Unsaturated glucuronyl hydrolase of Bacillus sp. GL1: novel enzyme prerequisite for metabolism of unsaturated oligosaccharides produced by polysaccharide lyases.
The bacterium Bacillus sp. GL1 assimilates two kinds of heteropolysaccharides, gellan and xanthan, by using extracellular gellan and xanthan lyases, respectively, and produces unsaturated saccharides as the first degradation products. A novel unsaturated glucuronyl hydrolase (glycuronidase), which was induced in the bacterial cells grown on either gellan or xanthan, was found to act on the tetrasaccharide of unsaturated glucuronyl-glucosyl-rhamnosyl-glucose produced from gellan by gellan lyase, and the enzyme and its gene were isolated from gellan-grown cells. The nucleotide sequence showed that the gene contained an ORF consisting of 1131 base pairs coding a polypeptide with a molecular weight of 42,859. The purified enzyme was a monomer with a molecular mass of 42 kDa and was most active at pH 6.0 and 45 degrees C. Because the enzyme can act not only on the gellan-degrading product by gellan lyase, but also on unsaturated chondroitin and hyaluronate disaccharides produced by chondroitin and hyaluronate lyases, respectively, it is considered that the unsaturated glucuronyl hydrolase plays specific and ubiquitous roles in the degradation of oligosaccharides with unsaturated uronic acid at the nonreducing terminal produced by polysaccharide lyases.
Hashimoto
W
W
Research Institute for Food Science, Kyoto University, Uji, 611-0011, Japan. hasimoto@food2.food.kyoto-u.ac.jp
Kobayashi
E
E
Nankai
H
H
Sato
N
N
Miya
T
T
Kawai
S
S
Murata
K
K
D000595:Amino Acid Sequence; D001407:Bacillus / Q000378:metabolism*; D001426:Bacterial Proteins / Q000378:metabolism*; D001483:Base Sequence; D006026:Glycoside Hydrolases / Q000378:metabolism*; D006867:Hydrolases / Q000378:metabolism*; D008969:Molecular Sequence Data; D011133:Polysaccharide-Lyases / Q000378:metabolism*; D011135:Polysaccharides, Bacterial / Q000378:metabolism; D013379:Substrate Specificity
D001426:Bacterial Proteins; D011135:Polysaccharides, Bacterial; C048288:gellan gum; D006867:Hydrolases; D006026:Glycoside Hydrolases; C120983:unsaturated glucuronyl hydrolase; D011133:Polysaccharide-Lyases; C002563:xanthan gum