Producción de alcanos en E.coli usando fotoenzimas: Diseño y simulación de un plásmido de alta expresión.
Investigación experimental se realizó en el laboratorio de biofísica de la Universidad de los Andes.
- Autores:
-
Osorio Vivas, Juan Sebastián
- Tipo de recurso:
- Trabajo de grado de pregrado
- Fecha de publicación:
- 2023
- Institución:
- Universidad de los Andes
- Repositorio:
- Séneca: repositorio Uniandes
- Idioma:
- spa
- OAI Identifier:
- oai:repositorio.uniandes.edu.co:1992/64205
- Acceso en línea:
- http://hdl.handle.net/1992/64205
- Palabra clave:
- E.coli
Fotoenzimas
Hidrocarburos
Alcanos
Plasmido
Biofísica
Bioquímica
Física
- Rights
- openAccess
- License
- Attribution-NoDerivatives 4.0 Internacional
| id |
UNIANDES2_14180c776811a739db4d9da40437e52e |
|---|---|
| oai_identifier_str |
oai:repositorio.uniandes.edu.co:1992/64205 |
| network_acronym_str |
UNIANDES2 |
| network_name_str |
Séneca: repositorio Uniandes |
| repository_id_str |
|
| dc.title.none.fl_str_mv |
Producción de alcanos en E.coli usando fotoenzimas: Diseño y simulación de un plásmido de alta expresión. |
| title |
Producción de alcanos en E.coli usando fotoenzimas: Diseño y simulación de un plásmido de alta expresión. |
| spellingShingle |
Producción de alcanos en E.coli usando fotoenzimas: Diseño y simulación de un plásmido de alta expresión. E.coli Fotoenzimas Hidrocarburos Alcanos Plasmido Biofísica Bioquímica Física |
| title_short |
Producción de alcanos en E.coli usando fotoenzimas: Diseño y simulación de un plásmido de alta expresión. |
| title_full |
Producción de alcanos en E.coli usando fotoenzimas: Diseño y simulación de un plásmido de alta expresión. |
| title_fullStr |
Producción de alcanos en E.coli usando fotoenzimas: Diseño y simulación de un plásmido de alta expresión. |
| title_full_unstemmed |
Producción de alcanos en E.coli usando fotoenzimas: Diseño y simulación de un plásmido de alta expresión. |
| title_sort |
Producción de alcanos en E.coli usando fotoenzimas: Diseño y simulación de un plásmido de alta expresión. |
| dc.creator.fl_str_mv |
Osorio Vivas, Juan Sebastián |
| dc.contributor.advisor.none.fl_str_mv |
Pedraza Leal, Juan Manuel |
| dc.contributor.author.none.fl_str_mv |
Osorio Vivas, Juan Sebastián |
| dc.contributor.jury.none.fl_str_mv |
Forero Shelton, Antonio Manu |
| dc.contributor.researchgroup.es_CO.fl_str_mv |
Biofísica Uniandes |
| dc.subject.keyword.none.fl_str_mv |
E.coli Fotoenzimas Hidrocarburos Alcanos Plasmido Biofísica Bioquímica |
| topic |
E.coli Fotoenzimas Hidrocarburos Alcanos Plasmido Biofísica Bioquímica Física |
| dc.subject.themes.es_CO.fl_str_mv |
Física |
| description |
Investigación experimental se realizó en el laboratorio de biofísica de la Universidad de los Andes. |
| publishDate |
2023 |
| dc.date.accessioned.none.fl_str_mv |
2023-01-25T21:33:44Z |
| dc.date.available.none.fl_str_mv |
2023-01-25T21:33:44Z |
| dc.date.issued.none.fl_str_mv |
2023-01-25 |
| dc.type.es_CO.fl_str_mv |
Trabajo de grado - Pregrado |
| dc.type.driver.none.fl_str_mv |
info:eu-repo/semantics/bachelorThesis |
| dc.type.version.none.fl_str_mv |
info:eu-repo/semantics/acceptedVersion |
| dc.type.coar.none.fl_str_mv |
http://purl.org/coar/resource_type/c_7a1f |
| dc.type.content.es_CO.fl_str_mv |
Text |
| dc.type.redcol.none.fl_str_mv |
http://purl.org/redcol/resource_type/TP |
| format |
http://purl.org/coar/resource_type/c_7a1f |
| status_str |
acceptedVersion |
| dc.identifier.uri.none.fl_str_mv |
http://hdl.handle.net/1992/64205 |
| dc.identifier.instname.es_CO.fl_str_mv |
instname:Universidad de los Andes |
| dc.identifier.reponame.es_CO.fl_str_mv |
