Salt-induced conformational change and hydrolase susceptibility of polysaccharide responsible for floc formation in Pseudomonas sp.
| Project/Area Number |
02660105
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
応用微生物学・発酵学
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| Research Institution | The University of Tokyo |
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Principal Investigator |
TAGO Yoshitaka The University of Tokyo Institute of Applied Microbiology Instructor, 応用微生物研究所, 助手 (70111573)
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| Project Period (FY) |
1990 – 1991
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| Project Status |
Completed (Fiscal Year 1991)
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| Budget Amount *help |
¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1991: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1990: ¥1,400,000 (Direct Cost: ¥1,400,000)
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| Keywords | Mucopolysaccharise / Mucopolysaccharide hydrolase / Conformation of polysaccharid / Bacterial aggregation / Ionic strength / 多糖の重合度と立体構造 / 多糖の酵素による加水分解パタ-ン |
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| Research Abstract |
The MPS hydrolase was purified with Immobiline-Canal-Isoelectric-Forcusing. The enzyme was shown one band by SDS-PAGE. The polysaccharide (MPS) purified as previously was fractionated with GPC regarding it's molecular size. Using these enzyme and polysaccharide, following results were obtained.
(1) The conformation of MPS was dependent on the ionic strength of solute. MPS was disordered state in water, and addition of salts promotes the conformational change of MPS from disordered state to a ordered state(the change of ionic strength 0 to 0.01)and packing of the form was occured during the increase of. ionic strength up to 0.05 and no change was detected more than 0.05.
(2) It was clarified by the enzyme kinetics study that the inhibition of the hydrolysis of MPS with MPS hydrolase under the ionic strength was not derived from the result of the inhibition of the enzyme, but from the results of the effects of the ionic strength to the substrate (MPS).
(3) The ordered form of MPS was seemed antiparallel arrangement and the form was accomplished in the ionic strength 0.01.
(4) The inhibition of hydrolysis of MPS with MPS hydrolase by the ionic strength was coincident with the formation of ordered form of MPS promoted with the ionic strength.
(4) The increasing of the reducing power was not detected for 20min-60min under high ionic strength using the MPS which molecular weight was higher than 10, 000-20, 000. But after that, the reducing power increased as the same rate as in the case of low ionic strength condition and low molecular weight MPS.
(5) By the enzyme products analysis by HPLC, the first product was oligo-MPS(molecular weight=10, 000-20, 000), and then, two repeating units oligosaccharide from oligo-MPS, and finally one repeating unit oligosaccharide from oligo-2 was obtained.
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Report
(3 results)
Research Products
(22 results)
