Tierärztl. Hochsch., Diss., 2004--Hannover.

Τα τελευταία 30 χρόνια η βιοτεχνολογία χρησιμοποιεί υλικά που προέρχονται από ανανεώσιμες και φυσικές πηγές και έχει εφαρμογή στη χημεία τροφίμων. Πολλά πειράματα διεξάγονται με στόχο τη βελτίωση των τεχνικών κυτταρικής ακινητοποίησης. Ο νέος τομέας λέγεται "λευκή βιοτεχνολογία". Εκτός από τη γενικότερη εφαρμογή στη χημεία πιο συγκεκριμένα περιλαμβάνει επίσης καινούριες μεθόδους προκειμένου να αυξηθεί ο ρυθμός αλκοολικής ζύμωσης. Χρησιμοποιούνται κύτταρα, ένζυμα και οργανίδια για την κατάλυση μιας χημικής αντίδρασης με αποτέλεσμα να επιταχύνεται. Στην παρούσα εργασία έγινε μελέτη ζυμώσεων σε χαμηλές θερμοκρασίες ( στους 5°C, 15°C και 25°C. Η πειραματική διαδικασία ήταν μια καινοτόμος και τροποποιημένη μέθοδος σύμφωνα με τη οποία χρησιμοποιήθηκε ένας βιοκαταλύτης. Παρασκευάστηκε από θερμικά ξηραμένα και ακινητοποιημένα κύτταρα Saccharomyces secerevisiae ενός αλκοολανθεκτικού και κρυοανθεκτικού στελέχους ( ΑΧΑΖ-1 ) και απο δυο διαφορετικά υποστρώματα. Η πηγή των φυσικών υλικών ήταν το ξύλο (πριονίδι) και το άμυλο κριθαριού σε σκόνη. Μετά από απολιγνοποίηση το κατεργασμένο πριονίδι (18g) προστέθηκε σε προπαρασκευασμένο υπέρκορο διάλυμα αμύλου και αναμίχθηκε μαζί με ποσότητα μικροοργανισμού (10g) και θρεπτικού υλικού (10mL) στους 40°C. Ύστερα αποθηκεύτηκε στους 15°C. Και οι δυο φορείς είναι πολυμερή γλυκόζης. Τα κύτταρα εγκλωβίστηκαν στο gel πολυμερούς και επίσης ακινητοποιήθηκαν με προσρόφηση στο απολιγνοποιημένο κυτταρινούχο υλικό. Το θρεπτικό υλικό είχε 12% αρχική συγκέντρωση σακχάρων ( γλυκόζης/φρουκτόζης) με αποτέλεσμα η πυκνότητα να είναι 12 °Be. Οι ζυμώσεις πραγματοποιήθηκαν σε οικιακό ψυγείο εκτός από εκείνη που έγινε σε θερμοκρασία δωματίου. Η βιομάζα ακινητοποιημένων κυττάρων εκτιμήθηκε έμμεσα με τη μέθοδο Kjeldahl. Η παρατήρηση της μορφολογίας των κυττάρων έγινε με ηλεκτρονικό μικροσκόπιο σάρωσης καθώς οδηγεί σε συμπεράσματα για τη βιωσιμότητα. Η κινητική των ζυμώσεων υπο τη επίδραση της θερμοκρασίας υπολογίστηκε μετα από καταγραφή των μετρήσεων που έγιναν με πυκνόμετρο σε τακτά χρονικά διαστήματα. Η παραγωγικότητα αιθανόλης και η συγκέντρωση μεθανόλης που περιέχονταν στα δείγματα προσδιορίστηκαν με αέρια χρωματογραφία (με ανιχνευτή φλόγας , FID). Τέλος, τα πτητικά παραπροιότα συμπεριλαμβάνομένων των ακεταλδεύδη, οξικού αιθυλεστέρα, 1-προπανόλη, ισοβουτανόλη και αμυλικών αλκοόλών προσδιορίστηκαν με την ίδια αναλυτική μέθοδο, όπως αναφέρεται νωρίτερα. / -

Thesis (MSc)--Stellenbosch University, 2015. / ENGLISH ABSTRACT: Grape must gives rise to various stress conditions for the yeast inoculated for alcoholic fermentation. These include hyperosmotic stress due to the high initial sugar concentration and redox imbalances due to the fast depletion of oxygen. Under these stress conditions, Saccharomyces cerevisiae tends to produce glycerol as an osmoprotectant and to regenerate reducing equivalents. However, the production of glycerol often leads to increased acetic acid production. According to literature, it seems that many non-Saccharomyces yeasts have a different metabolic response to the above-mentioned stress conditions, especially since it has been found that they produce low levels of acetic acid. Only recently non-Saccharomyces yeasts were researched to be used as starter cultures in wine fermentations. It is found that they can confer beneficial characteristics to the resulting wine. However, most of the non-Saccharomyces yeasts lead to stuck fermentations as confirmed by this study. Therefore, if the positive characteristics of these yeasts were to be exploited in wine making they need to be inoculated together with S. cerevisiae. When two yeasts are inoculated together, they affect each other and consequently the wine. In this context, the aim of this study was to investigate the metabolic response to hyperosmotic stress during wine fermentation of the following wine-related non-Saccharomyces yeasts: Lachancea thermotolerans, Torulaspora delbrueckii and Starmerella bacillaris. Fermentations were performed in a synthetic grape must medium with pure cultures of the mentioned strains as well as mixed cultures of each non-Saccharomyces yeast with S. cerevisiae. The fermentation behaviour was monitored and concentrations of various wine-related metabolites were determined. Concerning polyol concentrations, S. cerevisiae produced only glycerol while the non-Saccharomyces yeasts also produced other polyols. The low production of acetic acid in the non-Saccharomyces fermentations was confirmed especially in the case of L. thermotolerans. Moreover, this yeast produced high levels of the higher alcohols butanol and propanol. St. bacillaris produced significant levels of acetoin and isobutyric acid and T. delbrueckii produced an increased concentration of succinic acid. All these metabolites might play a role in maintaining intracellular redox balance. However, a more extensive systematic study is needed to investigate the extent of their involvement. The mixed cultures completed fermentation and had higher final glycerol levels than the control and lower acetic acid concentrations and therefore can contribute positively to the wine aroma. Furthermore, the mixed culture fermentations showed the potential of lowering the ethanol concentrations of wine. Furthermore it has been shown in literature that the yeasts present in the mixed culture can affect each other on gene expression level as well. However, there is little genetic information available on non-Saccharomyces yeasts. In this study, we sequenced the genes involved in glycerol and acetic acid biosynthesis of L. thermotolerans and T. delbrueckii. The gene sequences are fairly homologous with only a few differences. These gene sequences can be used to study gene expression of GPD1 and ALD6 from fermentation samples in order to determine to what extent the yeasts in a mixed culture influence the gene expression of one another. / AFRIKAANSE OPSOMMING: Druiwemos gee oorsprong aan verskeie strestoestande vir die gis wat vir alkoholiese fermentasie geïnokuleer word. Hierdie strestoestande sluit hiper-osmotiese stres, as gevolg van die hoë suiker konsentrasie, in asook redoks wanbalanse toegeskryf aan die vinnige afname in beskikbare suurstof. Tydens hierdie toestande is Saccharomyces cerevisiae geneig om gliserol as beskerming teen die osmotiese stres te produseer, sowel as vir die regenereering van reduserings ekwivalente. Die produksie van gliserol lei egter dikwels tot toenemende asynsuur produksie. Volgens literatuur kom dit voor asof menige nie-Saccharomyces giste 'n ander metabolise reaksie tot die bogenoemde stresse het, omdat daar gevind is dat hulle laer vlakke van asynsuur produseer. Eers onlangs is navorsing gedoen op die potensiële gebruik van nie-Saccharomyces giste in gemengde kulture tydens wynfermentasies. Daar is bevind dat hulle voordelige eienskappe aan die wyn kan verleen. Meeste van die nie-Saccharomyces giste lei egter tot onvolledige fermentasies soos bevesting deur hierdie studie. Dus, indien die positiewe eienskappe van hierdie giste sou benut word in wynmaak sal hulle saam met S. cerevisiae geïnokuleer moet word. Wanneer twee giste saam geïnokuleer word, beïnvloed hulle mekaar en gevolglik die wyn. In hierdie konteks was die doel van die betrokke studie om die metaboliese reaksie tot hiperosmotiese stress tydens wynfermentasies te ondersoek in die volgende wyn verwante nie-Saccharomyces giste: Lachancea thermotolerans, Torulaspora delbrueckii en Starmerella bacillaris. Fermentasies was in sintetiese druiwemos medium uitgevoer met rein kulture van die genoemde gisrasse, sowel as gemengde kulture van elke nie-Saccharomyces gis met S. cerevisiae. Die fermentasiegedarg is gemonitor en die konsentrasies van verskeie wyn verwante metaboliete is bepaal. Wat die poliol konsentrasies betref, het S. cerevisiae slegs gliserol geproduseer terwyl die nie-Saccharomyces giste additionele poliole ook geproduseer het. Die lae produksie van asynsuur in die nie-Saccharomyces fermentasies is bevestig, veral in die geval van L. thermotolerans. Verder produseer hierdie gis hoë vlakke van asetoïen en iso-bottersuur en T. delbrueckii produseer 'n hoër konsentrasie van suksiensuur. Al hierdie metaboliete mag 'n rol speel in die handhawing van intrasellulêre redoksbalans. 'n Meer uitgebreide, sistematiese studie is egter nodig om die mate van hul betrokkenheid te ondersoek. Die gemengde kulture het hul fermentasies voltooi en het hoër finale gliserol vlakke as die kontrole gehad, asook laer asynsuur konsentrasies en kan dus positief bydra tot die wyn aroma. Verder het die gemengde kultuur fermentasies die potensiaal om die etanol vlakke van wyn te verlaag, getoon. Daar is verder in die literatuur gevind dat die giste teenwoordig in die gemengde kultuur mekaar op geenuitdrukkings vlak ook kan beïnvloed. Daar is egter min genetiese inligting beskikbaar vir die nie-Saccharomyces giste. In hierdie studie het ons die gene betrokke by die produksie van gliserol en asynsuur van L. thermotolerans en T. delbrueckii se nukleotied volgordes bepaal. Die gevolglike nukleotied volgordes is redelik homoloog met net 'n paar verskille. Hierdie volgordes kan gebruik word om die geenuitdrukking van GPD1 en ALD6 vanaf fermentasie monsters te bestudeer om sodoende te bepaal tot watter mate die giste in 'n gemengde kultuur mekaar se geenuitdukking kan beïnvloed.

