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Development of Multi-enzyme bioreactors using coenzyme-collecting system.

Research Project

Project/Area Number 07456051
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field 応用微生物学・応用生物化学
Research InstitutionKogakuin University

Principal Investigator

OHTA Takahisa  Kogakuin University, Department of Applied Chemistry Professor, 工学部, 教授 (30011844)

Co-Investigator(Kenkyū-buntansha) SAKAGUCHI Masayoshi  Kogakuin University, Department of Applied Chemistry Assistant, 工学部, 助手 (80281351)
Project Period (FY) 1995 – 1996
Project Status Completed (Fiscal Year 1996)
Budget Amount *help
¥7,900,000 (Direct Cost: ¥7,900,000)
Fiscal Year 1996: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1995: ¥7,100,000 (Direct Cost: ¥7,100,000)
KeywordsBIOREACTOR / IMMOBILIZED ENZYMES / COENZYMES / NAD^+ / ATP / バイオイアクター / NAD
Research Abstract

I.Coenzyme collecting column system
We extablished optimal conditions for collecting coenzymes.
1. Optimal conditons for collecting NAD^+ or NADH
Adsorption : 50mM Tris-HCl, pH8.5, Elution : 50mM MES-NaOH,pH6.0
2. Optimal conditions for collecting ATP or AMP
Adsorption : 50mM GlyGly-NaOH,pH9.5, Elution : 50mM Citrate, pH4.0
3. Optimal conditions for collecting ADP
Adsorption : 50mM CHS-NaOH,pH9.5, Elution : 50mM Citrate, pH4.0
II.Model systems for consumption and regeneration of coenzymes.
We immobilized enzymes for consuming and regenerating NADH or ATP using AffiGE115.
1. Reaction system for NAD^+/NADH
Immobilizing condition for L-lactate dehydrogenase (LDH) : 100mMMOPS-NaOH,pH6.0
Immobilizing condition for formate dehydrogenase (FDH) : 100mM MOPS-NaOH,pH8.0
2. Reaction system for ATP/ADP
Immobilizing condition for hexokinase (HK) : 100mM MOPS-NaOH,pH6.0
Immobilizing condition for pyruvate kinase (PK) : 100mM HEPES,pH8.0
3. Stability of immobilized enzymes
Immobilized FDH had kept 80% of its activity after one month. Activities of other immobilized enzymes somewhat increased during storage.

Report

(3 results)
  • 1996 Annual Research Report   Final Research Report Summary
  • 1995 Annual Research Report

URL: 

Published: 1995-04-01   Modified: 2016-04-21  

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