583-586Metagenomic DNA was isolated from dairy effluent collected from a milk processing unit. The Hpa I digests of metagenome were cloned in pEZ BAC vector. The transformed DH5 colonies were selected by blue white screening on chloramphenicol (25μg/ml) containing plates. Functional screening of the library on skim milk agar showed a clone having protease activity. This clone was confirmed to have a metagenome insert of size approx. 9kb. The crude cell extract of this clone at 24th h of growth showed protease activity of 166.99U/ml (111.32U/mg protein). The protease was found to be alkaline and had maximum activity at 400C and pH 9.0