Cleavage of the xylosyl serine linkage between a core peptide and a glycosaminoglycan chain by cellulases.
We previously found that endo-beta-xylosidase from Patinopecten is an endo-type glycosidase that cleaves the xylosyl serine linkage between a glycosaminoglycan chain and its core protein (Takagaki, K., Kon, A., Kawasaki, H., Nakamura, T., Tamura, S., and Endo, M. (1990) J. Biol. Chem. 265, 854-860). Screening for endo-beta-xylosidase activity in several cellulases detected this activity in the enzymes from Aspergillus niger, Penicillium funiculosum, Trichoderma reesei, Trichoderma viride, and Irpex lacteus. The cellulase derived from A. niger was purified, and its molecular weight was determined to be 26,000 by SDS-PAGE. Examination of the specificity of the cellulase revealed that 1) the enzyme acts on the linkage region (xylosyl serine) between a core peptide and a glycosaminoglycan chain; 2) enzymatic activity is greater with shorter glycosaminoglycan chains; 3) the enzyme readily hydrolyzes the linkage in glycosaminoglycan peptides, but intact proteoglycan is cleaved only slowly; and 4) the activity is unaffected by the glycosaminoglycan component (chondroitin sulfate, dermatan sulfate, and heparan sulfate). Judging from these enzymatic characteristics, this cellulase is different from the endo-beta-xylosidase of Patinopecten. We believe that this cellulase will become a useful tool in the further development of glycotechnology, because, like the endo-beta-xylosidase of Patinopecten, it enables the release of intact glycosaminoglycans from glycosaminoglycan peptides.
Takagaki
Keiichi
K
Department of Biochemistry, Hirosaki University School of Medicine, 5 Zaifu-cho, Hirosaki , Japan.
Iwafune
Mito
M
Kakizaki
Ikuko
I
Ishido
Keinosuke
K
Kato
Yoji
Y
Endo
Masahiko
M
D002240:Carbohydrate Sequence; D002480:Cellulase / Q000737:chemistry / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D005658:Fungi / Q000201:enzymology; D006025:Glycosaminoglycans / Q000378:metabolism*; D006868:Hydrolysis; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D008970:Molecular Weight; D012694:Serine / Q000378:metabolism*; D014994:Xylose / Q000378:metabolism*
D006025:Glycosaminoglycans; D012694:Serine; D014994:Xylose; D002480:Cellulase