reponame:Repositorio Institucional Séneca |
| dc.identifier.repourl.es_CO.fl_str_mv |
repourl:https://repositorio.uniandes.edu.co/ |
| url |
http://hdl.handle.net/1992/64205 |
| identifier_str_mv |
instname:Universidad de los Andes reponame:Repositorio Institucional Séneca repourl:https://repositorio.uniandes.edu.co/ |
| dc.language.iso.es_CO.fl_str_mv |
spa |
| language |
spa |
| dc.relation.references.es_CO.fl_str_mv |
U.S. Energy Information Administration - EIA - independent statistics and analysis. U.S. energy facts explained - consumption and production - U.S. Energy Information Administration (EIA), May 2021. Hannah Ritchie and Max Roser. Energy use worldwide. Our World in Data, 2020. annah Ritchie. How long before we run out of fossil fuels? Our World in Data, Aug 2017 David E. Sadava, David M. Hillis, H. Craig Heller, and M. Berenbaum. Life: The science of biology. Sinauer, 2014 aul Singleton. Bacteria in biology, biotechnology, and medicine. John Wiley, 1999. Escherichia coli - Details - Public Health Image Library(PHIL). Centers for Disease Control and Prevention, 2006. Bruce Alberts, Rebecca Heald, Alexander Johnson, David Morgan, Martin Raff, Keith Roberts, and Peter Walter. Molecular biology of the cell. W. W. Norton amp; Companyf, 2022. Stanley N. Cohen, Annie C. Chang, Herbert W. Boyer, and Robert B. Helling. Construction of biologically functional bacterial plasmids in vitro. Proceedings of the National Academy of Sciences, 1973 Nabih A Baeshen, Mohammed N Baeshen, Abdullah Sheikh, Roop S Bora, Mohamed Morsi M Ahmed, Hassan A I Ramadan, Kulvinder Singh Saini, and Elrashdy M Redwan. Cell factories for insulin production. Microb Cell Fact, 13(1), oct 2014. Addgene. Plasmid cloning by restriction enzyme digest (aka subcloning). https://www. addgene.org/protocols/subcloning/, 2017. Damien Sorigué, Bertrand Légeret, Stéphan Cuiné, Stéphanie Blangy, Solène Moulin, Emmanuelle Billon, Pierre Richaud, Sabine Brugière, Yohann Couté, Didier Nurizzo, Pavel Müller, Klaus Brettel, David Pignol, Pascal Arnoux, Yonghua Li-Beisson, Gilles Peltier, and Fred Beisson. An algal photoenzyme converts fatty acids to hydrocarbons. Science, 357(6354):903-907, sep 2017 Mengshi Xiao, Derong Lin, Zhuohao Li, Jingjing Zhao, Xiaomei Long, and Zhijun Wu. Synthesis of biodiesel from waste cooking oil by one-step esterification and its structural characterization. Waste Biomass Valor, 11(5):2087-2100, nov 2018 Damien Sorigué, Bertrand Légeret, Stéphan Cuiné, Stéphanie Blangy, Solène Moulin, Emmanuelle Billon, Pierre Richaud, Sabine Brugière, Yohann Couté, Didier Nurizzo, Pavel Müller, Klaus Brettel, David Pignol, Pascal Arnoux, Yonghua Li-Beisson, Gilles Peltier, and Fred Beisson. An algal photoenzyme converts fatty acids to hydrocarbons. Science, 357(6354):903-907, sep 2017. Edda Klipp, Wolfram Liebermeister, Christoph Wierling, Axel Kowald, and Ralf Herwig. Systems biology a textbook. Wiley-VCH Verlag GmbH and Co. KGaA, 2016. Adrian J. Brown. Xxxvi. enzyme action. J. Chem. Soc., Trans., 81:373-388, 1902. Daniel T Gillespie. A general method for numerically simulating the stochastic time evolution of coupled chemical reactions. Journal of Computational Physics, 22(4):403-434, 1976. Malgorzta M. Klosek. Markov processes: An introduction for physical scientists (Daniel t. Gillespie). SIAM Review, 35(2):324-325, 1993. Daniel T. Gillespie. Stochastic simulation of chemical kinetics. Annual Review of Physical Chemistry, 58(1):35-55, 2007. Michael . Elowitz and Justin Bois. 12. Stochastic Simulation of Biological Circuits. http://be150.caltech.edu/2019/handouts/12_stochastic_ simulation_all_code.html. François Jacob and Jacques Monod. Genetic regulatory mechanisms in the synthesis of proteins. Journal of Molecular Biology, 3(3):318-356, 1961. James Keener. Chapter 10, volume I, page 427-470. Springer, 2009. Uri Alon. An introduction to systems biology: Design principles of biological circuits. CRC Press, 2020. Xiaobo Guo, Ao Xia, Feng Li, Yun Huang, Xianqing Zhu, Wuyuan Zhang, Xun Zhu, and Qiang Liao. Photoenzymatic decarboxylation to produce renewable hydrocarbon fuels: A comparison between whole-cell and broken-cell biocatalysts. Energy Conversion and Management, 255:115311, March 2020. Paul Santner, László Krisztián Szabó, Santiago Nahuel Chanquia, Aske Høj Merrild, Frank Hollmann, Selin Kara, and Bekir Engin Eser. Optimization and engineering of fatty acid photodecarboxylase for substrate specificity. ChemCatChem, 13(18):4038-4046, July 2021. |
| dc.rights.license.spa.fl_str_mv |
Attribution-NoDerivatives 4.0 Internacional |
| dc.rights.uri.*.fl_str_mv |
http://creativecommons.org/licenses/by-nd/4.0/ |
| dc.rights.accessrights.spa.fl_str_mv |
info:eu-repo/semantics/openAccess |
| dc.rights.coar.spa.fl_str_mv |
http://purl.org/coar/access_right/c_abf2 |
| rights_invalid_str_mv |
Attribution-NoDerivatives 4.0 Internacional http://creativecommons.org/licenses/by-nd/4.0/ http://purl.org/coar/access_right/c_abf2 |
| eu_rights_str_mv |
openAccess |
| dc.format.extent.es_CO.fl_str_mv |
68 páginas |
| dc.format.mimetype.es_CO.fl_str_mv |
application/pdf |
| dc.publisher.es_CO.fl_str_mv |
Universidad de los Andes |
| dc.publisher.program.es_CO.fl_str_mv |
Física |
| dc.publisher.faculty.es_CO.fl_str_mv |
Facultad de Ciencias |
| dc.publisher.department.es_CO.fl_str_mv |
Departamento de Física |
| institution |
Universidad de los Andes |
| bitstream.url.fl_str_mv |
https://repositorio.uniandes.edu.co/bitstreams/d5cf4aab-f0a5-47ca-a2f6-ed29f779ca3d/download https://repositorio.uniandes.edu.co/bitstreams/b757ccf0-ceb6-4927-8637-912b2d2315fc/download https://repositorio.uniandes.edu.co/bitstreams/6f10a295-04b9-4e0c-afcb-76aae1ec7f8c/download https://repositorio.uniandes.edu.co/bitstreams/0df22ff5-8431-4db2-8604-e1f323c806cf/download https://repositorio.uniandes.edu.co/bitstreams/2c0aab92-1165-4667-8a35-47d767ea73d3/download https://repositorio.uniandes.edu.co/bitstreams/c22be740-8d37-407a-af04-7b38b19a0b53/download https://repositorio.uniandes.edu.co/bitstreams/0133ce42-0776-42cd-bcd7-00d6d8e65161/download https://repositorio.uniandes.edu.co/bitstreams/58e28cdf-d442-4c2d-ab5b-4f80b9422c1e/download |
| bitstream.checksum.fl_str_mv |
077409021764c157fbe16a174b10359c 4491fe1afb58beaaef41a73cf7ff2e27 c35da838ea0d460b3d8b55c0c97ad42d 56d4845131377bb322568cdfcd0e7610 f7d494f61e544413a13e6ba1da2089cd 5aa5c691a1ffe97abd12c2966efcb8d6 2a023facfd521e3397922e6ebc67ffb7 84e348e0f421e14a56c0295d47e16e0b |
| bitstream.checksumAlgorithm.fl_str_mv |
MD5 MD5 MD5 MD5 MD5 MD5 MD5 MD5 |
| repository.name.fl_str_mv |
Repositorio institucional Séneca |
| repository.mail.fl_str_mv |
adminrepositorio@uniandes.edu.co |
| _version_ |
1812133960216477696 |
| spelling |
