A thermophilic fungus was isolated from composted horse manure. The organism was as a Chaetomium sp. by sequencing the highly conserved ITS region of the fungus and comparing to known regions in a genomic database and was referred to as TM-417. TM-417 was found to have an optimal growth temperature of 45 oC and an optimal pH of 7.0. An extracellular DNase and RNase was found to be produced by the isolate and were purified 145.58-fold and 127.6-fold respectively using a combination of size exclusion chromatography and a novel affinity membrane purification system. The extent of purification was determined electrophoretically using 4-15% gradient polyacrylamide gels. Both DNase and RNase were dependent on metal co-factors for activity. The metal ion Mg2+ was the preferred ion for the DNase, whereas for the RNase, Zn2+ and Mn2+ yielded an increase in enzyme activity over that with Mg2+. The purified DNase demonstrated maximum activity at pH 6.0 with no activity at pH 2.0 or 10.0. The RNase exhibited two peaks of maximum activity, on at pH 3.0 and the other at pH 7.0 with no activity at pH 2.0 or 10.0. The optimal temperature for the purified DNase was 65oC. The optimal temperature for the RNase was 70oC. The molecular of the DNase and RNase were determined to be 56 kDa and 69kDa respectively using a Sephadex G-75 column. A standard curve was generated using several standard proteins of known molecular weight.

Bacterial resistance to antibiotics has become a worldwide health problem, resulting in untreatable infections and escalating healthcare costs. Wastewater treatment plants are a critical point of control between anthropogenic sources of pathogens, antibiotic resistant bacteria (ARBs), antibiotic resistance genes (ARGs), and the environment through discharge of treated effluent and land application of biosolids. Recent studies observing an apparent resuscitation of pathogens and pathogen indicators and the widening realization of the importance of addressing environmental reservoirs of ARGs all lead toward the need for improved understanding of ARG fate and pathogen inactivation kinetics and mechanisms in sludge stabilization technologies. This research has investigated the fate of two pathogens, methicillin-resistant Staphylococcus aureus (MRSA) and Escherichia coli, and various ARGs under pasteurization, anaerobic digestion, biosolids storage, and land application conditions. Pathogen die-off occurs at a rate specific to each pathogen and matrix in ambient and mesophilic temperature environments. Viable but nonculturable (VBNC) states are initiated by thermal treatments, such as thermophilic digestion and possibly pasteurization, and allow the persistence of pathogen cells and any ARGs contained therein through treatment and into the receiving environment where resuscitation or transformation could occur. Raw sludge ARG content does affect digester effluent quality, although the predominant mechanisms of ARG persistence may be different in mesophilic versus thermophilic digestion. In both thermophilic and mesophilic digestion, a correlation was observed between raw sludge and digester ARGs associated with Class 1 integrons, possibly as a result of horizontal gene transfer. ARB survival was shown to contribute to ARG content in mesophilic digestion, but not thermophilic digestion. Thermophilic digestion may achieve a higher ARG reduction because of reduced microbial diversity compared to mesophilic digestion. However, it is evident that horizontal gene transfer still does occur, particularly with highly mobile integrons, so that complete reduction of all ARGs would not be possible with thermophilic digestion alone. Surprisingly, the experiments that introduced various concentrations of antibiotic sulfamethoxazole and antimicrobial nanosilver did not induce enhanced rates of horizontal gene transfer. Finally, ARG concentrations in biosolids increased during cold temperature storage suggesting that there is a stress induction of horizontal gene transfer of integron-associated ARGs. / Ph. D.

