In this work Rhizopus Delemar was grown on an apple pomace medium, a waste product from the juice industry. The apple pomace was pre-treated with the enzyme invertase to hydrolyse the sucrose available in the waste to glucose and fructose, which are digestible by the fungus. Combination of invertase pre-treatment and yeast extract supplementation, resulted in highest biomass growth which was 4.3 ± 0.5 g/l biomass. The fungal cell wall was separated from fungal biomass using an alkali treatment. A hydrogel was formed from the cell wall material and used for spinning of filaments using dry gel spinning. The average dry weight percentage of the gel was 11.6 ± 1.3 %. The gel was spun through a needle to a collecting rotating surface to make filaments. The filaments were easy to spin and to collect continuous fibres. The spun filaments had a rubber-like texture. All the tested filaments had an ultimate tensile strength approximately 2-3 MPa and 10 – 12% elongation at break. The conclusion of this work is that it is possible to produce fibers from fungi grown in apple waste and that it is possible to improve fungal growth using invertase and yeast extract. The tensile strength of the filaments needs further improvement to compete with other materials used in woven fabrics.

Banana peels account for about 61 million tons of waste each year globally. The aim of this project was to investigate the possibility of using banana peels as a substrate to cultivate edible filamentous fungi. The peels were subjected to physical and thermal pretreatments while variables such as changes in the medium pH, biomass concentration, fungal strain dependence, and protein content of the fungal biomass were analyzed. The experiments were carried out in three phases. The purpose of phase I was to identify which of the four fungal strains among Neurospora intermedia, Aspergillus oryzae, Rhizopus oryzae, and Rhizopus oligosporus could grow in a medium containing ball-milled banana peel powder (BPP) only. In phase II, the best performing strains from phase I in terms of biomass concentration, i.e., A. oryzae and R. oryzae, were cultivated using banana peel broth (BPB) obtained from thermal pretreatment of BPP. During this phase, the impact of medium supplementation with yeast extract was also assessed. The biomass yield for A. oryzae and R. oryzae 2.9 g/L and 1.6 g/L, respectively, yeast supplementation compared to 2.7 g/L and 0.7 g/L, respectively, without supplementation. In phase III, the experiments performed in phase II without yeast extract supplementation were scaled up, after which protein analysis was performed. A crude protein content of 8.82% was determined for A. oryzae, while in R. oryzae, a higher value of 21.1% was obtained. The protein content from both fungal strains was much higher than that present in the BPP, which was 4.8 g/L. The results showed the potential of using banana peel as a substrate to produce edible fungal biomass with higher protein content and thus has potential applications as animal feed or human food. Further studies are needed to optimize the process in order to raise the fungal biomass yield as well as increase the protein content of the biomass. In addition, comprehensive characterization of the fungal biomass would reveal other important components, such as the amino acid profile.

The circular economy is about rethinking the definition of waste into resource. Tempeh boiling water is cheap and would otherwise be washed into the river and pollute the water which would affect the environment badly. Tempeh boiling water is going to be used as substrate to produce mycoprotein with the fungi Rhizopus oligosporus. This study is about making a mycoprotein nugget and evaluate it with a sensory evaluation and to evaluate the protein content in the nugget and chemical oxygen demand of the boiling water. The sensory evaluation will have 65 panellists to assess the liking of the nugget with two control samples. The purpose of this project was to evaluate the potential of tempeh boiling water for the circular economy as substrate. To produce high mycoprotein nugget that will be accepted by the community. Assess the protein content in the mycoprotein nugget and assess the carbon used by the fungi with chemical oxygen demand analyse. The target group for evaluating the fungal nugget was students studying at Gadjah Mada University, Yogyakarta, Indonesia. The project was done in multiple following stages: Finding best formula of mycoprotein, mycoprotein production, sensory evaluation, and protein analyse with Kjeldahl method and COD analyse of the boiling water.The result of this study is that the mycoprotein nugget were not liked nor disliked with the average score of 3,9 out of 7. The overall characteristics (appearance, colour, texture, and taste) were 4,0 out of 7. The COD before and after fermentation were 6,6 g/L. The most COD were removed by pre-treatment of the boiling water from 172 to 121 g/L.The protein content of the mycoprotein nugget were 23,8%. The social aspect to produce healthy foods to a low cost at the same time improve water quality by removing foods for toxic microorganisms.

