| Project/Area Number |
06555239
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| Research Category |
Grant-in-Aid for Scientific Research (A)
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| Allocation Type | Single-year Grants |
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| Section | 試験 |
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| Research Field |
反応・分離工学
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| Research Institution | Waseda University |
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Principal Investigator |
HIRATA Akira Waseda University, School of Science and Engineering, Professor, 理工学部, 教授 (00063610)
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| Co-Investigator(Kenkyū-buntansha) |
OYAMA Kiyotaka Toso Co., Finechemical Department, Researcher, ファインケミカル事業部, 副理事(研究職)
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| Project Period (FY) |
1994 – 1996
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| Project Status |
Completed (Fiscal Year 1996)
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| Budget Amount *help |
¥8,600,000 (Direct Cost: ¥8,600,000)
Fiscal Year 1996: ¥1,700,000 (Direct Cost: ¥1,700,000)
Fiscal Year 1995: ¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1994: ¥4,800,000 (Direct Cost: ¥4,800,000)
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| Keywords | Biologically active peptide / enzymatic reaction / extraction / crystallization / soluble enzyme / continuous synthesis / extractive separation of product / aspartame precursor |
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| Research Abstract |
The novel method to synthesize biologically active peptides with a continuous use of soluble enzyme and a simultaneous operation of an enzymatic reaction with a product separation was developed in this study, and applied to the protease-catalyzed synthesis of the precursor of aspartame (Z-APM and F-APM)
A) Continuous synthesis of Z-APM
peptides are synthesized with enzyme at its optimal pH,though the enzymatic synthesis of peptides at low pH has a lot of advantages for an industrial-scaled production, such as, suppression of nonenzymatic decomposition of amino ester substrate and less requirement of NaOH for pH adjustment. We synthesize Z-APM at low pH in aqueous/organic biphasic system and obtained more than 95% of yield in this study. The mathe mathematical model to predict the continuous synthesis of Z-APM with extractive reaction was proposed and agreed well with the experimental results.
B) Continuous synthesis of F-APM
N-Benzyloxycarbonyl-L-aspartic acid (Z-L-Asp) and L-phenylalanine methyl ester (L-PheOMe) were, in general, used as the substrates for the enzymatic synthesis of aspartame precursor. N-Formyl group (F-group), which was cheaper and could be removed more easily than Z-group, was examined to be used for the protection of aspartic acid instead of Z-group to lower the total cost of the industrially continuous production of aspartame in this study. The F-APM yield was about 3% in aqueous monophsic system. The extremely high conversion, more than 95%, was, however, obtained for the thermolysin-catalyzed synthesis of F-APM in aqueous/TBP biphasic system.
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