Current antibiotics and chemotherapeutics are becoming ineffective because pathogenic bacteria and tumor cells have developed multiple drug resistance. Therefore, it is necessary to find new substances that can be used in treatment, either alone or as sensitizing molecules in combination with existing drugs. Peptaibols are bioactive, membrane-active peptides of non-ribosomal origin, mainly produced by filamentous fungi such as Trichoderma spp. This study focused on producing peptaibol-rich extracts from Trichoderma atroviride O1, cultivated on malt extract agar (MA) under circadian and constant darkness conditions for 13 days. Peptaibol production was detected by MALDI-TOF mass spectrometry after six days of cultivation. The extracts demonstrated antibacterial activity against Staphylococcus aureus strains, particularly the methicillin-resistant variant, but not against the Gram-negative Pseudomonas aeruginosa. Quorum sensing interference revealed that a peptaibol-rich extract suppressed Vibrio campbellii BAA-1119's AI-2 signaling system to a degree comparable with gentamycin. Beyond antibacterial properties, the extracts exhibited notable antiproliferative activity against human ovarian cancer cells and their adriamycin-resistant subline in both 2D and 3D models. Specifically, MA-derived extracts reduced ovarian cancer cell viability by 70% at 50 μg/mL, especially under light/dark regime of cultivation. Compared to previously published results for PDA-based extracts, MA cultivation shifted the biological effects of peptaibol-containing extracts toward anticancer potential. These findings support the idea that modifying fungal cultivation parameters, the bioactivity of secondary metabolite mixtures can be tailored for specific therapeutic applications.
- MeSH
- agar * chemie MeSH
- antibakteriální látky * farmakologie metabolismus MeSH
- Hypocreales MeSH
- kultivační média chemie MeSH
- lidé MeSH
- mikrobiální testy citlivosti MeSH
- nádorové buněčné linie MeSH
- peptaiboly * farmakologie metabolismus biosyntéza chemie MeSH
- proliferace buněk účinky léků MeSH
- protinádorové látky * farmakologie metabolismus MeSH
- Pseudomonas aeruginosa účinky léků MeSH
- Staphylococcus aureus účinky léků MeSH
- Trichoderma * metabolismus růst a vývoj chemie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- agar * MeSH
- antibakteriální látky * MeSH
- kultivační média MeSH
- peptaiboly * MeSH
- protinádorové látky * MeSH
The study evaluates the survivability and storage stability of seven Trichoderma strains belonging to the species: T. harzianum (1), T. atroviride (4), and T. virens (2) after the lyophilization of their solid state cultures on wheat straw. Biomass of Trichoderma strains was freeze-dried with and without the addition of maltodextrin. Furthermore, in order to determine the ability of tested Trichoderma strains to preserve selected technological features, the biosynthesis of extracellular hydrolases (cellulases, xylanases, and polygalacturonases) after a 3-month storage of lyophilizates was investigated. Strains of T. atroviride (except TRS40) and T. harzianum TRS85 showed the highest viability after lyophilization process (up to 100%). After 3 months of storage, T. atroviride TRS14 exhibited the highest stability (95.23%); however, the number of active conidia remained at high level of 106-107 cfu/g for all tested T. atroviride strains and T. harzianum TRS85. Interestingly, after a 3-month storage of lyophilized formulations, most of the tested Trichoderma strains exhibited higher cellulolytic and xylanolytic activities compared to the control, i.e., before freeze-drying process. The highest activities of these enzymes exhibited the following: T. atroviride TRS14-2.37 U/g and T. atroviride TRS25-21.47 U/g, respectively, whereas pectinolytic activity was weak for all tested strains, with the highest value of 0.64 U/g registered for T. virens TRS109.
- MeSH
- biomasa MeSH
- časové faktory MeSH
- fermentace MeSH
- hydrolasy metabolismus MeSH
- lyofilizace * MeSH
- mikrobiální viabilita * MeSH
- pšenice metabolismus MeSH
- skladování léků MeSH
- spory hub růst a vývoj MeSH
- Trichoderma klasifikace růst a vývoj fyziologie MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- hydrolasy MeSH
Efficient hydrolysis of holocellulose depends on a proper balance between cellulase (endoglucanase, exoglucanase, β-glucosidase) and xylanase activities. The present study aimed to induce the production of cellulases and xylanases using liquid cultures (one, two, three, and four fungal strains on the same bioreactor) of wild strains of Trichoderma harzianum, Aspergillus niger, and Fusarium oxysporum. The strains were identified by amplification and analysis of the ITS rDNA region and the obtained sequences were deposited in Genbank. Enzymes (endoglucanase, exoglucansae, β-glucosidase, and xylanase activities) and the profile of extracellular protein isoforms (SDS-PAGE) produced by different fungal combinations (N = 14) were analyzed by Pearson's correlation matrix and principal component analysis (PCA). According to our results, induction of endoglucanase (19.02%) and β-glucosidase (6.35%) were obtained after 4 days when A. niger and F. oxysporum were cocultured. The combination of A. niger-T. harzianum produced higher endoglucanase in a shorter time than monocultures. On the contrary, when more than two strains were cultured in the same reactor, the relationships of competition were established, trending to diminish the amount of enzymes and the extracellular protein isoforms produced. The xylanase production was sensible to stress produced by mixed cultures, decreasing their activity. This is important when the aim is to produce cellulase-free xylanase. In addition, exoglucanase activity did not change in the combinations tested.
