Proteases from actinomycetes interfere in solid media plate assays of hyaluronidase activity

Four hundred and fifteen actinomycete strains were screened for hyaluronidase activity in two plate assays media. In the first one, using hyaluronic acid as substrate and bovine serum albumin (BSA) to help precipitation of the nondegraded substrate, only strain 594 and hyaluronidase control were pos...

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Bibliographic Details
Published in:Journal of microbiological methods 2001-07, Vol.45 (3), p.207-212
Main Authors: de Azeredo, L.A.I, Leite, S.G.F, Freire, D.M.G, Benchetrit, L.C, Coelho, R.R.R
Format: Article
Language:English
Subjects:
Actinomycetales - enzymology
Actinomycetes
Bacteriological Techniques - methods
Bacteriology
Biological and medical sciences
Brazil
Culture Media
Endopeptidases - metabolism
Fundamental and applied biological sciences. Psychology
Hyaluronidase
Hyaluronoglucosaminidase - metabolism
Metabolism. Enzymes
Microbiology
Plate assays
Protease
Protein interference
Serum Albumin, Bovine
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Summary:Four hundred and fifteen actinomycete strains were screened for hyaluronidase activity in two plate assays media. In the first one, using hyaluronic acid as substrate and bovine serum albumin (BSA) to help precipitation of the nondegraded substrate, only strain 594 and hyaluronidase control were positive. In the second assay, plates with hyaluronic acid, but not BSA, gave the same results. For plates containing only BSA, proteinase activity was detected in strain 594. When hyaluronic acid was treated with pronase, the only clear zones, in the second assay without BSA, were those around hyaluronidase controls. Protease activity, commonly found in actinomycetes, was detected only in strain 594, among the 415 studied, when tested in hyaluronidase assay using hyaluronate plus BSA. This may be due to the composition of the growth medium, since media with different composition gave different results for protease activity in each of the 15 strains analyzed. These data suggest that proteases can affect an accurate detection of hyaluronidase in media containing proteins, not only from hyaluronate preparations, but also from other medium ingredients. Thus, for a correct interpretation of the method, they must be excluded. Commercial Hyaluronidase used as controls must be also tested for the presence of protease contamination.
ISSN:0167-7012
1872-8359
DOI:10.1016/S0167-7012(01)00251-2