Bioethanol production from lignocellulosic materials is hindered by the high costs of pretreatment and the enzymes. The present study aimed to evaluate whether co-cultivation of four selected cellulolytic fungi yields higher cellulase and xylanase activities compared to the monocultures and to investigate whether the enzymes from the co-cultures yield higher saccharification on selected plant materials without thermo-chemical pretreatment. The fungal isolates, Trichoderma reesei F118, Penicillium javanicum FS7, Talaromyces sp. F113, and Talaromyces pinophilus FM9, were grown as monocultures and binary co-cultures under submerged conditions for 7 days. The cellulase and xylanase activities of the culture filtrates were measured, and the culture filtrates were employed for the saccharification of sugarcane leaves, Guinea grass leaves, and water hyacinth stems and leaves. Total reducing sugars and individual sugars released from each plant material were quantified. The co-culture of Talaromyces sp. F113 with Penicillium javanicum FS7 and of T. reesei F118 with T. pinophilus FM9 produced significantly higher cellulase activities compared to the corresponding monocultures whereas no effect was observed on xylanase activities. Overall, the highest amounts of total reducing sugars and individual sugars were obtained from Guinea grass leaves saccharified with the co-culture of T. reesei F118 with T. pinophilus FM9, yielding 63.5% saccharification. Guinea grass leaves were found to be the most susceptible to enzymatic saccharification without pre-treatment, while water hyacinth stems and leaves were the least. Accordingly, the study suggests that fungal co-cultivation could be a promising approach for the saccharification of lignocellulosic materials for bioethanol production.
- Klíčová slova
- Bioethanol production, Cellulases, Fungal co-cultures, Lignocellulosic materials, Saccharification, Xylanases,
- MeSH
- celulasa * metabolismus MeSH
- endo-1,4-beta-xylanasy * metabolismus MeSH
- houby * metabolismus enzymologie růst a vývoj MeSH
- Hypocreales růst a vývoj metabolismus MeSH
- kokultivační techniky MeSH
- lignin * metabolismus MeSH
- listy rostlin metabolismus MeSH
- Penicillium metabolismus růst a vývoj enzymologie MeSH
- Saccharum metabolismus MeSH
- Talaromyces * enzymologie metabolismus růst a vývoj MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- celulasa * MeSH
- endo-1,4-beta-xylanasy * MeSH
- lignin * MeSH
- lignocellulose MeSH Prohlížeč
Mycoparasitism is a key feature of Trichoderma (Hypocreales, Ascomycota) biocontrol agents. Recent studies of intracellular signal transduction pathways of the potent mycoparasite Trichoderma atroviride revealed the involvement of Tmk1, a mitogen-activated protein kinase (MAPK), in triggering the mycoparasitic response. We previously showed that mutants missing Tmk1 exhibit reduced mycoparasitic activity against several plant pathogenic fungi. In this study, we identified the most robustly regulated targets that were governed by Tmk1 during mycoparasitism using transcriptome and proteome profiling. Tmk1 mainly exerts a stimulating function for T. atroviride during its mycoparasitic interaction with the fungal plant pathogen Rhizoctonia solani, as reflected by 89% of strongly differently responding genes in the ∆tmk1 mutant compared to the wild type. Specifically, 54% of these genes showed strong downregulation in the response with a deletion of the tmk1 gene, whereas in the wild type the same genes were strongly upregulated during the interaction with the fungal host. These included the gene encoding the mycoparasitism-related proteinase Prb1; genes involved in signal transduction pathways such as a candidate coding for a conserved 14-3-3 protein, and a gene coding for Tmk2, the T. atroviride cell-wall integrity MAP kinase; genes encoding a specific siderophore synthetase, and multiple FAD-dependent oxidoreductases and aminotransferases. Due to the phosphorylating activity of Tmk1, different (phospho-)proteomics approaches were applied and identified proteins associated with cellular metabolism, energy production, protein synthesis and fate, and cell organization. Members of FAD- and NAD/NADP-binding-domain proteins, vesicular trafficking of molecules between cellular organelles, fungal translational, as well as protein folding apparatus were among others found to be phosphorylated by Tmk1 during mycoparasitism. Outstanding downregulation in the response of the ∆tmk1 mutant to the fungal host compared to the wild type at both the transcriptome and the proteome levels was observed for nitrilase, indicating that its defense and detoxification functions might be greatly dependent on Tmk1 during T. atroviride mycoparasitism. An intersection network analysis between the identified transcripts and proteins revealed a strong involvement of Tmk1 in molecular functions with GTPase and oxidoreductase activity. These data suggest that during T. atroviride mycoparasitism this MAPK mainly governs processes regulating cell responses to extracellular signals and those involved in reactive oxygen stress.
