Effect of Flocculation Technology on Lipids, Carbohydrates and Proteins and Extraction from Chlorella vulgaris
Nowadays, Chlorella vulgaris has emerged as an attractive source of metabolites such as lipids, proteins and carbohydrates, which are widely applied to obtain high value products. This work is focused on evaluating the effect of flocculation on metabolites recovery from Chlorella vulgaris microalgae...
- Autores:
-
Sanguino, Paola Andrea
González-Delgado, Angel Darío
Barajas Solano, andres F
- Tipo de recurso:
- Article of journal
- Fecha de publicación:
- 2018
- Institución:
- Universidad Francisco de Paula Santander
- Repositorio:
- Repositorio Digital UFPS
- Idioma:
- eng
- OAI Identifier:
- oai:repositorio.ufps.edu.co:ufps/1602
- Acceso en línea:
- http://repositorio.ufps.edu.co/handle/ufps/1602
- Palabra clave:
- Microalgae
Carbohydrates
Proteins
Lipids
Flocculation
- Rights
- openAccess
- License
- Atribución 4.0 Internacional (CC BY 4.0)
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Effect of Flocculation Technology on Lipids, Carbohydrates and Proteins and Extraction from Chlorella vulgaris |
| title |
Effect of Flocculation Technology on Lipids, Carbohydrates and Proteins and Extraction from Chlorella vulgaris |
| spellingShingle |
Effect of Flocculation Technology on Lipids, Carbohydrates and Proteins and Extraction from Chlorella vulgaris Microalgae Carbohydrates Proteins Lipids Flocculation |
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Effect of Flocculation Technology on Lipids, Carbohydrates and Proteins and Extraction from Chlorella vulgaris |
| title_full |
Effect of Flocculation Technology on Lipids, Carbohydrates and Proteins and Extraction from Chlorella vulgaris |
| title_fullStr |
Effect of Flocculation Technology on Lipids, Carbohydrates and Proteins and Extraction from Chlorella vulgaris |
| title_full_unstemmed |
Effect of Flocculation Technology on Lipids, Carbohydrates and Proteins and Extraction from Chlorella vulgaris |
| title_sort |
Effect of Flocculation Technology on Lipids, Carbohydrates and Proteins and Extraction from Chlorella vulgaris |
| dc.creator.fl_str_mv |
Sanguino, Paola Andrea González-Delgado, Angel Darío Barajas Solano, andres F |
| dc.contributor.author.none.fl_str_mv |
Sanguino, Paola Andrea González-Delgado, Angel Darío Barajas Solano, andres F |
| dc.subject.proposal.eng.fl_str_mv |
Microalgae Carbohydrates Proteins Lipids Flocculation |
| topic |
Microalgae Carbohydrates Proteins Lipids Flocculation |
| description |
Nowadays, Chlorella vulgaris has emerged as an attractive source of metabolites such as lipids, proteins and carbohydrates, which are widely applied to obtain high value products. This work is focused on evaluating the effect of flocculation on metabolites recovery from Chlorella vulgaris microalgae biomass. Carbohydrates were extracted through acid and alkaline hydrolysis in order to identify the efficiency of these treatments for cell wall disruption. Protein quantification was carried out under alkaline conditions and lipids were recovered by solvent extraction with hexane. It was found that carbohydrate recovery was higher using centrifuged biomass (40.55%) than flocculated biomass (24.31%) by acid treatment, while alkaline treatment exhibited yield of 20.37% and 13.19%, respectively. In addition, efficiency of 53.75% and 32.27% in protein recovery was obtained from both centrifuged and flocculated biomass. The highest lipid extraction yield (54.2%) was achieved using acid hydrolysis and hexane solvent route when centrifuged biomass was used as metabolite source. These results suggested that flocculation affect significantly the efficiency of metabolites extraction. |
