Abstract
An extracellular lipase-producing fungus was isolated from oil-rich soil. This fungus belongs to the genus Rhizopus and clades with Rhizopus oryzae. Lipase was purified to homogeneity from this novel fungal source using ammonium sulphate precipitation followed by Q-Sepharose chromatography. The extracellular lipase was purified 8.6–fold, and enzymatic properties were studied. The molecular mass of the purified enzyme was estimated to be 17 kD by sodium dodecyl sulphate-polyacrylamide gel electrophoresis and 16.25 kD by matrix-assisted laser desorption ionization/time-of-flight analysis. The native molecular mass was estimated to be 17.5 kD by gel filtration, indicating the protein to be monomer. The optimum pH and temperature for the enzyme catalysis were 7.0 °C and 40 °C, respectively. Enzyme was stable in pH range 6.0–7.0 and retains 95–100% activity when incubated at 50 °C for 1 h. The pI of the purified lipase was 4.2. Enzyme was stable in the organic solvents such as ethanol, hexane and methanol for 2 h. Purified enzyme was used for transesterification of oleic acid in the presence of ethanol for production of oleic acid ethyl ester with a conversion efficiency of 66% after 24 h at 30 °C.
Similar content being viewed by others
References
Fukuda, H., Kondo, A., & Noda, H. (2001). Journal of Biosciences and Bioengineering, 92, 405–416.
Hasan, F., Shah, A. A., & Hameed, A. (2006). Enzyme and Microbial Technology, 39, 235–251.
Jaeger, K., & Reetz, M. T. (1998). Trends in Biotechnology, 16, 396–403.
Jaeger, K., & Eggert, T. (2002). Current Opinion in Biotechnology, 13, 390–397.
Essamri, M., Deyris, V., & Comeau, L. (1998). Journal of Biotechnology, 60, 97–103.
Kantak, J. B., Bagade, A. V., Mahajan, S. A., Pawar, S. P., Shouche, Y. S., & Prabhune, A. A. (2011). Applied Biochemistry and Biotechnology, 164, 969–978.
Rapp, P., & Backhaus, S. (1992). Enzyme and Microbial Technology, 14, 938–943.
Winkler, U. K., & Stuckmann, M. (1979). Journal of Bacteriology, 138, 663–670.
Lowry, O. H., Rosenbrough, N. J., Farr, A. L., & Randall, R. J. (1951). Journal of Biological Chemistry, 193, 265–275.
Laemmli, U. K. (1970). Nature, 227, 680–685.
Morrissey, J. H. (1981). Analytical Biochemistry, 117, 307–310.
Vesterberg, O. (1972). Biochimica et Biophysica Acta, 257, 11–19.
Andrews, P. (1964). Biochemical Journal, 91, 222–233.
Hiol, A., Jonzo, M. D., Rugani, N., Druet, D., Sarda, L., & Comeau, L. C. (2000). Enzyme and Microbial Technology, 26, 421–430.
Iwai, M., & Tsujisaka, Y. (1974). Agricultural and Biological Chemistry, 38, 1241–1247.
Sun, S. Y., Xu, Y., & Wang, D. (2009). Journal of Chemical Technology and Biotechnology, 84, 435–441.
Mateos Diaz, J. C., Rodriguez, J. A., Roussos, S., Cordova, J., Abousalham, A., & Carriere, F. (2006). Enzyme and Microbial Technology, 39, 1042–1050.
Fukumoto, J., Iwai, M., & Tsujisaka, Y. (1963). Journal of General and Applied Microbiology, 9, 353–361.
Liu, H. W., Beppu, T., & Arima, K. (1973). Agricultural and Biological Chemistry, 37, 157–163.
Nagaoka, K., & Yamada, Y. (1973). Agricultural and Biological Chemistry, 37, 2791–2796.
Kermasha, S., Safari, M., & Bisakowski, B. (1998). Journal of Agricultural and Food Chemistry, 46, 4451–4456.
Hoil, A., Marie, J. D., Danielle, D., & Comeau, L. (1999). Enzyme and Microbial Technology, 25, 80–87.
Yu, M., Qin, S., & Tan, T. (2007). Process Biochemistry, 42, 384–391.
Saxena, R. K., Davidson, W. S., Sheoran, A., & Giri, B. (2003). Process Biochemistry, 39, 239–247.
Salameh, M. D., & Wiegel, J. (2007). Applied and Environmental Microbiology, 73, 7725–7731.
Ateslier, Z. B. B., & Metin, K. (2006). Enzyme and Microbial Technology, 38, 628–635.
Shah, S., Sharma, S., & Gupta, M. N. (2004). Energy & Fuels, 18, 154–159.
Acknowledgments
We thank Dr. Mahesh Kulkarni, Centre for Material Characterization, National Chemical Laboratory, for help in MALDI-TOF-MS analysis. The financial support provided by the Council of Scientific and Industrial Research (CSIR), Govt. of India to Jayshree B. Kantak in the form of Senior Research Fellowships is duly acknowledged.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Kantak, J.B., Prabhune, A.A. Characterization of Smallest Active Monomeric Lipase from Novel Rhizopus Strain: Application in Transesterification. Appl Biochem Biotechnol 166, 1769–1780 (2012). https://doi.org/10.1007/s12010-012-9584-0
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12010-012-9584-0