DEVELOPMENT OF BIOREACTOR SYSTEM FOR OLIGOPEPTIDE SYNTHESIS COMBINED WITH A SIMULATED MOVING BED ADSORBER
| Project/Area Number |
07555563
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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 |
生物・生体工学
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| Research Institution | OKAYAMA UNIVERSITY |
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Principal Investigator |
NAKANISHI Kazuhiro OKAYAMA UNIV., ENGINEERING,PROFESSOR, 工学部, 教授 (90026584)
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| Co-Investigator(Kenkyū-buntansha) |
SHIOTA Katashi MITSUBISHI CHEMICAL CORP., INTERMEDIATE CHEMICALS DEPARTMENT,GENERAL MANAGER, 中間体事業部, 部長(研究職)
UTAGAWA Takashi AJINOMOTO CO., INC., PROCESS DEVELOPMENT LAB., DEPUTY GENERAL MANAGER, プロセス開発研究所, 副部長(研究職)
SIRAI Yoshihito KYUSHU INSTITUTE OF TECHNOLOGY,COMPUTER SCIENCE AND SYSTEM ENGINEERING,ASSOCIATE, 情報工学部, 助教授 (50175395)
TANAKA Takaaki OKAYAMA UNIV., ENGINEERING,ASSISTANT, 工学部, 助手 (00217043)
SAKIYAMA Takaharu OKAYAMA UNIV., ENGINEERING,LECTURER, 工学部, 講師 (70170628)
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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 |
¥1,700,000 (Direct Cost: ¥1,700,000)
Fiscal Year 1996: ¥1,700,000 (Direct Cost: ¥1,700,000)
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| Keywords | BIOREACTOR / IMMOBILIZED ENZYME / THERMOLYSIN / ASPARTAME / ORGANIC SOLVENT / SIMULATED MOVING BED / THERMAL STABILITY / OLIGOPEPTIDE / 酵素合成 / 自己消化 / イオンペア- |
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| Research Abstract |
In this study, we aimed to construct an effective bioreactor for synthesis of aspartame precursor, as a model oligopeptide in an organic solvent, using thermolysin immobilized onto Amberlite XAD-7. First, we investigated the factors affecting the synthesis of the asparatme precursor using an immobilized enzyme. We studied a synthetic rate, particularly in a mixed organic solvent with ethyl acetate and tert-amyl alcohol. Using a mixed solvent of 33% tert-amyl alcohol and 67% ethyl acetate, the activity and stability of the immobilized thermolysin were high enough to conduct a continuous reaction. With increasing the content of tert-amyl alcohol, the stability of the immobilized enzyme was enhanced. The reason for the high activity and stability of the immobilized thermolysin in the presence of tert-amyl alcohol was studied from the viepoint of stability of calcium bound on the enzyme molecule, a stabilizing factor for thermolysin. An optimum water content for the synthesis of asparatame
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precursor was found to be 4-6 % in the mixed organic solvent. The concentration of L-phenyllalanine methyl ester (PheOMe) and N- (benzy loxy carbonyl) -L-as partic acid (Z-Asp) was optimized. The concentration of PheOMe was set to 200 mM and that of Z-Asp was 40 mM.We could also conduct a continuous reaction in a column reactor at 40ﾟC.The conversion at the outlet of the reactor with respect of Z-Asp was around 99% with a space velocity of 4.9 (1/h). Based on this result, we aimed to develop a bioreactor system combined with a simulated moving bed absorber. Namely, at the outlet of the bioreactor, the Z-Asp concentration was negligible low with asparatame precursor Z-Asp PheOMe of around 40 mM and PheOMe of 160 mM.Thus, we could recycle PheOMe to the bioreactor with continuous separation of PheOMe and Z-AspPheOMe using a simulated mooving bed adsorber. We found Dowex MWA-1 as a suitable adsorbent and analyzed adsorption/desorption behaviors of PheOMe and Z-AspPheOMe. We also calculated a separation performance for a simulated moving bed adosorber. We also showed a possible bioreactor system which could be used for an industrial production. Finally, we briefly compared an enzymatic synthesis and chemical synthesis. Less
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Report
(3 results)
Research Products
(10 results)