Attribution-NoDerivatives 4.0 Internacionalhttp://creativecommons.org/licenses/by-nd/4.0/info:eu-repo/semantics/openAccesshttp://purl.org/coar/access_right/c_abf2Pedraza Leal, Juan Manuelvirtual::10223-1Osorio Vivas, Juan Sebastiáne27443e9-4d16-49f2-b014-6cee22a74690600Forero Shelton, Antonio ManuBiofísica Uniandes2023-01-25T21:33:44Z2023-01-25T21:33:44Z2023-01-25http://hdl.handle.net/1992/64205instname:Universidad de los Andesreponame:Repositorio Institucional Sénecarepourl:https://repositorio.uniandes.edu.co/Investigación experimental se realizó en el laboratorio de biofísica de la Universidad de los Andes.Se diseñó un plásmido de alta expresión para la producción de la fotoenzima CvFAP (Chlorella variabilis photoenzyme) que descarboxila ácidos grasos en alcanos. Se implementó un protocolo experimental que confirmó la producción de alcanos debido a la enzima en presencia de luz azul.FísicoPregrado68 páginasapplication/pdfspaUniversidad de los AndesFísicaFacultad de CienciasDepartamento de FísicaProducción de alcanos en E.coli usando fotoenzimas: Diseño y simulación de un plásmido de alta expresión.Trabajo de grado - Pregradoinfo:eu-repo/semantics/bachelorThesisinfo:eu-repo/semantics/acceptedVersionhttp://purl.org/coar/resource_type/c_7a1fTexthttp://purl.org/redcol/resource_type/TPE.coliFotoenzimasHidrocarburosAlcanosPlasmidoBiofísicaBioquímicaFísicaU.S. Energy Information Administration - EIA - independent statistics and analysis. U.S. energy facts explained - consumption and production - U.S. Energy Information Administration (EIA), May 2021.Hannah Ritchie and Max Roser. Energy use worldwide. Our World in Data, 2020.annah Ritchie. How long before we run out of fossil fuels? Our World in Data, Aug 2017David E. Sadava, David M. Hillis, H. Craig Heller, and M. Berenbaum. Life: The science of biology. Sinauer, 2014aul Singleton. Bacteria in biology, biotechnology, and medicine. John Wiley, 1999.Escherichia coli - Details - Public Health Image Library(PHIL). Centers for Disease Control and Prevention, 2006.Bruce Alberts, Rebecca Heald, Alexander Johnson, David Morgan, Martin Raff, Keith Roberts, and Peter Walter. Molecular biology of the cell. W. W. Norton amp; Companyf, 2022.Stanley N. Cohen, Annie C. Chang, Herbert W. Boyer, and Robert B. Helling. Construction of biologically functional bacterial plasmids in vitro. Proceedings of the National Academy of Sciences, 1973Nabih A Baeshen, Mohammed N Baeshen, Abdullah Sheikh, Roop S Bora, Mohamed Morsi M Ahmed, Hassan A I Ramadan, Kulvinder Singh Saini, and Elrashdy M Redwan. Cell factories for insulin production. Microb Cell Fact, 13(1), oct 2014.Addgene. Plasmid cloning by restriction enzyme digest (aka subcloning). https://www. addgene.org/protocols/subcloning/, 2017.Damien Sorigué, Bertrand Légeret, Stéphan Cuiné, Stéphanie Blangy, Solène Moulin, Emmanuelle Billon, Pierre Richaud, Sabine Brugière, Yohann Couté, Didier Nurizzo, Pavel Müller, Klaus Brettel, David Pignol, Pascal Arnoux, Yonghua Li-Beisson, Gilles Peltier, and Fred Beisson. An algal photoenzyme converts fatty acids to hydrocarbons. Science, 357(6354):903-907, sep 2017Mengshi Xiao, Derong Lin, Zhuohao Li, Jingjing Zhao, Xiaomei Long, and Zhijun Wu. Synthesis of biodiesel from waste cooking oil by one-step esterification and its structural characterization. Waste Biomass Valor, 11(5):2087-2100, nov 2018Damien Sorigué, Bertrand Légeret, Stéphan Cuiné, Stéphanie Blangy, Solène Moulin, Emmanuelle Billon, Pierre Richaud, Sabine Brugière, Yohann Couté, Didier Nurizzo, Pavel Müller, Klaus Brettel, David Pignol, Pascal Arnoux, Yonghua Li-Beisson, Gilles Peltier, and Fred Beisson. An algal photoenzyme converts fatty acids to hydrocarbons. Science, 357(6354):903-907, sep 2017.Edda Klipp, Wolfram Liebermeister, Christoph Wierling, Axel Kowald, and Ralf Herwig. Systems biology a textbook. Wiley-VCH Verlag GmbH and Co. KGaA, 2016.Adrian J. Brown. Xxxvi. enzyme action. J. Chem. Soc., Trans., 81:373-388, 1902.Daniel T Gillespie. A general method for numerically simulating the stochastic time evolution of coupled chemical reactions. Journal of Computational Physics, 