- Klíčová slova
- Aspergillus, Endoglucanase, Exoglucanase, Filamentous fungi, Fusarium, Trichoderma, β-Glucosidase,
- MeSH
- Ascomycota enzymologie růst a vývoj izolace a purifikace metabolismus MeSH
- Aspergillus niger enzymologie růst a vývoj izolace a purifikace metabolismus MeSH
- biomasa MeSH
- bioreaktory mikrobiologie MeSH
- celulasy biosyntéza metabolismus MeSH
- celulosa metabolismus MeSH
- fermentace MeSH
- fungální proteiny biosyntéza metabolismus MeSH
- Fusarium enzymologie růst a vývoj izolace a purifikace metabolismus MeSH
- kokultivační techniky * MeSH
- mikrobiální interakce fyziologie MeSH
- průmyslová mikrobiologie metody MeSH
- Trichoderma enzymologie růst a vývoj izolace a purifikace metabolismus MeSH
- xylosidasy biosyntéza metabolismus MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- celulasy MeSH
- celulosa MeSH
- fungální proteiny MeSH
- xylosidasy MeSH
Biofilms represent mixed communities present in a diverse range of environments; however, their utility as inoculants is less investigated. Our investigation was aimed towards in vitro development of biofilms using fungal mycelia (Trichoderma viride) as matrices and nitrogen-fixing and P-solubilizing bacteria as partners, as a prelude to their use as biofertilizers (biofilmed biofertilizers, BBs) and biocontrol agents for different crops. The most suitable media in terms of population counts, fresh mass and dry biomass for Trichoderma and Bacillus subtilis/Pseudomonas fluorescens was found to be Pikovskaya broth ± 1 % CaCO(3), while for Trichoderma and Azotobacter chroococcum, Jensen's medium was most optimal. The respective media were then used for optimization of the inoculation rate of the partners in terms of sequence of addition of partners, fresh/dry mass of biofilms and population counts of partners for efficient film formation. Microscopic observations revealed significant differences in the progress of growth of biofilms and dual cultures. In the biofilms, the bacteria were observed growing intermingled within the fungal mycelia mat. Further, biofilm formation was compared under static and shaking conditions and the fresh mass of biofilms was higher in the former. Such biofilms are being further characterized under in vitro conditions, before using them as inoculants with crops.
- MeSH
- biofilmy * MeSH
- kultivační média chemie metabolismus MeSH
- mycelium růst a vývoj fyziologie MeSH
- očkovadla agrotechnická genetika růst a vývoj fyziologie MeSH
- Trichoderma genetika růst a vývoj fyziologie MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- kultivační média MeSH
Conditions for conidiation of a natural isolate of Trichoderma atroviride during submerged cultivation in Erlenmeyer flasks and in a laboratory stirred-tank fermenter were optimized. From the simple sugars tested, cellobiose was the best substrate for conidia production while cellulose fines from paper mill waste proved to be a suitable cheap complex carbon source. Optimum temperature for conidiation was 24-26 degrees C, and the required dissolved oxygen level was > 40% saturation. After initial slight decrease during the 1st d after inoculation, the pH of the culture medium constantly increased throughout the sporulation period. Attempts to regulate the pH during fermentation did not improve the spore yields. The most intense formation of conidia took place between 2nd and 3rd d of growth and the overall volumetric productivity of conidia was 4.1-8.2 x 10(9) conidia per L/h.
- MeSH
- bioreaktory mikrobiologie MeSH
- fermentace fyziologie MeSH
- koncentrace vodíkových iontů MeSH
- metabolismus sacharidů fyziologie MeSH
- mykologie metody MeSH
- osmotický tlak MeSH
- spory hub fyziologie MeSH
- Trichoderma růst a vývoj metabolismus MeSH
- vysoká teplota MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Thirty-three isolates belonging to six species of the genus Trichoderma were tested for the ability to hydroxylate progesterone to 11alpha-, 11beta-, 11alpha,17alpha- and 6beta, 17alpha-derivatives, and epicortisol. T. aureoviride, T. harzianum, T. polysporum and T. pseudokoningii produced 11alpha-hydroxyprogesterone. T. harzianum and T. hamatum can form only the 11beta-isomer. T. koningii and T. hamatum produced 11alpha-, 11beta-, 11alpha,17alpha- and 6beta,11alpha-hydroxy derivatives. 11alpha, 11beta, 6beta,11alpha- and 11alpha,17alpha-hydroxyprogesterones and epicortisol are produced by T. aureoviride and T. pseudokoningii. Cortisol was produced only when the medium was fortified by 10 g/L peptone. This is the first record of conversion of progesterone to mono-, di- and trihydroxyprogesterones by these Trichoderma species.