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2018 |
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2018-03-03 |
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2021-11-30T23:32:16Z |
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2021-11-30T23:32:16Z |
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10.12988/ces.2018.8118 |
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eng |
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Contemporary Engineering Sciences ISSN: 1313-6569, 2018 vol:11 fasc: 6 págs: 283 - 291, DOI:https://doi.org/10.12988/ces.2018.8118 |
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Vol. 11, No. 6 (2018) |
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291 |
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6 (2018) |
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Sanguino-Barajas, P. A., Gonzalez-Delgado, A. D. y Barajas-Solano, A. F. (2018). Effect of flocculation technology on lipids, carbohydrates and proteins and extraction from Chlorella vulgaris. Contemporary Engineering Sciences, 11(6), 283–291. https://doi.org/10.12988/ces.2018.8118 |
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Contemporary Engineering Sciences |
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Atribución 4.0 Internacional (CC BY 4.0) |
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Atribución 4.0 Internacional (CC BY 4.0) http://purl.org/coar/access_right/c_abf2 |
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9 páginas |
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Contemporary Engineering Sciences |
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Bulgaria |
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Sanguino, Paola Andreaeea47e25d3d2679fcacfd40c55a39c62González-Delgado, Angel Darío0102ed6cc17aae314ed4ababae07d17c600Barajas Solano, andres F7237a5ca918751f9d045f15b62fd2f1f6002021-11-30T23:32:16Z2021-11-30T23:32:16Z2018-03-03http://repositorio.ufps.edu.co/handle/ufps/160210.12988/ces.2018.8118Nowadays, Chlorella vulgaris has emerged as an attractive source of metabolites such as lipids, proteins and carbohydrates, which are widely applied to obtain high value products. This work is focused on evaluating the effect of flocculation on metabolites recovery from Chlorella vulgaris microalgae biomass. Carbohydrates were extracted through acid and alkaline hydrolysis in order to identify the efficiency of these treatments for cell wall disruption. Protein quantification was carried out under alkaline conditions and lipids were recovered by solvent extraction with hexane. It was found that carbohydrate recovery was higher using centrifuged biomass (40.55%) than flocculated biomass (24.31%) by acid treatment, while alkaline treatment exhibited yield of 20.37% and 13.19%, respectively. In addition, efficiency of 53.75% and 32.27% in protein recovery was obtained from both centrifuged and flocculated biomass. The highest lipid extraction yield (54.2%) was achieved using acid hydrolysis and hexane solvent route when centrifuged biomass was used as metabolite source. These results suggested that flocculation affect significantly the efficiency of metabolites extraction.9 páginasapplication/pdfengContemporary Engineering SciencesBulgariaContemporary Engineering Sciences ISSN: 1313-6569, 2018 vol:11 fasc: 6 