Purification and characterization of beta-glucuronidase from Escherichia coli HGU-3, a human intestinal bacterium.
beta-Glucuronidase was purified 360-fold from Escherichia coli HGU-3, an human intestinal bacterium. The specific activity of the purified enzyme was 17.78 units/mg protein. The enzyme (M.W. 290000) is composed of four subunits (M.W. 72000) with a pI and optimal pH of 4.8 and 6-7, respectively. The apparent Km for p-nitrophenyl-beta-D-glucuronide was found to be 0.22 mM. The enzyme was inhibited by saccharic acid 1,4-lactone, glycyrrhizin, N-ethylmaleimide (NEM) and p-chloromercuriphenylsulfonic acid (PCMS). Using the bile containing bilirubin diglucuronide as a substrate, the purified beta-glucuronidase was able to hydrolyze it to bilirubin. This hydrolyzed bilirubin formed calcium bilirubinate with a reaction mixture containing CaCl2.
Kim
D H
DH
College of Pharmacy, Kyung-Hee University, Seoul, Korea.
Jin
Y H
YH
Jung
E A
EA
Han
M J
MJ
Kobashi
K
K
D001663:Bilirubin / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology*; D005966:Glucuronidase / Q000302:isolation & purification*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007422:Intestines / Q000382:microbiology*; D008970:Molecular Weight
D005966:Glucuronidase; D001663:Bilirubin