No description available.

"2003". / Thesis (PhD)--Macquarie University, Division of Environmental & Life Sciences, Department of Biological Sciences, 2004. / Bibliography: leaves 136-160. / Introduction: The Antarctic environment; Antarctic inhabitants; Microfungi; Identification of microfungi; Physiological factors affecting Antactic microfungi; Flow cytometry and microfungi; Hydrolytic enzymes of industrial interest; Isolation of genes from microfungi; Aims of this study -- Materials and methods: Fungal strains and cultivation conditions; Molecular identification of fungal isolates; Fungal physiology; Hydrolase activity of secreted proteins; Gene cloning and expression -- Results and discussion: Microfungal identification; Physiological factors affecting Antarctic microfungi; Activity in microfungi when grown on solid media; Characterisation of hemicellulases from selected Antarctic microfungi; Cloning of an Antarctic Penicillium allii lipase gene and its expression in Trichoderma reesei -- Conclusions and future prospects. / The Antarctic occupies that region of the planet that falls below the 60th parallel of South latitude. Although it has been frequented by adventurers, journeyman scientists and tourists for the past 100 years, the Continent has remained virtually unoccupied. The intense cold, the absence of human occupation and the limited range of local higher animal species have combined to create the impression that the Continent is virtually devoid of life. -- Although the microbiota of the Antarctic has attracted some small level of attention in the past, the examination of filamentous microfungi has been largely overlooked and fallen to a small group of dedicated investigators. In this study it will be shown that far from being an insignificant component of the Antarctic network, microfungi represent a potentially large and so far untapped bioresource. -- From just 11 bryophyte samples collected at four sites in the Ross Sea/Dry Valleys region of Southern Antarctica, some 30 microfungal isolates were recovered. Using molecular techniques, the internal transcribed spacer (ITS) region of the nuclear ribosomal DNA (nrDNA) was sequenced to reveal no less than nine unique microfungal species. For only two of these species did the ITS sequence data produce a 100% match with records held on the public databases. This investigation also highlighted the problems inherent in the traditional morphological identification system which are now being perpetuated in the molecular database records. -- A set of seven notionally identified isolates obtained from ornithogenic soil samples gathered in the Windmill Islands in Eastern Antarctica (offshore from the Australian Antarctic Division's Casey Station) were also subjected to molecular identification based on ITS sequence data. Each of the seven isolates was identified as a unique species; six were cosmopolitan in nature and the one remaining bore very little resemblance at the molecular level to any of the recorded species although it was provided with an epithet commonly used in the identification of Antarctic microfungal species. -- To evaluate their potential as a bioresource, samples of Antarctic microfungi were examined to determine if the same physiological factors common to mesophilic species also applied to their Antarctic analogues. It is known that when placed under stress, trehalose can act as a protectant against cold (cryoprotection) and dehydration in mesophilic yeasts and fungi. The level of trehalose produced by the Antarctic isolates and their mesophilic analogues when subjected to stress was compared. A similar comparison was made for the production of glycerol which is well established as a compatible solute providing protection to mesophilic species against osmotic stress. Only in the case of trehalose production by an Antarctic Embellisia was there any indication that either of these two compounds could play a significant role in providing protection to the Antarctic fungi against the rigours of their environment, which leaves open to question what in fact does. -- In the course of investigating the means by which Antarctic microfungi guard against the damage which can ensue when subjected to oxidative stress, flow cytometry was introduced as an investigatory tool. It was established that there is a window of opportunity during which flow cytometry can be used to undertake a detailed analysis of the early stages of fungal growth from germination through hyphal development. -- Of