Filiz Ekinci Sanal
Department of Biology, Faculty of Science and Art, Trakya University, 22080 Edirne, Turkey
Figen Ertan
Department of Biology, Faculty of Science and Art, Trakya University, 22080 Edirne, Turkey
Tulin Aktac
Department of Biology, Faculty of Science and Art, Trakya University, 22080 Edirne, Turkey
ABSTRACT
The effect of different carbon and nitrogen sources in the medium on inulinase production were investigated in this research. The enzyme activity was maximum in media containing 3% Jerusalem artichoke powder. NH4H2PO4 was found to be the better nitrogen source. NH4H2PO4 + NH4NO3 combination had higher inulinase yields than other combined nitrogen sources. The enzyme was purified 65.89 fold by using dialysis, concentrating, gel filtration (Sephadex G-150) and ion exchange chromatography (DEAE-cellulose) techniques. The Km and Vmax values of inulinase for inulin were 0.52 mM and 8.69 mmol mL 1 min 1, respectively. Optimum temperature and pH for the activity of the purified enzyme were found to be 30°C and 5.0 and the enzyme was found to be stable up to 40°C and in a pH range of 4.0 to 6.0. The enzyme was inhibited strongly by Mn2+ , KCN and DTNB. The molecular weight of the enzyme was estimated to be 66.000 and 68.000 Da by SDS polyacrylamide gel electrophoresis and gel filtration on sephadex G-200, respectively. The inulinase of A. alternata was hydrolised pure inulin and raw jerusalem artichoke powder in similar amounts. This results have industrial importance.
PDF References Citation
How to cite this article
Filiz Ekinci Sanal, Figen Ertan and Tulin Aktac, 2005. Production of Exo-inulinase from Alternaria alternata Growth on
Jerusalem Art choke and Some Biochemical Properties. Journal of Biological Sciences, 5: 497-505.
DOI: 10.3923/jbs.2005.497.505
URL: https://scialert.net/abstract/?doi=jbs.2005.497.505
DOI: 10.3923/jbs.2005.497.505
URL: https://scialert.net/abstract/?doi=jbs.2005.497.505
REFERENCES
- Gupta, A.K., P. Rathore, N. Kaur and R. Sing, 1990. Production, thermal stability and immobilization of inulinase from Fusarium oxysporum. J. Chem. Biotechnol., 47: 245-257.
CrossRefDirect Link - Gupta, A.K., D.P. Singh, N. Kaur and R. Singh, 1994. Production, purification and immobilization of inulinase from Kluyveromyces fragilis. J. Chem. Technol. Biotechnol., 59: 377-385.
CrossRefDirect Link - Gupta, A.K., A. Gill and N. Kaur, 1998. A HgCl2 insensitive and thermally stable inulinase from Aspergillus oryzae. Phytochemistry, 49: 55-58.
CrossRef - Nakamura, T., A. Shitara, S. Matsuda, T. Matsuo, M. Suiko and K. Ohta, 1997. Production, purification and properties of an endoinulinase of Penicillium sp. TN-88 that liberates inulotriose. J. Ferment. Bioeng., 84: 313-318.
CrossRef - Xiao, R., M. Tanida and S. Takao, 1988. Inulinase from Chrysosporium pannorum. J. Ferment. Technol., 66: 553-558.
CrossRef - Miller, G.L., 1959. Use of dinitrosalicylic acid reagent for determination of reducing sugar. Anal. Chem., 31: 426-428.
CrossRefDirect Link - Lowry, O.H., N.J. Rosebrough, A.L. Farr and R.J. Randall, 1951. Protein measurement with the folin phenol reagent. J. Biol. Chem., 193: 265-275.
CrossRefPubMedDirect Link - Arand, M., A.M. Golubev, J.R.B. Neto, I. Polikarpov and R. Wattiez et al., 2002. Purification, characterization, gene cloning and preliminary X-ray data of the exo-inulinase from Aspergillus awamori. Biochem. J., 362: 131-135.
Direct Link - Kushi, R.T., R. Monti and J. Contiero, 2000. Production, purification and characterization of an extracellular inulinase from Kluyveromyces marxianus var. bulgaricus. J. Ind. Microbiol. Biotechnol., 25: 63-69.
CrossRefDirect Link - Pessoni, R.A.B., R.L.C. Figueiredo-Ribeiro and M.R. Braga, 1999. Extracellular inulinases from Penicillium janczewskii, a fungus isolated from the rhizosphere of Vernonia herbacea (Asteraceae). J. Applied Microbiol., 87: 141-147.
CrossRef - Pandey, A., C.R. Soccol, P. Selvakumar, V.T. Soccol, N. Krieger and J.D. Fontana, 1999. Recent developments in microbial inulinases. Its production, properties and industrial applications. Applied Biochem. Biotechnol., 81: 35-52.
CrossRefPubMedDirect Link - Barthomeuf, C., F. Regerat and H. Pourrat, 1991. Production of inulinase by a new mold of Penicillium rugulosum. J. Ferment. Bioeng., 72: 491-494.
CrossRef - Yun, J.W., D.H. Kim, B.W. Kim and S.K. Song, 1997. Production of inulo-oligosaccharides from inulin by immobilised endoinulinase from Pseudomonas sp. J. Ferment. Bioeng., 84: 369-371.
Direct Link - Kim, D.H., Y.J. Choi, S.K. Song and J.W. Yun, 1997. Production of inulo-oligosaccharides using endo-inulinase from a Pseudomonas sp. Biotechnol. Lett., 9: 369-372.
CrossRefDirect Link - Gupta, A.K., B. Nagpal, N. Kaur and R. Singh, 1988. Mycelial and exracellular inulinases from Fusarium oxysporum grown on aqueous extract of Cichorium intybus roots. J. Chem. Technol. Biotechnol., 42: 69-76.
Direct Link