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Submitted by Divis??o de Documenta????o/BC Biblioteca Central (ddbc@ufam.edu.br) on 2018-04-11T13:11:41Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Tese_Neuza B. Barros.pdf: 2710465 bytes, checksum: 4f522332291442b7edb66a372b6259ce (MD5) Approved for entry into archive by Divis??o de Documenta????o/BC Biblioteca Central (ddbc@ufam.edu.br) on 2018-04-11T13:11:55Z (GMT) No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Tese_Neuza B. Barros.pdf: 2710465 bytes, checksum: 4f522332291442b7edb66a372b6259ce (MD5) Made available in DSpace on 2018-04-11T13:11:55Z (GMT). No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Tese_Neuza B. Barros.pdf: 2710465 bytes, checksum: 4f522332291442b7edb66a372b6259ce (MD5) Previous issue date: 2017-12-04 CAPES - Coordena????o de Aperfei??oamento de Pessoal de N??vel Superior Cutaneous leishmaniasis (CL) is endemic in several regions of the world. In Brazil, despite the wide variety of clinical manifestations, 75% of patients with CL display a typical clinical picture, characterized by one or more ulcers and high round edges with a central necrosis, usually in the lower extremities. Leishmania amazonensis is a major etiologic agent of a wide spectrum of clinical forms of leishmaniasis. The objective of this study was to investigate the in vitro and in vivo antileishmania amazonensis activity of phospholipase A2 (Asp49) isolated from Bothrops jararacussu venom encapsulated in liposomal system. The cytotoxicity of PLA2-Asp49-liposomal was determined by MTT method in J774 macrophages after 48h of incubation. Macrophages treated with PLA2-Asp49-liposomes had cell viability around 82% at the highest concentration (100 ??g/mL), while promastigotes treated with the same concentration resulted 78% of inhibition. On the other hand, peritoneal macrophages from Balb/c mice infected and treated with PLA2-Asp49-liposomes showed a 28% reduction in phagocytic index and a 55% reduction (p

Submitted by Divis??o de Documenta????o/BC Biblioteca Central (ddbc@ufam.edu.br) on 2018-04-11T13:55:32Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Tese_Kaynara D. Zaqueo.pdf: 8755372 bytes, checksum: d4cb41bc00bf604cbe13dca0bf8c1774 (MD5) Approved for entry into archive by Divis??o de Documenta????o/BC Biblioteca Central (ddbc@ufam.edu.br) on 2018-04-11T13:55:49Z (GMT) No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Tese_Kaynara D. Zaqueo.pdf: 8755372 bytes, checksum: d4cb41bc00bf604cbe13dca0bf8c1774 (MD5) Made available in DSpace on 2018-04-11T13:55:49Z (GMT). No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Tese_Kaynara D. Zaqueo.pdf: 8755372 bytes, checksum: d4cb41bc00bf604cbe13dca0bf8c1774 (MD5) Previous issue date: 2017-09-04 CAPES - Coordena????o de Aperfei??oamento de Pessoal de N??vel Superior 69-99258-0496 In the world, there are 7,726 species of amphibians described to date and the Anura order has the largest number of identified species, with approximately 6,784 species. The Brazilian anura fauna has 1,039 species, being the most representative. In terms of diversity, Rondonia state has several sub-sampled gaps and it has the highest rates of deforestation in the Amazon. Therefore, the study of the anurans of Pacaas Novos National Park (PNPN), located in Campo Novo of Rond??nia, propose to contribute with the biodiversity knowledge of this region. Two areas of the park, upland sites non-subject of flooding and the seasonally flooded areas were sampled in six expeditions carried out during the months of July 2014 to August 2016. Time Limited Visual Search (PVLT) methods, Reproductive Sites (ASR) and Pitfall Traps with Drift Fences (AIQ) were used. The teams used in PVLT and ASR samplings was formed by four or five researchers collecting for two hours/methodology/night, totaling 322 h/man in both ecosystems. Four AIQs (10 pails of 100 Liters) were used with two traps at each area, totaling 340 pails/day/area. During expeditions, 1,571 specimens belonging to 62 species of anurans from thirteen families were recorded: Aromobatidae (5), Bufonidae (4), Centrolenidae (1), Ceratophryidae (1), Craugastoridae (5), Dendrobatidae (2), Hylidae 23), Leptodactylidae (10), Microhylidae (5), Odontophrynidae (1), Phyllomedusidae (3), Pipidae (1) and Ranidae (1). The most frequent family in the park was Hylidae (37.1%). However, Pristimantis fenestratus of the Craugastoridae family was the most abundant species, with 222 specimens recorded (14.13%). The less abundant species (less than 1%) were: Allobates femoralis, Dendropsophus leucophyllatus, D. sarayacuensis, Pristimantis delius, P. diadematus, Teratohyla adenocheira and Trachycephalus cf. cunauaru. The new records for the state were Allobates flaviventris, A. cf. gasconi, Pristimantis delius and P. diadematus. Dendropsophus juliani represented the first record for Brazil. When comparing the richness of two areas, it can be stated that the richness in the seasonally flooded areas is higher in the PNPN, with 18 species that were only found in this area. The richness of PNPN is similar that found in Espig??o d'Oeste also located in in Rondonia. No mundo, foram descritas 7.726 esp??cies de anf??bios, sendo a ordem Anura a que possui maior n??mero de esp??cies identificadas com cerca de 6.784 esp??cies. O Brasil ?? o pa??s com a maior riqueza da ordem Anura, apresentando o total de 1.039 esp??cies. O estado de Rond??nia apresenta uma das maiores taxas de desmatamento na Amaz??nia, possuindo v??rias lacunas sub-amostradas quanto ?? sua diversidade. Portanto, o estudo dos anuros do Parque Nacional de Paca??s Novos, localizado em Campo Novo de Rond??nia, prop??em contribuir com o melhor conhecimento quanto a biodiversidade desta regi??o. Foram amostradas duas ??reas do parque, uma de floresta de terra firme e a outra de v??rzea, em seis expedi????es realizadas durante os meses de julho de 2014 a agosto de 2016, utilizando os m??todos de procura visual limitada por tempo (PVLT), amostragem em s??tios reprodutivos (ASR) e armadilha de intercepta????o e queda (AIQ). Para as amostragens de PVLT e ASR, equipe era formada quatro ou cinco pesquisadores coletando durante duas horas/metodologia/noite, totalizando 322 h/observador em ambos ecossistemas. Foram utilizadas quatro AIQ (10 baldes-100L) sendo duas armadilhas em cada base, totalizando 340 baldes/dia/base. Durante as expedi????es foram registrados 1.571 esp??cimes, pertencentes a 62 esp??cies de anuros de 13 fam??lias: Aromobatidae (5), Bufonidae (4), Centrolenidae (1), Ceratophryidae (1), Craugastoridae (5), Dendrobatidae (2), Hylidae (23), Leptodactylidae (10), Microhylidae (5), Odontophrynidae (1), Phyllomedusidae (3), Pipidae (1) e Ranidae (1). A fam??lia de maior ocorr??ncia no parque foi a Hylidae (37,1%). No entanto, Pristimantis fenestratus da fam??lia Craugastoridae foi a esp??cie mais abundante sendo registrados 222 esp??cimes (14,13%). As esp??cies menos abundantes (menos de 1%) foram: Allobates femoralis, Dendropsophus leucophyllatus, D. sarayacuensis, Pristimantis delius, P. diadematus, Teratohyla adenocheira, e Trachycephalus cf. cunauaru. Dentre os novos registros para o estado est??o Allobates flaviventris, A. cf. gasconi, Pristimantis delius e P. diadematus. Dendropsophus juliani representa o primeiro registro para o Brasil. Quando comparada a riqueza das duas ??reas, pode-se afirmar que a riqueza na ??rea de v??rzea ?? maior no PNPN, com 18 esp??cies que foram apenas encontradas nessa ??rea. A riqueza do PNPN ?? similar a encontrada em Espig??o d???Oeste tamb??m localizado em Rond??nia.

