1996 Fiscal Year Final Research Report Summary
Elucidation of sulfate-dependent acid phosphatase from Thiobacillus thiooxidans and determination of sulfate in waters using whole cells
| Project/Area Number |
07650955
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
生物・生体工学
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| Research Institution | Yamanashi University |
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Principal Investigator |
AMANO Yoshifumi Yamanashi Univ. , Fac.Eng. , Professor, 工学部, 教授 (70020401)
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| Co-Investigator(Kenkyū-buntansha) |
NAKAMURA Kazuo Yamanashi Univ. , Fac.Eng. , Research Assistant, 工学部, 助手 (80111780)
KUROSAWA Hiroshi Yamanashi Univ. , Fac.Eng. , Assistant Professsor, 工学部, 助教授 (10225295)
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| Project Period (FY) |
1995 – 1996
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| Keywords | Thiobacillus thiooxidans / Acid phosphatase / Sulfate |
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| Research Abstract |
We developed a microbial method to determine sulfate concentration in water based on a sulfate-dependent acid phosphatase (APase) in whole cells of Thiobacillus thiooxidans.
The activity of APase was colorimetrically determined by using p-nitrophenylphosphate as substrate. The APase was activated by sulfate. A linear relationship was obtained between the activity of APase and the concentration of sulfate in the range of to 0.6mM.Therefore, the concentration of sulfate was estimated from the activity of APase represented byabsorbance. The microbial method was applied to the determination of sulfate in water. The values measured by the microbial method almost agreed with those by a conventional barium chloranilate method.
A system composed of a microbial column packing the cells of T.thiooxidans and an oxygen electrode immobilizing glucose oxidase (GOD) was developed to perform a continuous determination of sulfate. In this system, the heat-treated cells were employed as APase because the activation effect by sulfate was increased when the cells were exposed to the heat-treatment (70ﾟC,30min), and glucose-6-phosphate (G-6-P) was used as substrate for APase. The APase in the heat-treated cells produced glucose from G-6-P and its activity was activated by sulfate. The concentration of glucose produced from G-6-P was determined by the oxygen electrode with GOD.The linear relationship was obtained between the response value of oxygen electrode corresponding to glucose concentration and the concentration of sulfate in the range of 0 to 1.0mM.Therefore, the concentration of sulfate was estimated from the response value of the oxygen electrode. The values measured by this system expressed a high reproducibility for 5 measurements of sample.
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