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Vieira (andre_fonseca.vieira@hotmail.com) on 2019-08-15T15:33:14Z No. of bitstreams: 5 Ata da defesa.pdf: 284002 bytes, checksum: 7048f66ead80bb068e518c363ef40be0 (MD5) Ficha catalogr??fica.pdf: 1885 bytes, checksum: 195830af898dd533dd3789152521f630 (MD5) PROJETO - VERS??O FINAL.pdf: 4234814 bytes, checksum: 5b7ccb4fa8875beccbfb27475a3bf52c (MD5) Carta assinada.pdf: 235455 bytes, checksum: 9fe59f99850d1643d70db80bc9fd0243 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Approved for entry into archive by PPGBIOTEC Biotecnologia (ppg_biotec.ufam@yahoo.com.br) on 2019-08-15T15:40:24Z (GMT) No. of bitstreams: 5 Ata da defesa.pdf: 284002 bytes, checksum: 7048f66ead80bb068e518c363ef40be0 (MD5) Ficha catalogr??fica.pdf: 1885 bytes, checksum: 195830af898dd533dd3789152521f630 (MD5) PROJETO - VERS??O FINAL.pdf: 4234814 bytes, checksum: 5b7ccb4fa8875beccbfb27475a3bf52c (MD5) Carta assinada.pdf: 235455 bytes, checksum: 9fe59f99850d1643d70db80bc9fd0243 (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 2019-08-19T12:53:43Z (GMT) No. of bitstreams: 5 Ata da defesa.pdf: 284002 bytes, checksum: 7048f66ead80bb068e518c363ef40be0 (MD5) Ficha catalogr??fica.pdf: 1885 bytes, checksum: 195830af898dd533dd3789152521f630 (MD5) PROJETO - VERS??O FINAL.pdf: 4234814 bytes, checksum: 5b7ccb4fa8875beccbfb27475a3bf52c (MD5) Carta assinada.pdf: 235455 bytes, checksum: 9fe59f99850d1643d70db80bc9fd0243 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Made available in DSpace on 2019-08-19T12:53:43Z (GMT). No. of bitstreams: 5 Ata da defesa.pdf: 284002 bytes, checksum: 7048f66ead80bb068e518c363ef40be0 (MD5) Ficha catalogr??fica.pdf: 1885 bytes, checksum: 195830af898dd533dd3789152521f630 (MD5) PROJETO - VERS??O FINAL.pdf: 4234814 bytes, checksum: 5b7ccb4fa8875beccbfb27475a3bf52c (MD5) Carta assinada.pdf: 235455 bytes, checksum: 9fe59f99850d1643d70db80bc9fd0243 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2019-06-25 CAPES - Coordena????o de Aperfei??oamento de Pessoal de N??vel Superior 92993458428 As triacylglycerol lipases, (E.C.3.1.1.3), are called lipases, they catalyze carboxylic ester reactions to triacylglycerol to form free fatty acids, have a characteristic of catalytic activity in aqueous hydrolysis medium. Outside the aqueous medium, esterification, interesterification or transesterification reactions occur. The potential biotechnological, lipolytic enzymes, and their industrial industry for their global leukopithetic lipopruticent, which in the industrial industry, which must be important in the importance of biocatalysis, favoring greater sale in the process. Justifying the study of new molecules with high biotechnological potential to supply new needs of the world market, especially those coming from the Amazon, the region seems richer in biodiversity. In order to enzymatically characterize a recombinant lipase in Pichia pastoris, the lipase coding gene was isolated from a library. The production of clones in the minimum in amino acids and the clones were detected in medium. Selection of recombinant clones was completed in BMMY containing 1% tributary solids in BMMY medium with positive clones grown in 0.5% methanol-induced submerged fermentation for lipase production. The structural region of the lipase gene was enlarged by 1200 bp. Degrading halos were observed around the colonies. The presence of the 50 kDa enzyme in the SDS PAGE gel was verified. Enzymatic characterization was important to verify the biotechnological possibilities of lipase through colorimetric assays, which were positively activated for enzymatic research of 50.80 U / mL with optimum temperature of 50 ?? C and pH of 7 being the best time of 5 minutes As triacilglicerol lipases, (E.C.3.1.1.3), usualmente chamadas de lipases, catalisam rea????es de ??steres carbox??licos em triacilglicerol formando ??cidos graxos livres, possuem a caracter??stica de exercer atividade catal??tica em meio aquoso de hidr??lise. Fora do meio aquoso, ocorrem rea????es, de esterifica????o, interesterifica????o ou transesterifica????o. Devido ao grande potencial biotecnol??gico, as enzimas lipol??ticas tem ganhado enorme destaque para uso industrial devido ao seu abrangente leque de aplica????es, constituindo um dos grupos de maior import??ncia em biocat??lise, favosrescendo maior efic??cia no processo. Justifica-se o estudo de novas mol??culas com alto potencial biotecnol??gico para suprir novas necessidades do mercado mundial, especialmente as que prov??m da Amaz??nia, regi??o ??mpar em diversidade biol??gica. Com o objetivo de caracterizar enzimaticamente uma lipase recombinante expressa em Pichia pastoris, o gene codificador de lipase foi isolado de uma biblioteca metagen??mica de terra preta de ??ndio por PCR e integrado no genoma da levedura P. pastoris no vetor de express??o pPIC9. A transforma????o ocorreu por eletropora????o com 10 ??g do cassete de express??o linearizado e clones foram selecionados em meio minimo em amino??cidos. A sele????o de clones recombinantes ocorreu em BMMY s??lido contendo 1% de tributirina em meio BMMY com clones positivos cultivados em fermenta????o submersa induzida com metanol 0,5% para a produ????o de lipase. A regi??o estrutural do gene de lipase foi amplifica com aproximadamente 1200 pb. Foram observados halos de degrada????o aos redores das col??nias. Averiguou-se a presen??a da enzima com 50 kDa no gel de SDS PAGE. A caracteriza????o enzim??tica foi ??ltimo passo para verificar as possibilidades biotecnol??gicas desta lipase por meio de ensaios colorim??tricos, que resultou positivamente para atividade enzim??tica de 50,80 U/mL temperatura ??tima de 50??C e pH de 7 sendo o melhor tempo de rea????o de 5 minutos