Purification and Substrate-Specificity of β-Xylosidase from Sycamore Cell (Acer-Pseudoplatanus L.): Application for Structural Analysis of Xylose-Containing N-Linked Oligosaccharides

Tezuka, Katsunari; Hayashi, Megumi; Ishihara, Hideko; Nishimura, Mariko; Onozaki, Kaori; Takahashi, Nobutaka

doi:10.1006/abio.1993.1258

Cited by 20 publications

(12 citation statements)

References 0 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Until now, only the purification and partial characterization of one b-D-xylosidase from dicotyledonous plant has been described (Tezuka et al, 1993) for sycamore cells (Acer pseudoplantanus). In this study, we describe the purification, enzymatic properties, sequence identity, and action on various substrates of these b-D-xylosidases.…”

Section: Discussionmentioning

confidence: 99%

“…One b-D-xylosidase from young barley (Hordeum vulgare) seedlings was recently purified, sequenced, and biochemically characterized (Lee et al, 2003). Other plant b-D-xylosidases have been purified and partially characterized in wheat flour (Cleemput et al, 1997), sycamore cell cultures (Tezuka et al, 1993), pea seedlings (O'Neill et al, 1989), and sugarcane (Chinen et al, 1982). Recently, a gene from Arabidopsis encoding a putative b-D-xylosidase was identified, and a role for this enzyme in secondary cell wall metabolism and plant development has been proposed (Goujon et al, 2003).…”

mentioning

confidence: 99%

See 1 more Smart Citation

Purification and Characterization of Enzymes Exhibiting β-d-Xylosidase Activities in Stem Tissues of Arabidopsis

et al. 2004

View full text Add to dashboard Cite

This work describes the purification and characterization of enzymes that exhibit b-D-xylosidase activity in stem tissues of Arabidopsis. This is the first detailed investigation that concerns the characterization of catalytic properties and sequence identity of enzymes with b-D-xylosidase activities in a dicotyledonous plant. Three different enzymes, ARAf, XYL4, and XYL1 with apparent molecular masses of 75, 67, and 64 kD, respectively, were purified to homogeneity. ARAf was identified as a putative a-L-arabinofuranosidase, and XYL4 and XYL1 as putative b-D-xylosidases using matrix-assisted laser-desorption ionization time of flight. ARAf belongs to family 51 and XYL4 and XYL1 to family 3 of glycoside hydrolases. ARAf and XYL1 have highest specificity for p-nitrophenyl-a-L-arabinofuranoside and XYL4 for p-nitrophenyl-b-D-xylopyranoside and natural substrates such as xylobiose and xylotetraose. XYL4 was shown to release mainly D-Xyl from oat spelt xylan, rye arabinoxylan, wheat arabinoxylan, and oligoarabinoxylans. ARAf and XYL1 can also release D-Xyl from these substrates but less efficiently than XYL4. Moreover, they can also release L-Ara from arabinoxylans and arabinan. Overall, the results indicate that XYL4 possesses enzymatic specificity characteristic for a b-D-xylosidase, while ARAf and XYL1 act as bifunctional a-L-arabinofuranosidase/b-D-xylosidases. Analysis of the activity of these three enzymes in stem tissues at different stages of development has shown that young stems possess the highest activities for all three enzymes in comparison to the activities of the enzymes present in stems at older stages of development. High enzyme activities are most likely related to the necessary modifications of cell wall structure occurring during plant growth. Plant cells are surrounded by an extracellular matrix known as the cell wall. The regulation of cell wall morphology and composition is important for the determination of cell size and shape, cellular interactions with the environment, mechanical resistance, and defense against pathogen attacks (Reiter, 2002). The cell wall is also involved in plant growth and development (Stolle-Smits et al., 1999;Obel et al., 2002). In addition, although cell walls consist mainly of polysaccharides broadly classified as celluloses, hemicelluloses, and pectins, their constitution varies, not only from plant to plant, but also in different tissues of the same plant (Heredia et al., 1995;Cosgrove, 1997;Popper and Fry, 2003). A coordinated series of biochemical processes occur during plant development resulting in the biosynthesis and degradation of cell wall components. Consequently, numerous enzymes must be implicated in these processes (Heredia et al., 1995;Cosgrove, 1997;Stolle-Smits et al., 1999;Obel et al., 2002;Reiter, 2002;Popper and Fry, 2003). However, to date, little information is available concerning such enzymes, their mode of action, and their physiological role. Increased research interest has focused on enzymes involved in the metabolism of hemicelluloses in ...

show abstract

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

Purification and Characterization of Enzymes Exhibiting β-d-Xylosidase Activities in Stem Tissues of Arabidopsis

et al. 2004

View full text Add to dashboard Cite

show abstract

“…The existence of several exoglycosidases in the sycamore cell culture medium was also demonstrated (41). Formation of Type I oligosaccharides in BG60 and the heterogeneity of the BG60 oligosaccharides may also be the result of degradative reactions rather than of imperfect biosyntheses.…”

Section: Fig 1 Hplc Of Pa-oligosaccharides Of Bg60 On Ods-and Amidementioning

confidence: 92%

“…The pyridylamino (PA) 1 derivatives of isomalto-oligosaccharides (4 -20 glucose residues) and of reference oligosaccharides (code numbers 2 000.1, 000.1FX, M2.1FX, and M1.1FX) were from Nakano Vinegar Co. (Handa City, Japan). The following oligosaccharides were prepared by the methods described previously: M1.1F by ␤-xylosidase digestion of M1.1FX (17), 000.1X from rice ␣-amylase (18), and 100.1FX from laccase of sycamore cells (19).…”

Section: Methodsmentioning

confidence: 99%

“…Exoglycosidase Digestion-Each oligosaccharide isolated on the ODS and amide columns was sequentially digested with exoglycosidases (␤-N-acetylhexosaminidase, ␣-mannosidase, and ␤-xylosidase) under the condition described previously (17,22). After each step of exoglycosidase digestion, the elution coordinates of the trimmed oligosaccharides were located on the two-dimensional map to verify the structural identity.…”

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation