RESUMO
The health benefits of polyunsaturated fatty acids (PUFAs) have encouraged the search for rich sources of these compounds. However, the supply chain of PUFAs from animals and plants presents environmental concerns, such as water pollution, deforestation, animal exploitation and interference in the trophic chain. In this way, a viable alternative has been found in microbial sources, mainly in single cell oil (SCO) production by yeast and filamentous fungi. Mortierellaceae is a filamentous fungal family world-renowned for PUFA-producing strains. For example, Mortierella alpina can be highlighted due to be industrially applied to produce arachidonic acid (20:4 n6), an important component of infant supplement formulas. Thus, the state of the art of strategies to increase PUFAs production by Mortierellaceae strains is presented in this review. Firstly, we have discussed main phylogenetic and biochemical characteristics of these strains for lipid production. Next, strategies based on physiological manipulation, using different carbon and nitrogen sources, temperature, pH and cultivation methods, which can increase PUFA production by optimizing process parameters are presented. Furthermore, it is possible to use metabolic engineering tools, controlling the supply of NADPH and co-factors, and directing the activity of desaturases and elongase to the target PUFA. Thus, this review aims to discuss the functionality and applicability of each of these strategies, in order to support future research for PUFA production by Mortierellaceae species.
Assuntos
Ácidos Graxos Insaturados , Mortierella , Animais , Filogenia , Ácidos Graxos Insaturados/metabolismo , Engenharia Metabólica , Saccharomyces cerevisiae/metabolismo , Carbono/metabolismo , Mortierella/genética , Mortierella/química , Ácidos Graxos/metabolismoRESUMO
Petroleum hydrocarbons and toxic metals are sources of environmental contamination and are harmful to all ecosystems. Fungi have metabolic and morphological plasticity that turn them into potential prototypes for technological development in biological remediation of these contaminants due to their ability to interact with a specific contaminant and/or produced metabolites. Although fungal bioinoculants producing enzymes, biosurfactants, polymers, pigments and organic acids have potential to be protagonists in mycoremediation of hydrocarbons and toxic metals, they can still be only adjuvants together with bacteria, microalgae, plants or animals in such processes. However, the sudden accelerated development of emerging technologies related to the use of potential fungal bioproducts such as bioinoculants, enzymes and biosurfactants in the remediation of these contaminants, has boosted fungal bioprocesses to achieve higher performance and possible real application. In this review, we explore scientific and technological advances in bioprocesses related to the production and/or application of these potential fungal bioproducts when used in remediation of hydrocarbons and toxic metals from an integral perspective of biotechnological process development. In turn, it sheds light to overcome existing technological limitations or enable new experimental designs in the remediation of these and other emerging contaminants.
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Petróleo , Animais , Biodegradação Ambiental , Ecossistema , Hidrocarbonetos , Compostos OrgânicosRESUMO
Biosurfactants are in demand by the global market as natural commodities that can be added to commercial products or use in environmental applications. These biomolecules reduce the surface/interfacial tension between fluid phases and exhibit superior stability to chemical surfactants under different physico-chemical conditions. Biotechnological production of biosurfactants is still emerging. Fungi are promising producers of these molecules with unique chemical structures, such as sophorolipids, mannosylerythritol lipids, cellobiose lipids, xylolipids, polyol lipids and hydrophobins. In this review, we aimed to contextualize concepts related to fungal biosurfactant production and its application in industry and the environment. Concepts related to the thermodynamic and physico-chemical properties of biosurfactants are presented, which allows detailed analysis of their structural and application. Promising niches for isolating biosurfactant-producing fungi are presented, as well as screening methodologies are discussed. Finally, strategies related to process parameters and variables, simultaneous production, process optimization through statistical and genetic tools, downstream processing and some aspects of commercial products formulations are presented.
