Enzymatic Conversion of Pheophorbide a to the Precursor of Pyropheophorbide a in Leaves of Chenopodium album
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概要
- 論文の詳細を見る
Soluble proteins extracted from leaves of Chenopodi-um album catalyzed the conversion of pheophorbide a to a precursor of pyropheophorbide a, putatively identified as C-13^2-carboxyl-pyropheophorbide a. The precursor was then decarboxylated non-enzymatically to yield pyropheophorbide a. Soluble proteins and pheophorbide a, as the substrate, were required for the formation of the precursor, and boiled proteins were enzymatically inactive. The maximum rate of conversion of pheophorbide a to the precursor occurred at pH 7.5. The K_m for pheophorbide a was 12.5 μM at pH 7.0. Both pheophorbide b and bacterio-pheophorbide a could serve as substrates, but protopheophorbide a could not. Formation of methanol was detected during the enzymatic reaction, an indication that the enzyme is an esterase. Among seven alcohol analogs tested, only methanol inhibited the enzymatic activity uncompetitively, with a K_i of 71.6 mM. Mass-spectrometric (MS) analysis of the precursor yield a peak at m/z 579 that indicated the release of a methyl group from pheophorbide a. It ap-pears therefore that the enzyme catalyzes the demethylation of the carbomethoxy group at C-13^2 of pheophorbide a by hydrolysis to yield methanol and the precursor, C-13^2-carboxyl-pyropheophorbide a, which is converted to pyropheophorbide a by spontaneous decarboxylation. We have tentatively designated the enzyme "pheophorbidase". The presence of the enzyme was dependent on plant species and it was expressed constitutively.
- 日本植物生理学会の論文
著者
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WATANABE Kenji
Department of Kampo Medicine, Keio University, School of Medicine
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SHIRANO Yumiko
Mitsui Plant Biotechnology Research Institute
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Shioi Y
Mitsui Plant Biotechnology Research Institute
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Shioi Yuzo
Department Of Biological Sciences Faculty Of Bioscience And Biotechnology Tokyo Institute Of Technol
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Takamiya Ken-ichiro
Department of Biological Sciences, Faculty of Bioscience and Biotechnology, Tokyo Institute of Techn
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Watanabe K
Tokyo Institute Of Technology Faculty Of Bioscience And Biotechnology Department Of Biological Scien
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Takamiya Ken-ichiro
Department Of Biological Sciences Faculty Of Bioscience And Biotechnology
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Shioi Yuzo
Department Of Biological Science Faculty Of Science Shizuoka University
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Watanabe Kenji
Department Of Gastroenterology Graduate School Of Medicine Osaka City University
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Watanabe Kenji
Department Of Anatomy Fukui Medical School
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Watanabe Kenji
Department Of Agricultural Chemistry College Of Agriculture And Veterinary Medicine Nihon University
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