Cette étude porte sur l'identification et la dispersion des bioaérosols générés sur les plates-formes de compostage et plus précisément lors du retournement des andains en cours de fermentation. L'analyse des bioaérosols émis sur cinq plates-formes par des inventaires moléculaires (ADNr 16S et ADNr 18S) a permis de montrer la dominance de deux phyla bactériens Firmicutes et Actinobacteria et du phylum fongique Ascomycota. En comparant la structure de la population microbienne des cinq bioaérosols, une signature a été identifiée. Dix phylotypes microbiens sont communs à au moins quatre bioaérosols. Deux sont présents dans les cinq bioaérosols : NA07 appartenant à l'espèce Saccharopolyspora rectivirgula et représentant 7% des séquences bactériennes totales et EQ07 affiliée à Thermomyces (49% des séquences fongiques). Un second phylotype bactérien, NC38, affilié à la famille des Thermoactinomycetaceae a été sélectionné du fait q u'il n'ait été identifié que dans le compost. Des systèmes de PCRq ont été développés pour quantifier ces trois indicateurs potentiels. Ces derniers ont été validés expérimentalement par des mesures sur sites industriels. La dispersion des bioaérosols a ensuite été caractérisée en utilisant plusieurs méthodologies, dont les trois indicateurs conçus et une technique d'empreinte moléculaire, la SSCP. Lors d'une activité de retournement, la concentration des indicateurs est supérieure à leur niveau basal dans l'air (bruit de fond). Les indicateurs présentent des profils de dispersion différents d'où l'intérêt de les coupler afin d'obtenir une meilleure vision de la dispersion des bioaérosols de compostage. / The aim of this work was to analyze the diversity and the dispersal of composting bioaerosol emitted during the turning of compost windrows in thermophilic phase on composting platforms. The study of the microbial diversity of aerosols emitted on five composting plants was realized by 16S and 18S rDNA molecular inventories. Two bacterial phyla Firmicutes and Actinobacteria and one fungal phylum Ascomycota dominated. A common microbial signature emerged from the five composting bioaerosols: ten microbial phylotypes (seven bacterial and three fungal) were common to at least four bioaerosols. Two have been identified in five bioaerosols: NA07 belonging to the species Saccharopolyspora rectivirgula, and representing 7% of total number of bacterial sequences, and EQ05,affiliated to Thermomyces (49% of total number of fungal sequences). A second bacterial phylotype, NC38, affiliated to the Thermoactinomycetaceae family, was selected because it was id entified only in the environmental source compost'. qPCR systems were then designed for each phylotype. Measurements performed on industrial composting sites validated the use of these microorganisms as indicators of composting bioaerosols. The dispersal of composting bioaerosols was then characterized using the three indicators developed and a fingerprint technique, the SSCP.

Avloppsslam innehåller näringsämnen, som bör återföras till naturen för att skapa ett så kallat näringskretslopp. Genom att optimera hygieniseringen av avloppsslam med effektivare behandlingsmetod kan avloppsslam bli mer lämpligt att avsättas ur ett bättre ekonomiskt- och miljömässigt perspektiv. Syftet med arbetet var att undersöka och bedöma om effektiviseringav befintlig slambehandling är möjlig för att uppnå tillräcklig hygienisering. Utifrån den effektiviserade slambehandlingsmetoden utreda möjliga avsättningsalternativ som lämpar sig bäst för avloppsslammet i Avesta kommun. En litteraturöversikt utfördes för att öka kunskapen och förståelsen av avloppsslam, desshantering och avsättning. Fallstudien är utformad med kvalitativ metod och informationen har inhämtats från vetenskapliga artiklar, myndigheter, rapporter, examensarbeten och personligkontakt som komplement för att erhålla en god förståelse. Uppgradering av mesofil rötning av avloppsslam vid 37 °C till termofil rötning, 55 °C – 65 °C innebär förhöjd temperatur; det kräver mer energi och ökade energikostnader. Komponenter av den mesofila rötningsprocessen kommer att behöva uppgraderas eller bytas ut och därför