major significance in determining the potential of Antarctic microfungi as a resource is their ability to produce new and novel enzymes and proteins. The microfungal isolates were screened for hydrolytic activity on solid media containing indicative substrates and proved to be a fruitful source of enzymes active over a range of temperatures. A detailed characterisation of two hemicellulases, β-mannanase and xylanase, secreted into a liquid medium by a subset of the Antarctic fungi and a high producing mesophilic reference strain permitted direct comparisons to be made. It was shown that the maximum hemicellulase activity of the Antarctic strains occurred at least 10°C and as much as 30°C lower than that of the reference strain and that mannanase activity for two of the Antarctic isolates exceeded 40% of their maximum at 0°C. These assay results highlight the potential of Antarctic microfungi to yield novel cold-active enzymes. -- As a final measure of the capacity of the Antarctic to yield novel enzymes from its microfungal stock, a lipase gene was selected as a target for isolation and expression in a heterologous fungal host. Using PCR techniques, the gene of interest was isolated from an Antarctic isolate of Penicillium allii, transformed into the mesophilic production host Trichoderma reesei and the active protein successfully produced in the growth medium. The recombinant lipase was assayed and found to exhibit novel characteristics consistent with a cold-adapted enzyme. / Mode of access: World Wide Web. / 186 leaves ill

in English β-N-Acetylhexosaminidase (EC 3.2.1.52) is exoglycosidase, which exhibits the unique properties in the filamentous fungi. Enzyme from these organisms are dimeric, inducible and secreted extracelluary. It is expresed as preproprotein, consists of a signal sequence, a large propeptid and a catalytic subunit. Although the enzyme is widely distributed, its structure differs in varies organisms. Bacteria have only monomeric hexosaminidase. Human enzymes are dimeric as well as fungal, but only hexosaminidase from filamentous fungi have the catalytic subunit noncovalently associated with the propeptide. Propeptide is a essential for the enzyme activity. It exists a homologues model of the catalytic subunit of β-N-acetylhexosaminidase from Penicillium oxalicum, but the structure of the propeptide has not yet been solved. The first part of this diploma thesis deals with the optimization of production and purification conditions. The second part deals with structural studies of β-N-acetylhexosaminidases from the filamentous fungi Penicillium oxalicum CCF 3438. These studies were carried out using chemical cross-linking and high resolution mass spectrometry. The combination of these methods revealed region of the noncovalent interaction of the catalytic subunit with the propeptide.

A B-lapachona é uma orto-naftoquinona consagrada por suas atividades farmacológicas, principalmente pela antitumoral, porém não há descrição de estudos de biotransformação microbiana da ?-lapachona. Tais estudos podem propiciar a obtenção de novos derivados dessa naftoquinona, além de fornecerem informações importantes sobre seu metabolismo. Muitos trabalhos descrevem que micro-organismos podem catalisar reações mimetizando enzimas humanas. Para o desenvolvimento dessa pesquisa, a ?-lapachona foi obtida por semissíntese a partir do lapachol. Nos processos de biotransformação foram utilizados os fungos filamentosos Mucor rouxii, Cunninghamella elegans, Cunninghamella echinulata, Penicillium crustosum e Papulaspora immersa e as bactérias gastrointestinais Escherichia coli, cultivada em aerobiose e anaerobiose, Lactobacillus acidophilus, Bifidobacterium sp. e cultura mista composta por Lactobacillus acidophilus, Bifidobacterium sp. e Streptococcus salivarius subesp. thermophilus. Com o intuito de estabelecer uma comparação entre o metabolismo microbiano da ?-lapachona com o do seu isômero ?-lapachona, estudos de biotransformação utilizando o fungo M. rouxii foram também conduzidos com a ?-lapachona. Sete derivados de biotransformação da ?-lapachona com o fungo M. rouxii foram identificados, sendo um inédito, cinco já descritos na literatura em um trabalho de metabolismo dessa naftoquinona utilizando sangue de mamíferos e humanos e uma espirobenzolactona relatada em um trabalho de síntese. Outros dois derivados inéditos da ?