Abstract
The localization of lipase in cells of the fungus Penicillium cyclopium was investigated. It was shown by differential centrifugation of a homogenate of mycelial cells that the activity of the enzyme is associated with the cell wall. A study of ultrathin sections of mycelium fixed using the method of Zvyagintseva in an electron microscope showed that the final products of lipolytic activity of the enzyme is localized on the cell wall. Antibodies were raised against the purified A and B lipases from P. cyclopium and their specificity was assessed by enzyme-linked immunosorbent assay. The antibody preparation was used in cytochemical investigation by immunogold labelling. This study permits the localization of cell-bound lipase mainly in the cell wall and in the periplasmic space. The identity of the cell-bound lipase with one of the two extracellular lipases is also demonstrated.
Similar content being viewed by others
References
Dargent R (1977) Cytodifferentiation et croissance apicale chez les champignons filamenteux. Thèse de Biologie végétale, Université de Toulouse. III
Davranov KD, Alimdzhanova MI, Bezborodov AM (1980) Lipase activity of Oospora lactis protoplasts. Mikrobiologiya 49:421–426
Druet D, El Abadi N, Comeau LC (1992) Purification and characterization of the extracellular and cell-bound lipases from a Penicillium cyclopium variety. Appl Microbiol Biotechnol 37:745–749
Glynn JA, Schaffel S, Mc Nicholas JM, Hulett FM (1977) Biochemical localization of the alkaline phosphatase of Bacillus licheniformis as a function of culture age. J Bacteriol 129:1010–1019
Engvall E, Perlamann P (1972) Enzyme-linked immunosorbent assay, ELISA III. Quantification of specific antibodies by enzyme-labeled anti-immunoglobulin in antigen-coated tubes. J Immunol Methods 109:129–135
Legier V, Comeau LC (1992) Continuous synthesis of esters by cell-bound fungal lipases in organic solvent. Appl Microbiol Biotechnol 37:732–736
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275
Luft JH (1961) Improvements in epoxy resin embedding methods. J Biophys Biochem Cytol 9:409–413
Pottu-Boumendil J (1989) Microscopie electronique: principes et méthodes de préparation. Ed l'INSERM, Paris, pp 191–217
Sabatini DD, Bensch K, Barrnnett RJ (1963) Cytochemistry and electron microscopy. The preservation of cellular ultrastructure and enzymatic activity by aldehyde fixation. J Cell Biol 17:19–58
Smith E, Roberts K, Butcher GW, Galfre G (1984) Monoclonal antibody screening: two methods using antigens immobilized on nitrocellulose. Anal Biochem 138:119–124
Reynolds ES (1963) The use of lead citrate at high pH as an electron opaque stain in electron microscopy. J Cell Biol 17:208–213
Vaitukaitis LJ (1981) Production of antisera with small dose of immunogen: multiple intradermal injection. Methods Enzymol 73:46–52
Watson ML (1958) Staining of tissue sections for electron microscopy with heavy metals. II Application of solutions containing lead and barium. J Biophys Biochem Cytol 4:727–730
Zvyagintseva IS, Dmitriev VV, Melikhovskii VA, Fikhte BA (1983) Localization of lipase in Penicillium cyclopium hyphae. Mikrobiologiya 52:263–267
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
El Abbadi, N., Druet, D. & Comeau, L.C. Immunocytochemical identification and localization of lipase in cells of the mycelium of Penicillium cyclopium variety. Appl Microbiol Biotechnol 42, 923–930 (1995). https://doi.org/10.1007/BF00191192
Received:
Revised:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00191192