págs: 283 - 291, DOI:https://doi.org/10.12988/ces.2018.8118Vol. 11, No. 6 (2018)2916 (2018)28311Sanguino-Barajas, P. A., Gonzalez-Delgado, A. D. y Barajas-Solano, A. F. (2018). Effect of flocculation technology on lipids, carbohydrates and proteins and extraction from Chlorella vulgaris. Contemporary Engineering Sciences, 11(6), 283–291. https://doi.org/10.12988/ces.2018.8118Contemporary Engineering Sciences2018 P.A. Sanguino-Barajas et al. This article is distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.info:eu-repo/semantics/openAccessAtribución 4.0 Internacional (CC BY 4.0)http://purl.org/coar/access_right/c_abf2http://www.m-hikari.com/ces/ces2018/ces5-8-2018/8118.htmlEffect of Flocculation Technology on Lipids, Carbohydrates and Proteins and Extraction from Chlorella vulgarisArtículo de revistahttp://purl.org/coar/resource_type/c_6501http://purl.org/coar/resource_type/c_2df8fbb1Textinfo:eu-repo/semantics/articlehttp://purl.org/redcol/resource_type/ARTinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/version/c_970fb48d4fbd8a85MicroalgaeCarbohydratesProteinsLipidsFlocculationR. Zhang, J. Chen, X. Zhang, Extraction of intracellular protein from Chlorella pyrenoidosa using a combination of ethanol soaking, enzyme digest, ultrasonication and homogenization techniques, Bioresource Technology, 247 (2018), 267–272. https://doi.org/10.1016/j.biortech.2017.09.087E. Blanco-Carvajal, E. Sánchez-Galvis, Á. González-Delgado, Janet Bibiana Garcia Martinez, Andres Fernando Barajas-Solano, Cultivation of Chlorella vulgaris in Aquaculture Wastewater for Protein Production, Contemporary Engineering Sciences, 11 (2018), 93–100. https://doi.org/10.12988/ces.2018.712203Á. González-Delgado, J. García-Martínez, Y.Y. Peralta-Ruíz, Evaluation of Two Pre-Treatments for Improving Lipid Extraction from Microalgae Navicula sp., Contemporary Engineering Sciences, 10 (2017), 851–859. https://doi.org/10.12988/ces.2017.78792L.K. Torres-carvajal, Á.D. González-Delgado, A.F Barajas-Solano, Jhon Hermogenes Suarez-Gelvez, Nestor Andres Urbina-Suarez, Astaxanthin Production from Haematococcus pluvialis : Effects of Light Wavelength and Salinity, Contemporary Engineering Sciences, 10 (2017), 1739–1746. https://doi.org/10.12988/ces.2017.711196Á. González-Delgado, J. García-Martínez, Y.Y. Peralta-Ruíz, Cell Disruption and Lipid Extraction from Microalgae Amphiprora sp . Using Acid HydrolysisSolvent Extraction Route, Contemporary Engineering Sciences, 10 (2017), 841–849. https://doi.org/10.12988/ces.2017.78791Y. Heoa, H. Leea, C. Lee, Juwon Kang, Joon-Woo Ahn, Young Min Lee, KyuYoung Kang, Yoon-E Choi, Jae-Jin Kim, An integrative process for obtaining lipids and glucose from Chlorella vulgaris biomass with a single treatment of cell disruption, Algal Research, 27 (2017), 286–294. https://doi.org/10.1016/j.algal.2017.09.022E. Saraylooa, S. Simseka, Y. Unlud, Gul Cevahir, Can Erkey, Ibrahim Halil Kavakli, Enhancement of the lipid productivity and fatty acid methyl ester profile of Chlorella vulgaris by two rounds of mutagenesis, Bioresource Technology, 250 (2018), 764–769. https://doi.org/10.1016/j.biortech.2017.11.105M. Chang, D. Li, W. Wang, Dongchu Chen, Yuyuan Zhang, Huawen Hu, Xiufang Ye, Comparison of sodium hydroxide and calcium hydroxide pretreatments on the enzymatic hydrolysis and lignin recovery of sugarcane bagasse, Bioresource Technology, 244 (2017), 1055–1058. https://doi.org/10.1016/j.biortech.2017.08.101M. Dubois, K.A. Gilles, J.K. Hamilton, P.A. Rebers, F. Smith, Colorimetric Method for