Betalains have potential as the natural colourant and functional food with pharmaceutical purposes in food applications. The total betalain content of Basella rubra L. grown at Kalaburagi region is evaluated for the first time. The present study is carried out to determine the pigment stability and antimicrobial activity of fruits of Basella rubra. Total betalain content of fruits was evaluated and it was found to contain a high amount of 48.15 mg betacyanins and sparing amount of 2.53 mg betaxanthins and total betalains account for 50.69mg/100g of fresh fruit pulp. The influence of different additives on the colour stability of pigment was evaluated. More than 80% pigment colour is stable in the presence of 5% sucrose at 4 °C under dark condition. The betalain extract of Basella rubra L fruits showed significant antimicrobial activity against S.typhi, Aspergillus flavus, Microphomia fungi and Rhizopus stolonifer. It is for the first time that the total betalain content and anti-microbial activity of Basella rubra cultivar grown in Hyderabad Karnataka region of Karnataka, India is reported.