Assuntos
Biotecnologia/métodos , Fungos/metabolismo , Tensoativos/metabolismo , Metabolismo dos Lipídeos , Tensão Superficial , TermodinâmicaRESUMO
The enhancement of enzyme complex produced by Penicillium echinulatum grown in several culture media components (bagasse sugarcane pretreated by various methods, soybean meal, wheat bran, sucrose, and yeast extract) was studied to increment FPase, xylanase, pectinase, and ß-glucosidase enzyme activities. The present results indicated that culture media composed with 10 g/L of the various bagasse pretreatment methods did not have any substantial influence with respect to the FPase, xylanase, and ß-glucosidase attained maximum values of, respectively, 2.68 FPU/mL, 2.04, and 115.4 IU/mL. On the other hand, proposed culture media to enhance ß-glucosidase production composed of 10 g/L steam-exploded bagasse supplemented with soybean flour 5.0 g/L, yeast extract 1.0 g/L, and sucrose 10.0 g/L attained, respectively, 3.19 FPU/mL and 3.06 IU/mL while xylanase was maintained at the same level. The proteomes obtained from the optimized culture media for enhanced FPase, xylanase, pectinase, and ß-glucosidase production were analyzed using mass spectrometry and a panel of GH enzyme activities against 16 different substrates. Culture medium designed to enhance ß-glucosidase activity achieved higher enzymatic activities values (13 measured activities), compared to the culture media for FPase/pectinase (9 measured activities) and xylanase (7 measured activities), when tested against the 16 substrates. Mass spectrometry analyses of secretome showed a consistent result and the greatest number of spectral counts of Cazy family enzymes was found in designed ß-glucosidase culture medium, followed by FPase/pectinase and xylanase. Most of the Cazy identified protein was cellobiohydrolase (GH6 and GH7), endoglucanase (GH5), and endo-1,4-ß-xylanase (GH10). Enzymatic hydrolysis of hydrothermally pretreated sugarcane bagasse performed with ß-glucosidase enhanced cocktail achieved 51.4 % glucose yield with 10 % w/v insoluble solids at enzyme load of 15 FPU/g material. Collectively the results demonstrated that it was possible to rationally modulate the GH activity of the enzymatic complex secreted by P. echinulatum using adjustment of the culture medium composition. The proposed strategy may contribute to increase enzymatic hydrolysis of lignocellulosic materials.
Assuntos
Meios de Cultura/farmacologia , Glicosídeo Hidrolases/metabolismo , Complexos Multienzimáticos/metabolismo , Penicillium/efeitos dos fármacos , Penicillium/enzimologia , Celulase/metabolismo , Celulose , Meios de Cultura/química , Meios de Cultura/metabolismo , Endo-1,4-beta-Xilanases/metabolismo , Glicosídeo Hidrolases/biossíntese , Hidrólise/efeitos dos fármacos , Complexos Multienzimáticos/química , Penicillium/metabolismo , Poligalacturonase/metabolismo , Saccharum , beta-Glucosidase/biossíntese , beta-Glucosidase/metabolismoRESUMO
The use of glycerol obtained as an intermediate of the biodiesel manufacturing process as carbon source for microbial growth is a potential alternative strategy for the production of enzymes and other high-value bioproducts. This work evaluates the production of cellulase enzymes using glycerol for high cell density growth of Trichoderma harzianum followed by induction with a cellulosic material. Firstly, the influence of the carbon source used in the pre-culture step was investigated in terms of total protein secretion and fungal morphology. Enzymatic productivity was then determined for cultivation strategies using different types and concentrations of carbon source, as well as different feeding procedures (batch and fed-batch). The best strategy for cellulase production was then further studied on a larger scale using a stirred tank bioreactor. The proposed strategy for cellulase production, using glycerol to achieve high cell density growth followed by induction with pretreated sugarcane bagasse, achieved enzymatic activities up to 2.27 ± 0.37 FPU/mL, 106.40 ± 8.87 IU/mL, and 9.04 ± 0.39 IU/mL of cellulase, xylanase, and ß-glucosidase, respectively. These values were 2 times higher when compared to the control experiments using glucose instead of glycerol. This novel strategy proved to be a promising approach for improving cellulolytic enzymes production, and could potentially contribute to adding value to biomass within the biofuels sector.