tillkommer extra kostnader som också behöver ses över. Fördelarna med termofil rötning inkluderar kortvariga uppehållstider, mindre rötkammare och slammet blir tillräckligt hygieniserat. Ett framtidsscenario är framtaget utifrån effektiviserad slambehandling där två avsättningsalternativ är mest lämpliga för avloppsslammet i Avesta kommun. Gödsel till produktiv mark är ett av de föreslagna alternativ som tillåter fosforn återvinnas och återanvändas. Om inte termofil rötning som effektivisering hade erhållit tillräcklig hygienisering borde andra komplementerande processer ha införts, exempelvis komposteringstrumma eller pastörisering. En fullständig kostnadsjämförelse mellan befintlig slambehandlingsmetod och mer effektiviserad slambehandlingsmetod är dock nästan omöjlig att utföra då alla kostnader beror på val av komponenter och faktorer. Denna fallstudie bidrar med ett underlag som kan ge större möjligheter för Avesta Vatten och Avfall AB att välja den mest lämpliga avsättningen för avloppsslammet i Avesta kommun i framtiden. / Sewage sludge contains nutrients, which should be returned to nature to create a so-called nutrient circulation. By optimizing sanitisation of sewage sludge with effective treatment methods, sewage sludge can be more appropriately disposed from a better economic- and environmental perspective. The aim of this work was to investigate and judge the efficiency of existing sludge treatment is possible to achieve a sufficent sanitation. Based on the streamlined sludge treatment method investigate the disposal options that is best suited for sewage sludge in Avesta town. A literature review was conducted to increase the knowledge and understanding of sewage sludge, its treatment and disposal. The case study is formed with qualitative method and information which has been collected from the scientific articles, authorities, reports, theses and personal contact as a complement to obtain a good understanding. Upgrading of mesophilic digestion of sludge at 37 ° C to thermophilic digestion at 55 ° C - 65 ° C means increased temperature; this requires more energy and hence increases energy costs. Components of the mesophilic digestion process need to be upgraded or replaced and therefore additional costs should also be considered. The advantages of the thermophilic digestion include short residence time, smaller digester and sludge becomes sufficiently disinfected. A future scenario is developed based on more efficient sludge treatment where two disposal options are most appropriate for the sewage sludge in Avesta town. Returning digestate to productive land is one of those suggested alternatives that allows phosphorus to be recovered and recycled. If not thermophilic digestion as streamlining is not effective enough for sufficient sludge sanitation, other complementary processes have to be introduced, for instance, composting drum or pasteurization. A full cost comparison between the existing sludge treatment and more efficient sludge treatment method is, however, almost impossible to be performed when all costs depend on the choice of components, factors and so on. This case study contributes with first-hand information that can provide greater opportunities for Avesta Vatten och Avfall AB to select the most appropriate options for sewage sludge disposal in Avesta town in the future.

O hidrogênio é estudado como alternativa ao uso de combustíveis fósseis para geração de energia, uma vez que é um combustível renovável. Entre os processos de produção de hidrogênio destaca-se o processo fermentativo que possibilita unir tratamento de efluente e geração de energia. Uma das alternativas para esta produção é a utilização do reator anaeróbio de leito fluidificado (RALF). Dentre os resíduos industriais que podem ser utilizados para a produção de hidrogênio está o soro de queijo que, se descartado incorretamente, pode causar danos ao meio ambiente. Neste sentido, o presente estudo teve como principal objetivo avaliar a capacidade de produção contínua de hidrogênio, sob condições termófilas (55°C), a partir de diferentes concentrações de soro de queijo e tempos de detenção hidráulica (TDH) em