Abstract
VARIOUS methods for the estimation of pectin-esterase and polygalacturonase have been used1,2; but for rapid examination of cultures of micro-fungi, a modification of the ‘cup-plate’ diffusion method is useful. The polygalacturonase assay is carried out on a medium containing 1.5 per cent agar and 1 per cent sodium pectate at pH 5.0; and after incubation at 37° C. for 18 hr., the plates are sprayed with acid, when the enzyme activity is revealed as clear circular zones on an opalescent background. For the pectin-esterase assay, buffer-free agar containing 1 per cent purified pectin and methyl red adjusted to pH 6.0 is used, and the activity is indicated as red zones on a yellow background, due to the lowering of the pH by demethylation of the pectin. With each enzyme there is a linear relationship between the diameter of the zone and the logarithm of the enzyme concentration. Other enzymes (amylase, cellulase, xylanase, arabanase) may be determined similarly3, and the activities calculated by the usual method4,5.
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Reid, W. W., Tech. Comm. No. 21, Commonwealth Bureau of Horticulture and Plantation Crops, chapter 11 (1950).
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Lineweaver, H., and Ballou, G. A., Arch. Biochem., 6, 373 (1945).
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REID, W. Estimation and Separation of the Pectin-Esterase and Polygalacturonase of Micro-fungi. Nature 166, 569 (1950). https://doi.org/10.1038/166569a0
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DOI: https://doi.org/10.1038/166569a0
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