Bioprospeccion de bacterias productoras de polihidroxialcanoatos (pha's) en el departamento de Nariño

  • Ivan Dario Otero Universidad de Nariño.
  • Pablo Fernandez Universidad de Nariæo.
Palabras clave: Suelos de Nariño, Laguna de la Cocha, Ro Pasto, Polihidroxibutirato, Polihidroxivalerato

Resumen

Los polihidroxialcanoatos (PHA's), gránulos intracelulares acumulados por bacterias en ambientes con desbalance Carbono/Nitrógeno, Carbono/Fósforo tienen propiedades físico-químicas similares a plásticos convencionales, pero son biodegradables. Sin embargo, los elevados costos de producción de los PHA's conllevan a la búsqueda de estrategias con la menor inversión posible; por ello, en esta investigación se exploraron diferentes ambientes del departamento de Nariño en búsqueda de bacterias productoras de PHA's y se seleccionaron aquellas con potencial industrial. La detección presuntiva de bacterias PHA's a partir de muestras de suelo y agua, se realizó utilizando Rojo Nilo, los aislamientos seleccionados se sometieron a una fermentación Batch y el polímero obtenido se caracterizó por cromatografía de gases y cromatografía de gases-espectrometría de masas. Aquellos aislamientos con mayor acumulación de polímero se caracterizaron mediante pruebas bioquímicas y la secuencia parcial del gen rRNA 16S.Finalmente, se evaluó la producción de PHA's utilizando como sustrato aguas residuales domésticas. Se obtuvo 189 aislamientos bacterianos que produjeron desde 0,01gL-1 hasta 5,24gL-1 del homopolimero Poli-3-hidroxibutirato o el copolímero Poli-3-hidroxibutirato-co-3-hidroxivalerato. Así mismo, se recuperó 0,111gL-1 de Poli-3-hidroxibutirato a partir de las aguas residuales. Desde esta perspectiva las bacterias silvestres productoras de PHA's del departamento de Nariño tienen potencial en procesos ambientales e industriales.

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Biografía del autor/a

Ivan Dario Otero, Universidad de Nariño.
Biologo, Investigador Grupo de Biotecnologa Microbiana,
Pablo Fernandez, Universidad de Nariæo.
Ph.D. Ciencias Biolgicas `rea Microbiologa. Director Grupo de Biotecnologa Microbiana,

Citas

ANDERSON, A. and DAWES, E. Occurrence, metabolism, metabolic role, and industrial uses of bacterial Polyhydroxyalkanoates. Rev. Microbiol., 54, 1990, p. 4450-4472.

RHU, D., LEE, W., KIM, J. and CHOI, E. Polyhydroxyalkanoate (PHA) production from waste. Water Science and Technology, 48 (8), 2003, p. 221228.

VOLOVA, T., GLADYSSHEV, I., TRUSOVA, Y. and ZHILA, N. Degradation of polyhydroxyalkanoates in eutrophic reservoir.Polymer Degradation and Stability, 92, 2007, p. 580 586.

RUIZ, A., ALMEIDA, A., LPEZ, I. y PETTINARI, J. BioplÆsticos: una alternativa ecolgica. RevistaQumicaViva,3, 2004, p. 122 133.

ANASTASSIOS, C., SISKA, H., CHOUHAN, B. and DIETER, J. Structural Basis of poly(3-Hidroxybutyrate) Hydrolysis by PhaZ7 Depolymerase from Paucimonaslemoignei. J. Mol. Biol., 382, 2008, p. 1184-1194.

LLANA, S. and JAY, D. Process design for microbial plastic factories: metabolic engineering of polyhydroxyalkanoates. Current Opinion in Biotechnology, 14, 2003, p.475483.B.

REHM, B. Biogenesis of microbial polyhydroxyalkanoate granules: a platform technology for the production of tailor made bioparticules. Curr. Issues Mol. Biol., 9, 2007, p. 41-42.

VALAPPIL, P., BOCCACCINI, R., BUCKE, C. and ROY, I. Polyhydroxyalkanoates in Gram positive bacteria: insights from the genera Bacillus and Streptomyces. Antonie van Leeuwenhoek, 91, 2007, p.117.

WU, Q., HUANG, H., HU, G., CHEN, J., HO, K. and CHEN, G.Production of poly-3-hydroxybutyrate by Bacillus sp. JMa5 cultivated in molasses media. Antonie van Leeuwenhoek,80, 2001, p. 111118.

BORAH, B., THAKUR, P. and NIGAM, J. The influence of nutritional and environmental conditions on the accumulation of poly-E-hydroxybutyrate in Bacillus mycoidesRLJ B-017.J ApplMicrobiol.,92, 2002, p. 776-783.