- MeSH
- Hypocreales * metabolismus MeSH
- mitogenem aktivované proteinkinasy genetika metabolismus MeSH
- proteom metabolismus MeSH
- regulace genové exprese u hub MeSH
- signální transdukce MeSH
- Trichoderma * metabolismus MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- mitogenem aktivované proteinkinasy MeSH
- proteom MeSH
Fighting resistance to antibiotics and chemotherapeutics has brought bioactive peptides to the fore. Peptaibols are short α-aminoisobutyric acid-containing peptides produced by Trichoderma species. Here, we studied the production of peptaibols by Trichoderma atroviride O1 and evaluated their antibacterial and anticancer activity against drug-sensitive and multidrug-resistant bacterium and cancer cell lines. This was substantiated by an analysis of the activity of the peptaibol synthetase-encoding gene. Atroviridins, 20-residue peptaibols were detected using MALDI-TOF mass spectrometry. Gram-positive bacteria were susceptible to peptaibol-containing extracts of T. atroviride O1. A synergic effect of extract constituents was possible, and the biolo-gical activity of extracts was pronounced in/after the peak of peptaibol synthetase activity. The growth of methicillin-resistant Staphylococcus aureus was reduced to just under 10% compared to the control. The effect of peptaibol-containing extracts was strongly modulated by the lipoteichoic acid and only slightly by the horse blood serum present in the cultivation medium. Peptaibol-containing extracts affected the proliferation of human breast cancer and human ovarian cancer cell lines in a 2D model, including the multidrug-resistant sublines. The peptaibols influenced the size and compactness of the cell lines in a 3D model. Our findings indicate the molecular basis of peptaibol production in T. atroviride O1 and the potential of its peptaibol-containing extracts as antimicrobial/anticancer agents.
- Klíčová slova
- Trichoderma spp., anticancer peptides, antimicrobial peptides, peptaibols,
- MeSH
- antibakteriální látky farmakologie MeSH
- bakteriální léková rezistence * MeSH
- fungální proteiny metabolismus MeSH
- Hypocreales enzymologie metabolismus MeSH
- koně MeSH
- lidé MeSH
- ligasy metabolismus MeSH
- methicilin rezistentní Staphylococcus aureus účinky léků MeSH
- MFC-7 buňky MeSH
- nádorové buněčné linie MeSH
- nádory farmakoterapie MeSH
- peptaiboly analýza metabolismus farmakologie MeSH
- protinádorové látky farmakologie MeSH
- spektrometrie hmotnostní - ionizace laserem za účasti matrice MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- antibakteriální látky MeSH
- fungální proteiny MeSH
- ligasy MeSH
- peptaiboly MeSH
- protinádorové látky MeSH
Fungal metabolic carbon acquisition and its subsequent partitioning between biomass production and respiration, i.e. the carbon-use efficiency (CUE), are central parameters in biogeochemical modeling. However, current available techniques for estimating these parameters are all associated with practical and theoretical shortcomings, making assessments unreliable. Gene expression analyses hold the prospect of phenotype prediction by indirect means, providing new opportunities to obtain information about metabolic priorities. We cultured four different fungal isolates (Chalara longipes, Laccaria bicolor, Serpula lacrymans and Trichoderma harzianum) in liquid media with contrasting nitrogen availability and measured growth rates and respiration to calculate CUE. By relating gene expression markers to measured carbon fluxes, we identified genes coding for 1,3-β-glucan synthase and 2-oxoglutarate dehydrogenase as suitable markers for growth and respiration, respectively, capturing both intraspecific variation as well as within-strain variation dependent on growth medium. A transcript index based on these markers correlated significantly with differences in CUE between the fungal isolates. Our study paves the way for the use of these markers to assess differences in growth, respiration and CUE in natural fungal communities, using metatranscriptomic or the RT-qPCR approach.
