Epiphytic bet-mannanase producing bacterial strains

Dry remains of the herbal species of the plantain (Plantago major ), the wormwood (Artemisia vulgaris ) and the reed grass (Calamagrostis acutiflora ) were used as a natural source for isolation of β- mannanase producing strains. They were isolated by using the carob gum as a single source of carbon and energy. Each chosen plant species was found to be colonized with a single dominant epiphytic group of microorganism, although the plants had been collected in the same location. Bacillus circulans was only found in P. major , Bacillus subtilis on A. vulgaris , whereas Pantoea sp. was found in C. acutiflora . Identification of the taxonomy affiliation of the isolated β-mannanase producers allowed using the formerly proposed primers for PCR cloning of β-mannanase genes previously occurred in the respective bacterial species. This approach let us cloning 330 bp fragment of β-mannanase genes from B. circulans and B. subtilis and 1000 bp fragment of β-mannanase gene from Pantoea sp. Testing the enzymatic activity of the isolated strains by staining the carob gum hydrolysis zones on the plates with Congo Red was carried out. As a result, the maximum activity was found in Pantoea sp.