KATIRCIOGLU, H., ASLIM, B., YKSEKDA—, Z., MERCAN, N. and BEYATLI, Y.Production of polyE-hydroxybutyrate (PHB) and differentiation of putative Bacillus mutant strains by SDS-PAGE of total cell protein. Afr. J.Biotechnol.,2, 2003, p.147-149.

YILMAZ, M., SORAN, H. and BEYATLI, Y. Determination of poly-E-hydroxybutyrate (PHB) production by some Bacillus sp. World J.Microbiol. Biotechnol.,21, 2005, p. 565-566.

PORWAL, S.H., KUMAR, T., LAL, S., RANI, A., KUMAR, S., CHEEMA, S., PUROHIT, H., SHARMA, R., KUMAR, S., PATEL, S. and KALIA, V. Hy

drogen and polyhydroxybutyrate producing abilities of microbes from diverse habitats by dark fermentative process.Bioresource Technology, 99, 2008, p. 2444-2451.

LEE, S.Y., CHOI, J., and LEE, S.H. Production of Polyhydroxyalkanoates by fermentation of bacteria. Macromol. Symp., 159, 2000, p. 259-266.

LABUZEK, S. and RADECKA I. Biosynthesis of PHB tercopolymer by Bacilllus cereus UW85.J. Appl.Microbiol.,90, 2001, p.353-357.

VALAPPIL, P., RAI, R., BUCKE, C. and ROY, I. Polyhydroxyalkanoate biosynthesis in Bacillus cereus SPV under varied limiting conditions and an insight into the biosynthetic genes involved. J. Appl.Microbiol.,104, 2008, p. 1624-1635.

FOSTER, L., SAUFI, A. and HOLDEN, P. Environmental concentrations of polyhydroxyalkanoates and their potential as bioindicators of pollution. Biotechnology Letters, 23, 2001, p. 893-898.

MOSTACEDO, B. y FREDERICKSEN, T. Manual de mØtodos bÆsicos de muestreo y anÆlisis en ecologa vegetal. Santa Cruz(Bolivia):BOLFOR, 2000, p. 3 19. [19] AMERICAN PUBLIC HEALTH ASSOCIATION, AMERICAN WATER WORKS ASSOCIATION, WATER ENVIRONMENT FEDERATION (APHA, AWWA, WEF). Standard Methods for the examination of water & wastewater. 21 ed. 2005, 900 p.

FERN`NDEZ, P., ORTIZ, F., BURBANO, O., GUERRERO, M. y ESPAA, J. Influencia de las fuentes de carbono y nitrgeno en el crecimiento bacteriano y sntesis de polihidroxialcanoato de una cepa de BacillusMicoides FBL2. Revista Centro de Estudios en Salud, 1 (7), 2006, p. 34-42.

OSTLE, A. and HOLT, J. Nile Blue A as a Fluorescent Satn forPoly-E-Hydroxybutyrate. Applied and Environmental Microbiology, 44 (1), 1982, p. 238-241.

SPIEKERMANN, P., REHM, B., KALSCHEUER, R., BAUMEISTER, D.andSTEINBUCHEL, A. Sensitive, Viable Colony Staining Method Using Nile Red for Direct Screening of Bacteria That Accumulate Polyhydroxyalkanoic Acids and Other Lipid Storage Compounds. Arch.Microbiol., 171, 1999, p. 73-80.

TORTORA, G. FUNKE, B. and CASE, C.H. Microbiology an introduction. 10 ed. Boston (USA): Pearson Education, 2010, 960 p.

SEGURA, J. Notas de diseæos experimentales. Facultad de medicina veterinaria y Zootecnia. YucatÆn (MØxico): Universidad Autnoma de YucatÆn, 2000, 59 p.20Biotecnologa en el Sector Agropecuario y Agroindustrial Edicin Especial No. 2 (12- 20)

BRAUNEGG, G., LEFEBVRE, G. and GENSER, K. Polyhydroxyalkanoates, Biopolyesters from renewable resources: Physiological and engineering aspects. Journal of Biotechnology. 65, 1998, p. 127-161.

RIVERA, I., CHOWDHURY, M., HUQ, A., JACOBS, D., MARTINS, M. and COLWELL, R. Enterobacterial Repetitive Intergenic Consensus Sequences and the PCR to Generate Fingerprints of Genomic DNAs from Vibrio cholerae O1, O139, and NonO1 Strains.Appl. Environ. Microbiol. 61, 1995, p. 2898-2904.

SAMBROK, J., FRITSCH, E. and MANIATIS, T. Molecular cloning: A laboratory manual. 2 ed. New York (USA): Cold Spring Harbor Laboratory Press, 1989, 1448 p.

LANE, D. 16S/23S rRNA sequencing. In: STACKEBRANDT, E. e OODFELLOW, M. (Ed.). Nucleic acid techniques in bacterial systematics. England: John Wiley & Sons, Chichester, 1991, p.115163.