Thesis (MSc)--Stellenbosch University, 2015. / ENGLISH ABSTRACT: The use of non-Saccharomyces yeasts together with the yeast S. cerevisiae in multistarter wine fermentations has emerged as a useful tool to modulate wine aroma and/or to decrease the concentration of undesirable compounds. However, upon inoculation, these yeast species do not co-exist passively, but interact in various ways. While competition for nutrients and the excretion of killer toxins in an antagonistic relationship are obvious and well established types of interactions, some studies have suggested the existence of other forms of cellular or molecular interactions. One of these includes physical cell-cell contact and to our knowledge, only one previous study has confirmed its existence in wine yeasts. Yeast interactions are also influenced by other factors, such as ethanol concentration, however some studies have highlighted the role that dissolved oxygen plays on the survival of non-Saccharomyces yeasts and their ability to compete for space with S. cerevisiae and little research has focused on this. This study aimed to investigate the occurrence of a physical cell-cell and/or metabolic interaction between S. cerevisiae and L. thermotolerans in mixed culture fermentations of synthetic grape must. For this purpose, fermentations in a Double Compartment Bioreactor (DCB) which separates yeast population through the use of a membrane were compared to mixed fermentations in the absence of the membrane, using the same reactor. Furthermore, the impact of oxygen supply on yeast behaviour was also assessed. Following mixed culture fermentations in a DCB, it was observed that the presence of S. cerevisiae led to a significant decline in viability in L. thermotolerans. This decline was significantly less prominent in mixed cultures where the cells were in indirect contact. Together, the data provided evidence for both cell-cell and metabolic interactions whereby S. cerevisiae had a strong negative influence on the growth of L. thermotolerans. However, it was also observed that L. thermotolerans had some negative impact on the growth of S. cerevisiae, leading to a reduction in biomass (when in indirect contact) and a reduced maximum CFU/mL compared to pure cultures. The data also suggest that direct physical contact may increase the production of glycerol and propanol, but this needs further investigation. By decreasing the frequency at which oxygen pulses were provided, a reduction in biomass and increase in fermentation duration was observed for all fermentations. However, this effect was somewhat reduced in mixed cultures. Here, no impact on fermentation duration was observed and the decrease in biomass was less compared to pure cultures. The impact of these oxygen pulses was also greater on L. thermotolerans. In the latter yeast’s pure culture a slight increase in glycerol was observed when less oxygen was provided and in general there appeared to be no impact on acetic acid production. Furthermore, there was little or no impact on volatile production, however, more repeats might reveal different results and therefore more research is needed to confirm these results. To our knowledge, this is the first study of its kind to confirm a physical cell-cell interaction between the yeast pair S. cerevisiae and L. thermotolerans. / AFRIKAANSE OPSOMMING: Die gebruik van nie-Saccharomyces gis saam met die gis S. cerevisiae in multi-inokuleringskulture het die afgelope paar jaar as n goeie hulpmiddel na vore gekom om wyn aroma te moduleer en/of om die konsentrasie van ongewensde verbindings te verminder. Sodra inokulasie plaasgevind het, het hierdie gis die potensiaal om op verskeie maniere teenoor mekaar te reageer. Kompetisie vir nutriente en die afskeiding van toksiese verbindings in n antagonistiese verhouding is alreeds goed beskryf in die literatuur. Somige studies het, alhoewel, die bestaan van ander vorme van sellulêre of molekulêre interaksies voorgestel. Een van hierdie sluit in n fisiese sell-sell interaksie en so ver as wat ons kennis strek, het nog net een studie van tevore so ‘n interaksie bevestig tussen wyn giste. Gis interaksies word ook beïnvloed deur ander faktore, soos byvoorbeeld etanol konsentrasie. Terwyl sommige studies die rol wat opgelosde suurstof speel in die oorlewing van nie-Saccharomyces gis en hulle vermoë om te kompeteer vir spasie met S. cerevisiae alreeds beklemtoon, het min navorsing al hierop gefokus. Hierdie studie het gestreef om die voorkoms van n fisiese sell-sell en/of metaboliese interaksie tussen S. cerevisie en L. thermotolerans in gemengde