Determination of Sugars and Related Substances, Analytical Chemistry, 28 (1956), 350–356. https://doi.org/10.1021/ac60111a017O.H. Lowry, N.J. Rosebrough, A.L. Farr and RJ. Randall, Protein measurement with the Folin phenol reagent, The Journal of Biolical Chemistry, 193 (1951), 265–275.R. Robert, R-M. Knuckey, D-M-F. Frampton, M-R. Brown, Production of microalgal concentrates by flocculation and their assessment as aquaculture feeds, Aquacultural Engineering, 35 (2006), 300–313. https://doi.org/10.1016/j.aquaeng.2006.04.001Y.I. Mandik, B. Cheirsilp, P. Boonsawang, P. Prasertsan, Optimization of flocculation efficiency of lipid-rich marine Chlorella sp. biomass and evaluation of its composition in different cultivation modes, Bioresource Technology, 182 (2015), 89–97. https://doi.org/10.1016/j.biortech.2015.01.125M.R. Talukder, P. Das, J.C. Wu, Microalgae (Nannochloropsis salina) biomass to lactic acid and lipid, Biochemical Engineering Journal, 68 (2012), 109–113. https://doi.org/10.1016/j.bej.2012.07.001S. Bellou, G. Aggelis, Biochemical activities in Chlorella sp. And Nannochloropsis salina during lipid and sugar synthesis in a lab-acale open pond simulating reactor, Journal of Biotechnology, 164 (2013), 318–329. https://doi.org/10.1016/j.jbiotec.2013.01.010S.P. Cuellar-Bermudez, I. Aguilar-Hernandez, D.L. Cardenas-Chavez, N. Ornelas-Soto, M. A. Romero-Ogawa and Roberto Parra-Saldivar, Extraction and purification of high-value metabolites from microalgae: essential lipids, astaxanthin and phycobiliproteins, Microbial Biotechnology, 8 (2014), 190– 209. https://doi.org/10.1111/1751-7915.12167ORIGINALEffect of Flocculation Technology on Lipids, Carbohydrates and Proteins and Extraction from Chlorella vulgaris.pdfEffect of Flocculation Technology on Lipids, Carbohydrates and Proteins and Extraction from Chlorella vulgaris.pdfapplication/pdf549663https://repositorio.ufps.edu.co/bitstream/ufps/1602/1/Effect%20of%20Flocculation%20Technology%20on%20Lipids%2c%20Carbohydrates%20and%20Proteins%20and%20Extraction%20from%20Chlorella%20vulgaris.pdf85f8e2f937df2b73bb321aa7eb2a14a5MD51open accessLICENSElicense.txtlicense.txttext/plain; charset=utf-814828https://repositorio.ufps.edu.co/bitstream/ufps/1602/2/license.txt2f9959eaf5b71fae44bbf9ec84150c7aMD52open accessTEXTEffect of Flocculation Technology on Lipids, Carbohydrates and Proteins and Extraction from Chlorella vulgaris.pdf.txtEffect of Flocculation Technology on Lipids, Carbohydrates and Proteins and Extraction from Chlorella vulgaris.pdf.txtExtracted texttext/plain17388https://repositorio.ufps.edu.co/bitstream/ufps/1602/3/Effect%20of%20Flocculation%20Technology%20on%20Lipids%2c%20Carbohydrates%20and%20Proteins%20and%20Extraction%20from%20Chlorella%20vulgaris.pdf.txt62a6ca0b93e0da4cb809a453a90f5506MD53open accessTHUMBNAILEffect of Flocculation Technology on Lipids, Carbohydrates and Proteins and Extraction from Chlorella vulgaris.pdf.jpgEffect of Flocculation Technology on Lipids, Carbohydrates and Proteins and Extraction from Chlorella vulgaris.pdf.jpgGenerated Thumbnailimage/jpeg12399https://repositorio.ufps.edu.co/bitstream/ufps/1602/4/Effect%20of%20Flocculation%20Technology%20on%20Lipids%2c%20Carbohydrates%20and%20Proteins%20and%20Extraction%20from%20Chlorella%20vulgaris.pdf.jpgfd3aa5d34ef71d9e772834bd6b70363bMD54open accessufps/1602oai:repositorio.ufps.edu.co:ufps/16022022-05-23 10:39:13.24open accessRepositorio Universidad Francisco de Paula 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 incorporada en las Obras Colectivas.

