ß-Galactosidase assay using Lab On Paper Platform
Anushree Lokur,
Associate Professor,
Ramnarain Ruia College
[FROM SPEAKER] Enzyme ß-Galactosidase has varied applications. It has applications in food industry for manufacturing of products that are free of lactose and for production of galactosylated food products. Presence of ß-Galactosidase in food samples indicates presence of coliforms which in turn indicates unhygienic conditions while preparing foods. Also, ß-gal gene is an important reporter of gene expression in molecular biology and hence its assay is routinely required in laboratories.
Lab on Paper is an ideal platform for detection of enzymes with chromogenic substrates. Detection can be carried out on- site, is convenient, cost effective and does not require any sophisticated instruments.
In the current work we have standardised ß-Galactosidase assay on paper. The method uses chromogenic yellow Orange substrate Chlorophenol Red-ß-D-Galactopyranoside (CPRG), which gives a red coloured chomophore namely Chlorophenol Red. The colour produced can be quantified using Image -J software. This can be used to calculate enzyme activity and in turn can be correlated to number of cells producing the enzyme.
The method requires only a few micro litres of sample. Results can be obtained within 3-4 hours. The method is specific, sensitive & can detect enzyme produced by up to 103 -104 cells /ml on paper.
Spiked & Raw food samples were tested using this assay and was found to give good correlation to standard methods of coliform detection.
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