kultuur fermentasies van sintetiese druiwe sap te ondersoek. Vir hierdie doeleinde was fermentasies uitgevoer met behulp van ‘n Dubbel Kompartement Bioreaktor (DKB) wat gis populasies skei deur middel van ‘n membraan en hierdie was vergelyk met gemengde kultuur fermentasies sonder die membraan in dieselfde reaktor sisteem. Verder was die impak van suurstof toevoer op gis gedrag ook geassesseer. Na afloop van gemengde kultuur fermentasies in ‘n DKB, was daar waargeneem dat die teenwoordigheid van S. cerevisiae gelei het tot ‘n betekenisvolle afname in lewensvatbaarheid in L. thermotolerans. Hierdie afname was aansienlik minder in gemengde kulture waar die gis in indirekte kontak was. Saam verskaf hierdie data bewyse vir n sell-sell asook metaboliese interaksie waardeur S. cerevisiae ‘n sterk, negatiewe invloed op die groei van L. thermotolerans gehad het. Daar was egter ook waargeneem dat L. thermotolerans tot ‘n mindere mate ‘n negatiewe impak op die groei van S. cerevisiae gehad het en dat dit gelei het tot ‘n verlaging in biomassa (toe die gis in indirekte kontak was) en ‘n verlaagde maksimum CFU/mL in vergelyking met suiwer kulture. Die data dui ook aan dat fisiese kontak kon gelei het tot ‘n verhoging in gliserol en propanol produksie, maar hierdie kort verdere ondersoek. Deur die frekwensie te verminder waardeur suurstof pulse aan die fermentasies verskaf was, was ‘n verlaging in biomassa produksie en ‘n verlenging in fermentasie tydperk waargeneem. Hierdie tendense was waargeneem in almal, behalwe die gemengde kultuur fermentasies. Die effek van suurstof puls verlaging was minder op hierdie fermentasies aangesien daar geen impak op fermentasie tydperk was nie en die verlaging in biomassa minder was. Die impak van hierdie suurstof pulse was ook groter op L. thermotolerans. ‘n Klein toename in gliserol produksie was waargeneem in laasgenoemde gis se suiwer kultuur toe minder suurstof beskikbaar was en oor die algemeen was asynsuur onveranderd. Verder was daar ‘n klein of geen impak op vlugtige verbindings nie, alhoewel, meer herhalings mag verskillende resultate lewer en daarom is meer navorsing nodig om hierde resultate te bevestig. So ver as wat ons kennis strek is hierdie die eerste studie van sy soort om ‘n fisiese sell-sell interaksie tussen die gispaar S. cerevisiae en L. thermotolerans te bevestig.

Resumo: A principal meta deste trabalho foi a de realizar estudos com leveduras industriais brasileiras utilizadas em usinas, na tentativa de viabilizar o emprego da Tecnologia de Fermentação de Mostos com Altos Teores de Açúcares Fermentecíveis, na produção de etanol combustível. Inicialmente, foram realizados experimentos para obtenção de informações relativas as linhagens, na fermentação de sacarose, maltose e glicose, em meios contendo uma base nitrogenada e suplementado com fontes de nitrogênio com diferentes complexidades estruturais. Os estudos mostraram que as linhagens industriais apresentam diferentes perfis de crescimento nos meios suplementados com diferentes fontes de nitrogênio, que variaram de um simples sal de amônio, a um hidrolisado ácido de proteínas (casaminoácidos) e hidrolisado enzimático de proteínas (peptona). Entretanto, a maior capacidade de acúmulo de biomassa pode não ter um reflexo direto na capacidade de utilização de açúcares e a consequente produção de etanol pelas leveduras industriais. Além da natureza estrutural da fonte de nitrogênio e do tipo de açúcar, a presença do oxigênio, em maior quantidade nos cultivos agitados, interferiu sobremaneira no desempenho fermentativo das leveduras industriais, e até no acúmulo de trealose. No geral, foi a suplementação com peptona que propiciou maior produção de biomassa, preservação da viabilidade celular e consumo mais eficiente da fonte de carbono, em cultivos agitados e não agitados. Os estudos com alta densidade celular tiveram como objetivo definir condições experimentais para a condução da Fermentação de Mosto com Altos Teores de Açúcares Fermentescíveis, e que pudessem propiciar ao final do processo a completa utilização da sacarose, associada á preservação da viabilidade... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: In this work studies with Brazilian yeast for ethanol production were conducted in order to verify the possibility of utilization of brazilian yeasts for ethanol production in an industrial process known as Very High Gravity Sucrose Fermentation Technology, to produce wine with high ethanol contends. This technology has the advantage to reduce production costs by increasing fermentation yields. Initially, studies were conducted with sucrose, maltose and glucose fermetation in medium containing YNB, supplemented with a nitrogen source varying from a single ammonium salt (ammonium sulfate) to free amino acids (casamino acids) and peptides (peptone). Data suggest that yeast strains vary in their response to nitrogen source complex structure, kind of sugar and to oxygen availabity. In general, under peptone supplementation all strains, in shaking and static conditions, showed higher biomass accumulation, efficient sugar utilization and yeast viability was preserved. Sugar utilization by industrial strains not always was directly correlated with higher biomass accumulation. Trehalose accumulation was also influenced by the structural complexity of nitrogen sources, the kind of sugar and the presence of oxygen. Studies with high cell density were conducted to define experimental parameters for Very High Gravity Sucrose Fermentation, in order to induce at the end of process full sugar exhaustion together cell viability preservation. Complete sucrose utilization was detected only in media with 22 and 25% (w/v) sucrose. In the presence of higher sucrose contends (30% (w/v)), total sucrose exhaustion was obtained in a sugar cane based medium, supplemented with 2% (w/v) peptone. The results described in this thesis suggest that industrial yeasts show differing nitrogen demand, and the utilization of Very High Gravity Sucrose Fermentation technology could be carried only after finding the appropriated nutritional and fermentation conditions / Orientador: José Roberto Ernandes / Banca: João Atilio Jorge / Banca: Jonas Contiero / Banca: Sandra Helena da Cruz / Banca: Luis Henrique Souza Guimarães / Doutor

Submitted by Lucienne Costa (lucienneferreira@ufcg.edu.br) on 2018-03-23T20:07:03Z No. of bitstreams: 1 MARCIO JOSÉ VASCONCELOS DA SILVA – TESE (PPGEP) 2017.pdf: 2886790 bytes, checksum: 4e2912ac8b6df6d1df2aeb12d63b2646 (MD5) / Made available in DSpace on 2018-03-23T20:07:03Z (GMT). No. of bitstreams: 1 MARCIO JOSÉ VASCONCELOS DA SILVA – TESE (PPGEP) 2017.pdf: 2886790 bytes, checksum: 4e2912ac8b6df6d1df2aeb12d63b2646 (MD5) Previous issue date: 2017-12-05 / Na busca por fontes de energia renováveis e alternativas que reduzam a emissão de gases nocivos ao meio ambiente, diminuindo assim o uso de combustíveis fósseis em face da conscientização global, os biocombustíveis são uma fonte alternativa de energia, sendo principal destaque o etanol de 1ª e 2ª geração. Objetivou-se neste trabalho avaliar o potencial de diferentes cultivares de sorgo doce (Sorghum bicolor (L.) Moench.) para produção de etanol de 1ª geração a partir da fermentação do caldo dos colmos e etanol de 2ª geração a partir do substrato (bagaço). Para obtenção dos açúcares fermentescíveis a partir do bagaço, utilizou-se pré-tratamento ácido seguido de básico com auxílio de uma matriz de planejamento experimental 2³+3 pontos centrais para avaliar os efeitos das variáveis de entrada, tempo, temperatura e concentrações de ácido e base, verificando a solubilização da hemicelulose e lignina. Para avaliar a hidrólise enzimática verificou-se, por meio da aplicação de uma matriz de planejamento experimental 2²+3 pontos centrais, o efeito da carga enzimática e a razão massa seca de bagaço de sorgo e extrato enzimático. Foram usadas as enzimas comerciais Celluclast 1.5L da Novozyme e betaglicosidase da Prozyn na hidrólise enzimática para obtenção de açúcares fermentescíveis. A pesquisa foi desenvolvida no laboratório de engenharia bioquímica – LEB, na Universidade Federal de Campina Grande – UFCG, no estado da Paráiba e no Instituto Agronômico de Pernambuco – IPA, ambas localizadas na região do nordeste Brasileiro, a colheita das progens foi realizada na Estação Experimental do IPA no município de Vitória de Santo Antão, localizado na Zona da Mata de Pernambuco (lat: 35° 22' W, long: 08° 08' S, altitude: 146 m) no período de março/2013 a setembro/2017. Os resultados referentes aos dados agronômicos apresentaram diferenças significativas para um intervalo de confiança de 95% pelo teste Tukey. Dentre as sete cultivares pesquisadas as que obtiveram maiores teores de sólidos solúveis totais em campo foram entre BRS506, IPA2502, IPA EP17, IPA P222, IPA P134, IPA P228 e IPA SF15 com valores entre 12,3±0,08; 5,1±0,10; 16,8±0,40; 14,8±0,11; 7,1±0,10; 9,2±0,10 e 13,6±0,27 ºBrix, respectivamente. Os resultados referentes a fração sacarina (caldo do sorgo sacarino) foram analisados usando o teste de Tukey para um intervalo de confiança de 95%. Nos dados encontrados para concentração de etanol de 1ª geração, obtidos na fermentação dos