Purification and Characterization of a Novel Lactonohydrolase from Agrobacterium tumefaciens

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概要

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• A novel lactonohydrolase, catalyzing the stereospecific hydrolysis of L-pantoic acid, was purified 2,400-fold to apparent homogeneity with a 1.96% overall recovery from Agrobacterium tumefaciens AKU 316 through a purification procedure including ammonium sulfate fractionation, and column chromatographies on DEAE-Sephacel, phenyl-Sepharose CL-4B, Sephacryl S-200,Mono-Q and alkyl-Superose. The relative molecular mass of the native enzyme estimated on high-pressure gel permeation chromatography was 62,000 Da, and the subunit molecular mass was estimated to 26,500 Da on SDS-polyacrylamide gel electrophoresis. The enzyme hydrolyzes several aromatic lactones, such as 3,4-dihydrocoumarin and homogentisic acid lactone, other than L-pantoyl lactone. The K_m and V_max for L-pantoyl lactone were 3.59 mM and 13.7μmol/min/mg, respectively. The enzymatic activity was inhibited by several chelating reagents, Fe^<2+>, Sn^<2+>, Pb^<2+>, and Fe^<3+>.

• 社団法人日本農芸化学会の論文
• 2000-06-23

著者

• Shimizu Sakayu

  Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University

• Kataoka Michihiko

  Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University

• Kataoka M

  Division Of Applied Life Sciences Graduate School Of Agriculture Kyoto University

• Kataoka Michihiko

  Div. Of Applied Life Sci. Graduate School Of Agriculture Kyoto Univ.

• Kataoka Michihiko

  Division Of Applied Life Sciences Graduate School Of Agriculture Kyoto University

• Shimizu Sakayu

  Division Of Applied Life Sciences Graduate School Of Agriculture Kyoto University

• Shimizu Sakayu

  Division Of Applied Life Science Graduate School Of Agriculture Kyoto University

• Sakamoto Keiji

  Graduate School Of Natural Science And Technology Okayama University

• Nose K

  Department Of Microbiology Showa University Of Pharmaceutical Sciences

• Nose K

  Showa Univ. Pharmaceutical Sci. Tokyo Jpn

• Nose Kiyoshi

  Division Of Applied Life Sciences Graduate School Of Agriculture Kyoto University

• Kataoka Michihiko

  Divisin Of Applied Life Sciences Graduate School Of Life And Environmental Sciences Osaka Prefecture

• NOMURA Jun-ichi

  Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University

• SHINOHARA Makoto

  Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University

• SAKAMOTO Keiji

  Research Institute, Fuji Chemical Industries Ltd.

• Nose Kiyoshi

  Department Of Microbiology Showa University Of Pharmaceutical Sciences

• Nomura Jun-ichi

  Division Of Applied Life Sciences Graduate School Of Agriculture Kyoto University

• Sakamoto Keiji

  Research Institute Daiichi Fine Chemical Co.

• Shinohara Makoto

  Division Of Applied Life Sciences Graduate School Of Agriculture Kyoto University

• Kataoka Michihiko

  Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University:Division of Applied Life Sciences, Graduate School of Life and Environmental Sciences, Osaka Prefecture University

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