Assuntos
Reatores Biológicos , Celulase/biossíntese , Celulose/metabolismo , Glicerol/metabolismo , Trichoderma/crescimento & desenvolvimento , Trichoderma/metabolismo , Biocombustíveis , Biomassa , Celulose/farmacologia , Glucose/metabolismo , Glucose/farmacologia , Glicerol/farmacologia , Saccharum/química , Trichoderma/citologia , Trichoderma/enzimologia , beta-Glucosidase/metabolismoRESUMO
The mangrove ecosystem plays a crucial role in preserving the biodiversity of plants, animals, and microorganisms that are essential for materials cycles. However, the exploration of endophytic fungi isolated from mangroves, particulary in Santa Catarina (SC, Brazil), remains limited. Therefore, the purpose of this study was to assess the biodiversity of endophytic fungi found in Avicennia schaueriana, Laguncularia racemosa, Rhizophora mangle, and Spartina alterniflora from two mangroves on the Island of Santa Catarina: one impacted by anthropic action (Itacorubi mangrove) and the other environmentally preserved (Ratones mangrove). Samplings were carried out between January 2020 and May 2021. Fungi were isolated from leaves, stems, and roots, identified, and clustered into groups through morphological characteristics. Further, a representative strain of each group was identified through ITS1 sequencing. A total of 373 isolates were obtained from plant tissues, of which 96 and 277 isolates were obtained from Itacorubi and Ratones mangroves, respectively. Molecular identification showed that the endophytic fungal community comprised at least 19 genera. The data on fungal community diversity revealed comparable diversity indices for genera in both mangroves. However, we observed differences in the total frequency of fungal genera between impacted (27.38%) and non-impacted (72.62%) mangroves. These findings suggest that anthropic activities in and around the Santa Catarina mangroves have had negative impact on the frequency of endophytic fungi. This emphasizes the reinforcing the significance of preserving these environments to ensure the maintenance of fungal community diversity.
Assuntos
Biodiversidade , Endófitos , Fungos , Filogenia , Rhizophoraceae , Áreas Alagadas , Endófitos/classificação , Endófitos/isolamento & purificação , Endófitos/genética , Brasil , Fungos/classificação , Fungos/isolamento & purificação , Fungos/genética , Rhizophoraceae/microbiologia , Avicennia/microbiologia , Ilhas , Raízes de Plantas/microbiologia , Micobioma , Folhas de Planta/microbiologiaRESUMO
Natural pigments have received special attention from the market and industry as they could overcome the harm to health and the environmental issues caused by synthetic pigments. These pigments are commonly extracted from a wide range of organisms, and when added to products they can alter/add new physical-chemical or biological properties to them. Fungi from extreme environments showed to be a promising source in the search for biomolecules with antimicrobial and antiparasitic potential. This study aimed to isolate fungi from Antarctic soils and screen them for pigment production with antimicrobial and antiparasitic potential, together with other previously isolated strains A total of 52 fungi were isolated from soils in front of the Collins Glacier (Southeast border). Also, 106 filamentous fungi previously isolated from the Collins Glacier (West border) were screened for extracellular pigment production. Five strains were able to produce extracellular pigments and were identified by ITS sequencing as Talaromyces cnidii, Pseudogymnoascus shaanxiensis and Pseudogymnoascus sp. All Pseudogymnoascus spp. (SC04.P3, SC3.P3, SC122.P3 and ACF093) extracts were able to inhibit S. aureus ATCC6538 and two (SC12.P3, SC32.P3) presented activity against Leishmania (L.) infantum, Leishmania amazonensis and Trypanossoma cruzii. Extracts compounds characterization by UPLC-ESI-QToF analysis confirmed the presence of molecules with biological activity such as: Asterric acid, Violaceol, Mollicellin, Psegynamide A, Diorcinol, Thailandolide A. In conclusion, this work showed the potential of Antartic fungal strains from Collins Glacier for bioactive molecules production with activity against Gram positive bacteria and parasitic protozoas.
