New method for determination of agarase enzyme from bacterial origin [electronic resource].

Includes references.

This context concentrated mainly on determination of a new method for agarse enzyme assay by so called agar cup plate clearing zone(ACZ) technique. Bacillus macernas 8M produced constitutive extracellular agarase, and could grow without using agar in its culture medium. Optimization of this new technique controlled by using the most important parameters of enzymes assay. An optimum gum Arabic concentration was at 1.5%(w/v); optimum substrate (agar) concentration was detected at 1 %; the best temperature for enzyme assay in its reaction mixture was detected at 50°C; the best time to get the highest rate of enzyme activity determined after 48h. In attempt to determine the most applicable buffer at its identical pH value to get the highest rate of agarase enzyme activity, different buffer have been carried out; Tris-HCI was represented with optimum pH value of 7.2; boric acid borate buffer (8.8); Phosphate buffer (6.8); Citrate-phosphate buffer (5.0); Citrate buffer (5.8). This technique may be used fairly in molecular microbiology and biotechnology, also from the essential application of this technique for differentiation between enzymes assay produced by the same organism that degrade not only agar but also various plant polysaccharides, i.e., cellulose, pectin, starch, and xylan. Key words: Agarase determination, New technique, Bacillus macernas SM.

Summary in Arabic.