Dioscorea opposita Thunb. a-mannosidase belongs to the glycosyl hydrolase family 38

• Autorzy: Uno Y. , Hashidume S. , Kurita O. , Fujiwara T. , Nomura K.
• Języki publikacji: EN

Abstrakty

EN

α-Mannosidase (EC 3.2.1.24) was purified from ‘Iseimo’, a native variety of Dioscorea opposita Thunb. Before purification, we investigated the composition of a viscous polysaccharide that interferes with column chromatography procedures. The polysaccharide consisted mainly of mannose, and also contained uronic acid. We used the cationic detergent cetylpyridinium chloride (CPC) to remove the polysaccharide. CPC treatment decreased viscosity without affecting α-mannosidase activity. We purified α-mannosidase 2,650-fold. The optimal pH of the enzyme was 6.0 and the optimum temperature was 65℃. The Km value for p-nitrophenyl-α-D-mannopyranoside was 0.35 ± 0.03 mM. Activity was inhibited by swainsonine but not kifunensine. The enzyme cleaved α-1,2 linkages preferentially to α-1,6 and α-1,3 linkages. The Mr of purified α-mannosidase was estimated to be 250–260 kDa by gel filtration and native-PAGE. Four polypeptides (86, 83, 80, and 28 kDa) were detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. It is unclear whether the polypeptides are encoded by one gene or multiple genes. However, N-terminal sequence analysis suggested that post-translational cleavage and/or glycosylation resulted in the three large fragments, if these amino acids were encoded by the same gene. Homology searches and characterization of the enzyme’s properties indicated that Iseimo α-mannosidase belongs to the glycoside hydrolase family 38 proteins, and to the Class II mannosidase group.

• Wydawca: -
• Czasopismo: Acta Physiologiae Plantarum
• Rocznik: 2010
• Tom: 32
• Numer: 4
• Opis fizyczny: p.713-718,fig.,ref.

Twórcy

autor

Uno Y.

• Graduate School of Agricultural Science, Kobe University, 1-1 Rokkodai-cho, Nada-ku, Kobe 657-8501, Japan

autor

Hashidume S.

• Graduate School of Agricultural Science, Kobe University, 1-1 Rokkodai-cho, Nada-ku, Kobe 657-8501, Japan

autor

Kurita O.

• Industrial Research Division, Mie Prefectural Science and Technology Promotion Center, 5-5-45 Takachaya, Tsu 514-0819, Japan

autor

Fujiwara T.

• Industrial Research Division, Mie Prefectural Science and Technology Promotion Center, 5-5-45 Takachaya, Tsu 514-0819, Japan

autor

Nomura K.

• Graduate School of Agricultural Science, Kobe University, 1-1 Rokkodai-cho, Nada-ku, Kobe 657-8501, Japan

Bibliografia

• Bensadoun A, Weinstein D (1976) Assay of proteins in the presence of interfering material. Anal Biochem 70:241–250
• Bitter T, Muir HM (1962) A modified uronic acid carbazole reaction. Anal Biochem 4:330–334
• Curdel A, Petek F (1980) Purification and properties of α-D-mannosidase from the germinated seeds of Medicago sativa (Alfalfa). Biochem J 185:455–462
• Daniel PF, Winchester B, Warren CD (1994) Mammalian α-mannosidases—multiple forms but a common purpose? Glycobiology 4:551–566
• Kaushal GP, Szumilo T, Pastuszak I, Elbein AD (1990) Purification and homogeneity and properties of mannosidase II from mung bean seedlings. Biochemistry 29:2168–2176
• Kishimoto T, Hori J, Takano D, Nakano Y, Watanabe M, Mitsui T (2001) Rice α-mannosidase digesting the high mannose glycopeptides of glutelin. Physiol Plant 112:15–24
• Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685
• Misaki R, Fujiyama K, Yokoyama H, Ido Y, Miyauchi K, Yoshida T, Seki T (2003) Characterization of almond α-mannosidase and its application for structure analysis of sugar chain. J Biosci Bioeng 96:187–192
• Moremen KW, Trimble RB, Herscovics A (1994) Glycosidases of the asparagines-linked oligosaccharide processing pathway. Glycobiology 3:113–125
• Nakagawa H, Enomoto N, Asakawa M, Ueda Y (1988) Purification and characterization of α-mannosidase and β-N-acetylhexosaminase from watermelon fruit. Agric Biol Chem 52:2223–2230
• Ohtani K, Misaki A (1983) Purification and characterization of β-D-galactosidase and α-D-mannosidase from papaya (Carica papaya) seeds. Agric Biol Chem 47:2441–2451
• Rabouille C, Kuntz DA, Lockyer A, Watson R, Signorelli T, Rose DR, van den Heuval M, Roberts DB (1999) The Drosophila GMII gene encodes a Golgi alphα-mannosidase II. J Cell Sci 112:3319–3330
• Strasser R, Schoberer J, Jin C, Glo¨ssl J, Mach L, Steinkellner H (2006) Molecular cloning and characterization of Arabidopsis thaliana Golgi α-mannosidase II, a key enzyme in the formation of complex N-glycans in plants. Plant J 45:789–803
• Tsukui M (2003) Analysis of properties and chemical structure of mucilage from Yam (Dioscorea opposite Thumb.). Nippon Shokuhinhozou Kagakukaishi 29:229–236 (in Japanese)
• Woo KK, Miyazaki M, Hara S, Kimura M, Kimura Y (2004) Purification and characterization of a Co(II)-sensitive α-mannosidase from Ginkgo biloba seeds. Biosci Biotechnol Biochem 68:2547–2556