Screening and Optimization of Polyhydroxyalkanoate Production from Bacillus spp.
Abstract
Polyhydroxyalkanoates (PHA) are produced and accumulated as granules in cytoplasm of various bacteria for carbon and energy storages. This work aimed to screen the high PHA-producing Bacillus bacteria and optimize culture conditions for PHA production. The results revealed that among 200 bacterial isolates, Bt109-16 is the most efficient PHA producing bacterium (0.45 g/L, 34.31% of dry cell weight (DCW)). The maximum PHA production of 1.86 g/L (50.86% DCW) was achieved when culturing the Bt109-16 in the optimized medium containing 2% (w/v) sucrose, 0.3% (w/v) tryptone, pH 7.0 and cultured at 37°C for 72 h. Identification using 16S rRNA gene sequencing revealed that the Bt109-16 was Bacillus thuringiensis. Keywords : Bacillus, bioplastic, polyhydroxyalkanoates, carbon source, nitrogen sourceReferences
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Anderson, A. J. and Dawes E. A. (1990). Occurrence, metabolism, metabolic role, and industrial uses of bacterial polyhydroxyalkanoates. Microbiology and Molecular Biology Reviews, 54(4), 450-472.
Charen, T., Vaishali, P., Kaushalya, M., Amutha, K., Ponnusami, V. and Gowdhaman, D. (2014). Isolation and identification of polyhydroxybutyrate producing bacterial strain (Bacillus thuringiensis GVP) from chlorine contaminated soil. International Journal of ChemTech Research, 6(5), 3197-3202.
Desouky, S.E., Shiekh, H.H., Elabd, M.A. and Shehab, A.M. (2014). Screening, optimization and extraction of polyhydroxyalkanoates (PHAs) from Bacillus thuringienesis. Journal of Advances in Biology & Biotechnology, 1(1), 40-54.
Gowda, V. and Shivakumar S. (2013). Poly (3) hydroxybutyrate (PHB) production in Bacillus thuringiensis IAM 12077 under varied nutrient limiting conditions and molecular detection of class IV PHA synthase gene by PCR. International Journal of Pharmaceutical and Biological Sciences, 4(1), 794–802.
Luengo, J.M., Garcia, B., Sandoval, A., Naharre, G. and Olivera, E.R. (2003). Bioplastics from microorganisms. Current Opinion in Microbiology, 6, 251-260.
Mohamed, T., Shaaban, M., Azza, T. and Mowafy, E.I. (2012). Production of some biopolymers by some selective Egyptian soil bacterial isolates. Journal of Applied Sciences Research, 8(1), 94-105.
Murray, R.G.E., Doetsch, R.N. and Robinow, C.F. (1994). Determinative and cytological light microscopy. In P. Gerhardt, R.G.E. Murray, W.A. Wood and N.R. Krieg (Eds.), Manual of methods for general microbiology. (pp. 21-24). American Society for Microbiology.
Pal, A., Prabhu, A., Kumar, A.A., Rajagopal, B., Dadhe, K., Ponnamma V. and Shivakumar, S. (2009). Optimization of process parameters for maximum poly-β-hydroxybutyrate (PHB) production by Bacillus thuringiensis IAM 12077. Polish Journal of Microbiology, 58, 149-154.
Rai, R. Keshavarz, T., Roether, J.A., Boccaccini, A.R., Roy, I. (2011). Medium chain length polyhydroxyalkanoates, promising new biomedical materials for the future. Materials Science and Engineering: R: Reports, 72, 29–47.
Santhanam, A. and Sasidharan, S. (2010). Microbial production of polyhydroxy alkanote (PHA) from Alcaligens spp. and Pseudomonas oleovorans using different carbon sources. African Journal of Biotechnology, 9, 3144-3150.
Singhaboot, P. and Kaewkannetra, P. (2015). A higher in value biopolymer product of polyhydroxyalkanoates
(PHAs) synthesized by Alcaligenes latus in batch/repeated batch fermentation processes of sugar cane juice. Annals Microbiology, 65, 2081-2089.
Slepecky, R.A. and Law, J.H. (1960). A rapid spectrophotometric assay of alpha, beta-unsaturated acids and beta-hydroxy acids. Analytical Chemistry, 32(12), 1697-1699.
Suriyamongkol, P., Weselake, R., Narine, S., Moloney, M. and Sha, S.H. (2007). Biotechnological approaches for the production of polyhydroxyalkanoates in microorganisms and plants. Biotechnology Advances, 25, 148-175.
