Abstract
A protocol is described which allows the detection of transglycosylase isoenzyme activity. Crude extracts of cauliflower (Brassica oleracea var. botrytis) florets were treated with 1 M sodium chloride, filtered through cellulose acetate, dialysed, concentrated and subjected to isoelectric focusing under non-denaturing conditions in a polyacrylamide gel. After electrophoresis, the gel was overlaid with a test paper impregnated with a mixture of high-M(r) xyloglucan and a sulphorhodamine-conjugated oligosaccharide of xyloglucan. Transglycosylation caused the production of sulphorhodamine-labelled polysaccharide, which remained bound to the test paper after prolonged washing in water, revealing eight fluorescent bands corresponding to XET isoenzymes with pI values of 8.8, 8.6, 7.8, 7.6, 7.3, 7.1, 6.7 and 6.6.
| Original language | English |
|---|---|
| Pages (from-to) | 238-240 |
| Number of pages | 3 |
| Journal | Phytochemical Analysis |
| Volume | 10 |
| Issue number | 5 |
| DOIs | |
| Publication status | Published - 1999 |
Keywords
- Hydrolase
- Isoelectric focusing
- Native polyacrylamide gel electrophoresis
- Transglycosylase
- Xyloglucan
- Zymogram
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