Ammonia Assimilation in Klebsiella pneumoniae F-5-2 That Can Utilize Ammonium and Nitrate Ions Simultaneously : Purification and Characterization of Glutamate Dehydrogenase and Glutamine Synthetase

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概要

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• The two ammonia-assimilating enzymes glutamate dehydrogenase (GDH; EC 1.4.1.4) and glutamine synthetase (GS; EC 6.3.1.2) were synthesized steadily during the cell growth of Kiebsiella pneumoniae F-5-2 that can utilize NH^+_4 and NO^-_3 simultaneously under aerobic conditions. The enzymes were purified to homogeneity from cell extracts and characterized. The molecular mass of the purified GDH was 300 kDa with six identical 52-kDa subunits. GDH showed its maximal activity (aminating) at pH 8.0 and was stable between pHs 5.5 and 11.5. The enzyme was NADP-specific and strongly inhibited by Ag^+. It catalyzed the amination of 2-ketovalerate, 2-ketoadipate, and 2-ketobutyrate, in addition to 2-ketoglutarate. The purified GS has a molecular mass of 470 kDa with eight identical 60-kDa subunits. GS showed its maximal activity at pH 8.0 and was stable between pHs 6.0 and 7.0. The enzyme was strongly inhibited by Fe^<3+>, Hg^<2+>, and Cu^<2+>.

• 社団法人日本生物工学会の論文
• 2002-06-20

著者

• Takenaka Shinji

  Laboratory of Applied Microbiology, Department of Biofunctional Chemistry, Faculty of Agriculture, K

• Murakami Shuichiro

  Laboratory of Applied Microbiology, Department of Biofunctional Chemistry, Faculty of Agriculture, K

• Aoki Kenji

  Laboratory of Applied Microbiology, Department of Biofunctional Chemistry, Faculty of Agriculture, K

• Aoki K

  Department Of Applied Biological Chemistry Graduate School Of Agricultural Science Kobe University

• Takenaka Shinji

  Department of Agrobioscience, Graduate School of Agricultural Science, Kobe University

• Kim Young-ju

  Division Of Science Of Biological Resources Graduate School Of Science And Technology Kobe Universit

• YOSHIZAWA MAKOTO

  Laboratory of Applied Microbiology Department of Biofunctional Chemistry Faculty of Agriculture, Kob

• Yoshizawa Makoto

  Laboratory Of Applied Microbiology Department Of Biofunctional Chemistry Faculty Of Agriculture Kobe

• Murakami S

  Kikkoman Corp. Chiba Pref. Jpn

• Murakami Shuichiro

  Dep. Of Bioscience Biotechnology And Agrochemistry Fac. Of Agriculture Meiji Univ.

• Takenaka Shinji

  Department Of Agrobioscience Graduate School Of Agricultural Science Kobe University

• Aoki K

  Department Of Agricultural Chemistry Faculty Of Agriculture Kobe University

• Aoki Kenji

  Department Of Agricultural Chemistry Faculty Of Agriculture Kobe University

• Kim Young-Ju

  Division of Life Science, The Graduate School of Science and Technology, Kobe University

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