Submitted by Divis??o de Documenta????o/BC Biblioteca Central (ddbc@ufam.edu.br) on 2017-03-16T14:11:57Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Reprodu????o N??o Autorizada.pdf: 47716 bytes, checksum: 0353d988c60b584cfc9978721c498a11 (MD5) Approved for entry into archive by Divis??o de Documenta????o/BC Biblioteca Central (ddbc@ufam.edu.br) on 2017-03-16T14:12:14Z (GMT) No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Reprodu????o N??o Autorizada.pdf: 47716 bytes, checksum: 0353d988c60b584cfc9978721c498a11 (MD5) Approved for entry into archive by Divis??o de Documenta????o/BC Biblioteca Central (ddbc@ufam.edu.br) on 2017-03-16T14:15:32Z (GMT) No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Reprodu????o N??o Autorizada.pdf: 47716 bytes, checksum: 0353d988c60b584cfc9978721c498a11 (MD5) Made available in DSpace on 2017-03-16T14:15:32Z (GMT). No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Reprodu????o N??o Autorizada.pdf: 47716 bytes, checksum: 0353d988c60b584cfc9978721c498a11 (MD5) Previous issue date: 2016-06-24 CAPES - Coordena????o de Aperfei??oamento de Pessoal de N??vel Superior The Bothrops asper is one of the main species of Central America because of its medical importance in countries such as Panam?? and Costa Rica, where they cause a high number of accidents. A great number of basic phopholipase A2 (PLA2s, EC 3.1.1.4) this species have been characterized, but a few number of studies have been carried out with acidic isoforms. Moreover, as the snake venom is a rich source of bioactive, the investigation of the biotechnological potential is required. Therefore, from the B. asper venom, this study shows three main lines of research: the caracterization of novel PLA2s isoforms; the study of PLA2s in poisoning pathophysiology and the study of potential antiparasitic of PLA2s. The snake venom was fractionated on three chromatographic methods that have proved effective in purification: ion exchange, hydrophobic interaction and reversed phase, giving rise to nine PLA2, four acidic (BaspAc-I, BaspAc-II, BaspAc-III e BaspAc-IV) and five basic (BaspB-I, BaspB-II, BaspB-III, BapsB-IV e BaspB-V). The molecular weight was determined by mass spectrometry (MALDI-TOF) showed values ranging between 13.8 and 14.3 kDa. The isoelectric point of the acidic PLA2 were determined by 2D SDS-PAGE, showing pI values between 4.6 and 5.0. The N-terminal sequence of nine PLA2 was determined, and 05 are new [four Asp49 (BaspAc-I, BaspAc-II, BaspAc-III and BaspAc-IV) and one Lys49 (BaspB-II)]. The enzymatic activity and the evaluation of the preferred substrates by PLA2 Asp49 was performed using fluorescent phospholipids which was observed that, among the tested phospholipids, acid PLA2 catalyzed all substrates, however, the basic PLA2 showed the highest specificity for phosphatidyl glycerol and phosphatidic acid. A discussion about the role of acidic PLA2 in case of poisoning by snake bite is performed based on myotoxicity assay. Moreover, it is demonstrated the antiparasitic action of these molecules, since the acidic PLA2 (BaspAc-I, BaspAc-II, BaspAc-III and BaspAc-IV) showed effects against epimastigote and pomastigotes forms of Trypanossoma cruzi and Leishmania infantum respectively. On the other hand, the basic PLA2, BaspB-II and IV-BaspB showed activity against Plasmodium falciparum. Furthermore, for the first time it demonstrated the synergistic action between Lys49 and Asp49 PLA2s against the malaria parasite, demonstrating that these molecules might be important targets in the development of new therapeutics. Keywords: Snake Venoms, Bothrops Asper, Phospholipase A2, Venomics, Synergism in Parasiticide Action A serpente Bothrops asper ?? uma das esp??cies mais importantes da Am??rica Central, principalmente, por sua import??ncia m??dica em pa??ses como Panam?? e Costa Rica, onde causam um n??mero elevado de acidentes. Diversas fosfolipases A2 (PLA2s, EC 3.1.1.4) b??sicas