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Gibberellic acid production by Gibberella fujikuroi under solid-state fermentation of cassava bagasse and rice hull
Abstracts / Journal of Biotechnology 136S (2008) S356–S401 Ni, Y., Xu, J.H., 2002. Asymmetric reduction of aryl ketones with a new isolate Rhodotorula sp. AS2.2241. J. Mol. Catal. B: Enzym. 18, 233–241. Yang, W., Xu, J.H., Xie, Y., Xu, Y., Zhao, G., Lin, G.Q., 2006. Asymmetric reduction of ketones by employing Rhodotorula sp. AS2.2241 and synthesis of the betablocker (R)-nifenalol. Tetrahedron: Asymmetry 17, 1769–1774.
S371
V3-P-042 Synthesis of a novel fatty amide derivative of 7,10-dihydroxy8(E)-octadecenoic acid by lipase-catalyzed amidation reaction Sunil K. Khare 1,∗ , Tsung Min Kuo 2 , N.G. Ramesh 1
doi:10.1016/j.jbiotec.2008.07.852 V3-P-041 Gibberellic acid production by Gibberella fujikuroi under solidstate fermentation of cassava bagasse and rice hull Ángela M. Otálvaro ∗ , Germán D. Gutiérrez, Diego A. Pierotty, Fabián A. Parada, Néstor A. Algecira Departamento de Ingeniería Química, Universidad Nacional de Colombia, Bogotá, Colombia E-mail address: [email protected] (Á.M. Otálvaro). The study purpose was to produce gibberellic acid from cassava bagasse (CB) and rice hull (RH) to improve this agricultural residue in one product of high added value. In the solid-state fermentation (SSF) using Gibberella fujikuroi (ATCC 12616), four variables were evaluated: substrate (CB, RH), pH (4.0, 4.5 and 5.0), temperature (28, 30 and 32 ◦ C), and moisture (50, 60 and 70%). A simple factorial analysis was used to compare the fermentation conditions and each test was repeated by triplicate. In each case the substrate was added with salts solution with micro- and macronutrients (Kumar and Lonsane, 1989). Gibberellic acid was quantified by high-performance liquid chromatography (HPLC) using a RP (Li-Chrosorb) column and UV detector at 206 nm. For this analysis it was necessary to generate methanolic extracts of solid matrices. The fermentation time was established in 8 days. The results indicate that it is possible to obtain this substance under solid-state fermentation and it is clear that the CB is the best substrate to obtain it. The higher amount of gibberellic acid extracted of this substrate was 1.58 mg/g of dry substrate obtained at 28 ◦ C, pH 4.5 and 70% of moisture. At the end of this study it was propose the possibility to establish another techniques to produce gibberellic acid with G. fujikuroi, one of the is its use as supported biocatalysts on porous matrices, and the other possibility studied was the liquid state fermentation using medium added with cassava bagasse as carbon source (Gelmi et al., 2000; Escamilla et al., 2000; Pierotty et al., 2006). References ˜ I., Parra, R., De la Torre, M., 2000. OptimizaEscamilla, E.M., Dendooven, L., Magana, tion of gibberellic acid production by immobilized Gibberella fujikuroi mycelium in fluidized bioreactors. J. Biotechnol. 76, 147–155. Gelmi, C., Pérez, C.R., Gonzáles, M., Agosin, E., 2000. Solid substrate cultivation of Gibberella fujikuroi on an inert support. Process Biochem. 35, 1227–1233. Kumar, P.K.R., Lonsane, B.K., 1989. Microbial production of gibberellins: state of the art. Adv. Appl. Microbiol. 34, 29–138. Pierotty, D.A., Otálvaro, A.M., Algecira, N.E., 2006. Producción de ácido giberélico por fermentación utilizando biocatalizadores soportados en matrices porosas. Revista Universitas Scientiarium 11 (2), 41–50.
doi:10.1016/j.jbiotec.2008.07.853
1
Department of Chemistry, Indian Institute of Technology, New Delhi, India 2 Microbial Genomics and Bioprocess Research Unit, National Centre for Agricultural Utilization Research, Peoria, IL 61604, USA E-mail addresses: [email protected] [email protected] (T.M. Kuo).
(S.K.