caldos das cultivares que se destacaram foram IPA SF15, BRS506, IPA EP17, IPA P222 e IPA P134 com valores entre 55,72 ± 0,24; 45,46 ± 1,5; 44,6 ± 0,18; 33,78 ± 0,57 e 31,78 ± 0,28 g.L-1 respectivamente. As frações (bagaço) para avaliação na produção de etanol de 2ª geração, as cultivares IPA EP17 e BRS506, após pré-tratamento ácido seguido de básico, mostraram-se eficientes em concentrar a celulose, pela solubilização da hemicelulose e lignina, acarretando um aumento da celulose de 31,70 ± 0,49 para 60,42 ± 0,91% na variedade IPA-EP17 e para variedade BRS506 de 40,6 ± 1,1 para 64,52 ± 0,82% respectivamente. Os resultados encontrados no processo de hidrólise enzimática dos bagaços das variedades IPA EP17 para produção de glicose foi de 26,23 g L-1, nas condições de carga enzimática e razão (relação massa seca de bagaço) por volume de meio reacional (25 FPU/g e 2%) no tempo de 36 h de hidrólise e rotação constante de 150 RPM, e para BRS506 a produção de glicose foi de 25,17 g L-1, nas condições de carga enzimática e razão (relação massa seca de bagaço) por volume de meio reacional (25 FPU/g e 2%) no tempo de 36 h de hidrólise. / In the search for renewable energy sources and alternatives that reduce the emission of harmful gases to the environment, thus reducing the use of fossil fuels in the face of global awareness, biofuels present as an alternative, the main highlight being ethanol. The objective of this work was to evaluate the potential of different cultivars of sweet sorghum (Sorghum bicolor (L.) Moench.) For the production of ethanol of 1st generation from the fermentation of the broth the 2st generation ethanol from the substrate (bagasse). To obtain the fermentable sugars from the bagasse, acid pretreatment followed by base with the aid of experimental planning 2³ + 3 central points was applied to evaluate the effects of the variables of entry, time, temperature and acid and base concentrations, verifying the solubilization of hemicellulose and lignin. In order to evaluate the enzymatic hydrolysis, the effect of the enzymatic loading and the dry mass ratio of sorghum bagasse and enzyme extract wereverified using experimental planning design 2² + 3 central points. The commercial enzymes Celluclast 1.5L from Novozyme and beta-glucosidase from Prozyn wereused in the enzymatic hydrolysis to obtain fermentable sugars. The research was carried out at the biochemical-engineering laboratory - LEB, at the Federal University of Campina Grande - UFCG, in the state of Paráiba and at the Agronomic Institute of Pernambuco - IPA, both located in the northeast region of Brazil. The progenies were collected at the IPA Experimental Station in the city of Vitória de Santo Antão, located in the Zona da Mata of Pernambuco (lat: 35 ° 22 'W, long: 08 ° 08' S, altitude: 146 m) from March / 2013 to September / 2017. The results concerning agronomic data presented significant differences for a 95% confidence interval by the Tukey test. Among the seven cultivars studied, the highest soluble solids in the field were BRS506, IPA2502, IPA EP17, IPA P222, IPA P134, IPA P228 and IPA SF15 with values between 12,3 ± 0,08; 5,1 ± 0,10; 16,8 ± 0,40; 14,8 ± 0,11; 7,1 ± 0,10; 9,2 ± 0,10 and 13,6 ± 0,27 °Brix, respectively. Results for saccharin fraction (sorghum broth) were analyzed using the Tukey's test for a 95% confidence interval. In the data obtained for the concentration of ethanol of 1st generation, obtained in the fermentation of the broths of the cultivars that stood out were IPA SF15, BRS506, IPA EP17, IPA P222 and IPA P134 with values between 55,72 ± 0,24; 45,46 ± 1,5; 44,6 ± 0,18; 33,78 ± 0,57 and 31,78 ± 0,28 g.L-1 respectively. The fractions (bagasse) for evaluation in the production of second generation ethanol, IPA EP17 and BRS506, after acid pretreatment followed by basic, were efficient in concentrating the cellulose by the solubilization of hemicellulose and lignin, resulting in an increase of cellulose from 31,70 ± 0,49 to 60,42 ± 0,91% in the variety IPA-EP17 and for the BRS506 variety from 40,6 ± 1,1 to 64,52 ± 0,82% respectively. The results obtained in the enzymatic hydrolysis process for the IPA EP17 variety in the glucose production were 26,23 g L-1, for enzyme loading conditions and ratio (bagasse dry mass ratio) per reaction medium volume (25 FPU / and 2%) in the time of 36 hours of hydrolysis and constant rotation of 150 RPM, for BRS506 variety the glucose production was 25,17 g.L-1, for the conditions of enzymatic loading and ratio (bagasse dry mass ratio) by volume of reaction medium (25 FPU / g and 2%) in the time of 36 hours of hydrolysis.