Assuntos
Antiparasitários , Pigmentos Biológicos , Regiões Antárticas , Pigmentos Biológicos/farmacologia , Pigmentos Biológicos/biossíntese , Antiparasitários/farmacologia , Anti-Infecciosos/farmacologia , Anti-Infecciosos/metabolismo , Fungos/efeitos dos fármacos , Fungos/metabolismo , Fungos/classificação , Microbiologia do Solo , Bactérias/efeitos dos fármacos , Bactérias/classificação , Bactérias/metabolismo , Bactérias/isolamento & purificação , Bactérias/genética , Testes de Sensibilidade Microbiana , Animais , Staphylococcus aureus/efeitos dos fármacosRESUMO
BACKGROUND: There is an imperative necessity for alternative sources of energy able to reduce the world dependence of fossil oil. One of the most successful options is ethanol obtained mainly from sugarcane and corn fermentation. The foremost residue from sugarcane industry is the bagasse, a rich lignocellulosic raw material uses for the production of ethanol second generation (2G). New cellulolytic and hemicellulytic enzymes are needed, in order to optimize the degradation of bagasse and production of ethanol 2G. RESULTS: The ability to produce hemicellulases and related enzymes, suitable for lignocellulosic biomass deconstruction, was explored using 110 endophytic fungi and 9 fungi isolated from spoiled books in Brazil. Two initial selections were performed, one employing the esculin gel diffusion assay, and the other by culturing on agar plate media with beechwood xylan and liquor from the hydrothermal pretreatment of sugar cane bagasse. A total of 56 isolates were then grown at 29°C on steam-exploded delignified sugar cane bagasse (DEB) plus soybean bran (SB) (3:1), with measurement of the xylanase, pectinase, ß-glucosidase, CMCase, and FPase activities. Twelve strains were selected, and their enzyme extracts were assessed using different substrates. Finally, the best six strains were grown under xylan and pectin, and several glycohydrolases activities were also assessed. These strains were identified morphologically and by sequencing the internal transcribed spacer (ITS) regions and the partial ß-tubulin gene (BT2). The best six strains were identified as Aspergillus niger DR02, Trichoderma atroviride DR17 and DR19, Alternaria sp. DR45, Annulohypoxylon stigyum DR47 and Talaromyces wortmannii DR49. These strains produced glycohydrolases with different profiles, and production was highly influenced by the carbon sources in the media. CONCLUSIONS: The selected endophytic fungi Aspergillus niger DR02, Trichoderma atroviride DR17 and DR19, Alternaria sp. DR45, Annulohypoxylon stigyum DR47 and Talaromyces wortmannii DR49 are excellent producers of hydrolytic enzymes to be used as part of blends to decompose sugarcane biomass at industrial level.
Assuntos
Alternaria/enzimologia , Aspergillus niger/enzimologia , Glicosídeo Hidrolases/biossíntese , Talaromyces/enzimologia , Trichoderma/enzimologia , Celulase/metabolismo , Celulases/metabolismo , Celulose/química , DNA Fúngico/genética , Etanol/metabolismo , Fermentação , Glicosídeo Hidrolases/metabolismo , Hidrólise , Saccharum/metabolismo , Saccharum/microbiologia , Análise de Sequência de DNA , Resíduos , Zea mays/metabolismo , Zea mays/microbiologia , beta-Glucosidase/metabolismoRESUMO
The spread of organic farming and the development of resistance to anthelmintics by parasites, especially in small ruminants, have necessitated the search for alternative methods of nematode control. Biological control using nematophagous fungi is one option; however, few studies have been conducted with native strains. The present study was divided into two phases. In the first phase, we aimed to isolate, identify, and assess the in vitro predatory activity of nematophagous fungi that had been isolated on Trichostrongylidae third-instar larvae. In the second phase, the isolate with superior predatory activity in vitro was molecularly characterized, and its morphological plasticity was observed using scanning electron microscopy (SEM) on Haemonchus third-instar larvae. Of the 56 soil samples from different regions of Paraná State, Brazil, 57 fungal strains were recovered, of which four exhibited predatory activity. Two pure isolates were obtained: the CED and LIN strains. After demonstrating 96.35 % predatory activity for the CED strain, this strain was selected and characterized using molecular criteria by sequencing the rDNA internal transcribed spacer and was identified as Arthrobotrys conoides (GenBank ID: JN191309). Morphological patterns in this strain during the interaction between the fungus and the nematode were revealed by SEM, in which two extensions of the infection bulb that was used to pierce the nematode's cuticle were clearly visible.