-lapachona, os quais são regioisômeros conjugados com glicose, foram produzidos após formação de hidroquinona no processo coduzido com o fungo C. elegans. O fungo P. immersa forneceu duas lactonas isoméricas também obtidas com a biotransformação com o fungo M. rouxii. Houve resultados positivos, com detecção de possíveis produtos de biotransformação da ?-lapachona por CLAE-DAD, com as bactérias E. coli em aerobiose e Bifidobacterium sp. No entanto, esses processos apresentaram um baixo rendimento, sendo possível a identificação de apenas um derivado com a E. coli, que também foi obtido com a biotransformação com o fungo M. rouxii. Um derivado glicosilado da ?-lapachona foi produzido após 24 horas de incubação no processo desenvolvido com o fungo M. rouxii, sendo posteriormente convertido em hidroxilapachol, que por sua vez originou a ?-lapachona novamente e também a ?-lapachona, a qual foi metabolizada também. O derivado glicosilado majoritário, obtido da biotransformação com a ?-lapachona com o fungo C. elegans, foi submetido à avaliação da atividade citotóxica frente à linhagem de câncer de mama humano SKBR-3 apresentando IC50 igual a 312,5 ?M, sendo o IC50 da ?-lapachona frente à mesma linhagem igual a 5,6 ?M. O derivado majoritário não apresentou citotoxidade frente à linhagem de fibroblastos normais humanos GM07492-A, enquanto a ?-lapachona foi altamente citotóxica (IC50 igual a 7,25 ?M). Esse mesmo derivado inédito foi também produzido em pequena quantidade no processo desenvolvido com o fungo C. echinulata. Na metabolização microbiana da ?-lapachona ocorreram tanto reações de fase I como de fase II, havendo mimetização do metabolismo de mamíferos, inclusive de humanos, como relatado em trabalhos da literatura. / B-lapachone is considered an important ortho-naphthoquinone by their pharmacological activities, mainly antitumor, but there is no description of microbial biotransformation studies of ?-lapachone. These researches may furnish new derivatives and significant information on its metabolism. Many studies describe that microorganisms can catalyze reactions mimicking human enzymes. ?-lapachone was obtained by semisynthetic procedure from lapachol. Biotransformation processes were carried out using the filamentous fungi Mucor rouxii, Cunninghamella elegans, Cunninghamella echinulata, Penicillium crustosum and Papulaspora immersa and the gastrointestinal bacteria Escherichia coli grown aerobically and anaerobically, Lactobacillus acidophilus, Bifidobacterium sp. and mixed culture with Lactobacillus acidophilus, Bifidobacterium sp. and Streptococcus salivarius subsp. thermophilus. In order to establish a comparison between ?-lapachone microbial transformation and those of its isomer ?-lapachone, biotransformation studies of ?-lapachone were also carried out using M. rouxii. Seven derivatives of ?-lapachone were produced in the process performed by M. rouxii, including one unpublished, five already described in a study of metabolism by mammalian and human blood and one spirobenzolactone reported in a syntetic study. Other two unpublished derivatives of ?-lapachone, which are regioisomers conjugated with glucose, were produced after formation of hydroquinone in the process carried out by C. elegans. P. immersa provided two isomeric lactones also obtained by biotransformation with M. rouxii. Possible biotransformation products were detected by using HPLC-DAD in the processes carried out by the bacteria E. coli under aerobic condition and Bifidobacterium sp. However, these processes exhibited a low yield, and it was possible to identify only one derivative produced by E. coli, which was also obtained in the process performed by M. rouxii. A glycosylated derivative of ?-lapachone was produced by biotransformation with M. rouxii after 24 hours of incubation and subsequently was converted in hydroxylapachol, which in turn gave rise to ?-lapachone again and also to ?-lapachone, which was also metabolized. The major derivative produced in the process carried out by C. elegans was submitted to cytotoxic activity evaluation using human breast cancer cell line SKBR3 showing IC50 312.5 ?M, being the ?-lapachone IC50 5.6 ?M against the same cell line. The major derivative did not show cytotoxicity to normal human fibroblast GM07492-A cell line, while ?-lapachone was highly cytotoxic (IC50 7.25 ?M). The same major derivative was also produced in smaller yield in the process performed by C. echinulata. In the ?-lapachone microbial transformation studies occurred phase I and phase II reactions, mimicking the metabolism of mammals, including humans, as reported in literature.