- Klíčová slova
- carbon-use efficiency, fungi, gene markers, growth, metatranscriptomics, respiration,
- MeSH
- Ascomycota genetika metabolismus MeSH
- Basidiomycota genetika MeSH
- biologické markery * analýza MeSH
- fungální proteiny * genetika metabolismus MeSH
- houby * genetika metabolismus MeSH
- Hypocreales genetika metabolismus MeSH
- Laccaria genetika metabolismus MeSH
- transkriptom * MeSH
- Trichoderma genetika metabolismus MeSH
- uhlík * metabolismus MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- biologické markery * MeSH
- fungální proteiny * MeSH
- uhlík * MeSH
Unlike any protein studied so far, the active site of bilirubin oxidase from Myrothecium verrucaria contains a unique type of covalent link between tryptophan and histidine side chains. The role of this post-translational modification in substrate binding and oxidation is not sufficiently understood. Our structural and mutational studies provide evidence that this Trp396-His398 adduct modifies T1 copper coordination and is an important part of the substrate binding and oxidation site. The presence of the adduct is crucial for oxidation of substituted phenols and it substantially influences the rate of oxidation of bilirubin. Additionally, we bring the first structure of bilirubin oxidase in complex with one of its products, ferricyanide ion, interacting with the modified tryptophan side chain, Arg356 and the active site-forming loop 393-398. The results imply that structurally and chemically distinct types of substrates, including bilirubin, utilize the Trp-His adduct mainly for binding and to a smaller extent for electron transfer.
- MeSH
- bilirubin metabolismus MeSH
- Hypocreales metabolismus MeSH
- konformace proteinů MeSH
- molekulární modely * MeSH
- oxidace-redukce MeSH
- oxidoreduktasy působící na CH-CH vazby metabolismus MeSH
- transport elektronů fyziologie MeSH
- vazba proteinů fyziologie MeSH
- vazebná místa MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- bilirubin oxidase MeSH Prohlížeč
- bilirubin MeSH
- oxidoreduktasy působící na CH-CH vazby MeSH
Formosan subterranean termites (FST) were exposed to strains of Beauveria pseudobassiana (Bpb) and Isaria fumosorosea (Ifr) to determine virulence of the fungi. Once lethality was determined, sublethal doses of Bpb were combined with enzymes capable of degrading the insect cuticle to measure the potential to enhance fungal infection. Bpb applied to FST in combination with proteinases and a chitinase caused increased mortality over the fungus alone. Mortality was enhanced when Ifr was applied to FST in combination with a chitinase isolated from Serratia marcesans. A lipase isolated from Pseudomonas cepacia, when combined with Ifr, also resulted in greater mortality than all control treatments. FST were also exposed to the eicosanoid biosynthesis inhibitors (EBIs) dexamethasone (DEX), ibuprofen (IBU), and ibuprofen sodium salt (IBUNA), in combination with Ifr. Combining Ifr with IBUNA caused significantly increased mortality on days 6, 7, and 9. Cuticle-degrading enzymes and EBIs may have potential to enhance the pathogenic effect of a fungal control agent against the Formosan subterranean termite.