22(4):403-434, 1976.Malgorzta M. Klosek. Markov processes: An introduction for physical scientists (Daniel t. Gillespie). SIAM Review, 35(2):324-325, 1993.Daniel T. Gillespie. Stochastic simulation of chemical kinetics. Annual Review of Physical Chemistry, 58(1):35-55, 2007.Michael . Elowitz and Justin Bois. 12. Stochastic Simulation of Biological Circuits. http://be150.caltech.edu/2019/handouts/12_stochastic_ simulation_all_code.html.François Jacob and Jacques Monod. Genetic regulatory mechanisms in the synthesis of proteins. Journal of Molecular Biology, 3(3):318-356, 1961.James Keener. Chapter 10, volume I, page 427-470. Springer, 2009.Uri Alon. An introduction to systems biology: Design principles of biological circuits. CRC Press, 2020.Xiaobo Guo, Ao Xia, Feng Li, Yun Huang, Xianqing Zhu, Wuyuan Zhang, Xun Zhu, and Qiang Liao. Photoenzymatic decarboxylation to produce renewable hydrocarbon fuels: A comparison between whole-cell and broken-cell biocatalysts. Energy Conversion and Management, 255:115311, March 2020.Paul Santner, László Krisztián Szabó, Santiago Nahuel Chanquia, Aske Høj Merrild, Frank Hollmann, Selin Kara, and Bekir Engin Eser. Optimization and engineering of fatty acid photodecarboxylase for substrate specificity. ChemCatChem, 13(18):4038-4046, July 2021.201714174Publicationhttps://scholar.google.es/citations?user=x8-YWMsAAAAJvirtual::10223-10000-0002-1802-3337virtual::10223-1https://scienti.minciencias.gov.co/cvlac/visualizador/generarCurriculoCv.do?cod_rh=0000001817virtual::10223-1a4c0056f-ab75-4234-9297-925380d7633avirtual::10223-1a4c0056f-ab75-4234-9297-925380d7633avirtual::10223-1TEXTTesisJuanOsorioFinal.pdf.txtTesisJuanOsorioFinal.pdf.txtExtracted texttext/plain73709https://repositorio.uniandes.edu.co/bitstreams/d5cf4aab-f0a5-47ca-a2f6-ed29f779ca3d/download077409021764c157fbe16a174b10359cMD55FormatoAceptacionTesis.pdf.txtFormatoAceptacionTesis.pdf.txtExtracted texttext/plain1163https://repositorio.uniandes.edu.co/bitstreams/b757ccf0-ceb6-4927-8637-912b2d2315fc/download4491fe1afb58beaaef41a73cf7ff2e27MD57ORIGINALTesisJuanOsorioFinal.pdfTesisJuanOsorioFinal.pdfTesis de grado de Juan Sebastian Osorio Vivas.application/pdf3191276https://repositorio.uniandes.edu.co/bitstreams/6f10a295-04b9-4e0c-afcb-76aae1ec7f8c/downloadc35da838ea0d460b3d8b55c0c97ad42dMD53FormatoAceptacionTesis.pdfFormatoAceptacionTesis.pdfHIDEapplication/pdf751865https://repositorio.uniandes.edu.co/bitstreams/0df22ff5-8431-4db2-8604-e1f323c806cf/download56d4845131377bb322568cdfcd0e7610MD54CC-LICENSElicense_rdflicense_rdfapplication/rdf+xml; charset=utf-8799https://repositorio.uniandes.edu.co/bitstreams/2c0aab92-1165-4667-8a35-47d767ea73d3/downloadf7d494f61e544413a13e6ba1da2089cdMD52LICENSElicense.txtlicense.txttext/plain; charset=utf-81810https://repositorio.uniandes.edu.co/bitstreams/c22be740-8d37-407a-af04-7b38b19a0b53/download5aa5c691a1ffe97abd12c2966efcb8d6MD51THUMBNAILTesisJuanOsorioFinal.pdf.jpgTesisJuanOsorioFinal.pdf.jpgIM Thumbnailimage/jpeg8607https://repositorio.uniandes.edu.co/bitstreams/0133ce42-0776-42cd-bcd7-00d6d8e65161/download2a023facfd521e3397922e6ebc67ffb7MD56FormatoAceptacionTesis.pdf.jpgFormatoAceptacionTesis.pdf.jpgIM Thumbnailimage/jpeg15751https://repositorio.uniandes.edu.co/bitstreams/58e28cdf-d442-4c2d-ab5b-4f80b9422c1e/download84e348e0f421e14a56c0295d47e16e0bMD581992/64205oai:repositorio.uniandes.edu.co:1992/642052024-03-13 14:08:00.127http://creativecommons.org/licenses/by-nd/4.0/open.accesshttps://repositorio.uniandes.edu.coRepositorio institucional Sénecaadminrepositorio@uniandes.edu.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 |