As estatinas são os agentes mais eficazes para a redução de colesterol no tratamento de doenças cardiovasculares, e algumas destas moléculas podem ser produzidas através de processos biológicos como o cultivo semi-sólido de fungos filamentosos. O objetivo deste estudo foi determinar a capacidade de produção de estatinas e compostos antimicrobianos por cinco cepas de fungos isolados de Cana de Açúcar. Para isso, extratos obtidos a partir dos cultivos foram analisados por métodos analíticos como CLAE e RMN para determinar a presença de estatinas; adicionalmente, os extratos foram testados contra diferentes modelos biológicos incluindo bactérias, leveduras, fungos filamentosos, células de ovário de hamster chinês, e parasitas. De acordo com os resultados obtidos, os cinco fungos avaliados não produzem estatinas, e em relação ao biomonitoramento dos extratos foi observado um efeito biológico sobre os parasitas e as células de mamífero, no entanto, é possível que o efeito obtido seja uma resposta dos compostos do substrato dos cultivos (Farelo de trigo). / Statins are the most effective cholesterol lowering agents for the treatment of cardiovascular disease, and some of these molecules can be produced through biological process such as the solid state fermentation. The aim of this study was determinate the capability of production of statins and antimicrobials compounds by five strains of fungi isolated from Brazilian sugar cane. For this purpose, extracts were obtained from the cultures and analyzed through analytical methods as HPLC and NMR in order to determinate the presence of statins; in addition, the extracts were tested against different biological models including bacteria, yeast, filamentous fungi, chinese hamster ovary cells, and parasites. According to the results obtained, the five fungal strains tested did not produce statins, and the extracts produced a biological effect against the parasites and mammalian cells, nevertheless it is possible that this effect observed was a response of the compounds from the culture substrate (wheat bran).

Snabb och korrekt artidentifiering är avgörande för effektiv behandling av svampinfektioner, särskilt bland immunsupprimerade patienter. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) används rutinmässigt på kliniska laboratorier för identifiering av karaktäristiska proteinmönster hos bakterier och jästsvampar genom tolkning av proteinspektra i en masspektradatabas för korrekt artidentifiering. Mögelsvamparnas hårda cellvägg och heterogena växtsätt med varierande proteinuttryck beroende på mognadsstadie, försvårar identifiering med MALDI-TOF MS. Metodens tänkbara fördelar mot traditionella metoden mikroskopering är förkortade svarstider, säkrare artidentifiering av fler arter och mindre beroende av subjektiv morfologisk bedömning. Studiens syfte var att undersöka om MALDI-TOF MS kunde anpassas och användas för identifieringen av mögelsvamp i klinisk rutindiagnostik. Fyra referensstammar (Aspergillus niger, A. fumigatus, A.terreus, A.flavus) och ett kliniskt isolat (A.terreus) undersöktes. Preparationsmetoderna (I) fullständig myrsyraextraktion, (II) direktapplicering och (III) suspension i destillerat vatten användes för analys av sporer och frontmycel hos yngre och äldre mögelkulturer. Två olika masspektradatabaser för artidentifiering jämfördes; rutindatabasen BDAL och den specialiserade mögeldatabasen Filamentous Fungi Library. Även plocktekniken av mögelmaterial inför analys med MALDI-TOF MS utvärderades. Vid vissa tillfällen förbättrades artidentifieringen efter extraktion av mögelkulturerna, medan i andra fall var direktapplicering fullt tillräcklig. Mögelmaterial med mycket sporer tenderade ge något fler artidentifieringar i BDAL oavsett kulturernas ålder.  Filamentous Fungi Library tenderade i vissa fall ge bättre resultat jämfört med BDAL för yngre kulturer. Fler studier krävs för att utvärdera och optimera MALDI-TOF MS som metod för artidentifiering av mögelsvamp. / Rapid and accurate species identification is crucial for successful treatment of fungal infections, especially among immunosuppressed patients. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) is used routinely at clinical laboratories to identify characteristic protein patterns of bacteria and yeast by the interpretation of protein spectra in a database for accurate species identification. The hard cell wall of the mold and the heterogeneous growth with varying protein expression due to maturation, complicates identification with MALDI-TOF MS. The potential benefits of this method compared to microscopy as traditional method are shortened turn-around times, safer species identification of more species that is independent on subjective morphological assessment. The purpose of the study was to investigate whether MALDI-TOF MS could be adapted and used for the identification of molds in clinical routine diagnostics. Four reference strains (Aspergillus niger, A.fumigatus, A.terreus, A.flavus) and a clinical isolate (A.terreus) were examined. The preparation methods (I) complete formic acid extraction, (II) direct application and (III) suspension in distilled water were used for analysis of spores and frontmycelium from younger and older mold cultures. Two different masspektradatabases for species identification were compared; routine database BDAL and the specialized mold database, Filamentous Fungi Library. Also the collecting technique of mold prior to analysis with MALDI-TOF MS was evaluated. Sometimes, the species identification improved after extraction of mold cultures, while in other cases direct application was sufficient. Cultures with a lot of spores tended to give slightly more species identifications in BDAL regardless of the age of cultures. Filamentous Fungi Library, in some cases, tended to improve the performance compared to BDAL for younger cultures. More studies are required to evaluate and optimize MALDI-TOF MS as a method of mold identification.

As estatinas são os agentes mais eficazes para a redução de colesterol no tratamento de doenças cardiovasculares, e algumas destas moléculas podem ser produzidas através de processos biológicos como o cultivo semi-sólido de fungos filamentosos. O objetivo deste estudo foi determinar a capacidade de produção de estatinas e compostos antimicrobianos por cinco cepas de fungos isolados de Cana de Açúcar. Para isso, extratos obtidos a partir dos cultivos foram analisados por métodos analíticos como CLAE e RMN para determinar a presença de estatinas; adicionalmente, os extratos foram testados contra diferentes modelos biológicos incluindo bactérias, leveduras, fungos filamentosos, células de ovário de hamster chinês, e parasitas. De acordo com os resultados obtidos, os cinco fungos avaliados não produzem estatinas, e em relação ao biomonitoramento dos extratos foi observado um efeito biológico sobre os parasitas e as células de mamífero, no entanto, é possível que o efeito obtido seja uma resposta dos compostos do substrato dos cultivos (Farelo de trigo). / Statins are the most effective cholesterol lowering agents for the treatment of cardiovascular disease, and some of these molecules can be produced through biological process such as the solid state fermentation. The aim of this study was determinate the capability of production of statins and antimicrobials compounds by five strains of fungi isolated from Brazilian sugar cane. For this purpose, extracts were obtained from the cultures and analyzed through analytical methods as HPLC and NMR in order to determinate the presence of statins; in addition, the extracts were tested against different biological models including bacteria, yeast, filamentous fungi, chinese hamster ovary cells, and parasites. According to the results obtained, the five fungal strains tested did not produce statins, and the extracts produced a biological effect against the parasites and mammalian cells, nevertheless it is possible that this effect observed was a response of the compounds from the culture substrate (wheat bran).