Purification and characterization of intracellular alpha-L-rhamnosidase from Pseudomonas paucimobilis FP2001.
alpha-L-Rhamnosidase was extracted and purified from the cells of Pseudomonas paucimobilis FP2001 with a 19.5% yield. The purified enzyme, which was homogeneous as shown by SDS-PAGE and isoelectric focusing, had a molecular weight of 112,000 and an isoelectric point of 7.1. The enzyme activity was accelerated by Ca2+ and remained stable for several months when stored at -20 C. The optimum pH was 7.8; the optimum temperature was 45 degrees C. The Km, V(max) and k(cat) for p-nitrophenyl alpha-L-rhamnopyranoside were 1.18 mM, 92.4 microM x min(-1) and 117,000 x min(-1), respectively. Examination of the substrate specificity using various synthetic and natural L-rhamnosyl glycosides showed that this enzyme had a relatively broader substrate specificity than those reported so far.
Miake
F
F
Microbiology Laboratory, Faculty of Pharmaceutical Sciences, Fukuoka University, Japan. fmiake@psat.fukuoka-u.ac.jp
Satho
T
T
Takesue
H
H
Yanagida
F
F
Kashige
N
N
Watanabe
K
K
D006026:Glycoside Hydrolases / Q000302:isolation & purification* / Q000378:metabolism; D011549:Pseudomonas / Q000201:enzymology*; D013379:Substrate Specificity
D006026:Glycoside Hydrolases; C022525:alpha-L-rhamnosidase