Purification and Properties of Squalene-2,3-epoxide Cyclases from Pea Seedlings

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概要

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• Dramatic changes in the activities of squalene-2,3-epoxide : cycloartenol cyclase and β-amyrin cyclase were observed in germinating pea seeds. By taking advantage of this phenomenon, the two cyclases were purified from pea seedlings. The cyclases were purified to homogeneity by solubilization with Triton X-100,chromatography on hydroxylapatite and diethylaminoethyl (DEAE)-cellulose, isoelectric focusing and gel filtration. Cycloartenol cyclase was purified 471-fold to a specific activity of 167 pkat/mg protein, while β-amyrin cyclase was purified 4290-fold to a specific activity of 28 pkat/mg protein. They each showed a single band on sodium dodecyl sulfate polyacrylamide gel electrophoresis with a molecular mass of 55 and 35 kilodaltons (kDa), respectively. The apparent K_m values for (3S)-squalene-2,3-epoxide were estimated to be 25 and 50 μM, respectively. The cyclases required Triton X-100 or deoxycholate for their highest activity and each showed a broad pH optimum within the range of pH 6.5-7.5. Inhibition by p-chloromercuribenzene sulfonic acid and N-ethylmaleimide suggested involvement of an SH group at the active site of each enzyme.

• 社団法人日本薬学会の論文
• 1992-07-25

著者

• 三川 潮

  Faculty of Pharmaceutical Sciences, University of Tokyo

• 海老塚 豊

  Faculty of Pharmaceutical Sciences, The University of Tokyo

• 三川 潮

  富山医薬大薬

• 三川 潮

  Faculty Of Pharmaceuical Sciences The University Of Tokyo

• 三川 潮

  Faculty Of Pharmaceutical Sciences University Of Tokyo

• 三川 潮

  東大 薬

• 阿部 郁郎

  Faculty of Pharmaceutical Sciences, The University of Tokyo

• Sankawa U

  International Research Center For Traditional Medicine

• Sankawa Ushio

  (present Address)faculty Of Pharmaceutical Sciences Toyama Medical And Pharmaceutical University

• Sankawa Ushio

  Faculty Of Pharmaceutical Sciences Toyama Medical And Pharmaceutical University

• 阿部 郁郎

  Faculty Of Pharmaceutical Sciences The University Of Tokyo

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