Abstract: In the present study, the four bacterial strains were isolated from soil and water at Rajshahi region. The strains were isolated on xylan agar media and screened by xylanolysis method. The bacterial strains (St1, St2, St3 and St4) were characterized and identified based on morphological, biochemical and physiological characters. On the basis of these tests, the four strains were initially identified as belonging to the genus Bacillus. Bacillus strains were capable of growing in xylan medium at pH 7.0 and produced 55 IU (μ mol xylose min-1 ml-1) of xylanase when cultivated in neutral medium at pH 7.0. Maximal enzyme activity was obtained by cultivation in oat spelt xylan, but high enzyme production was also obtained on wheat bran. With cultivation at pH 7.0 and assays performed in phosphate buffer, maximal activity was observed at pH 7.0; with phosphate buffer, maximal activity was between pH 6 and 7. The xylanase temperature optimum at pH 7.0 was 50°C. The enzyme could hydrolyze the oat spelt xylan and birch wood xylan effectively but did not hydrolyze cellulose, carboxymethyl-cellulose and starch. Bacillus strains also showed drug resistance to ampicilline, amoxycilline, cotromosilline and chlorophenical and sensitive to tetracycline and erythromycin. Plasmid DNA was isolated from the xylanase-producing multi-drug resistant strain. Agarose gel electrophoresis of the isolated plasmid DNA showed the presence of a plasmid of about 32 kb in size.