- MeSH
- analýza přežití MeSH
- chitinasy metabolismus MeSH
- dexamethason metabolismus MeSH
- dezinsekce metody MeSH
- Hypocreales růst a vývoj metabolismus MeSH
- ibuprofen metabolismus MeSH
- ikosanoidy antagonisté a inhibitory MeSH
- insekticidy metabolismus MeSH
- Isoptera metabolismus mikrobiologie fyziologie MeSH
- lipasa metabolismus MeSH
- proteasy metabolismus MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- chitinasy MeSH
- dexamethason MeSH
- ibuprofen MeSH
- ikosanoidy MeSH
- insekticidy MeSH
- lipasa MeSH
- proteasy MeSH
A new simple ultra-high-performance liquid chromatography method with diode array detection (UHPLC-DAD) was developed for chemical fingerprinting analysis of extracellular metabolites in fermentation broth of Geosmithia spp. The SPE method employing Oasis MCX strong cation-exchange mixed-mode polymeric sorbent was chosen for extraction of the metabolites. The analyses were performed on an Acquity UPLC BEH C18 column (100 × 2.1 mm i.d.; particle size, 1.7 μm; Waters) using a gradient elution program with an aqueous solution of trifluoroacetic acid and acetonitrile as the mobile phase. The applicability of the method was proved by analysis of 38 strains produced by different species and isolated from different sources (hosts). The results revealed the correlation of obtained UHPLC-DAD fingerprints with taxonomical identity.
Geosmithia fungi are little known symbionts of bark beetles. Secondary metabolites of lilac colored species G. lavendula and other nine Geosmithia species were investigated in order to elucidate their possible role in the interactions of the fungi with environment. Hydroxylated anthraquinones (yellow, orange, and red pigments), were found to be the most abundant compounds produced into the medium during the submerged cultivation. Three main compounds were identified as 1,3,6,8-tetrahydroxyanthraquinone (1), rhodolamprometrin (1-acetyl-2,4,5,7-tetrahydroxyanthraquinone; 2), and 1-acetyl-2,4,5,7,8-pentahydroxyanthraquinone (3). Compounds 2 and 3 (representing the majority of produced metabolites) inhibited the growth of G+-bacteria Staphylococcus aureus and Bacillus subtilis with minimum inhibitory concentration of 64-512 microg/mL. Anti-inflammatory activity detected as inhibition of cyclooxygenase-2 was found only for compound 3 at 1 and 10 microg/mL. Compound 2 interfered with the morphology, compound 3 with cell-cycle dynamics of adherent mammalian cell lines.
- MeSH
- anthrachinony chemie metabolismus farmakologie MeSH
- antibakteriální látky chemie metabolismus farmakologie MeSH
- antiflogistika nesteroidní chemie metabolismus farmakologie MeSH
- Bacillus subtilis účinky léků MeSH
- biologické pigmenty chemie metabolismus farmakologie MeSH
- biotechnologie metody MeSH
- buněčný cyklus účinky léků MeSH
- Ficus parazitologie MeSH
- HeLa buňky MeSH
- hydroxylace MeSH
- Hypocreales růst a vývoj metabolismus MeSH
- inhibitory cyklooxygenasy chemie metabolismus farmakologie MeSH
- lidé MeSH
- nosatcovití mikrobiologie fyziologie MeSH
- spory hub růst a vývoj metabolismus MeSH
- Staphylococcus aureus účinky léků MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- 1-acetyl-2,4,5,7-tetrahydroxyanthraquinone MeSH Prohlížeč
- 1-acetyl-2,4,5,7,8-pentahydroxyanthraquinone MeSH Prohlížeč
- 1,3,6,8-tetrahydroxyanthraquinone MeSH Prohlížeč
- anthrachinony MeSH
- antibakteriální látky MeSH
- antiflogistika nesteroidní MeSH
- biologické pigmenty MeSH
- inhibitory cyklooxygenasy MeSH