Khare),
Fatty acid amides are of considerable interest due to their wide ranging applications in detergents, shampoo, cosmetics, surfactant and other allied industries (Kuo and Kaneshiro, 2002; Sharma et al., 2005). Desired amides are produced from fatty acids by reacting with anhydrous ammonia at approximately 200 ◦ C and 345–690 kPa pressure. Candida antarctica lipase B (CALB) is known to catalyze direct amidation of carboxylic acid in organic solvent (Gotor-Fernández et al., 2006). CALB mediated production of secondary amide from novel hydroxyl fatty acid—7,10-dihydroxy8(E)-octadecenoic acid (DOD) (Kuo et al., 2003) using methyl ethanolamine as amino group donor was investigated. A new peak corresponding to secondary amide of DOD (D2AM) was detected in the HPLC. Amidation was also confirmed by IR spectrum. Ninetyfive percent yield was obtained after 72 h of CALB catalyzed reaction with 50 mM DOD, 100 mM methyl ethanolamine in isoamyl alcohol by 100 IU enzyme activity. The product D2AM was further investigated for its biological activity. It displayed potent antimicrobial activity towards Gram positive (Bacillus subtilis and Staphylococcus aureus) and Gram negative bacteria (Proteus vulgaris and Klebsiella pneumonae) with minimal inhibition concentration (MIC) of 15.0, 6.0, 10.0 and 2.0 g/ml, respectively. D2Am also exhibited weak DPPH radical scavenging antioxidant activity. Acknowledgements Dedicated to great scientist Dr. Tsung Min Kuo under whose guidance the work was done. It is with great sadness to write that Dr. Tsung Min Kuo died on 10 June 2008 after battling with cancer for last couple of years. The financial support by DBT and IIT Delhi, India and facilities provided by NCAUR, USDA, Peoria, USA to SKK are gratefully acknowledged. References Gotor-Fernández, V., Busto, E., Gotor, V., 2006. Candida antarctica lipase B: an ideal biocatalyst for the preparation of nitrogenated organic compounds. Adv. Synthesis Catal. 348, 797–812. Kuo, T.M., Kaneshiro, T.H., 2002. Microbiological conversions of fatty acids to valueadded products. In: Kuo, T.M., Gardner, H.W. (Eds.), Lipid Biotechnology. Marcel Dekker Inc., NY. Kuo, T.M., Ray, K.J., Manthey, L.K., 2003. A facile process for production of 7,10dihydroxy 8-(E)-octadecenoic acid from oleic acid conversion by Pseudomonas aeruginosa. Biotechnol. Lett. 25, 29–33. Sharma, J., Batovsaka, D., Kuwamori, Y., Asano, Y., 2005. Enzymatic chemoselective synthesis of secondary amide surfactant from N-methylethanol amine. J. Biosci. Bioeng. 100, 662–666.
doi:10.1016/j.jbiotec.2008.07.854
S371
V3-P-042 Synthesis of a novel fatty amide derivative of 7,10-dihydroxy8(E)-octadecenoic acid by lipase-catalyzed amidation reaction Sunil K. Khare 1,∗ , Tsung Min Kuo 2 , N.G. Ramesh 1
doi:10.1016/j.jbiotec.2008.07.852 V3-P-041 Gibberellic acid production by Gibberella fujikuroi under solidstate fermentation of cassava bagasse and rice hull Ángela M. Otálvaro ∗ , Germán D. Gutiérrez, Diego A. Pierotty, Fabián A. Parada, Néstor A. Algecira Departamento de Ingeniería Química, Universidad Nacional de Colombia, Bogotá, Colombia E-mail address: [email protected] (Á.M. Otálvaro). The study purpose was to produce gibberellic acid from cassava bagasse (CB) and rice hull (RH) to improve this agricultural residue in one product of high added value. In the solid-state fermentation (SSF) using Gibberella fujikuroi (ATCC 12616), four variables were evaluated: substrate (CB, RH), pH (4.0, 4.5 and 5.0), temperature (28, 30 and 32 ◦ C), and moisture (50, 60 and 70%). A simple factorial analysis was used to compare the fermentation conditions and each test was repeated by triplicate. In each case the substrate was added with salts solution with micro- and macronutrients (Kumar and Lonsane, 1989). Gibberellic acid was quantified by high-performance liquid chromatography (HPLC) using a RP (Li-Chrosorb) column and UV detector at 206 nm. For this analysis it was necessary to generate methanolic extracts of solid matrices. The fermentation time was established in 8 days. The results indicate that it is possible to obtain this substance under solid-state fermentation and it is clear that the CB is the best substrate to obtain it. The higher amount of gibberellic acid extracted of this substrate was 1.58 mg/g of dry substrate obtained at 28 ◦ C, pH 4.5 and 70% of moisture. At the end of this study it was propose the possibility to establish another techniques to produce gibberellic acid with G. fujikuroi, one of the is its use as supported biocatalysts on porous matrices, and the other possibility studied was the liquid state fermentation using medium added with cassava bagasse as carbon source (Gelmi et al., 2000; Escamilla et al., 2000; Pierotty et al., 2006). References ˜ I., Parra, R., De la Torre, M., 2000. OptimizaEscamilla, E.M., Dendooven, L., Magana, tion of gibberellic acid production by immobilized Gibberella fujikuroi mycelium in fluidized bioreactors. J. Biotechnol. 76, 147–155. Gelmi, C., Pérez, C.R., Gonzáles, M., Agosin, E., 2000. Solid substrate cultivation of Gibberella fujikuroi on an inert support. Process Biochem. 35, 1227–1233. Kumar, P.K.R., Lonsane, B.K., 1989. Microbial production of gibberellins: state of the art. Adv. Appl. Microbiol. 34, 29–138. Pierotty, D.A., Otálvaro, A.M., Algecira, N.E., 2006. Producción de ácido giberélico por fermentación utilizando biocatalizadores soportados en matrices porosas. Revista Universitas Scientiarium 11 (2), 41–50.