b.	Distribuir copias o fonogramas de las Obras, exhibirlas públicamente, ejecutarlas públicamente y/o ponerlas a disposición pública, incluyéndolas como incorporadas en Obras Colectivas, según corresponda.

c.	Distribuir copias de las Obras Derivadas que se generen, exhibirlas públicamente, ejecutarlas públicamente y/o ponerlas a disposición pública.
Los derechos mencionados anteriormente pueden ser ejercidos en todos los medios y formatos, actualmente conocidos o que se inventen en el futuro. Los derechos antes mencionados incluyen el derecho a realizar dichas modificaciones en la medida que sean técnicamente necesarias para ejercer los derechos en otro medio o formatos, pero de otra manera usted no está autorizado para realizar obras derivadas. Todos los derechos no otorgados expresamente por el Licenciante quedan por este medio reservados, incluyendo pero sin limitarse a aquellos que se mencionan en las secciones 4(d) y 4(e).

4. Restricciones.
La licencia otorgada en la anterior Sección 3 está expresamente sujeta y limitada por las siguientes restricciones:

a.	Usted puede distribuir, exhibir públicamente, ejecutar públicamente, o poner a disposición pública la Obra sólo bajo las condiciones de esta Licencia, y Usted debe incluir una copia de esta licencia o del Identificador Universal de Recursos de la misma con cada copia de la Obra que distribuya, exhiba públicamente, ejecute públicamente o ponga a disposición pública. No es posible ofrecer o imponer ninguna condición sobre la Obra que altere o limite las condiciones de esta Licencia o el ejercicio de los derechos de los destinatarios otorgados en este documento. No es posible sublicenciar la Obra. Usted debe mantener intactos todos los avisos que hagan referencia a esta Licencia y a la cláusula de limitación de garantías. Usted no puede distribuir, exhibir públicamente, ejecutar públicamente, o poner a disposición pública la Obra con alguna medida tecnológica que controle el acceso o la utilización de ella de una forma que sea inconsistente con las condiciones de esta Licencia. Lo anterior se aplica a la Obra incorporada a una Obra Colectiva, pero esto no exige que la Obra Colectiva aparte de la obra misma quede sujeta a las condiciones de esta Licencia. Si Usted crea una Obra Colectiva, previo aviso de cualquier Licenciante debe, en la medida de lo posible, eliminar de la Obra Colectiva cualquier referencia a dicho Licenciante o al Autor Original, según lo solicitado por el Licenciante y conforme lo exige la cláusula 4(c).

b.	Usted no puede ejercer ninguno de los derechos que le han sido otorgados en la Sección 3 precedente de modo que estén principalmente destinados o directamente dirigidos a conseguir un provecho comercial o una compensación monetaria privada. El intercambio de la Obra por otras obras protegidas por derechos de autor, ya sea a través de un sistema para compartir archivos digitales (digital file-sharing) o de cualquier otra manera no será considerado como estar destinado principalmente o dirigido directamente a conseguir un provecho comercial o una compensación monetaria privada, siempre que no se realice un pago mediante una compensación monetaria en relación con el intercambio de obras protegidas por el derecho de autor.

c.	Si usted distribuye, exhibe públicamente, ejecuta públicamente o ejecuta públicamente en forma digital la Obra o cualquier Obra Derivada u Obra Colectiva, Usted debe mantener intacta toda la información de derecho de autor de la Obra y proporcionar, de forma razonable según el medio o manera que Usted esté utilizando: (i) el nombre del Autor Original si está provisto (o seudónimo, si fuere aplicable), y/o (ii) el nombre de la parte o las partes que el Autor Original y/o el Licenciante hubieren designado para la atribución (v.g., un instituto patrocinador, editorial, publicación) en la información de los derechos de autor del Licenciante, términos de servicios o de otras formas razonables; el título de la Obra si está provisto; en la medida de lo razonablemente factible y, si está provisto, el Identificador Uniforme de Recursos (Uniform Resource Identifier) que el Licenciante especifica para ser asociado con la Obra, salvo que tal URI no se refiera a la nota sobre los derechos de autor o a la información sobre el licenciamiento de la Obra; y en el caso de una Obra Derivada, atribuir el crédito identificando el uso de la Obra en la Obra Derivada (v.g., "Traducción Francesa de la Obra del Autor Original," o "Guión Cinematográfico basado en la Obra original del Autor Original"). Tal crédito puede ser implementado de cualquier forma razonable; en el caso, sin embargo, de Obras Derivadas u Obras Colectivas, tal crédito aparecerá, como mínimo, donde aparece el crédito de cualquier otro autor comparable y de una manera, al menos, tan destacada como el crédito de otro autor comparable.

