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Purification of an α‐L‐arabinofuranosidase from carrot cell cultures and its involvement in arabinose‐rich polymer degradation
Author(s) -
Konno Haruyoshi,
Yamasaki Yoshiki,
Katoh Kenji
Publication year - 1987
Publication title -
physiologia plantarum
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.351
H-Index - 146
eISSN - 1399-3054
pISSN - 0031-9317
DOI - 10.1111/j.1399-3054.1987.tb09217.x
Subject(s) - sephadex , chemistry , chromatography , daucus carota , size exclusion chromatography , isoelectric point , polyacrylamide gel electrophoresis , arabinose , gel electrophoresis , enzyme , biochemistry , biology , xylose , fermentation , botany
Konno, H., Yamasaki, Y. and Katoh, K. 1987. Purification of an α‐L‐arabinofurano‐sidase from carrot cell cultures and its involvement in arabinose‐rich polymer degradation. An α‐L‐arabinofuranosidase (α‐L‐arabinofuranoside arabinofuranohydrolase, EC 3.2.1.55) was isolated from a homogenate of cell suspension cultures of carrot ( Daucus carota L. cv. Kintoki). The buffer‐soluble enzyme was purified to homogeneity by a procedure involving ammonium sulfate fractionation, chromatography on DEAE‐Sephadex A‐50, Sephadex G‐150, Con A‐Sepharose 4B and CM‐Sephadex C‐50, and preparative polyacrylamide gel electrophoresis. The size of this enzyme as determined by polyacrylamide gel electrophoresis in the presence of sodium laurylsulfate and by Sephadex G‐200 gel filtration was 94 and 110 kDa, respectively. The isoelectric point was at pH 4.7. The K m and V max values for p‐nitrophenyl α‐L‐arabinofuranoside were 1.33 mM and 20.2 μimol (mg protein) ‐1 h ‐1 , respectively. The optimal activity occurred at pH 4.2 with Mcllvaine buffer. The enzyme was stimulated by Ca 2+ and Zn 2+ , whereas it was strongly inhibited by Cu 2+ , Ag 2+ , Hg 2+ , p‐chloromercuri‐benzoate and L‐arabono‐l,4‐lactone. The enzyme acted on beet arabinan in an exo‐fashion. Furthermore, the enzyme was partially involved in the hydrolysis of the ara‐binogalactan and pectic polymer purified from carrot cell walls.