Made available in DSpace on 2014-06-11T19:32:56Z (GMT). No. of bitstreams: 0 Previous issue date: 2011-12-16Bitstream added on 2014-06-13T21:05:19Z : No. of bitstreams: 1 silva_zf_dr_jabo.pdf: 288941 bytes, checksum: c6a44e855aa5751be49544caeb8b11a1 (MD5) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / Fundação de Amparo à Pesquisa e ao Desenvolvimento Científico do Maranhão (FAPEMA) / Avaliou-se efeito da inclusão de 0, 15 e 30% de glicerina bruta (GB) na matéria seca (MS) de dietas em substituição ao milho grão moído sobre o consumo de MS, produção de leite (PL), fermentação ruminal e digestibilidade da MS (DMS) em vacas leiteiras. Foram utilizadas seis vacas multíparas, da raça Holandesa providas de cânula permanente de 4” no rúmen, estando no início do experimento com peso médio de 587 ± 39 kg, com 114 ± 29 dias em lactação e produzindo média de 20 ± 1,5 kg/leite em duas ordenhas diárias. As vacas foram distribuídas em dois quadrados latinos 3 x 3 com períodos de 21 dias, sendo 14 de adaptação. As dietas continham silagem de milho, milho grão, farelo de girassol, glúten de milho, uréia, vitaminas, minerais e GB (86% de glicerol, 95% MS, 6% sais e <100 g/kg de metanol) e foram oferecidas ad libitum, duas vezes ao dia, na forma de dieta total. A dieta controle - ausência de GB, continha 36% de milho; a dieta com 15% e 30% de GB continham, respectivamente, 19,3 e 2,8% de milho. A utilização da GB na dieta reduziu a PL (P=0,10), o consumo de MS (P=0,08), os tempos gastos com atividade mastigatória (P<0,03), a digestibilidade da FDN (P<0,04) e a produção de bactérias associadas à fase líquida do rúmen (P=0,04). Não foram observadas diferenças quanto a cinética de degradação e passagem. Porém, houve aumento na concentração de metano quando a GB foi incluída na dieta. A utilização de glicerina bruta (86% de glicerol) em 15% da matéria seca da dieta de vacas em lactação deprime produção de bactérias de fase líquida, digestibilidade da fibra e desempenho. A sua inclusão em até 10% parece ser o limite ótimo / The purpose of this work was to evaluate the effect of the inclusion of 0, 15 and 30% crude glycerin (CG) in the MS diet in replacement of corn grain (diet control) on the ground on the dry matter intake (DMI), milk production (MY), rumen fermentation and dry matter digestibility in dairy cows . We used six multiparous Holstein cows rumen cannulated, being at the experiment with 587±39 kg BW, 114±29 DIM and with average 20±1.5 kg/d MY in two daily milkings, distributed in two 3x3 Latin Square with 21-days periods. The diets contained corn silage (45% DM), corn grain, sunflower meal, corn gluten meal, urea, vitamins, minerals and CG (86% glycerol, 95% DM, salts 6% and <0.1ppm of methanol). Cows were individually fed on total mixed ration in twice daily. The control diet - no glycerin, containing 36% corn, the diet with 15% and 30% contained GB, respectively, 19.3 and 2.8% of the corn. The use of the CG in the diet reduced the MY (P=0.10), DMI (P=0.08), the time chewing activities (P<0.03), NDF digestibility (P=0.04) and the liquid-associated bacteria content (P=0,04). There were no differences in the kinetics of degradation and passage. However, increased methane output when GB was in the diet. The use of crude glycerin (86% glycerol) in 15% of the diet dry matter for lactating cows depresses production of bacteria in the liquid phase, fiber digestibility and performance. Inclusion in up to 10% seems to be the optimum limit

Made available in DSpace on 2014-06-11T19:32:58Z (GMT). No. of bitstreams: 0 Previous issue date: 2004-07-30Bitstream added on 2014-06-13T19:22:53Z : No. of bitstreams: 1 coalho_mr_dr_botfmvz.pdf: 157837 bytes, checksum: a8e4ecfef81594485e4f922fe2a6be19 (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Universidade Estadual Paulista (UNESP) / O experimento foi realizado no Departamento de Zootecnia da Faculdade de Zootecnia e Engenharia de Alimentos da Universidade de São Paulo no Campus da USP, em Pirassununga-SP. Foram utilizados 4 bovinos da raça Nelore, castrados, com peso corporal médio de 550 kg e providos de cânulas ruminais, alimentados com dietas contendo diferentes níveis de sais de cálcio de ácidos graxos: 0, 1, 2, e 4%, em um delineamento em quadrado latino (4x4). Amostras de líquido ruminal foram colhidas e submetidas a mensurações de pH, variação da amônia ruminal e coletas de sangue para a determinação da uréia plasmática. O pH não foi influenciado pelos níveis de sais de cálcio de ácidos graxos (P>0,05), bem como, para o efeito tempo (P>0,05); indicando ser pouco provável ter ocorrido dissociação de sais de cálcio de ácidos graxos em relação aos níveis de inclusão. Os efeitos de tratamento e de tempo não apresentaram diferenças estatísticas significativas (P>0,05) na concentração de amônia no rúmen, demonstrando que os sais de cálcio de ácidos graxos não interferiu na fermentação ruminal. Observou-se que os valores de uréia plasmática (NUS) não foram significativos nos efeitos de tratamento e tempo (P>0,05) pelos níveis de sais de cálcio de ácidos graxos. / The experiment was carried out at the Faculdade de Zootecnia e Engenharia de Alimentos of the Universidade de São Paulo. Four crossbred Nerolle steers with 550 kg of live weight and fitted with rumen cannulas were fed diets with 0, 1, 2 or 4% of calcium salts of fatty acids, in 4 x 4 a latin square design. Rumen liquid sampled was for pH, rumen ammonia concentration and plasma urea measurements. The pH of the rumen liquid was not influenced by the levels of calcium salts of fatty acids (P>0,05), as well as, there was no influence of sampling time on pH, indicating that probably there was no dissociation of the calcium salts of fatty acids. The sampling time and treatment also had no effect (P>0,05) on the ammonia concentration in the rumen, demonstrating that the calcium salts of fatty acids had no effect in the ruminal fermentation. It was not observed significant effects of calcium salts of fatty acids level or time on plasmatic urea values (NUS) (P>0,05).