NUBEL, U., ENGELEN, B., FELSKE, A., SNAIDR, J., WIESHUBER, A., AMANN, R., LUDWIG, W. and BACKHAUS, H. Sequence Heterogeneities of Genes Encoding 16S rRNAs in Paenibacilluspolymyxa Detected by Temperature Gradient Gel Electrophoresis. J.Bacteriol., 178, 1996 .p. 5636-5643.

SOUZA, C., BURBANO-ROSERO, E., ALMEIDA, B., BARRETO, C. and RIVERA, I. Chitinolytic Bacteria Diversity Isolated from Marine Ecosystems with Different Levels of Anthropogenic Activity in Sªo Paulo State, Brazil. Sao Paulo (Brasil): The ISME Journal (In press), 2012.

COLE, J., WANG, Q., CARDENAS, E., FISH, J., CHAI, B., FARRIS, R., KULAM-SYED-MOHIDEEN, A., MCGARRELL, D., MARSH, T., GARRITY, G. and TIEDJE, J. The Ribosomal Database Project: improved alignments and new tools for rRNA analysis. Nucleic Acids Res., 37, 2009, p. D141D145.

DAFFONCHIO, D., BORIN, S., FROVA, G., GALLO, R., MORI, E., FANI, R. and SORLINI, C. A Randomly Amplified Polymorphic DNA Marker Specific for the Bacillus cereus Group Is Diagnostic for Bacillus anthracis. Appl. Environ. Microbiol., 65 (3), 1999, p. 1298 1303.

DI FRANCO, C., BECCARI, E., SANTINI, T., PISANESCHI, G., and TECCE, G. Colony shape as a genetic trait in the pattern-forming Bacillus mycoides. BMC microbiology, 2 (33), 2002. p. 1-15.

ASLIM, B., YUKSEKDAG, Z. and BEYATLI, Y. Determination of PHB growth quantities of certain Bacillus species isolated from soil. Turkish Electronic Journal of Biotechnology, 2002, p. 24-30.

SINGH, M., PATEL, S. and KALIA, V. Bacillus subtilisas potential producer for polyhydroxyalkanoates.Microbial cell factories. BioMed Central, 2009, p. 1-11.

BELLO, G. y BRANDL, H. Seleccin de bacterias productoras de polihidroxibutirato. Instituto Cubano de Investigaciones de los Derivados de la Caæa de Azœcar (ICIDCA). Redalyc, 41 (2), 2007, p. 10-18.

PORWAL, S.H., KUMAR, T., LAL, S., RANI, A., KUMAR, S., CHEEMA, S., PUROHIT, H., SHARMA, R., KUMAR, S., PATEL, S. and KALIA, V. Hydrogen and polyhydroxybutyrate producing abilities of microbes from diverse habitats by dark fermentative process. Bioresource Technology, 99. 2008, p. 5444-2451.

TAMDOGAN, N. and SIDAL, U. Investigation of poly-E-hydroxybutyrate (PHB) production by Bacillus subtilisATCC 6633 under different conditions. KafkasUniv Vet FakDerg, 17, 2011, p. 173-176.

DIONISI, D., CARUCCI, G., PETRANGELI, M., PAPINI, P., RICCARDI, C., MAJONE, M.andCARRASCO, F., Olive oil mill effluents as a feedstock for production of biodegradable polymers.Water, 10, 2005, p. 20762084.

ALBUQUERQUE, M., EIROA, M., TORRES, C., NUNES, B., andREIS, M. Strategies for the development of a side stream process for polyhydroxyalkanoate (PHA) production from sugar cane molasses. J. Biotechnol., 130, 2007, p. 411421.

COATSB, E., LOGE, F., SMITH, W., THOMPSON, D.andWOLCOTT, M. Functional stability of a mixed microbial consortium producing PHA from waste carbon sources. Applied Biochemistry and Biotechnology, 136, 2007, p. 909 - 926

BENGTSSON, S., WERKER, A., CHRISTENSSON, M., and WELANDER, T. Production of polyhydroxyalkanoates by activated sludge treating a paper mill wastewater.Bioresour.Technol., 99, 2008, p. 509516.

ALBUQUERQUE, M., CONCAS, S., BENGTSSON, S. andREIS, M. Mixed culture polyhydroxyalkanoates production from sugar molasses: the use of a 2-stage CSTR system for culture selection. Bioresource Technology, 101, 2010, p. 7112 712

Cómo citar
Otero, I. D., & Fernandez, P. (2013). Bioprospeccion de bacterias productoras de polihidroxialcanoatos (pha’s) en el departamento de Nariño. Biotecnología En El Sector Agropecuario Y Agroindustrial, 11(1), 12-20. Recuperado a partir de https://revistas.unicauca.edu.co/index.php/biotecnologia/article/view/1221
Publicado
2013-11-01
Sección
Artículos de Investigaciòn