Superprodu????o de ramnolip??deo em Saccharomyces cerevisiae: Constru????o de vetores de express??o

• Main Author: Campos, Rayane Luzia Viegas Campos
• Other Authors: Parachin, N??dia Skorupa
• Format: Others
• Language: Portuguese
• Published: Universidade Cat??lica de Bras??lia 2016
• Subjects: Biossurfactante; Ramnolip??deo; Produ????o; Larga escala; Saccharomyces cerevisiae; Gen??tica; Genoma; CIENCIAS BIOLOGICAS::GENETICA
• Online Access: https://bdtd.ucb.br:8443/jspui/handle/tede/1997

Submitted by Kelson Anthony de Menezes ([e-mail]) on 2016-12-19T18:06:48Z No. of bitstreams: 1 RayaneLuziaViegasCamposDissertacao2015.pdf: 2888012 bytes, checksum: 8f9962ac7690fc08e072b81ccd8516db (MD5) === Made available in DSpace on 2016-12-19T18:06:48Z (GMT). No. of bitstreams: 1 RayaneLuziaViegasCamposDissertacao2015.pdf: 2888012 bytes, checksum: 8f9962ac7690fc08e072b81ccd8516db (MD5) Previous issue date: 2015-03-31 === Biosurfactants are metabolites that contain both hydrophobic and hydrophilic domains and are produced by various microorganisms. Due to its characteristics such molecules localize preferable in the interface of polar and non-polar solvents. Biosurfactants are widely used in pharmaceutic, cosmetic, food and agriculture industries. Moreover, they are considered advantageous when compared to chemical ones because they are biodegradable and non-toxic. Currently one of the best known biosurfactant is the rhamnolipids. However, their utilization in large scale are still not possible due to its high production costs. Rhamnolipids are produced mainly by Pseudomonas aeruginosa and are composed by one or two molecules of L-rhamnose attached to a fatty acid molecule. This study aimed the construction of yeast expression vectors that will be used to construct a recombinant strain of Saccharomyces cerevisiae efficient in the production of L-rhamnose and mono-rhamnolipid using sucrose as substrate. For the construction of these vectors, six different genes, responsible for encoding both L-rhamnose and mono-ramnolipideos biosynthetic pathways enzymes described in P. aeruginosa (rmlA, rmlB, rmlC, rmlD, rhlA and rhlB), and one gene, responsible for the synthesis of a substrate break enzyme, described in Pelomonas sacchraphyla (sucrose phosphorylase) were utilized. The genetic engineering proposed in this study is innovative, and has never been done before. The production of rhamnolipid in large-scale at low costs will be highly beneficial for the development of national technologies for production of high value-added chemical. === Biossurfactantes s??o metab??litos produzidos por diversos microrganismos. Estes cont??m um dom??nio hidrof??lico e outro hidrof??bico, estando preferencialmente na interface de compostos polares e apolares. Devido ??s suas propriedades, os biosurfactantes s??o capazes de reduzir a tens??o de superf??cies tendo uma ampla aplica????o na ind??stria farmac??utica, cosm??tica, aliment??cia e agr??cola. O uso de biossurfactante em compara????o com os de origem qu??mica s??o considerados vantajosos, uma vez que s??o biodegrad??veis e n??o t??xicos. Atualmente um dos biossurfactantes mais conhecido e dispon??vel comercialmente s??o os ramnolip??deos. No entanto, a utiliza????o destas mol??culas em larga escala ?? prejudicada devido aos seus elevados custos de produ????o. Ramnolip??deos s??o produzidos principalmente por Pseudomonas aeruginosa e s??o constitu??dos por uma ou duas mol??culas de L-ramnose ligada a uma mol??cula de ??cido graxo. Assim, este estudo teve como objetivo a constru????o de vetores de express??o em levedura para posterior constru????o de cepas recombinantes de Saccharomyces cerevisiae visando a eficiente produ????o de L-ramnose e monoramnolip??deo a partir de sacarose. Para a constru????o desses vetores, foram utilizados seis genes, que codificam as enzimas respons??veis pelas vias de bioss??ntese de Lramnose e mono-ramnolipideos descritos em P. aeruginosa (rmlA, rmlB, rmlC, rmlD, rhlA e rhlB), e um gene, respons??vel pela s??ntese da enzima de quebra do substrato, descrito em Pelomonas sacchraphyla (sacarose fosforilase). A engenharia gen??tica proposta neste estudo ?? inovadora, e pela primeira vez realizada. Com a produ????o do ramnolip??deo em larga escala ?? um baixo custo de produ????o, beneficiar?? o desenvolvimento de tecnologias nacionais para produ????o de qu??micos de alto valor agregado.