RALF. Foram utilizados dois reatores, denominados R1 e R2 , no qual R1 manteve-se com concentração fixa de 5 gDQO.L-1, com variações de TDH de 8, 6, 4, 2, 1 e 0,5 hora e em R2 o TDH manteve-se fixo em 6 horas e variações de concentrações de 3, 5, 7 e 10 gDQO.L-1. Foi observado em R1 o comportamento de elevação de produção volumétrica de H2 (PVH) a partir da diminuição do TDH. A máxima PVH obtida foi de 2,9 L.h-1.L-1 em R1 para o TDH de 0,5 hora. No entanto, verificou-se a diminuição do rendimento de hidrogênio (HY) com a aplicação de TDHs reduzidos, alcançando valor máximo de 3,3 molH2.mol-1 lactose para o TDH de 4 horas. Para R2, o valor máximo de PVH de 0,854 L.h-1.L-1 para concentração de substrato de 7 g.L-1. Já para HY, o maior valor encontrado foi de 2,6 molH2.mol-1 lactose para a concentração de substrato de 3 g.L-1, uma vez que o aumento da concentração causou a diminuição de HY. O conteúdo de hidrogênio no biogás manteve-se na faixa de 48,2-54,2% e 39,8-48,4% para R1 e R2 respectivamente. Em ambos os reatores, os principais metabólitos encontrados nos efluentes foram ácido acético e ácido butírico. / Hydrogen is studied as an alternative to fossil fuels for power generation, since it is a renewable fuel. Between the processes of hydrogen production stands out the fermentation process that allows to merge wastewater treatment and power generation. An alternative to this production is the use of anaerobic fluidized bed reactor (AFBR). Among the industrial wastes can be used for hydrogen production is cheese whey that if improperly discarded, may cause damage to the environment. In this sense, the present study aimed to evaluate the ability of continuous hydrogen production under thermophilic conditions (55°C), from different concentrations of cheese whey and hydraulic retention time (HRT) in AFBR. Two reactors were used, denominated R1 and R2, in which R1 was maintained at a fixed concentration of 5gCOD.L-1 with variations in HRT of 8, 6, 4, 2, 1 and 0.5 hour and R2 kept fixed HRT at 6 hours, and changes in concentration of 3, 5, 7 and 10 gCOD.L-1. In R1, it was observed a volumetric H2 production (HPR) increase behavior from the HRT decrease. The maximum HPR obtained was 2.9 L.h-1.L-1 at HRT of 0.5 hour. However, there was a decrease in H2 yield (HY) with the application of reduce HRT, reaching maximum value of 3.3 molH2.mol-1 lactose at HRT of 4 hours. In R2, the maximum value of HPR was 0,854 L.h-1.L-1 for substrate concentration of 7 gCOD.L-1. The maximum HY obtained was 2.6 molH2.mol-1 lactose for concentration of 3 gCOD.L-1, once the value of HY decrease behavior from the concentration of cheese whey increase. The H2 content in biogas kept in range of 48.2-54.2% and 39.8-48.4% for R1 and R2, respectively. In both reactors, the main metabolites found were acetic and butyric acids.

Um sistema combinado anaeróbio-aeróbio composto de um filtro anaeróbio ascendente seguido de reator aeróbio de lodos ativados em batelada, foi continuamente alimentado com água residuária sintética da indústria de polpa celulósica não branqueada, durante o período de trezentos e sessenta e três dias, com o objetivo de verificar o desempenho dos reatores operados sob condições termofílicas. Os reatores foram inoculados com lodo mesofílico com gradual adaptação para a condição termofílica e operados em duas fases distintas. A primeira e segunda fases contemplam os períodos em que os reatores foram operados em temperatura termofílica de 55 e 65 graus Celsius, respectivamente. O filtro anaeróbio trabalhou com tempo de detenção hidráulica de 48 horas em fluxo contínuo e o reator aeróbio operado em bateladas seqüenciais (RBS) com tempos de ciclo de 12, 18 e 24 horas. A melhor eficiênica de remoção de DQO do sistema combinado anaeróbio-aeróbio foi de 65%, devida, em maior parte ao reator anaeróbio. O filtro anaeróbio operado em temperatura de 55 e 65 graus demonstrou eficiência na capacidade de retenção de biomassa termofílica, evitando a perda no efluente. O RBS, quando operado em temperatura de 55 graus e tempo de ciclo de 18 horas, demonstrou melhor eficiência para pós-tratamento de água residuária sintética. Os resultados