Abstract
Aspergillus niger ORS-4.410, a mutant of A. niger ORS-4, was generated by repeated ultraviolet (UV) irradiation. Analysis of the UV treatment dose on wild-type (WT) A. niger ORS-4, conidial survival, and frequency of mutation showed that the maximum frequency of positive mutants (25.5%) was obtained with a 57% conidial survival rate after the second stage of UV irradiation. The level of glucose oxidase (GOX) production from mutant A. niger ORS-4.410 thus obtained was 149% higher than that for WT strain A. niger ORS-4 under liquid culture conditions using hexacyanoferrate (HCF)-treated sugarcane molasses (TM) as a cheaper carbohydrate source. When subcultured monthly for 24 mo, the mutant strain had consistent levels of GOX production (2.62±0.51 U/mL). Mutant A. niger ORS-4.410 was markedly different from the parent strain morphologically and was found to grow abundantly on sugarcane molasses. The mutant strain showed 3.43-fold increases in GOX levels (2.62±0.51 U/mL) using HCF-TM compared with the crude form of cane molasses (0.762±0.158 U/mL).
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Witteveen, C. F. B., Vande Vondervoort, P., Swart, K., and Visser, J. (1990) Appl. Microbiol. Biotechnol. 33, 683–686.
Roehr, M., Kubicek, C. P., and Kominek, J. (1996), in Biotechnology, Product of Primary Metabolism (Rehm, H. J. and Reed, G. eds.), Verlag Chemie, Weinheim, pp. 347–362.
Hatzinikolaou, D.G., Hansen, O.C., Macris, B. T., Tingey, A., Kekos, D., Goodenough, P., and Stougaard, P. (1996), Appl. Microbiol. Biotechnol. 46, 371–381.
Kiess, M., Hecht, H. J., and Kalisz, H. M. (1998), Eur. J. Biochem., 252, 90–99.
Park, E. H., Shin, Y. M., Lim, Y.Y., Kwon, T.H., Kim, D.H., and Yang, M. S. (2000), J. Biotechnol. 81, 35–44.
Liu, J. Z., Hung, Y. Y., Liu, J., Weng, L. P., and Ji, L. N. (2001), Lett. Appl. Microbiol. 32, 16–19.
Malherbe, D. F., Du Toit, M., Cordero Otero, R. R., van Rensburg, P., and Pretorius, I. S. (2003), Appl. Microbiol. Biotechnol., 61, 502–511.
Witteveen, C. F. B., Van de Vondervoort, P. J. I., Van den Broeck, H. C., et al. (1993), Curr. Genet. 24, 408–416.
Petruccioli, M., Piccioni, P., Federict, F., and Polsinelli, M. (1995), FEMS Microbiol. Lett. 128, 107–112.
Kapat, A., Jung J. K., and Park, Y. H., (1999), Biotechnol. Lett. 20, 683–686.
Kapat, A., Jung, J. K., and Park, Y. H. (2001) J. Appl. Microbiol. 90, 216–222.
Fiedurek, J., Gromada, A., and Prelecki, J., (1998), Acta Microbiol. Pol. 47, 355–364.
Kona, R. P., Qureshi, N., and Pai, J. S. (2001) Bioresour. Technol. 78, 123–126.
Luque, R., Orejas, M., Perotti, N. I., Ramon, D., and Lucca, M. E. (2004), J. Appl. Microbiol. 97, 332–337.
Singh, O. V. (2000), PhD thesis, Indian Institute of Technology (IIT), (formerly University of Roorkee), Roorkee, India.
Bergmeyer, H. U. (1974), in Methods of Enzymatic Analysis, Bergmeyer, H. U., ed., Academic, New York, pp. 457–460.
Lowry, O. H., Rosebrough, N. J., Farr, A. L., and Randall, R. J. (1951), J. Biol. Chem. 193, 265–275.
Miller, G. L. (1959), Anal. Chem. 31, 426–428.
Mann, F. G., and Saunders, B. C. (1960), in Practical Organic Chemistry, 4th ed., Longmans, London, pp. 458–461.
Bagdasaryan, Z. N., Aleksanvan, G. A., Mirzoyan, A. M., Roseiro, J. C., and Bagdasaryan, S. N. (2005), Appl. Biochem. Biotechnol., 125, 113–126.
Shukla, V. B., Zhou, S., Yomano, L. P., Shanmugam, K. T., Preston, J. F., and Ingram, L. O. (2004), Biotechnol. Lett. 26, 689–693.
Fiedurek, J. and Ilczuk, Z. (1992), Acta Microbiol. Pol. 41, 179–186.
Fiedurek, J. and Gromada, A. (2000), J. Appl. Microbiol. 89, 85–89.
Gromada, A., and Fiedurek, J. (1996), Acta Microbiol. Pol. 45, 37–43.
Tahoun, M. K. (1993), Appl. Biochem. Biotechnol., 39–40, 289–295.
Markwell, J., Frakes, L. G., Brott, E. C., Osterman, J., and Wagner, F. W. (1989), Appl. Microbiol. Biotechnol. 30, 166–169.
Ko, J. H., Hahm, M. S., Kang, H. A., Nam, S. W., and Chung, B. H. (2002), Protein Expr. Purif. 25, 488–493.
Lomkatsi, E. T., Radiami, T. S., Shkolni, A. T., et al. (1990), Acta Biotechnol. 10, 377–381.
Jefferson, W. R. Jr. (1967), Biochemistry 6, 3479–3484.
Panda, T., Kundu, S., and Majumdar, S. K., (1984), Microbiol. J. 52, 61–66.
Lu, T., Peng, X., Yang, H., and Ji, L. (1996), Enzyme Microb. Technol., 19, 339–342.
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The results reported herein were obtained while the author was working at the Department of Biotechnology, Indian Institute of Technology, Roorkee-247667, India.
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Singh, O.V. Mutagenesis and analysis of mold Aspergillus niger for extracellular glucose oxidase production using sugarcane molasses. Appl Biochem Biotechnol 135, 43–57 (2006). https://doi.org/10.1385/ABAB:135:1:43
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DOI: https://doi.org/10.1385/ABAB:135:1:43