Tan, G.Y. A., Chen, C.L., Li, L., Ge, L., Wang, L., Razaad, I.M.N., Li, Y., Zhao, L., Mo, Y., and Wang, Ji.Y. (2014). Start a research on biopolymer polyhydroxyalkanoate (PHA): A Review. Polymers, 6,
706-754.
Taran, M. (2011). Utilization of petrochemical wastewater for the production of poly (3-hydroxybutyrate) by Haloarcula sp. IRU1. Journal of Hazardous Materials, 188, 26-28.
Anderson, A. J. and Dawes E. A. (1990). Occurrence, metabolism, metabolic role, and industrial uses of bacterial polyhydroxyalkanoates. Microbiology and Molecular Biology Reviews, 54(4), 450-472.
Charen, T., Vaishali, P., Kaushalya, M., Amutha, K., Ponnusami, V. and Gowdhaman, D. (2014). Isolation and identification of polyhydroxybutyrate producing bacterial strain (Bacillus thuringiensis GVP) from chlorine contaminated soil. International Journal of ChemTech Research, 6(5), 3197-3202.
Desouky, S.E., Shiekh, H.H., Elabd, M.A. and Shehab, A.M. (2014). Screening, optimization and extraction of polyhydroxyalkanoates (PHAs) from Bacillus thuringienesis. Journal of Advances in Biology & Biotechnology, 1(1), 40-54.
Gowda, V. and Shivakumar S. (2013). Poly (3) hydroxybutyrate (PHB) production in Bacillus thuringiensis IAM 12077 under varied nutrient limiting conditions and molecular detection of class IV PHA synthase gene by PCR. International Journal of Pharmaceutical and Biological Sciences, 4(1), 794–802.
Luengo, J.M., Garcia, B., Sandoval, A., Naharre, G. and Olivera, E.R. (2003). Bioplastics from microorganisms. Current Opinion in Microbiology, 6, 251-260.
Mohamed, T., Shaaban, M., Azza, T. and Mowafy, E.I. (2012). Production of some biopolymers by some selective Egyptian soil bacterial isolates. Journal of Applied Sciences Research, 8(1), 94-105.
Murray, R.G.E., Doetsch, R.N. and Robinow, C.F. (1994). Determinative and cytological light microscopy. In P. Gerhardt, R.G.E. Murray, W.A. Wood and N.R. Krieg (Eds.), Manual of methods for general microbiology. (pp. 21-24). American Society for Microbiology.
Pal, A., Prabhu, A., Kumar, A.A., Rajagopal, B., Dadhe, K., Ponnamma V. and Shivakumar, S. (2009). Optimization of process parameters for maximum poly-β-hydroxybutyrate (PHB) production by Bacillus thuringiensis IAM 12077. Polish Journal of Microbiology, 58, 149-154.
Rai, R. Keshavarz, T., Roether, J.A., Boccaccini, A.R., Roy, I. (2011). Medium chain length polyhydroxyalkanoates, promising new biomedical materials for the future. Materials Science and Engineering: R: Reports, 72, 29–47.
Santhanam, A. and Sasidharan, S. (2010). Microbial production of polyhydroxy alkanote (PHA) from Alcaligens spp. and Pseudomonas oleovorans using different carbon sources. African Journal of Biotechnology, 9, 3144-3150.
Singhaboot, P. and Kaewkannetra, P. (2015). A higher in value biopolymer product of polyhydroxyalkanoates
(PHAs) synthesized by Alcaligenes latus in batch/repeated batch fermentation processes of sugar cane juice. Annals Microbiology, 65, 2081-2089.
Slepecky, R.A. and Law, J.H. (1960). A rapid spectrophotometric assay of alpha, beta-unsaturated acids and beta-hydroxy acids. Analytical Chemistry, 32(12), 1697-1699.
Suriyamongkol, P., Weselake, R., Narine, S., Moloney, M. and Sha, S.H. (2007). Biotechnological approaches for the production of polyhydroxyalkanoates in microorganisms and plants. Biotechnology Advances, 25, 148-175.
Tan, G.Y. A., Chen, C.L., Li, L., Ge, L., Wang, L., Razaad, I.M.N., Li, Y., Zhao, L., Mo, Y., and Wang, Ji.Y. (2014). Start a research on biopolymer polyhydroxyalkanoate (PHA): A Review. Polymers, 6,
706-754.
Taran, M. (2011). Utilization of petrochemical wastewater for the production of poly (3-hydroxybutyrate) by Haloarcula sp. IRU1. Journal of Hazardous Materials, 188, 26-28.
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2017-08-01
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บทความวิจัยจากการประชุมวิชาการระดับชาติ"วิทยาศาสตร์วิจัย"ครั้งที่ 9
