Cleavage of the Xylosyl Serine Linkage between a Core Peptide and a Glycosaminoglycan Chain by Cellulases

Abstract

We previously found that endo-␤-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-␤-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-␤-xylosidase of Patinopecten. We believe that this cellulase will become a useful tool in the further development of glycotechnology, because, like the endo-␤-xylosidase of Patinopecten, it enables the release of intact glycosaminoglycans from glycosaminoglycan peptides.

Insect cell conditioned medium contains an endoglycosidase able to liberate chondroitin sulfate chains from aggrecan

“…An endo-h-xylosidase has been characterized from the mid-gut of the mollusk Patnopecten (Takagaki et al, 1990). In addition, recent studies have shown that fungal cellulases cleave the xylosyl -serine linkage between chondroitin sulfate and the core protein (Takagaki et al, 2002), and the Aspergillus niger enzyme has been used as the basis for glycome analysis (Iwafune et al, 2004). The present observations suggest that a similar activity exists in insect cells.…”

Matrix Protein Biglycan Induces Osteoblast Differentiation through Extracellular Signal-Regulated Kinase and Smad Pathways

“…7) Recent studies have found that the heparan sulfate mediates osteoblasts toward osteogenesis, and it regulates the anabolic activity of osteoblasts by triggering transient increase in Erk1/2 phosphorylation and activating Runx2. 30,31) Chondroitin sulfates involved in the osteogenic progression of osteoprogenitors are through Erk pathway.…”

Irpex lacteus, a white-rot fungus with biotechnological potential — review

“…The multidomain structure of the enzyme was characterized and En-1 showed a significant similarity to other endoglucanases belonging to Family 5 of glycosyl hydrolases (Toda et al 2005). The cellulase complex of I. lacteus was also shown to include an endo-β-xylosidase activity (Takagaki et al 2002). Cellulolytic mode of action of combined exo-and endocellulases of I. lacteus and Aspergillus niger on different types of pure cellulose was studied (Kanda et al 1986;Hoshino et al 1992Hoshino et al , 1993aHoshino et al ,b, 1994aHoshino et al ,b, 1997.…”