Purification and characterization of intracellular alpha-L-rhamnosidase from Pseudomonas paucimobilis FP2001

alpha-L-Rhamnosidase was extracted and purified from the cells of Pseudomon as paucimobilis FP2001 with a 19.5% yield. The purified enzyme, which was h omogeneous as shown by SDS-PAGE and isoelectric focusing, had a molecular w eight of 112,000 and an isoelectric point of 7.1. The enzyme activity was a ccelerated by Ca2+ and remained stable for several months when stored at -2 0 degrees C. The optimum pH was 7.8; the optimum temperature was 45 degrees C. The K-m, V-max, and k(cat) for p-nitrophenyl alpha L-rhamnopyranoside w ere 1.18 mM . 92.4 mu M min(-1) and 117,000 . min(-1), respectively. Examin ation of the substrate specificity using various synthetic and natural L-rh amnosyl glycosides showed that this enzyme had a relatively broader substra te specificity than those reported so far.