desta esp??cie j?? foram caracterizadas, por??m poucos estudos foram realizados com as isoformas ??cidas. Al??m disso, sendo o veneno de serpente uma rica fonte de bioativos, faz-se necess??ria a investiga????o de seu potencial biotecnol??gico. Desta forma, a partir do veneno de B. asper, o presente trabalho teve objetivos principais: a caracteriza????o de novas isoformas de PLA2s, o estudo das PLA2s na fisiopatologia do envenenamento e o estudo do potencial antiparasit??rio das PLA2s. Para tanto, o veneno foi fracionado em tr??s m??todos cromatogr??ficos que se mostraram eficientes na purifica????o: troca i??nica, intera????o hidrof??bica e fase reversa, dando origem a nove PLA2 isoladas, quatro ??cidas (BaspAc-I, BaspAc-II, BaspAc-III e BaspAc-IV) e cinco b??sicas (BaspB-I, BaspB-II, BaspB-III, BapsB-IV e BaspB-V). As massas moleculares foram determinadas por espectrometria (MALDI TOF) com valores variando entre 13,8 e 14,3 kDa. Os pontos isoel??tricos das PLA2s ??cidas foram determinados por 2D-SDS-PAGE com valores de pI variando entre 4,6 e 5,0. A sequ??ncia N-terminal das nove PLA2s foram determinadas, sendo observado que dentre as nove, 05 s??o in??ditas [quatro Asp49 (BaspAc-I, BaspAc-II, BaspAc-III e BaspAc-IV) e uma Lys49 (BaspB-II)]. A atividade enzim??tica e a avalia????o dos substratos preferenciais das PLA2s Asp49 foram realizadas utilizando-se fosfolip??deos fluorescentes e observou-se que, dentre os fosfolip??deos testados, as PLA2s ??cidas catalisaram a degrada????o de todos os substratos, entretanto, para a PLA2 b??sica houve prefer??ncia para o fosfatidilglicerol e ??cido fosfat??dico. Uma discuss??o a respeito do papel das PLA2s ??cidas no envenenamento of??dico ?? conduzida com base no ensaio de miotoxicidade. Ademais, ?? demostrado a a????o antiparasit??ria dessas mol??culas, j?? que as PLA2s ??cidas (BaspAc-I, BaspAc-II, BaspAc-III e BaspAc- IV) tiveram a????o contra as formas epimastigotas de T. cruzi e formas promastigotas de L. infantum e as PLA2s b??sicas BaspB-II e BaspB-IV apresentaram atividade contra Plasmodium falciparum. Posto isto, pela primeira vez, foi demonstrado a a????o sin??rgica entre uma PLA2 Lys49 e uma PLA2 Asp49 contra o parasita da mal??ria, demonstrando que estas mol??culas podem ser importantes alvos no desenvolvimento de novos terap??uticos.

Submitted by Divis??o de Documenta????o/BC Biblioteca Central (ddbc@ufam.edu.br) on 2017-10-19T13:15:55Z No. of bitstreams: 2 Tese - Edailson A. Correa.pdf: 7761566 bytes, checksum: 1872d5e4f92092f139b776f75cc18e5e (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Approved for entry into archive by Divis??o de Documenta????o/BC Biblioteca Central (ddbc@ufam.edu.br) on 2017-10-19T13:16:10Z (GMT) No. of bitstreams: 2 Tese - Edailson A. Correa.pdf: 7761566 bytes, checksum: 1872d5e4f92092f139b776f75cc18e5e (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Made available in DSpace on 2017-10-19T13:16:10Z (GMT). No. of bitstreams: 2 Tese - Edailson A. Correa.pdf: 7761566 bytes, checksum: 1872d5e4f92092f139b776f75cc18e5e (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2016-06-08 Venom components from snakes of the Bothrops genus have a wide range of biological actions that are not yet fully identified or understood. Of these compounds, phospholipases A2 have shown different potential for biotechnological applications. The objectives of this study were to isolate and biochemically and biologically characterize a Lys49 phospholipase A2 from Bothrops neuwiedi urutu venom. The protein was obtained with an elevated degree of purity after two chromatographic steps, the first being anion exchange (DEAE column) and the second reverse phase in a C18 column. The degree of purity and relative molecular mass were assessed by SDS-PAGE, obtaining a typical molecular mass of an svPLA2 of approximately 14 kDa, which was later confirmed by Mass Spectrometry, obtaining a mass value (m/z) of 13,733 Da. Analysis