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PMID: 16346881 Published · ppublish English Journal Article

Activity stain for rapid characterization of pectic enzymes in isoelectric focusing and sodium dodecyl sulfate-polyacrylamide gels.

Applied and environmental microbiology ·Vol. 50 ·No. 3 ·1985-09-00 ·Pages 615-22

Ried JL, Collmer A

Abstract

A system was developed for the rapid characterization of microbial pectic enzyme complexes and then tested on Erwinia chrysanthemi and Sclerotium rolfsii. Pectic enzymes in minute samples of crude culture filtrates were resolved by ultrathin-layer polyacrylamide gel isoelectric focusing and sodium dodecyl sulfate-polyacrylamide gel electrophoresis and then assayed with an ultrathin pectate-agarose overlay stained with ruthenium red. The simple procedure can be completed within 30 min after isoelectric focusing, can detect extremely low levels of pectate lyase (6.4 x 10 mumol of product per min), and is sufficiently sensitive to determine the pectate lyase isozyme profile of a single bacterial colony with a diameter of 4 mm. Pectate lyases and polygalacturonases can be distinguished by altering buffer conditions in the overlays. The assay system revealed additional isozymes not resolved by classical techniques and generally corroborated the previously published isoelectric points and molecular weights of the pectate lyase isozymes and exo-poly-alpha-d-galacturonosidase produced by E. chrysanthemi and the endopolygalacturonase and exopolygalacturonase produced by S. rolfsii.

Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Ried J L
Department of Botany, University of Maryland, College Park, Maryland 20742.
Collmer A
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Article Info
Journal
Applied and environmental microbiology
Abbr.
Appl Environ Microbiol
ISSN
0099-2240
DOI
10.1128/aem.50.3.615-622.1985
Published
1985-09-00
Pages
615-22
Language
English
Region
United States
NLM ID
7605801
PMCID
PMC238678
External Links
PubMed source DOI: 10.1128/aem.50.3.615-622.1985 PMC Full Text View journal details
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