biologia plantarum

International journal on Plant Life established by Bohumil Němec in 1959

Biologia plantarum 39:475-477, 1997 | DOI: 10.1023/A:1001194704203

A simple procedure for the detection of plant extracellular proteolytic enzymes

J. Stano1,*, P. Kovács1,2, I. Šafařík1,3, D. Kákoniová1,4, M. Šafaříková1,3
1 Garden of Medical Plants, Faculty of Pharmacy, Comenius University, Bratislava, Slovak Republic
2 Department of Cell and Molecular Biology of Drugs, Faculty of Pharmacy, Comenius University, Bratislava, Slovak Republic
3 Laboratory of Biochemistry and Biotechnology, Institute of Landscape Ecology, Academy of Sciences of the Czech Republic, České Budějovice, Czech Republic
4 Institute of Chemistry, Slovak Academy of Sciences, Bratislava, Slovak Republic

A simple procedure for the detection of extracellular plant proteolytic enzymes using insoluble dye stained gelatin substrates incorporated into an appropriate culture medium is described. Extracellular proteinases produced by the tested plant cells (callus culture and cell suspension) hydrolyzed the substrates and dyed peptide fragments were released. Dyed zones around and under the proteinase-producing callus cultures were formed on the agar medium. Similarly, coloration of the culture media using proteinase-producing cell suspensions was observed.

Keywords: extracellular proteolytic activity; insoluble chromolytic substrate; callus cultures
Subjects: callus (tissue) culture, proteolytic activity; extracellular proteolytic activity, detection; proteolytic enzymes, extracellular, detection

Published: November 1, 1997  Show citation

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Stano, J., Kovács, P., Šafařík, I., Kákoniová, D., & Šafaříková, M. (1997). A simple procedure for the detection of plant extracellular proteolytic enzymes. Biologia plantarum, 39(3), 475-477. doi: 10.1023/A:1001194704203
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    References

    1. Frazier, W.C., Rupp, P.: Studies on the proteolytic bacteria of milk. I. A medium for the direct isolation of caseolytic milk bacteria.-J. Bacteriol. 16: 57-63, 1928. Go to original source...
    2. Rust, L., Messing, C.R., Iglewski, B.H.: Elastasc assays.-Meth. Enzymol. 235: 554-62, 1994. Go to original source...
    3. Stano, J., Nemec, P., Kákoniová, D., Kovács, P., Neubert, K., Lišková, D.: Alpha-galactosidase in immobilized cells of Cucumis sativus L.-Biologia 50: 279-281, 1995.
    4. Šafařík, I.: An insoluble chromolytic substrate for the determination of proteolytic activity.-J. Biochem. biophys. Meth. 17: 193-198, 1988. Go to original source...
    5. Šafařík, I.: Spectrophotometric determination of proteolytic activity in coloured solutions.-J. Biochem. biophys. Meth. 19: 201-206, 1989. Go to original source...
    6. Šafařík, I.: Dye-stained gelatin microcarriers as insoluble chromolytic substrates for the determination of proteolytic activity.-Clin. chim. Acta 187, 149-154, 1990. Go to original source...
    7. Šafařík, I., Šafaříková, M.: A modified procedure for the detection of microbial producers of extracellular proteolytic enzymes.-Biotechnol. Techniques 8: 627-628, 1994. Go to original source...
    8. Šafařík, I., Stejskalová, R., Králová, B.: Bacterial producers of fibrinolytic enzymes isolated from soil.-Sbnorník VŠCHT Praha E 55: 75-86, 1983.
    9. Whaley, D.N., Dowel, V.R., Jr., Wanderlinder, L.M., Lomnard, G.L.: Gelatin agar medium for detecting gelatinase production by anaerobic bacteria.-J. Clin. Microbiol. 16: 224-229, 1982. Go to original source...
    10. Wunderwald, P.: Proteinases, Proteins as substrates.-In: Bergmeyer, H.U. (ed.): Methods of Enzymatic Analysis. Vol. 5. Pp. 258-270. Verlag Chemie, Weinheim 1984.

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