Purification and Comparison of Triosephosphate Isomerases from Ammonia-Oxidizing Bacteria Isolated from Terrestrial and Marine Environments

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概要

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• Triosephosphate isomerases[TIMs, EC 5.3.1.1]were purified from two ammonia-oxidizing bacteria: Nitrosomonas sp.K1(K1), Nitrosomonas sp.TNO632(TNO). The molecular masses of the native enzymes were estimated to be about 53.6(K1-TIM)and 51.9 kDa(TNO-TIM)by gel filtration, whereas SDS-PAGE produced one band for each enzyme with M_r values of 27.1(K1-TIM)and 26.4 kDa(TNO-TIM), respectively, suggesting that the enzymes consist of identical subunits. The apparent K_m values for _D-glyceraldehyde-3-phosphate(GAP)and dihydroxyacetone phosphate(DHAP)were about 1.19 and 4.78 mM(K1-TIM), and 0.41 and 6.01 mM(TNO-TIM), respectively. The two TIMs had different pH-activity curves with an optimum pH range of 6.5(K1-TIM)and 8.0(TNO-TIM). The temperature optima of K1-TIM and TNO-TIM were 50-60 and 60-65℃, respectively. Both enzymes were strongly inhibited by 5, 5′-ditiobis at 1.0 mM. The N-terminal amino acid sequences of K1-TIM and TNO-TIM were MRAGFVAGNWKMHG(K1-TIM)and MVRTGLVAGNWKMNG(TNO-TIM). A homology of 74.1% was observed between K1-TIM and TNO-TIM.

• 公益社団法人日本生物工学会の論文
• 2001-06-25

著者

• Takahashi Reiji

  Department Of Agricultural And Biological Chemistry College Of Bioresource Sciences Nihon University

• Tokuyama Tatsuaki

  Department Of Agricultural And Biological Chemistry College Of Bioresource Sciences Nihon University

• Miyamoto Rie

  Department Of Agricultural And Biological Chemistry College Of Bioresource Sciences Nihon University

• IWAHARA KAZUNOBU

  Department of Agricultural and Biological Chemistry, College of Bioresource Sciences, Nihon Universi

• NAOMI TATSUYA

  Department of Agricultural and Biological Chemistry, College of Bioresource Sciences, Nihon Universi

• KIDA MAKOTO

  Department of Agricultural and Biological Chemistry, College of Bioresource Sciences, Nihon Universi

• Kida Makoto

  Department Of Agricultural And Biological Chemistry College Of Bioresource Sciences Nihon University

• Naomi Tatsuya

  Department Of Agricultural And Biological Chemistry College Of Bioresource Sciences Nihon University

• Iwahara Kazunobu

  Department Of Agricultural And Biological Chemistry College Of Bioresource Sciences Nihon University

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