Characterization of Isofenphos Hydrolases from Arthrobacter sp. : Strain B-5
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概要
- 論文の詳細を見る
Two organophosphorus compound hydrolases constitutively expressed in Arthrobacter sp. strain B-5 that was capable of degrading various organophosphorus insecticides were purified from the cells' cytosol. The enzymes were similarly composed of a single subunit with a molecular weight of 45,000 Da, but had different isoelectric points of 3.6 and 3.9. In addition, the hydrolases were found to possess identical N-terminal amino acid sequences, which showed 51.6% amino acid identity with the N-terminus of the aryldialkylphosphatase from Nocardia sp. strain B-1 and revealed no significant homology for any other proteins. The pH and temperature optima for the activity of the enzyme mixture were 9.5 and 45℃, respectively. The B-5 hydrolases were remarkably activated by ammonium ions and thiols such as 2-mercaptoethanol and dithiothreitol, and significantly inactivated by divalent cations such as CuCl_2. Of all organophosphorus compounds tested, only isofenphos was rapidly hydrolyzed by the enzyme mixture, which cleaved an aryl phosphoester bond in isofenphos and produced its hydrolysates, O-ethyl isopropyl phosphoramidothioate and isopropyl salicylate.The K_m and V_<max> values for isofenphos were 1.8 μM and 9.4 U/mg of protein, respectively. Since the other substrated possessing the phosphoramide bond such as butamifos, amiprophos-methyl, and acephate were not hydrolyzed very much by the enzymes, the bond would not play an important role on the hydrolytic activity.Thus, the high activity for isofenphos and a little hydrolysis of other substrates revealed that the enzymes from Arthrobacter sp. strain B-5 are a novel type of organophosphorus compound hydrolase on substrate specificity.
- 社団法人日本生物工学会の論文
- 1997-03-25
著者
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OHSHIRO Kazufumi
Department of Structural Pathology, Institute of Nephrology, Faculty of Medicine, Niigata University
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Ono Tsuyoshi
Department of Cardiology, Toho university Ohashi medical center
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HOSHINO Tsutomu
Department of Applied Biological Chemistry, Faculty of Agriculture and Graduate School of Science an
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Uchiyama Takehiko
Research And Development Yaizu Suisankagaku Industry Co. Ltd.
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Ohshiro Kazufumi
Department Of Applied Biological Chemistry Faculty Of Agriculture Niigata University
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Hoshino T
Niigata Univ. Niigata Jpn
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Uchiyama Takeo
Department of Agricultural Chemistry, Faculty of Agriculture, Niigata University
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Uchiyama Takeo
Department Of Agricultural Chemistry Faculty Of Agriculture Niigata University
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Hoshino Tsutomu
Department Of Applied Biological Chemistry Faculty Of Agriculture And Graduate School Of Science And
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Hoshino T
Research Institute Of Genome-based Biofactory National Institute Of Advanced Industrial Science And
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Ono Tsuyoshi
Department Of Applied Biological Chemistry Faculty Of Agriculture
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Hoshino Tsutomu
Department Of Agricultural Chemistry Faculty Of Agriculture Niigata University
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Hoshino Tsutomu
Department Of Applied Biological Chemistry Faculty Of Agriculture And Graduate School Of Science And
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Hoshino Tsutomu
Graduate School Of Science And Technology Faculty Of Agriculture Niigata University:(present Address
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Hoshino Tsutomu
Department Of Applied Biological Chemistry Faculty Of Agriculture And Graduate School Of Science And
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Hoshino Tsutomu
Graduate School Of Science And Technology And Department Of Applied Biolobical Chemistry Faculty Of
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