Purification and characterization of flavone-specific β-glucuronidase from callus cultures of Scutellaria baicalensis Georgi

Satoshi Morimoto, Takahiro Harioka, Yukihiro Shoyama

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

β-Glucuronidase from callus cultures of Scutellaria baicalensis Georgi was purified to apparent homogeneity by fractionated ammonium-sulfate precipitation and chromatography on diethylaminoethyl-cellulose, hydroxylapatite and baicalin-conjugated Sepharose 6B. A 650-fold purification was obtained by this purification system. When subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis the purified protein migrated as a single band with a molecular mass of 55 kDa. We determined that the native enzyme has a molecular mass of 230 kDa using gel-filtration chromatography. These results suggested that the enzyme exists as a homotetramer composed of four identical 55-kDa subunits. The enzyme showed a broad pH optimum between 7.0 and 8.0. The Km values were 9 μM, 10 μM, 30 μM and 40 μM for luteolin 3′ -O-β-d-glucuronide, baicalin, wogonin 7-O-β-d-glucoronide and oroxlin 7-O-β-d-glucuronide, respectively. The enzyme was most active with flavone 7-O-β-d-glucuronides.

Original languageEnglish
Pages (from-to)535-540
Number of pages6
JournalPlanta
Volume195
Issue number4
DOIs
Publication statusPublished - Feb 1 1995

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flavone
Scutellaria baicalensis
Glucuronidase
Bony Callus
callus culture
flavones
Glucuronides
Enzymes
enzymes
molecular weight
Luteolin
luteolin
Ammonium Sulfate
Durapatite
ammonium sulfate
Cellulose
Sodium Dodecyl Sulfate
Sepharose
agarose
Gel Chromatography

All Science Journal Classification (ASJC) codes

  • Genetics
  • Plant Science

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Purification and characterization of flavone-specific β-glucuronidase from callus cultures of Scutellaria baicalensis Georgi. / Morimoto, Satoshi; Harioka, Takahiro; Shoyama, Yukihiro.

In: Planta, Vol. 195, No. 4, 01.02.1995, p. 535-540.

Research output: Contribution to journalArticle

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AB - β-Glucuronidase from callus cultures of Scutellaria baicalensis Georgi was purified to apparent homogeneity by fractionated ammonium-sulfate precipitation and chromatography on diethylaminoethyl-cellulose, hydroxylapatite and baicalin-conjugated Sepharose 6B. A 650-fold purification was obtained by this purification system. When subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis the purified protein migrated as a single band with a molecular mass of 55 kDa. We determined that the native enzyme has a molecular mass of 230 kDa using gel-filtration chromatography. These results suggested that the enzyme exists as a homotetramer composed of four identical 55-kDa subunits. The enzyme showed a broad pH optimum between 7.0 and 8.0. The Km values were 9 μM, 10 μM, 30 μM and 40 μM for luteolin 3′ -O-β-d-glucuronide, baicalin, wogonin 7-O-β-d-glucoronide and oroxlin 7-O-β-d-glucuronide, respectively. The enzyme was most active with flavone 7-O-β-d-glucuronides.

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