| Project/Area Number |
07455315
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
反応・分離工学
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| Research Institution | KYUSHU UNIVERSITY |
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Principal Investigator |
NAKASHIO Fumiyuko Kyushu Univ., Chem. Sci & Technol., Professor, 工学部, 教授 (70037729)
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| Co-Investigator(Kenkyū-buntansha) |
KUBOTA Fukiko Kyushu Univ., Chem. Sci & Technol., Associate Profe, 工学部, 教務員
GOTO Masahiro Kyushu Univ., Chem. Sci & Technol., Associate Profe, 工学部, 助教授 (10211921)
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| Project Period (FY) |
1995 – 1996
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| Project Status |
Completed (Fiscal Year 1996)
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| Budget Amount *help |
¥6,800,000 (Direct Cost: ¥6,800,000)
Fiscal Year 1996: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 1995: ¥5,700,000 (Direct Cost: ¥5,700,000)
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| Keywords | Reversed micelle / Protein / extraction / separation / surfactant / refolding / bioreactor / molecular aggregate / バイオテクノロジー / 酵素 / マイクロエマルション |
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| Research Abstract |
A separation technique using reversed micelles provides an attractive alternative to conventional procedures for the separation and the purification of bio-active materials. However, in most of works reported on the protein extraction by reversed micelles, the conventional syrfactant AOT has been used and little attention has been paid to search for a suitable micellar system for the protein extraction. The point we wish to emphasize in this study is that the surfactant plays an important role in the separation process. Most people who study the reversed micellar extraction realize the importance and recognize that few surfactants are available for a protein separation. From the view point, we have developed a new reversed micellar system. In this study, we have proposed a mixed reversed micellar system of AOT and DOLPA (dioleyl phosphoric acid), and extraction behavior of some proteins was studied with the mixed reversed micellar system. Proteins are easily extracted from aqueous phas
… More
e into organic iso-octane containing 10 mM of AOT and DOLPA (4 : 1). The extraction ability of the mixed reversed micelle of 10 mM is almost correspond to that of 200 mM AOT.The extraction of proteins with low concentrations of surfactants is advantageous from a processing standpoints because amphiphilic molecules like surfactants are very difficult to remove in downstream processing. The results of extraction indicated that the AOT-DOLPA mixed reversed micelles are very available for separating and enriching the protein. Back-extraction of chymotrypsin from the organic phase is also accomplished by adding a small amount of alcohol. The activity of chymotrypsin recovered from the mixed reversed micelles was completely kept the original enzymatic activity. The residual activity in the 10 mM AOT-DOLPA mixed reversed micelles was much higher than that of 200 mM AOT reversed micelles. This result means that the mixed reversed micellar system appears to do little harm to the protein. Reducing the concentration of surfactants will make the use of liquid-liquid protein extraction increasingly cost effective. Further, we demonstrate that reversed micelles are useful for refoding of denatured proteins. Less
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