2018 Fiscal Year Final Research Report
Studies on Asymmetric Nitroaldol Reaction using mutant albumins
| Project/Area Number |
16K05706
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Multi-year Fund |
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| Section | 一般 |
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| Research Field |
Organic chemistry
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| Research Institution | Meisei University |
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Principal Investigator |
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| Co-Investigator(Kenkyū-buntansha) |
須賀 則之 明星大学, 理工学部, 准教授 (00396219)
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| Project Period (FY) |
2016-04-01 – 2019-03-31
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| Keywords | アルブミン / 不斉ニトロアルドール / クローニング / 光学活性ビアリール化合物 |
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| Outline of Final Research Achievements |
We applied the reaction to prepare optically active biaryl-containing compounds. The reaction of 4-phenylbenzaldehyde with nitromethane was carried out using human serum albumin to afford the corresponding (R)-alcohol. The NMR analysis of the reaction mixture indicated that the substrate apparently interacted with albumin under the reaction conditions.
In order to obtain highly purified HSA, it is necessary to express and secrete a lot of HSA. Therefore, we conducted a construction of the PichiaPink Expression System (Invitrogen). Six kinds of plasmids were constructed by the combination of pPink-HC and pPink-LC plasmids and three secretion signals. These plasmids were introduced into Pichia pink strains 1 to 4 to construct a total of 141 HSA-secreting Pichia strains. As a result, a high expression clone was obtained which expressed and secreted 0.32 mg of HSA per 1 ml of culture supernatant.
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| Free Research Field |
有機合成化学
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| Academic Significance and Societal Importance of the Research Achievements |
アルブミンというありふれたタンパク質の新機能を用いて、現在注目される水中不斉合成を試みた本研究は、有機合成化学・タンパク質化学の境界領域の研究テーマとして極めてユニークといえる。また、本反応の最適化を達成し、光学活性ビアリール化合物を効率的に得られる新手法を明示したことにより、有機化学的にも大きな意味がある。一方で、HSAを高度に精製するために、HSAを分泌発現する高発現クローンを構築することができ、アルブミン変異体ライブラリー作製のノウハウを築けたことは、分子生物化学的にも医学的にも意義深いといえる。
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