doi:10.1016/j.jbiotec.2008.07.853
1
Department of Chemistry, Indian Institute of Technology, New Delhi, India 2 Microbial Genomics and Bioprocess Research Unit, National Centre for Agricultural Utilization Research, Peoria, IL 61604, USA E-mail addresses: [email protected] [email protected] (T.M. Kuo).
(S.K.
Khare),
Fatty acid amides are of considerable interest due to their wide ranging applications in detergents, shampoo, cosmetics, surfactant and other allied industries (Kuo and Kaneshiro, 2002; Sharma et al., 2005). Desired amides are produced from fatty acids by reacting with anhydrous ammonia at approximately 200 ◦ C and 345–690 kPa pressure. Candida antarctica lipase B (CALB) is known to catalyze direct amidation of carboxylic acid in organic solvent (Gotor-Fernández et al., 2006). CALB mediated production of secondary amide from novel hydroxyl fatty acid—7,10-dihydroxy8(E)-octadecenoic acid (DOD) (Kuo et al., 2003) using methyl ethanolamine as amino group donor was investigated. A new peak corresponding to secondary amide of DOD (D2AM) was detected in the HPLC. Amidation was also confirmed by IR spectrum. Ninetyfive percent yield was obtained after 72 h of CALB catalyzed reaction with 50 mM DOD, 100 mM methyl ethanolamine in isoamyl alcohol by 100 IU enzyme activity. The product D2AM was further investigated for its biological activity. It displayed potent antimicrobial activity towards Gram positive (Bacillus subtilis and Staphylococcus aureus) and Gram negative bacteria (Proteus vulgaris and Klebsiella pneumonae) with minimal inhibition concentration (MIC) of 15.0, 6.0, 10.0 and 2.0 g/ml, respectively. D2Am also exhibited weak DPPH radical scavenging antioxidant activity. Acknowledgements Dedicated to great scientist Dr. Tsung Min Kuo under whose guidance the work was done. It is with great sadness to write that Dr. Tsung Min Kuo died on 10 June 2008 after battling with cancer for last couple of years. The financial support by DBT and IIT Delhi, India and facilities provided by NCAUR, USDA, Peoria, USA to SKK are gratefully acknowledged. References Gotor-Fernández, V., Busto, E., Gotor, V., 2006. Candida antarctica lipase B: an ideal biocatalyst for the preparation of nitrogenated organic compounds. Adv. Synthesis Catal. 348, 797–812. Kuo, T.M., Kaneshiro, T.H., 2002. Microbiological conversions of fatty acids to valueadded products. In: Kuo, T.M., Gardner, H.W. (Eds.), Lipid Biotechnology. Marcel Dekker Inc., NY. Kuo, T.M., Ray, K.J., Manthey, L.K., 2003. A facile process for production of 7,10dihydroxy 8-(E)-octadecenoic acid from oleic acid conversion by Pseudomonas aeruginosa. Biotechnol. Lett. 25, 29–33. Sharma, J., Batovsaka, D., Kuwamori, Y., Asano, Y., 2005. Enzymatic chemoselective synthesis of secondary amide surfactant from N-methylethanol amine. J. Biosci. Bioeng. 100, 662–666.
doi:10.1016/j.jbiotec.2008.07.854