of the phospholipase activity, carried out on the substrate 4N3OBA showed that the isolated PLA2 is enzymatically inactive. The analyses using the Edman degradation method and that of the peptide fragments allowed for the identification of 108 of the toxin???s amino acid residues; high identity was observed with other phospholipases present in the venom of other species of snakes of the Bothrops genus. The data confirmed that it is a novel PLA2-Lys49 isoform from B. n. urutu venom, called BnuTX-I. The modeling in the dimeric form of the molecular structure of the toxin showed about 99.15% homology with PLA2-homologue 1 from Bothrops jararacussu (PDB code: 3I03) and identified the tertiary structure and active site. Biological assays in murine models revealed that both BnuTX-I and the venom were able to induce edema and myotoxic responses. Hemorrhagic activity was only observed for the venom. The antimicrobial and parasitic assays with the venom and BnuTX-I showed, mostly, the growth inhibition of P. aeruginosa strains and L. amazonensis promastigote forms with a CC50 of 22.91 ??g/mL and a selectivity index (SI) of 0.44. As for the cytotoxic effects of BnuTX-I in murine macrophages, the toxin presented cytotoxicity at different concentrations (??g/mL), and the production of IL 1?? cytokines was observed. The results allowed for the identification and characterization of a new myotoxin isoform with potential for future biotechnological applications. Os componentes do veneno de serpentes do g??nero Bothrops apresentam uma variada gama de a????es biol??gicas ainda n??o completamente identificadas ou esclarecidas. Dos compostos, as fosfolipases A2 v??m demonstrando diferentes potenciais para aplica????es biotecnol??gicas. Os objetivos deste estudo foram isolar e caracterizar bioqu??mica e biologicamente uma fosfolipase A2 Lys49 do veneno da serpente Bothrops neuwiedi urutu. A prote??na foi obtida com elevado grau de pureza ap??s duas etapas cromatogr??ficas, sendo a primeira de troca ani??nica (coluna DEAE) e a segunda de fase reversa em coluna C18. O grau de pureza e a massa molecular relativa foram avaliados por SDS-PAGE, obtendo-se uma massa molecular t??pica de svPLA2 de aproximadamente 14 kDa, que foi posteriormente confirmada por Espectrometria de Massa obtendo-se o valor de massa (m/z) de 13.733 Da. A an??lise da atividade fosfolip??sica, realizada sob o substrato 4N3OBA, mostrou que a PLA2 isolada ?? enzimaticamente inativa. As an??lises pelo m??todo de degrada????o de Edman e dos fragmentos pept??dicos permitiram a identifica????o de 108 res??duos de amino??cidos da toxina, sendo observada elevada identidade com outras fosfolipases presentes no veneno de outras esp??cies de serpentes do g??nero Bothrops. Os dados confirmaram tratar-se de uma isoforma in??dita de PLA2-Lys49 do veneno de B. n. urutu, denominada BnuTX-I. O modelamento, na forma dim??rica, da estrutura molecular da toxina mostrou homologia com cerca de 99,15% com a PLA2-hom??loga 1 de Bothrops jararacussu (c??digo PDB: 3I03), bem como a identifica????o das estruturas terci??ria e do s??tio ativo. Os ensaios biol??gicos mostraram, em modelos murinos, que tanto a BnuTX-I quanto o veneno foram capazes de induzir respostas edematog??nica e miot??xica. A atividade hemorr??gica foi observada somente para o veneno. Os ensaios antimicrobianos e parasit??rios realizados com veneno e BnuTX-I mostraram majoritariamente a inibi????o do crescimento das cepas de P. aeruginosa e de formas promastigota de L. amazonensis com CC50 de 22,91 ??g/mL e com ??ndice de seletividade (IS) de 0,44. Quanto aos efeitos citot??xicos, da BnuTX-I sobre macr??fagos murinos, observou-se que a toxina apresentou citotoxidade em diferentes concentra????es (??g/mL) e produ????o de citocinas IL 1??. Os resultados possibilitaram a identifica????o e caracteriza????o de uma nova isoforma de miotoxina com pontecial para futuras aplica????es biotecnol??gicas.

