title: A protease/peptidase from culture medium of Flammulina velutipes that acts on arabinogalactan-protein.
abstract: Arabinogalactan-proteins (AGPs) are highly diverse plant proteoglycans found on the plant cell surface. AGPs have large arabinogalactan (AG) moieties attached to a core-protein rich in hydroxyproline (Hyp). The AG undergoes hydrolysis by various glycoside hydrolases, most of which have been identified, whereas the core-proteins is presumably degraded by unknown proteases/peptidases secreted from fungi and bacteria in nature. Although several enzymes hydrolyzing other Hyp-rich proteins are known, the enzymes acting on the core-proteins of AGPs remain to be identified. The present study describes the detection of protease/peptidase activity toward AGP core-proteins in the culture medium of winter mushroom (Flammulina velutipes) and partial purification of the enzyme by several conventional chromatography steps. The enzyme showed higher activity toward Hyp residues than toward proline and alanine residues and acted on core-proteins prepared from gum arabic. Since the activity was inhibited in the presence of Pefabloc SC, the enzyme is probably a serine protease.
authors.affiliation: a Graduate School of Science and Engineering , Saitama University , Saitama , Japan.
authors.forename: Yoshihisa
authors.initials: Y
authors.lastname: Yoshimi
authors.affiliation: b Faculty of Science, Department of Biochemistry and Molecular Biology , Saitama University , Saitama , Japan.
authors.forename: Yumi
authors.initials: Y
authors.lastname: Sugawara
authors.affiliation: c Research Faculty of Agriculture , Hokkaido University , Sapporo , Japan.
authors.forename: Chiaki
authors.initials: C
authors.lastname: Hori
authors.affiliation: d Graduate School of Agricultural and Life Sciences , University of Tokyo , Tokyo , Japan.
authors.forename: Kiyohiko
authors.initials: K
authors.lastname: Igarashi
authors.affiliation: e Faculty of Agriculture , University of the Ryukyus , Nishinohara-cho , Japan.
authors.forename: Satoshi
authors.initials: S
authors.lastname: Kaneko
authors.affiliation: a Graduate School of Science and Engineering , Saitama University , Saitama , Japan.
authors.forename: Yoichi
authors.initials: Y
authors.lastname: Tsumuraya
authors.affiliation: a Graduate School of Science and Engineering , Saitama University , Saitama , Japan.|f Institute for Environmental Science and Technology , Saitama University , Saitama , Japan.
authors.forename: Toshihisa
authors.initials: T
authors.lastname: Kotake
mesh_terms: D003470:Culture Media / Q000737:chemistry; D055393:Flammulina / Q000166:cytology / Q000201:enzymology*; D005656:Fungal Proteins / Q000302:isolation & purification / Q000378:metabolism*; D005685:Galactans / Q000378:metabolism*; D006170:Gum Arabic / Q000737:chemistry; D010447:Peptide Hydrolases / Q000302:isolation & purification / Q000378:metabolism*; D011480:Protease Inhibitors / Q000494:pharmacology; D011509:Proteoglycans / Q000737:chemistry / Q000378:metabolism*; D013379:Substrate Specificity
chemical_list: D003470:Culture Media; D005656:Fungal Proteins; D005685:Galactans; D011480:Protease Inhibitors; D011509:Proteoglycans; D006170:Gum Arabic; D010447:Peptide Hydrolases; C005653:arabinogalactan
keywords: Flammulina velutipes; arabinogalactan-protein; core-protein; plant proteoglycan; serine protease