Pectolytic enzyme detection media

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Contents 1 Purpose 2 Ingredients 3 Instructions 4 Notes 5 References 6 Contributed by

Purpose

Ingredients

Ingredient	300 ml
Cold DH2O	300 ml
Polygalacturonic A	3 g
Yeast extract	3 g
cresol red solution	3 ml
agar	4.5 g

Instructions

1. Pour cold dH2O in 1 L beaker.
2. Slowly add polygalacturonic A--be careful not to clump.
3. Add remaining ingredients in same way.
4. Heat to nearly boiling while stirring continuously.
5. Adjust to pH 7.3 with 1 N NaOH.
6. Autoclave 15 minutes at 15 psi and 120 °C.
7. Bring pH to 7.8 with 1 N NaOH (turns pink), then cool.
8. Grow cultures 48 hours.
9. Flood with 2 N HCl or 1% aqueous solution hexadecyl-trimethyl ammonium bromide
10. Clear zones indicate presence of pectolytic enzymes.

Notes

References

• Cuppels, D. and Kelman, A. 1974. Evaluation of selective media for isolation of soft-rot bacteria from soil and plant tissue. Phytopathology 64: 468-475; modified by Y. B. Di and A. Kelman, and subsequently by M.C.M. Perombelon and E. M. Burnett, unpublished data.

Contributed by

From the Virginia Tech Mediabook; Orignially created by Robert Wick; contributed by Mary Ann Hansen.