Durante a germinação das sementes, proteínas de reserva são acumuladas e degradadas para dar suporte aos principais eventos metabólicos envolvidos na construção de novas células e tecidos. A mobilização proteica ocorre a partir da atividade de proteases e, pode ser controlada por inibidores de proteases, evitando a hidrólise prematura das proteínas de reserva. O objetivo desta pesquisa foi investigar a atividade de proteases e inibidores de proteases durante a germinação e em períodos pré e pós-germinativos de sementes de Parkia multijuga, bem como purificar inibidores de tripsina. Para tanto, as sementes, após quebra da dormência, foram semeadas em bandejas plásticas contendo vermiculita e acondionadas em câmaras de germinação à 25ºC. Os estádios de coleta foram: sementes quiescentes (SQ), após 24 h de embebição (EM), emissão da radícula (RA), expansão do nó cotiledonar (NO) e emissão da parte aérea (PA). As proteínas foram extraídas em NaCl 0,15 M. As atividades de serinoproteases e de inibidores de tripsina e quimotripsina foram avaliadas utilizando-se como substratos o BAPNA e a azocaseína e, a atividade das cisteínoproteases e de inibidores de papaína e bromelaína, utilizando-se o BANA e azocaseína, respectivamente. Os inibidores de tripsina foram purificados a partir de cromatografias em tripsina-Sepharose 4B e HiTrap DEAE FF. O monitoramento da purificação e da mobilização das proteínas foi realizado por meio eletroforeses em SDS-PAGE e géis 2D. Durante a germinação das sementes de P. multijuga observou-se decréscimo expressivo no conteúdo de proteínas após RA, permanecendo até PA, sendo confirmado por géis 2D, demonstrando a degradação de proteínas na faixa de 35 a 75 kDa durante PA, ao passo que, proteínas de 9 kDa foram sintetizadas durante EM. Quanto às atividades inibitórias, verificou-se estabilidade na inibição das serinoproteases em todos os estádios estudados, sendo a atividade anti-tríptica vinte vezes maior quando comparada à da quimotripsina. A inibição de cisteínoproteases foi mais variável, verificando-se máxima inibição da papaína e da bromelaína em EM e NO, respectivamente. As atividades de serino e cisteínoproteases apresentaram-se constantes até NO, com acréscimo em PA. A purificação dos inibidores de tripsina envolveu duas etapas cromatográficas, onde os perfis obtidos foram similares em todos os estádios estudados, indicando a purificação das mesmas moléculas ou de isoinibidores nos diferentes estádios estudados. A partir da purificação dos inibidores de tripsina (PmTI1 e PmTI2) de SQ, essas proteínas foram caracterizadas por apresentarem especificidade inibitória contra a tripsina, alta estabilidade quanto à variação de temperatura e pH e, homologia à sequência de aminoácidos de inibidores do tipo Bowman-Birk de outras espécies de Fabaceae. Neste sentido, inibidores de serino e cisteínoproteases devem participar da regulação da atividade de proteases durante a germinação e nos eventos pré e pós-germinativos de sementes de P. multijuga, estando os inibidores de tripsina do tipo Bowman-Birk presentes durante todo o processo de formação da plântula. During seed germination, storage proteins are accumulated and degraded to support the major metabolic events involved in development new cells and tissues. The mobilization of proteins occurs through the activity of proteases, which can be regulated through the presence of proteases inhibitors that prevent premature hydrolysis of the storage proteins. The objective of this research was to investigate the activity of proteases and protease inhibitors during germination and pre-and postgermination of seeds of Parkia multijuga and purify trypsin inhibitors. Thereby, the seeds after breaking dormancy, they were germinated in plastic pots containing vermiculite and packed in germination chambers at 25ºC. The collection stages were quiescent seeds (SQ), after 24 h of imbibition (EM), radicle protrusion (RA), expansion of cotyledon node (NO) and issue shoot (PA). Proteins were extracted into 0.15 M NaCl. The activities of serine proteinases and trypsin and chymotrypsin inhibitors were performed using BAPNA and azocasein as substrates. The activity of cysteine proteases and papain and bromelain inhibitors using BANA and azocasein, respectively. Trypsin inhibitors were purified by affinity chromatography usin trypsin- Sepharose 4B and ion exchange on HiTrap DEAE FF. The purification and the mobilization of proteins was performed by electrophoresis on SDS-PAGE and 2D gels. During seed germination of P. multijuga observed a significant decrease in the protein content after RA remaining until PA was confirmed by 2D gels, wich show proteins degradation in the range of 35 to 75 kDa in PA, while 9 kDa proteins were synthesized during EM. Regarding the inhibitory activity, serine protease inhibition showed high stability during all stages studied, of which trypsin inhibition was 20 times higher than chymotrypsin. Inhibition of cysteine proteases is more variable, observing for maximum inhibition of papain and bromelain in EM and NO, respectively. The activities of serine and cysteine proteases presented themselves to constant NO, with an increase in PA. The purification of trypsin inhibitors involved two chromatographic steps, which were similar to the profiles obtained in all stages studied, indicating the purification of the same molecule or isoinhibitors in different stages studied. From the purification of trypsin inhibitors (PmTI1 and PmTI2) SQ, these proteins are characterized by presenting against trypsin inhibitory specificity, high stability against variation of temperature and pH and amino acid sequence homology to the Bowman-type inhibitors Birk from other Fabaceae species. Thus, serine and cysteine proteases inhibitors could participate in the regulation of protease activity during germination and events pre and post-germination of seeds of P. multijuga, with the trypsin inhibitors Bowman-Birk type present throughout the process of formation of the seedling.