Purification and Properties of Bacillus sectorramus FERM-P8973 Pullulanase

Abstract

A pullulanase ([EC 3.2.1.41] pullulan 6-glucanohydrolase) from Bacillus sectorramus FERMP8973 (Amano Pharmaceutical Co . Ltd., ) was purified using Mono Q anion-exchange chromatography. Purified enzyme showed a single band on PAGE and SDS-PAGE, and a single peak on HPLC on a TSK-gel G3000SW . The molecular weight (95 kD), optimum pH (5-5.5), optimum temperature (55°C), pH stability and thermal stability were similar to the results of Mori et al. (Denpun Kagaku, 38, 9-16, 1991), but the isoelectric point (5 .3) was different from their result (4.7). Its amino acid composition was similar to those of other amylases. Its Nterminal amino acid sequence was YSNSTGVIVHTHRDSNYTN-, and thus did not resemble those of Klebsiella pneumoniae W70 pullulanase having a collagen-like sequence.