β-1,3-1,4-glucanase (CtGlu12A) from the thermophilic fungus Chaetomium sp. CQ31 was identified and biochemically characterized. CtGlu12A was most active at pH 7.5 and 65 °C, respectively, and exhibited a high specific activity of 999.9 U mg−1 towards lichenin. It maintained more than 80% of its initial activity in a wide pH range of 5.0–11.0, and up to 60 °C after incubation at 55 °C for 60 min. Moreover, the crystal structures of CtGlu12A with gentiobiose and tetrasccharide were resolved. CtGlu12A had a β-jellyroll fold, and performed retaining mechanism with two glutamic acids severing as the catalytic residues. In the complex structure, cellobiose molecule showed two binding modes, occupying subsites −2 to −1 and subsites + 1 to + 2, respectively. The concave cleft made mixed β-1,3-1,4-glucan substrates maintain a bent conformation to fit into the active site. Overall, this study is not only helpful for the understanding of the substrate-binding model and catalytic mechanism of GH 12 β-1,3-1,4-glucanases, but also provides a basis for further enzymatic engineering of β-1,3-1,4-glucanases.

The current study was conducted to evaluate the antiproliferative and oxidative damage protection potential of endophytic fungi Aspergillus fumigatus and Chaetomium globosum isolated from Moringa oleifera. The chloroformic extract (CE) of both the fungi showed dose dependent antiproliferative activity against human prostate adenocarcinoma (PC-3) cell line with (IC50) value of 0.055 mg/ml and 0.008 mg/ml, respectively. Further, CE of both the fungi was studied for their ability to induce apoptosis in PC-3 cell line. Various deformities in the cancerous cells treated with CE of both the fungi have been observed by confocal microscopy which indicates the cell death by apoptosis. Further apoptosis inducing ability of CE of both the fungi was observed using various flow cytometric studies. The chloroformic extract of both the fungi showed slight increase in the level of reactive oxygen species to induce apoptosis. It also showed arrest of cancerous cells at G0/G1 phase of cell cycle to induce apoptosis. The externalization of phosphatidylserine (PS) to induce apoptosis was also observed when analysed using Annexin V-FITC/PI double staining assay where the CE of A. fumigatus and C. globosum showed the total apoptosis of 94.2% and 90.3%, respectively, at the highest tested concentration of GI70. The CE of both the fungi further showed the protective behaviour for plasmid DNA pBR322, when tested for their effect against the oxidative stress caused by the Fenton’s reagent. Thus, the studies demonstrated a good antiproliferative and oxidative damage protection potential of the endophytic fungi.

This study investigated the effect of molecular weight on antibacterial activity of polysaccharides. Results showed that low molecular weight (3.105 × 104 Da) polysaccharide (GCP-2) had higher inhibitory effects against Escherichia coli and Staphylococcus aureus than high molecular weight (5.340 × 104 Da) polysaccharide (GCP-1). Meanwhile, antibacterial activities of GCP-2 and GCP-1 against S. aureus were higher than those of E. coli. Minimum inhibitory concentrations (MICs) of GCP-1 against E. coli and S. aureus were 2.0 mg/mL and 1.2 mg/mL, and MICs of GCP-2 against E. coli and S. aureus were 1.75 mg/mL and 0.85 mg/mL, respectively. Antibacterial mechanisms investigation revealed that GCP-2 and GCP-1 influenced cell membrane integrity, Ca2+-Mg2+-ATPase activity on cell membrane and calcium ions in cytoplasm of E. coli and S. aureus, but not cell wall. Present work provided important implications for future studies on development of antibacterial polysaccharides based on molecular weight feature.

Exploration of feruloyl esterase (FAE) with the resistance to heat and alkali conditions in biobleaching process to improve the separation efficiency of lignocellulose is the key to achieving green papermaking. Herein, we expressed FAEB of C. thermophilum and obtained a thermostable alkaline FAE that can effectively promote the removal of lignin from pulp. The faeB gene was successfully obtained through genomic Blast strategy and high-efficiency expressed under the control of strong alcohol oxidase promoter in Pichia pastoris. The recombinant CtFAEB has an optimal temperature of 65 °C and pH of 7.0. After treated at 65 °C for 1 h, CtFAEB can still retain 63.21 % of its maximum activity, showing a good thermal stability. In addition, the recombinant CtFAEB has broad pH stability and can retain about 56 % of the maximum activity even at pH 11.0. Compared with the effect of mesophilic FAE, pretreatment with thermostable CtFAEB can promote the delignification by laccase and alkaline hydrogen peroxide from the pulp at 70 °C and pH 9.0. Alignment of the protein sequences of CtFAEB and mesophilic FAE suggested that the percentage of amino acids that easily form alpha helix in CtFAEB increases, which enhances its structural rigidity and thereby improves its thermal stability and alkali tolerance. Our study provides an effective method to obtain thermostable and alkaline FAEs, which will promote its application in biobleaching and other biorefining industries.

