Epiphytic bet-mannanase producing bacterial strains

Abstract

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.