Assuntos
Ascomicetos/isolamento & purificação , Ascomicetos/patogenicidade , Trichostrongyloidea/microbiologia , Animais , Ascomicetos/genética , Ascomicetos/ultraestrutura , Brasil , Análise por Conglomerados , DNA Fúngico/química , DNA Fúngico/genética , DNA Espaçador Ribossômico/química , DNA Espaçador Ribossômico/genética , Larva/microbiologia , Microscopia Eletrônica de Varredura , Dados de Sequência Molecular , Filogenia , Análise de Sequência de DNA , Microbiologia do SoloRESUMO
The Antarctic continent is an extreme environment recognized mainly by its subzero temperatures. Fungi are ubiquitous microorganisms that stand out even among Antarctic organisms, primarily due to secondary metabolites production with several biological activities. Pigments are examples of such metabolites, which mainly occur in response to hostile conditions. Various pigmented fungi have been isolated from the Antarctic continent, living in the soil, sedimentary rocks, snow, water, associated with lichens, mosses, rhizospheres, and zooplankton. Physicochemical extreme environments provide a suitable setup for microbial pigment production with unique characteristics. The biotechnological potential of extremophiles, combined with concerns over synthetic pigments, has led to a great interest in natural pigment alternatives. Besides biological activities provided by fungal pigments for surviving in extreme environments (e.g., photoprotection, antioxidant activity, and stress resistance), it may present an opportunity for biotechnological industries. This paper reviews the biotechnological potential of Antarctic fungal pigments, with a detailed discussion over the biological role of fungal pigments, potential industrial production of pigments from extremophilic fungi, pigments toxicity, current market perspective and published intellectual properties related to pigmented Antarctic fungi.
Assuntos
Biotecnologia , Fungos , Regiões AntárticasRESUMO
Shrimp antilipopolysaccharide factors (ALFs) form a multifunctional and diverse family of antimicrobial host defense peptides (AMPs) composed of seven members (groups A to G), which differ in terms of their primary structure and biochemical properties. They are amphipathic peptides with two conserved cysteine residues stabilizing a central ß-hairpin that is understood to be the core region for their biological activities. In this study, we synthetized three linear (cysteine-free) peptides based on the amino acid sequence of the central ß-hairpin of the newly identified shrimp (Litopenaeus vannamei) ALFs from groups E to G. Unlike whole mature ALFs, the ALF-derived peptides exhibited an α-helix secondary structure. In vitro assays revealed that the synthetic peptides display a broad spectrum of activity against both Gram-positive and Gram-negative bacteria and fungi but not against the protozoan parasites Trypanosoma cruzi and Leishmania (L.) infantum. Remarkably, they displayed synergistic effects and showed the ability to permeabilize bacterial membranes, a mechanism of action of classical AMPs. Having shown low cytotoxicity to THP-1 human cells and being active against clinical multiresistant bacterial isolates, these nature-inspired peptides represent an interesting class of bioactive molecules with biotechnological potential for the development of novel therapeutics in medical sciences.
Assuntos
Antibacterianos , Anti-Infecciosos , Humanos , Antibacterianos/farmacologia , Conformação Proteica em alfa-Hélice , Lipopolissacarídeos/farmacologia , Bactérias Gram-Negativas , Bactérias Gram-Positivas , Anti-Infecciosos/química , Peptídeos Catiônicos Antimicrobianos/farmacologia , Peptídeos Catiônicos Antimicrobianos/química , Testes de Sensibilidade MicrobianaRESUMO
The discovery of biomolecules has been the subject of extensive research for several years due to their potential to combat harmful pathogens that can lead to environmental contamination and infections in both humans and animals. This study aimed to identify the chemical profile of endophytic fungi, namely Neofusicoccum parvum and Buergenerula spartinae, which were isolated from Avecinnia schaueriana and Laguncularia racemosa. We identified several HPLC-MS compounds, including Ethylidene-3,39-biplumbagin, Pestauvicolactone A, Phenylalanine, 2-Isopropylmalic acid, Fusaproliferin, Sespendole, Ansellone, Calanone derivative, Terpestacin, and others. Solid-state fermentation was conducted for 14-21 days, and methanol and dichloromethane extraction were performed to obtain a crude extract. The results of our cytotoxicity assay revealed a CC50 value > 500 µg/mL, while the virucide, Trypanosoma, leishmania, and yeast assay demonstrated no inhibition. Nevertheless, the bacteriostatic assay showed a 98% reduction in Listeria monocytogenes and Escherichia coli. Our findings suggest that these endophytic fungi species with distinct chemical profiles represent a promising niche for further exploring new biomolecules.
