The enzyme ẞ-galactosidase catalyses the breakdown of the compound ONPG (o-nitrophenyl 1-D-galactopyranoside) to the compound ONP (o-nitrophenyl), as shown in Fig. 1.1. ẞ-galactosidase ONPG ONP + galactose (colourless substrate) (yellow product) Fig. 1.1 As ONP is produced, the colour of the reaction mixture changes to yellow. The intensity of the yellow colour produced is proportional to the concentration of ONP. A colorimeter is used to measure the absorbance of the reaction mixture. Absorbance is a measure of the light absorbed by a coloured solution. In the reaction shown in Fig. 1.1, the more intense the yellow colour, the higher the absorbance.
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