d.	Para evitar toda confusión, el Licenciante aclara que, cuando la obra es una composición musical:

i.	Regalías por interpretación y ejecución bajo licencias generales. El Licenciante se reserva el derecho exclusivo de autorizar la ejecución pública o la ejecución pública digital de la obra y de recolectar, sea individualmente o a través de una sociedad de gestión colectiva de derechos de autor y derechos conexos (por ejemplo, SAYCO), las regalías por la ejecución pública o por la ejecución pública digital de la obra (por ejemplo Webcast) licenciada bajo licencias generales, si la interpretación o ejecución de la obra está primordialmente orientada por o dirigida a la obtención de una ventaja comercial o una compensación monetaria privada.

ii.	Regalías por Fonogramas. El Licenciante se reserva el derecho exclusivo de recolectar, individualmente o a través de una sociedad de gestión colectiva de derechos de autor y derechos conexos (por ejemplo, los consagrados por la SAYCO), una agencia de derechos musicales o algún agente designado, las regalías por cualquier fonograma que Usted cree a partir de la obra (“versión cover”) y distribuya, en los términos del régimen de derechos de autor, si la creación o distribución de esa versión cover está primordialmente destinada o dirigida a obtener una ventaja comercial o una compensación monetaria privada.

e.	Gestión de Derechos de Autor sobre Interpretaciones y Ejecuciones Digitales (WebCasting). Para evitar toda confusión, el Licenciante aclara que, cuando la obra sea un fonograma, el Licenciante se reserva el derecho exclusivo de autorizar la ejecución pública digital de la obra (por ejemplo, webcast) y de recolectar, individualmente o a través de una sociedad de gestión colectiva de derechos de autor y derechos conexos (por ejemplo, ACINPRO), las regalías por la ejecución pública digital de la obra (por ejemplo, webcast), sujeta a las disposiciones aplicables del régimen de Derecho de Autor, si esta ejecución pública digital está primordialmente dirigida a obtener una ventaja comercial o una compensación monetaria privada.

5. Representaciones, Garantías y Limitaciones de Responsabilidad.
A MENOS QUE LAS PARTES LO ACORDARAN DE OTRA FORMA POR ESCRITO, EL LICENCIANTE OFRECE LA OBRA (EN EL ESTADO EN EL QUE SE ENCUENTRA) “TAL CUAL”, SIN BRINDAR GARANTÍAS DE CLASE ALGUNA RESPECTO DE LA OBRA, YA SEA EXPRESA, IMPLÍCITA, LEGAL O CUALQUIERA OTRA, INCLUYENDO, SIN LIMITARSE A ELLAS, GARANTÍAS DE TITULARIDAD, COMERCIABILIDAD, ADAPTABILIDAD O ADECUACIÓN A PROPÓSITO DETERMINADO, AUSENCIA DE INFRACCIÓN, DE AUSENCIA DE DEFECTOS LATENTES O DE OTRO TIPO, O LA PRESENCIA O AUSENCIA DE ERRORES, SEAN O NO DESCUBRIBLES (PUEDAN O NO SER ESTOS DESCUBIERTOS). ALGUNAS JURISDICCIONES NO PERMITEN LA EXCLUSIÓN DE GARANTÍAS IMPLÍCITAS, EN CUYO CASO ESTA EXCLUSIÓN PUEDE NO APLICARSE A USTED.

