title: Purification and comparison of triosephosphate isomerases from ammonia-oxidizing bacteria isolated from terrestrial and marine environments.
abstract: 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) 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 degrees C, 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.
authors.affiliation: Department of Agricultural and Biological Chemistry, College of Bioresource Sciences, Nihon University, Kameino, Fujisawa, Kanagawa 252-8510, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Iwahara
authors.forename: R
authors.initials: R
authors.lastname: Takahashi
authors.forename: T
authors.initials: T
authors.lastname: Naomi
authors.forename: M
authors.initials: M
authors.lastname: Kida
authors.forename: R
authors.initials: R
authors.lastname: Miyamoto
authors.forename: T
authors.initials: T
authors.lastname: Tokuyama