Microbial diversity and enzymatic activity of bacterial strains isolated from water samples, bottom sediments and biofilms of lake were investigated and identified by PCR and sequencing of the 16S rRNA gene. Screening procedures included decimal dilutions performing with incubation at 37°C, isolation of pure non-pathogenic bacterial cultures, isolate cultivation on agar in Petri plates at 30°C and tests on enzyme production. Enzymatic activity of 65 selected strains (Bacillus, Planococcus, Micrococcus, Paenibacillus, Arthrobacter, Lysinibacillus, Bhargavaea, Peribacillus, Kocuria) was determined used tests on plates and by spectrophotometric approach. The isolates found to be a promising candidate for agriculture, textile, food and pharmaceutical application by showed enzyme-producing of amylase, lipase, cellulase, especially catalase.