PURIFICATION AND SOME PROPERTIES OF CYCLODEXTRIN-HYDROLYZING ENZYME FROM BACILLUS-SPHAERICUS

An intracellular cyclodextrin-hydrolyzing enzyme from Bacillus sphaericus E-244 isolated from soil was purified to a homogeneous state by means of Triton X-100 extraction, DEAE-Sepharose column chromatography, hydrophobic and molecular-sieve HPLC. The enzyme was estimated to have an Mr of 72 000 by sodium dodecyl sulfate polyacrylamide gel electrophoresis and 144 000 by HPLC gel filtration on TSK gel G 3000 SW. It had a pH optimum of 8.0, and the enzyme, stable at 25°C and pH 5.5-9.5 for 24 h, was inactivated at 50°C for 10 min. The enzyme hydrolyzed β-cyclodextrin more effectively than linear maltooligosaccharides such as maltopentaose, maltohexaose and maltoheptaose or polysaccharides such as starch, amylopectin, amylose and pullulan. © 1990.