RESUMO
Cladophialophora exuberans is a filamentous fungus related to black yeasts in the order Chaetothyriales. These melanized fungi are known for their 'dual ecology', often occurring in toxic environments and also being frequently involved in human infection. Particularly Cladophialophora exuberans, C. immunda, C. psammophila, and Exophiala mesophila have been described with a pronounced ability to degrade aromatic compounds and xenobiotic volatiles, such as benzene, toluene, ethyl-benzene, and xylene, and are candidates for bioremediation applications. The objective of the present study is the sequencing, assembly, and description of the whole genome of C. exuberans focusing on genes and pathways related to carbon and toxin management, assessing the tolerance and bioremediation of lead and copper, and verifying the presence of genes for metal homeostasis. Genomic evaluations were carried out through a comparison with sibling species including clinical and environmental strains. Tolerance of metals was evaluated via a microdilution method establishing minimum inhibitory (MIC) and fungicidal concentrations (MFC), and agar diffusion assays. Heavy metal bioremediation was evaluated via graphite furnace atomic absorption spectroscopy (GFAAS). The final assembly of C. exuberans comprised 661 contigs, with genome size of 38.10 Mb, coverage of 89.9X and a GC content of 50.8%. In addition, inhibition of growth was shown at concentrations of 1250 ppm for copper and at 625 ppm for lead, using the MIC method. In the agar tests, the strain grew at 2500 ppm of copper and lead. In GFAAS tests, uptake capacities were observed of 89.2% and 95.7% for copper and lead, respectively, after 21 experimental days. This study enabled the annotation of genes involved in heavy metal homeostasis and also contributed to a better understanding of the mechanisms used in tolerance of and adaptation to extreme conditions.
Assuntos
Ascomicetos , Metais Pesados , Humanos , Biodegradação Ambiental , Benzeno/metabolismo , Cobre/metabolismo , Ágar/metabolismo , Ascomicetos/genética , Ascomicetos/metabolismo , Hidrocarbonetos/metabolismo , Metais Pesados/metabolismo , EcossistemaRESUMO
Mangroves are ecosystems with unique characteristics due to the high salinity and amount of organic matter that house a rich biodiversity. Fungi have aroused much interest as they are an important natural source for the discovery of new bioactive compounds, with potential biotechnological and pharmacological interest. This review aims to highlight endophytic fungi isolated from mangrove plant species and the isolated bioactive compounds and their bioactivity against protozoa, bacteria and pathogenic viruses. Knowledge about this type of ecosystem is of great relevance for its preservation and as a source of new molecules for the control of pathogens that may be of importance for human, animal and environmental health.
RESUMO
Listeria monocytogenes is one of the foodborne pathogens of most concern for food safety. To limit its presence in foods, bacteriocins have been proposed as natural bio-preservatives. Herein, a bacteriocin was produced on hemicellulose hydrolysate of sugarcane bagasse by Pediococcus pentosaceous ET34, whose genome sequencing revealed an operon with 100% similarity to that of pediocin PA-1. ET34 grown on hydrolysate-containing medium led to an increase in the expression of PA-1 genes and a non-optimized purification step sequence resulted in a yield of 0.8 mg·L-1 of pure pediocin (purity > 95%). Culture conditions were optimized according to a central composite design using temperature and hydrolysate % as independent variables and validated in 3-L Erlenmeyers. Finally, a process for scaled-up implementation by sugar-ethanol industry was proposed, considering green chemistry and biorefinery concepts. This work stands up as an approach addressing a future proper sugarcane bagasse valorisation for pediocin production.
Assuntos
Bacteriocinas , Saccharum , Celulose , Pediocinas , Pediococcus , Pediococcus pentosaceus , PolissacarídeosRESUMO
Paecilomyces lilacinus has potential for pests control. We aimed to analyze mycelial growth and spore production in P. lilacinus strains in several agro-industrial residues and commercial media. This study suggests alternative nutrient sources for fungi production and that the biotechnological potential of agro-industrial refuses could be employed in byproducts development.