gerais mostram uma melhor eficiência dos reatores operados em temperatura de 55 graus Celsius. / A combined anaerobic-aerobic system, composed of an upflow anaerobic filter followed by an aerobic reactor of activated sludge in batch, was continuously feed with synthetic wastewater from unbleached cellulose pulp plant during a period of three hundred and sixty three days, aiming to verify the performance of the reactors operating under thermophilic conditions. The reactors were inoculated with mesophilic sludge having a gradual adaptation to the thermophilic condition and they were operated in two distinct phases. The first and second phases contemplate the periods in which the reactors were operated in a thermophilic temperature of 55 and 65 Celsius degrees, respectively. The anaerobic filter worked with hydraulic detention time of 48 hours in a continuous flow, and the aerobic reactor was operated in a sequencing batch (SBR) with a cycle time of 12, 18 and 24 hours. The best efficiency of the combined anaerobic-aerobic system to remove DQO was 65% due to, in most part, the anaerobic reactor. The anaerobic filter, operating in temperatures of 55 and 65 degrees, has demonstrated efficiency in the capacity to retain thermophilic biomass, avoiding effluent lost. The SBR reactor, when operated in temperature of 55 degrees and a cycle time of 18 hours, has demonstrated better efficiency in the post-treatment of synthetic wastewater. The general results have showed a better efficiency of the reactors operating in a thermophilic temperature of 55 Celsius degrees.

O hidrogênio é estudado como alternativa ao uso de combustíveis fósseis para geração de energia, uma vez que é um combustível renovável. Entre os processos de produção de hidrogênio destaca-se o processo fermentativo que possibilita unir tratamento de efluente e geração de energia. Uma das alternativas para esta produção é a utilização do reator anaeróbio de leito fluidificado (RALF). Dentre os resíduos industriais que podem ser utilizados para a produção de hidrogênio está o soro de queijo que, se descartado incorretamente, pode causar danos ao meio ambiente. Neste sentido, o presente estudo teve como principal objetivo avaliar a capacidade de produção contínua de hidrogênio, sob condições termófilas (55°C), a partir de diferentes concentrações de soro de queijo e tempos de detenção hidráulica (TDH) em RALF. Foram utilizados dois reatores, denominados R1 e R2 , no qual R1 manteve-se com concentração fixa de 5 gDQO.L-1, com variações de TDH de 8, 6, 4, 2, 1 e 0,5 hora e em R2 o TDH manteve-se fixo em 6 horas e variações de concentrações de 3, 5, 7 e 10 gDQO.L-1. Foi observado em R1 o comportamento de elevação de produção volumétrica de H2 (PVH) a partir da diminuição do TDH. A máxima PVH obtida foi de 2,9 L.h-1.L-1 em R1 para o TDH de 0,5 hora. No entanto, verificou-se a diminuição do rendimento de hidrogênio (HY) com a aplicação de TDHs reduzidos, alcançando valor máximo de 3,3 molH2.mol-1 lactose para o TDH de 4 horas. Para R2, o valor máximo de PVH de 0,854 L.h-1.L-1 para concentração de substrato de 7 g.L-1. Já para HY, o maior valor encontrado foi de 2,6 molH2.mol-1 lactose para a concentração de substrato de 3 g.L-1, uma vez que o aumento da concentração causou a diminuição de HY. O conteúdo de hidrogênio no biogás manteve-se na faixa de 48,2-54,2% e 39,8-48,4% para R1 e R2 respectivamente. Em ambos os reatores, os principais metabólitos encontrados nos efluentes foram ácido acético e ácido butírico. / Hydrogen is studied as an alternative to fossil fuels for power generation, since it is a renewable fuel. Between the processes of hydrogen production stands out the fermentation process that allows to merge wastewater treatment and power generation. An alternative to this production is the use of anaerobic fluidized bed reactor (AFBR). Among the industrial wastes can be used for hydrogen production is cheese whey that if improperly discarded, may cause damage to the environment. In this sense, the present study aimed to evaluate the ability of continuous hydrogen production under thermophilic conditions (55°C), from different concentrations of cheese