- MeSH
- biotransformace MeSH
- hydroxylace MeSH
- progesteron chemie metabolismus MeSH
- Trichoderma růst a vývoj izolace a purifikace metabolismus MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- progesteron MeSH
Laccase activity in Trichoderma harzianum and in our own isolate Trichoderma atroviride was correlated with the production of the green pigment in conidial spores. The laccases of the two fungal species exhibit comparable kinetic parameters, pH optima and thermal sensitivity but differed in physiological properties, such as their catalytic activity during growth.
- MeSH
- kinetika MeSH
- koncentrace vodíkových iontů MeSH
- lakasa MeSH
- oxidoreduktasy chemie metabolismus MeSH
- spory hub enzymologie růst a vývoj MeSH
- teplota MeSH
- Trichoderma enzymologie růst a vývoj MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- lakasa MeSH
- oxidoreduktasy MeSH
The growth and conidiation of the aged Trichoderma viride culture grown in the dark, and after an induction by a light pulse, was examined in the presence of selected mono-, di(tri)saccharides, amino acids and alcohols as sole carbon sources. Hexoses and disaccharides, but not pentoses and amino acids, promoted proportionally both growth and conidiation induced by aging or light. All compounds but pentoses promoted the conidiation in aged cultures and photoconidiation in a close correlation. Ethanol, glycerol and ethylene glycol supported both growth and conidiation but these processes were not supported equally. Conidia formation with hexoses and amino acids as sole carbon sources seems to be a function of growth promotion, rather than of growth restriction (starvation, stress, aging). With glucose as sole carbon source the conidiation was not triggered by nutrient limitation, nor by the accumulation of waste metabolites. The aging-induced conidiation can be considered to be triggered by the genetic program of the microorganism rather than by its nutrient status.
- MeSH
- alkoholy MeSH
- aminokyseliny MeSH
- disacharidy MeSH
- kultivační média MeSH
- monosacharidy MeSH
- světlo * MeSH
- tma MeSH
- Trichoderma růst a vývoj fyziologie MeSH
- trisacharidy MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- srovnávací studie MeSH
- Názvy látek
- alkoholy MeSH
- aminokyseliny MeSH
- disacharidy MeSH
- kultivační média MeSH
- monosacharidy MeSH
- trisacharidy MeSH
A Trichoderma sp. isolate, hereafter called T6, produces a 46-kDa endochitinase (CHIT 46) which had been shown to drastically affect in vitro the cell walls of the phytopathogens Sclerotium rolfsii and Rhizoctonia solani. We attempted to gain insight into its properties. The CHIT 46 N-terminal amino acid sequence shares a very high homology with other fungal chitinases. Western blot analysis using polyclonal antibodies anti-CHIT 46 revealed that this enzyme is immunologically distinct from other proteins produced by the same Trichoderma isolate T6, but is immunologically identical with proteins having equivalent molar mass, probably chitinases, produced by other Trichoderma spp. isolates. In addition, the antibodies revealed also that a substantial amount of this enzyme is secreted into the culture medium 2 d after the Trichoderma isolate T6 comes into contact with chitin.
- MeSH
- buněčná stěna účinky léků MeSH
- chitinasy chemie klasifikace genetika imunologie farmakologie MeSH
- elektroforéza v polyakrylamidovém gelu MeSH
- fungální proteiny chemie klasifikace genetika imunologie farmakologie MeSH
- myši MeSH
- Rhizoctonia účinky léků MeSH
- sekvenční homologie MeSH
- Trichoderma enzymologie růst a vývoj MeSH
- western blotting MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- chitinasy MeSH
- fungální proteiny MeSH
An express method of testing the cellulolytic activity of Trichoderma viride in media with cellulose-containing substrates by the rate of glucose production has been elaborated. The optimum temperature selections and duration of sample incubation (1 h, 50 degrees C) have been described, and the temperature coefficient for the scale-down of the enzyme complex from 50 to 30 degrees C (0.77) has been determined.
- MeSH
- celulasa analýza metabolismus MeSH
- hydrolýza MeSH
- kultivační média MeSH
- mykologie metody MeSH
- Trichoderma enzymologie růst a vývoj metabolismus MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- celulasa MeSH
- kultivační média MeSH