RESUMO
The coprophilic ascomycete fungus Podospora anserina was cultivated on three different plant biomasses, i.e. cotton seed hulls (CSH), soybean hulls (SBH) and acid-pretreated wheat straw (WS) for four days, and the potential of the produced enzyme mixtures was compared in the enzymatic saccharification of the corresponding lignocellulose feedstocks. The enzyme cocktail P. anserina produced after three days of growth on SBH showed superior capacity to release reducing sugars from all tested plant biomass feedstocks compared to the enzyme mixtures from CSH and WS cultures. Detailed proteomics analysis of the culture supernatants revealed that SBH contained the most diverse set of enzymes targeted on plant cell wall polymers and was particularly abundant in xylan, mannan and pectin acting enzymes. The importance of lytic polysaccharide monooxygenases (LPMOs) in plant biomass deconstruction was supported by identification of 20 out of 33 AA9 LPMOs in the SBH cultures. The results highlight the suitability of P. anserina as a source of plant cell wall degrading enzymes for biotechnological applications and the importance of selecting the most optimal substrate for the production of enzyme mixtures.
Assuntos
Biomassa , Glycine max/metabolismo , Podospora/enzimologia , Podospora/crescimento & desenvolvimento , Biotecnologia , Gossypium/anatomia & histologia , Gossypium/metabolismo , Hidrólise , Lignina/metabolismo , Caules de Planta/metabolismo , Podospora/metabolismo , Glycine max/anatomia & histologia , Triticum/anatomia & histologia , Triticum/metabolismoRESUMO
Abstract: Cerrado is the second largest biome in Brazil and majorly contributes to the country's grain production. Previous studies on soil metagenomics from the Cerrado revealed an outstanding microbial diversity. In this study, the abundance of pathogenic fungi was analyzed using metagenomic sequences of the Cerrado soils under native vegetation, and under agriculture with no-tillage and conventional tillage. In total, 128,627 sequences of fungi were identified, with 43,439 representing pathogenic fungi and were distributed as follows: native 17,301 (40%), no-tillage 13,780 (32%), and conventional tillage 12,358 (28%). We identified 41 pathogenic fungal species associated with human and animal infections. The data analysis revealed that the native soils had a higher relative abundance of fungal sequences, similar to pathogenic species sequences, in relation to the total eukaryotic sequences, than the conventional tillage and no-tillage treatments, which observed a reduction in fungal abundance because of anthropogenic activities.
RESUMO
Efficient polysaccharide degradation depends on interaction between enzymes acting on the main chain and the side chains. Previous studies demonstrated cooperation between several enzymes, but not all enzyme combinations have been explored. A better understanding of enzyme cooperation would enable the design of better enzyme mixtures, optimally profiting from synergistic effects. In this study, we analyzed the cooperation of several enzymes involved in the degradation of xylan, glucan, xyloglucan and crude plant biomass from Aspergillus nidulans by single and combined incubations with their polymeric substrate. Positive effects were observed between most enzymes, although not always to the same extent. Moreover, the tailor made cocktails formulated in this study resulted in efficient release of glucose from plant biomass. This study also serves as an example for the complex cooperation that occurs between enzymes in plant biomass saccharification and how expression in easily-accessible hosts, such as Pichia pastoris, can help in revealing these effects.
Assuntos
Aspergillus nidulans/enzimologia , Proteínas Fúngicas/metabolismo , Plantas/química , Polissacarídeos/química , Aspergillus nidulans/genética , Biomassa , Proteínas Fúngicas/genética , Glucanos/química , Hidrólise , Pichia/genética , Pichia/metabolismo , Saccharum/química , Xilanos/químicaRESUMO
Abstract Xylan degradation is an important step in different industries, such as in biorefinery for biomass hydrolysis. Talaromyces wortmannii is a known fungus due to second metabolite production but only few works showed the xylanolytic potential of this fungus. In this way, the aim of this study was to evaluate the production of xylanolytic enzymes from T. wortmannii DR49 on industrial agro wastes. Cultivation in shake flask showed highest xylanase titration (10.3 U/mL; 9.5 U/mL) for wheat bran (WB) and hydrothermal pretreated sugar cane bagasse (HB); in β-xylosidase production WB and xylose were the best carbon sources (0.57 U/mL; 0.34 U/mL) respectively. STR cultivation revealed that 29°C and pH 6.0 were the best conditions for xylanase (14.5 U/mL) and β-xylosidase (1.7 U/mL) production. T. wortmannii DR49 showed to be a potential candidate for xylanolytic enzymes production using agro wastes in bioreactors, which has never been previously reported in this fungus.