Studies on the enzymatic chemistry involved in the inorganic sulfur metabolism and its application
| Project/Area Number |
22580375
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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 |
Boundary agriculture
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| Research Institution | Okayama University |
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Principal Investigator |
KANAO Tadayoshi 岡山大学, 大学院・環境生命科学研究科, 准教授 (40379813)
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| Co-Investigator(Renkei-kenkyūsha) |
KAMIMURA Kazuo 岡山大学, 大学院・環境生命科学研究科, 教授 (80294445)
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| Project Period (FY) |
2010 – 2012
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| Project Status |
Completed (Fiscal Year 2012)
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| Budget Amount *help |
¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000)
Fiscal Year 2012: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000)
Fiscal Year 2011: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000)
Fiscal Year 2010: ¥2,730,000 (Direct Cost: ¥2,100,000、Indirect Cost: ¥630,000)
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| Keywords | 遺伝子資源 / 無機硫黄化合物 / 硫黄代謝 / 硫黄酸化細菌 / 酵素化学 / 好酸性細菌 / 酵素の結晶化 / refolding / 構造解析 |
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| Research Abstract |
This study has been performed to progress our knowledge about the enzymes involved in the dissimilatory inorganic sulfur metabolism in the sulfur-oxidizing bacteria. We have isolated and identified the gene encodingtetrathionate hydrolase (Af-tth) which is the key enzyme in the metabolism in Acidithiobacillus ferrooxidans. Although Af-tthwas cloned and expressed in recombinant Escherichia coli, the recombinant proteins were synthesized in inclusion bodies as an inactive form. However, we have developed a new method to obtain the recombinant Af-Tth in an active form by refolding under acidic conditions. The enzyme was purified to homogeneity and crystallized. X-ray crystallographic analysis showed that the crystal diffracted to 2.15 A resolution and belongs to space group P31or P32with unit-cell parameters a = b= 92.1, c = 232.6 A. Further analyses such as multiple/single anomalous dispersion methods are required to determine the Af-Tth structure.
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Report
(4 results)
Research Products
(30 results)