Enhanced covalent immobilization of xylanase from Chaetomium globosum (XylCg) onto SiO2 nanoparticles was achieved by the modification of surface residues. The mutation of surface residues to lysine by site-directed mutagenesis increased the immobilization efficiency (IE) and immobilization yield (IY). The immobilized mutant XylCg (N172K-H173K-S176K-K133A-K148A) exhibited an IY of 99.5% and IE of 135%, which were 1.8- and 4.3-fold higher than immobilized wildtype (WT). Regarding the catalytic properties, the kcat and kcat/Km values were 1850 s−1 and 2030 mL mg−1 s−1 for the immobilized mutant, and 331 s−1 and 404 mL mg−1 s−1 for the immobilized WT, respectively. Additionally, the immobilized mutant exhibited four times higher thermal stability than the immobilized WT at 60 °C. These results suggest that surface-mutated lysine residues confer good stability and orientation on the support matrix, thus improving the overall performance of xylanase.

Summary: The pyruvate dehydrogenase complex (PDHc) is a giant enzymatic assembly involved in pyruvate oxidation. PDHc components have been characterized in isolation, but the complex’s quaternary structure has remained elusive due to sheer size, heterogeneity, and plasticity. Here, we identify fully assembled Chaetomium thermophilum α-keto acid dehydrogenase complexes in native cell extracts and characterize their domain arrangements utilizing mass spectrometry, activity assays, crosslinking, electron microscopy (EM), and computational modeling. We report the cryo-EM structure of the PDHc core and observe unique features of the previously unknown native state. The asymmetric reconstruction of the 10-MDa PDHc resolves spatial proximity of its components, agrees with stoichiometric data (60 E2p:12 E3BP:∼20 E1p: ≤ 12 E3), and proposes a minimum reaction path among component enzymes. The PDHc shows the presence of a dynamic pyruvate oxidation compartment, organized by core and peripheral protein species. Our data provide a framework for further understanding PDHc and α-keto acid dehydrogenase complex structure and function.

The plant residues of tomato bring pressures to the environment and composting provides a feasible method to treat such agricultural waste. However, little is known about the succession and associations of the dominant lignocellulose degraders in the compost system. To further accelerate the process by inoculating key functional microorganisms, a compost pile composed of tomato stalk with maize straw addition was constructed, and the whole community structure and functions of the dominant were investigated by applying the integrated mata-omics. Results showed that Actinobacteria, Firmicutes, and Ascomycota dominated and drove the assembly of the co-occurrence network. In the thermophilic stage, Thermobifida was the exclusive degrader of cellulose, and Thermobifida fusca was the most important cellulolytic actinomycete. Saccharomonospora viridis, Planifilum fulgidum, Thermobacillus sp. and the dominant ascomycota of Aspergillus sclerotialis participated in hemicellulose decomposing. In the cooling phase, functional microorganisms became more diverse, with Nocardiopsis flavescens, Glycomyces artemisiae, Glycomyces sambucus, Streptomyces rubrolavendulae and Streptomyces vietnamensis joining the cellulose-degrading rank, and Chaetomium thermophilum emerging as the main hemicellulose degrader. More than two thirds of the bacteria-bacteria interactions and all the fungi-fungi associations were positive, while, both competition (for the same substrate of hemicellulose) and synergy (preference for cellulose and hemicellulose) coexisted in the bacteria-fungi interactions. In conclusion, these findings provide useful information for understanding the biodegradation of tomato plant residues better, and effects of the functional agents identified on composting process should be further studied.

Background: Thermophilic fungi have recently emerged as a promising source of thermostable enzymes. Superoxide dismutases are key antioxidant metalloenzymes with promising therapeutic effects in various diseases, both acute and chronic. However, structural heterogeneity and low thermostability limit their therapeutic efficacy. Objective: Although several studies from hypethermophilic superoxide dismutases (SODs) have been reported, information about Cu,Zn-SODs from thermophilic fungi is scarce. Chaetomium thermophilum is a thermophilic fungus that could provide proteins with thermophilic properties. Methods: The enzyme was expressed in Pichia pastoris cells and crystallized using the vapor-diffusion method. X-ray data were collected, and the structure was determined and refined to 1.56 Å resolution. Structural analysis and comparisons were carried out. Results: The presence of 8 molecules (A through H) in the asymmetric unit resulted in four different interfaces. Molecules A and F form the typical homodimer which is also found in other Cu,Zn- SODs. Zinc was present in all subunits of the structure while copper was found in only four subunits with reduced occupancy (C, D, E and F). Conclusion: The ability of the enzyme to form oligomers and the elevated Thr:Ser ratio may be contributing factors to its thermal stability. Two hydrophobic residues that participate in interface formation and are not present in other CuZn-SODs may play a role in the formation of new interfaces and the oligomerization process. The CtSOD crystal structure reported here is the first Cu,Zn-SOD structure from a thermophilic fungus.