whey and hydraulic retention time (HRT) in AFBR. Two reactors were used, denominated R1 and R2, in which R1 was maintained at a fixed concentration of 5gCOD.L-1 with variations in HRT of 8, 6, 4, 2, 1 and 0.5 hour and R2 kept fixed HRT at 6 hours, and changes in concentration of 3, 5, 7 and 10 gCOD.L-1. In R1, it was observed a volumetric H2 production (HPR) increase behavior from the HRT decrease. The maximum HPR obtained was 2.9 L.h-1.L-1 at HRT of 0.5 hour. However, there was a decrease in H2 yield (HY) with the application of reduce HRT, reaching maximum value of 3.3 molH2.mol-1 lactose at HRT of 4 hours. In R2, the maximum value of HPR was 0,854 L.h-1.L-1 for substrate concentration of 7 gCOD.L-1. The maximum HY obtained was 2.6 molH2.mol-1 lactose for concentration of 3 gCOD.L-1, once the value of HY decrease behavior from the concentration of cheese whey increase. The H2 content in biogas kept in range of 48.2-54.2% and 39.8-48.4% for R1 and R2, respectively. In both reactors, the main metabolites found were acetic and butyric acids.

No description available.

Bakgrund: Med industrialiseringen har de traditionella yoghurtkulturerna med en mångfald av samverkande mjölksyrabakterier fått ge plats åt de mer standardiserade. En gårdskultur framställs genom att obehandlad mjölk spontanfermenteras av gårdens egen bakterieflora och får därigenom en unik karaktär. Bakteriekulturen kan sedan ympas vidare för tillverkning av yoghurt.Syfte: Syftet med studien är framställning och vidareympning samt sensorisk och mikrobiologisk karaktärisering av termofil gårdskultur från obehandlad mjölk. Vidareympningen avser framställning av yoghurt fermenterad av gårdens egen bakterieflora.Metod: Gårdskulturen framställdes och ympades till yoghurt. Yoghurten undersöktes genom mikrobiologisk karaktärisering, antibiotikaresistens och sensorisk profilering samt jämfördes med industriell kultur och en traditionell heirloomkultur.Resultat: Resultatet visade att gårdskulturen skiljde sig både mikrobiologisk och sensoriskt. Gårdskulturen innehöll stammar av enterokocker vilka inte visade på resistens mot analyserade antibiotika.Slutsats: Det är möjligt att framställa en gårdskultur av godtagbar mikrobiologisk och sensorisk kvalitet. Gårdskulturen ger en differentierad mikrobiologisk och sensorisk karaktär i jämförelse med en industriell kultur och en traditionell heirloomkultur Metoden kan vara riskfylld och kulturen bör analyseras med avseende på patogen tillväxt. En unik gårdsyoghurt kan vara en metod för gårdsmejerister att i sitt varumärke bygga på terroir och platsens unicitet. / Background: With industrialization, the traditional yoghurt cultures with a multitude of lactic acid bacteria had to make way for the more standardized. An artisanal farm culture is produced by raw milk spontaneously fermented by the farm's own bacterial flora and thus develops a unique character. The bacterial culture can then be inoculated for the production of yoghurt.Purpose: The pupose of the study is to produce and inoculate as well as sensory and microbiological characterization of a thermophilic artisanal farm culture from raw milk. The inoculation relates to the production of yoghurt fermented by the farm's own bacterial flora.Method: The artisanal farm culture was produced and inoculated into yoghurt which was assessed by microbiological characterization, antibiotic resistance, sensory profiling and then compared with industrial culture and a traditional heirloom culture.Result: The result showed that the artisanal farm culture differed both microbiologically and with regard to sensory paramters. The farm culture contained strains of enterococci which did not show resistance to analyzed antibiotics.Conclusion: It is possible to produce an artisanal farm culture of good microbiological and sensory quality. The artisanal farm culture provides a differentiated microbiological and sensory character in comparison to an industrial culture and a traditional heirloom culture The method may be risky and the culture should be analyzed for pathogens. A unique farm yoghurt can be a method for artisan farm dairies to build their brand based on terroir.