Orientador: Jonas Contiero / Banca: Henrique Ferreira / Banca: Danielle Biscaro Pedrolli / Banca: Antonio José Gonçalves da Cruz / Banca: Luiz Carlos Basso / Resumo: O glicerol de biodiesel pode ser utilizado para produção de 1,3 propanodiol por bactérias. Nesse estudo sobre a produção de 1,3 propanodiol por Klebsiella pneumoniae GLC29 foi possível otimizar o meio de cultura para a produção de 1,3-propanodiol utilizando técnicas estatísticas de planejamento experimental e superfície de resposta, onde foram experimentados 11 possíveis variáveis no meio de cultura e verificou-se a influência de apenas 5 (glicerol, extrato de levedura, sulfato de amônio, vitamina B12 e Fumarato). Posteriormente, verificou-se a produção desse composto em bioreatores de 0,75 litros, tanto em batelada como batelada alimentada exponencial, onde foi possível atingir 23.6 g/L de 1,3-propanodiol em batelada usando glicerol puro, 27 g/L de 1,3-propanodiol em batelada usando glicerol bruto de biodiesel e 29.9 g/L de 1,3-propanodiol usando glicerol bruto de biodiesel por batelada alimentada exponencial. Além da otimização, foi possível amplificar e clonar todos os genes responsáveis à produção de 1,3-propanodiol de Klebsiella pneumoniae GLC29 em um plasmídeo usando a técnica Gibson Assembly, um método recente de montagem isotermal. Os genes foram expressos em cepas de Escherichia coli, e foi então verificada a produção de 1,3-propanodiol em frascos e bioreatores, onde foi possível atingir 11 g/L de 1,3-propanodiol produzido por E. coli SZ63 contendo o plasmídeo pSB1C3-dhaB123TFG em batelada alimentada / Abstract: Biodiesel glycerol can be used for the production of 1,3-propanediol by bacteria. In this study on the production of 1,3-propanediol by Klebsiella pneumoniae GLC29 was possible to optimize media culture the production of 1,3-propanediol using statistical experimental design techniques and response surface methodology, where 11 variables were tested and only 5 were found significant (glycerol, yeast extract, ammonium sulfate, vitamin B12 and fumarate). Subsequently, production of 1,3-propanediol was verified in 0,75 liter reactors, both in batch and exponential fed-batch. It was possible to achieve 23.6 g/L of 1,3- propanediol in batch cultures using pure glycerol, 27 g/L of 1,3-propanediol in batch cultures using biodiesel glycerol, and 29.9 g/L of 1,3-propanediol in exponential fed-batch using biodiesel glycerol. Also, it was possible to amplify and clone all the genes responsible for the production of 1,3-propanediol by Klebsiella pneumoniae GLC29 in a plasmid, using Gibson assembly, a recent isothermal assembly methodology. The genes were expressed in Escherichia coli strains, and it was then verified the production of 1,3-propanediol in flasks and bioreactors, in which it was possible to reach 11 g/L 1,3-propanediol produced by E. coli SZ63 with the plasmid pSB1C3-dhaB1234TFG in fed-batch cultures / Doutor

This Mater´s Degree Thesis focuses on the sensory analysis of the chosen products of the non-perishable meat products in the dependence on the production technology. Its aim is to compare what kind of the products is prefered by the consuments using the sensory analysis and then statistic evaluation. The theoretical part summarizes the findings of the sensory analysis of the products, it also includes the conditions of the analysis and the statistic evaluation, sense perception in general and the production technology of meat products. In the practical part is described the method and the results of the hedonic test performed. In the thesis were used for evaluation the samples of the non-perishable meat products of the leading Czech producers of non-perishable meat products distributed and available in the shops in the Czech Republic.