Abstract
Previously, we used a simple, sensitive agar plate method to screen lipase activity from 1229 selected cultures, including 508 bacteria, 479 yeasts, 230 actinomycetes and 12 fungi, that covered many genera and species. About 25% of the cultures tested were lipase-positive. We also expanded our screening method to focus specifically on the pH dependence and thermostability of these lipase activities. In this report, we have characterized 25 yeast lipases, obtained from our screening program, on the basis of their positional specificity against triglycerides. Lipase was produced by growing cultures on nutrient medium in the presence of vegetable oil at 25°C for 4 d. Of the 25 new yeast lipases analyzed, 19 showed 1,3-positional specificity and 6 showed random specificity. No 2-positional specific lipases were found. Among those cultures with highest lipase activity are: Candida silvicola NRRL YB-2846 (random); Candida sp. 55 (random); Candida sp. 125 (random); Pichia americana NRRL Y-2156 (1,3-specific); P. muscicola NRRL Y-7005 (random); P. petersanii NRRL YB-3808 (1,3-specific); and Yarrowia lipolytica NRRL YB-423 (random). Characterization of Candida sp. strain 55 lipase on its substrate preference showed that this enzyme hydrolyzed soybean oil triglyceride species LLLn, LLL, LLO, and LLP more readily than LOO, LOP, OOO, LOS, and POO, where L=linoleic, Ln=linolenic, O=oleic, P=palmitic, and S=stearin.
Similar content being viewed by others
References
Klibanov, A.M., Enzymes That Work in Organic Solvent, Chem. Technol. 16:354–359 (1986).
Tsujisaka, Y., and M. Iwai, Comparative Study on Microbial Lipases, Kagaku To Kogyo 58:60–69 (1984).
Casy, J., and A. Macrae, Biotechnology and the Oleochemical Industry, INFORM 3:203–207 (1992).
Sih, C.J., Q.-M. Gu, G. Fulling, S.-H. Wu, and D.R. Reddy, The Use of Microbial Enzymes for the Synthesis of Optically Active Pharmaceuticals, Dev. Ind. Microbiol. 29:221–229 (1988).
Patel, R.N., A. Banergee, and L.J. Szarka, Biocatalytic Synthesis of Some Chiral Pharmaceutical Intermediates by Lipases, J. Am. Oil. Chem. Soc. 73:1363–1375 (1996).
Kokusho, Y., H. Machida, and S. Iwasaki, Studies on Alkaline Lipase: Isolation and Identification of Lipase Producing Microorganisms, Agric. Biol. Chem. 46:1159–1164 (1982).
Machida, H., T. Higashi, and Y. Kokusho, Industrial Production and Application of Microbial Lipases, Nippon Nogei Kagaku Kaishi 58:799–804 (1984).
Wang, Y., and B.C. Saha, Purification and Characterization of Thermophilic and Alkalophilic Tributyrin Esterase from Bacillus strain A30-1 (ATCC 53841), J. Am. Oil Chem. Soc. 70:1135–1138 (1993).
Hou, C.T., and T.M. Johnston, Screening of Lipase Activities with Cultures from ARS Culture Collection, Ibid.:1088–1097 (1992).
Hou, C.T., Screening of Microbial Enzyme for Asymmetric Hydrolysis of 2-Ethylhexyl Butyrate, J. Ind. Microbiol. 11:73–81 (1993).
Hou, C.T., pH Dependence and Thermostability of Lipases from Cultures from ARS Culture Collection, Ibid.:242–248 (1994).
Anderson, M.M., and R.E. McCarty, Rapid and Sensitive Assay for Free Fatty Acids Using Rhodamine 6G. Anal. Biochem. 45:260–270 (1972).
Zeitoun, M.A.M., W.E. Neff, E. Selke, and T.L. Mounts, Analyses of Vegetable Oil Triglyceride Molecular Species by Reversed-Phase High-Performance Liquid Chromatography, J. Liq. Chromatog. 14:2685–2698 (1991).
Byrdwell, W.C., E.A. Emken, W.E. Neff, and R.O. Adlof, Quantitative Analysis of Triglycerides Using Atmospheric Pressure Chemical Ionization-Mass Spectrometry, Lipids 31:919–935 (1996).
Author information
Authors and Affiliations
Corresponding author
About this article
Cite this article
Hou, C.T. Characterization of new yeast lipases. J Amer Oil Chem Soc 74, 1391–1394 (1997). https://doi.org/10.1007/s11746-997-0242-6
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/s11746-997-0242-6