Orientador: Eleni Gomes / Banca: Luiz Juliano Neto / Banca: João Ruggiero Neto / Resumo: Fungos termofílicos têm despertado grande interesse acadêmico e industrial por produzirem uma variedade de enzimas termoestáveis com potenciais aplicações em processos biotecnológicos como biocatálise nas indústrias de couro, farmacêutica, têxtil e alimentícia, e na preparação de produtos de limpeza e cosméticos. Particularmente, as proteases, além de participarem de inúmeros processos fisiológicos vitais como vias metabólicas, hemostasia e sinalização celular, também representam hoje cerca de 60% do mercado mundial de enzimas. Neste trabalho, descrevemos a produção, purificação e caracterização bioquímica de uma serino-protease produzida por um fungo termofílico do gênero Myceliophthora. As taxas de atividade proteolítica foram avaliadas através de fermentação em meio sólido (FES) e submerso (FSM) e observou-se um rendimento na atividade proteolítica 4,5 vezes maior para o meio sólido. A enzima bruta obtida por ambos os procedimentos (FES e FSM) exibiu a mesma temperatura ótima de 50 ºC, porém em relação ao pH ótimo houve um deslocamento de 7 (FSM) para 9 (FES) sugerindo que o perfil enzimático do fungo difere de acordo com suas condições de fermentação. Baseado nesses resultados prosseguiu-se os estudos com o extrato bruto obtido por FES. A imobilização da enzima bruta em esferas de alginato de cálcio resultou no aumento da temperatura ótima e na estabilidade térmica quando comparado com a enzima livre. O extrato bruto obtido por FES foi, então, fracionado por métodos cromatográficos como exclusão molecular e troca-iônica que resultaram na protease pura com peso molecular de 28,2 kDa determinado por espectrometria de massa. A protease pura demonstrou pH ótimo de 9,0 e temperatura ótima de 45 °C que corroboram... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Thermophilic fungi have attracted great academic and industrial interest because they produce a variety of thermostable enzymes with potential applications in biotechnological processes such as biocatalysis in the industries of leather, pharmaceutical, textile and food, and the preparation of detergents and cosmetics. In particular, proteases not only participate in many vital physiological processes such as metabolic pathways, cell signaling and homeostasis, but also currently represent about 60% of the world market of enzymes. In this work, we describe the production, purification and biochemical characterization of a serine protease produced by a thermophilic fungus of the genus Myceliophthora. The levels of proteolytic activity were evaluated either by solid fermentation (SSF) and submerged (SmF). The crude enzyme obtained by both procedures (SSF and SmF) exhibited similar optimum temperature of around 50 ºC, but in relation to the optimum pH was shifted of 7 (SmF) to 9 (SSF), suggesting that the enzymatic profile of the fungus differs from according to its fermentation conditions. Based on these results, the studies were followed with crude extract obtained by SSF. The immobilized enzyme on beads of calcium alginate resulted in increased optimum temperature and thermal stability when compared to the free enzyme. The crude extract obtained by SSF was then fractionated by chromatographic methods including molecular exclusion and ion-exchange that resulted in the pure protease with molecular weight of 28.2 kDa as determined by mass spectrometry. The pure protease showed optimum pH of 9.0 and optimum temperature of 45 °C that corroborate to the preliminary characterization of the crude extract. Inhibition tests resulted in complete inhibition by PMSF, a canonical... (Complete abstract click electronic access below) / Mestre