6. Limitación de responsabilidad.
A MENOS QUE LO EXIJA EXPRESAMENTE LA LEY APLICABLE, EL LICENCIANTE NO SERÁ RESPONSABLE ANTE USTED POR DAÑO ALGUNO, SEA POR RESPONSABILIDAD EXTRACONTRACTUAL, PRECONTRACTUAL O CONTRACTUAL, OBJETIVA O SUBJETIVA, SE TRATE DE DAÑOS MORALES O PATRIMONIALES, DIRECTOS O INDIRECTOS, PREVISTOS O IMPREVISTOS PRODUCIDOS POR EL USO DE ESTA LICENCIA O DE LA OBRA, AUN CUANDO EL LICENCIANTE HAYA SIDO ADVERTIDO DE LA POSIBILIDAD DE DICHOS DAÑOS. ALGUNAS LEYES NO PERMITEN LA EXCLUSIÓN DE CIERTA RESPONSABILIDAD, EN CUYO CASO ESTA EXCLUSIÓN PUEDE NO APLICARSE A USTED.

7. Término.

a.	Esta Licencia y los derechos otorgados en virtud de ella terminarán automáticamente si Usted infringe alguna condición establecida en ella. Sin embargo, los individuos o entidades que han recibido Obras Derivadas o Colectivas de Usted de conformidad con esta Licencia, no verán terminadas sus licencias, siempre que estos individuos o entidades sigan cumpliendo íntegramente las condiciones de estas licencias. Las Secciones 1, 2, 5, 6, 7, y 8 subsistirán a cualquier terminación de esta Licencia.

b.	Sujeta a las condiciones y términos anteriores, la licencia otorgada aquí es perpetua (durante el período de vigencia de los derechos de autor de la obra). No obstante lo anterior, el Licenciante se reserva el derecho a publicar y/o estrenar la Obra bajo condiciones de licencia diferentes o a dejar de distribuirla en los términos de esta Licencia en cualquier momento; en el entendido, sin embargo, que esa elección no servirá para revocar esta licencia o que deba ser otorgada , bajo los términos de esta licencia), y esta licencia continuará en pleno vigor y efecto a menos que sea terminada como se expresa atrás. La Licencia revocada continuará siendo plenamente vigente y efectiva si no se le da término en las condiciones indicadas anteriormente.

8. Varios.

a.	Cada vez que Usted distribuya o ponga a disposición pública la Obra o una Obra Colectiva, el Licenciante ofrecerá al destinatario una licencia en los mismos términos y condiciones que la licencia otorgada a Usted bajo esta Licencia.

b.	Si alguna disposición de esta Licencia resulta invalidada o no exigible, según la legislación vigente, esto no afectará ni la validez ni la aplicabilidad del resto de condiciones de esta Licencia y, sin acción adicional por parte de los sujetos de este acuerdo, aquélla se entenderá reformada lo mínimo necesario para hacer que dicha disposición sea válida y exigible.

c.	Ningún término o disposición de esta Licencia se estimará renunciada y ninguna violación de ella será consentida a menos que esa renuncia o consentimiento sea otorgado por escrito y firmado por la parte que renuncie o consienta.

d.	Esta Licencia refleja el acuerdo pleno entre las partes respecto a la Obra aquí licenciada. No hay arreglos, acuerdos o declaraciones respecto a la Obra que no estén especificados en este documento. El Licenciante no se verá limitado por ninguna disposición adicional que pueda surgir en alguna comunicación emanada de Usted. Esta Licencia no puede ser modificada sin el consentimiento mutuo por escrito del Licenciante y Usted.
0000-0001-8100-88880102ed6cc17aae314ed4ababae07d17c6000000-0003-2765-91317237a5ca918751f9d045f15b62fd2f1f600 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