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Development of industrial recombinant cell fermentation and effective productions of gene products.

Research Project

Project/Area Number 04453125
Research Category

Grant-in-Aid for General Scientific Research (B)

Allocation TypeSingle-year Grants
Research Field 発酵工学
Research InstitutionNagoya University

Principal Investigator

KOBAYASHI Takeshi  Dept.of Biotechnology, Nagoya University, Professor, 工学部, 教授 (10043324)

Co-Investigator(Kenkyū-buntansha) HONDA Hiroyuki  Dept.of Biotechnology, Nagoya University, Associate Professor, 工学部, 助教授 (70209328)
朴 龍洙  名古屋大学, 工学部, 助手 (90238246)
Project Period (FY) 1992 – 1993
Project Status Completed (Fiscal Year 1993)
Budget Amount *help
¥6,300,000 (Direct Cost: ¥6,300,000)
Fiscal Year 1993: ¥2,800,000 (Direct Cost: ¥2,800,000)
Fiscal Year 1992: ¥3,500,000 (Direct Cost: ¥3,500,000)
KeywordsFermentation / Yeast / Online measurement system / High cell density culture / Recombinant cell / Growth-inhibitive metabolite / Fuzzy control / Bacillus sps.
Research Abstract

1. High cell density fermentation of recombinant Bacillus sps. is difficult because they produce growth-inhibitive metabolites under enough nutrient condition. The growth-inhibitive metabolites contain organic acids such as acetic acid, lactic acid and so on. Due to reduction of the organic acid production using an online control system, and effective fermentation of high cell density fermentation and high expression of gene products were realized.
2. We researched most effective host-promoter system for gene product. S.cerevisiae20B-12 and S.cerevisiaeYNN27 were used for host strains ans PGK, SUC2 nd GAL7 promoters were used. From the results of comparison between these recombinant cells, S.cerevisiae20B-12/PGK system was the best in growth rate and yeild of product.
3. In the case of heterologous protein production using recombinant yeasts, glucose concentrarion and ethanol concentration of medium are important factors of these effective produciton. In this study, we researched an effect of ethanol concentration. The heterologous protein production was inhibited by ethanol higher than 5 g/l. It was clear that a maintenance of low ethanol concentration was effective for fermentation process.
4. Online control system was developed for regulating both glucose and ethanol concentration. Cell, dissolved oxygen, glucose and ethanol concentrations were used for operating variables and a fuzzy rule of these variables was used for control of the fermentation system. The production of the gene product in this fuzzy control system was twice as much compared with a conventional fermentation.

Report

(3 results)
  • 1993 Annual Research Report   Final Research Report Summary
  • 1992 Annual Research Report
  • Research Products

    (14 results)

All Other

All Publications (14 results)

  • [Publications] Yong Soo Park: "Comparison of Three Different Promoter System for Secretory α-Amylase Production in Fed-batch Culture of Recombinant Saccharomyces cerevisia" Biotechnology and Bioengineering. 41. 854-861 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] 甲斐建一: "膜型バイオリアクターを用いたphoプロモーターからの遺伝子発現の誘導と物質生産の効率化" 生物工学会誌. 71. 317-323 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Chantal Cayuela: "Insecticide Production by Recombinant Bacillus subtilis 1A96 in Fed-Batch Culture with Control of Glucose Concentration" Journal of Fermentation and Bioengineering. 75. 383-386 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Yong Soo Park, Sumihisa Shiba, Shinji Iijima and Takeshi Kobayashi: "Comparison of three different promoter systems for secretory alpha-amylase production in fed-batch culture of recombinant Saccharomyces cerevisiae" Biotechnology and Bioengineering. 41. 854-861 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Kenichi Kai, Toshiro Minami, Sachifumi Imaizumi, Masao Kondo, Koji Yoda, Makari Yamasaki, Shinji Iijima and Takeshi Kobayashi: "Effective production of recombinant protein by using inducible pho promoter" Seibutsu-kogaku Kaishi. 71(5). 317-323 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Chantal Caynela, Kenichi Kai, Yong Soo Park, Shinji Iijima and Takeshi Kobayashi: "Insecticide production by recombinant Bacillus subtilis 1A96 in fed-batch culture with control of glucose concentration" Journal of Fermentation and Bioengineering. 75 (5). 383-386 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Yong Soo Park: "Comparison of Three Different Promoter System for Secretory α-Amylase Production in Fed-batch Culture of Recombinant Saccharomyces Cerevisiae" Biotechnology and Bioengineering. 41. 854-861 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] 甲斐建一: "膜型バイオリアクターを用いたphoプロモーターからの遺伝子発現の誘導と物質生産の効率化" 生物工学会誌. 71. 317-323 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Chantal Cayuela: "Insecticide Production by Recombinant Bacillus subtilis 1A96 in Fed-Batch Culture with Control of Glucose Concentration" Journal of Fermentation and Bioengineering. 75. 383-386 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Takao Saito: "Overproduction of thermostable β-galactosidase in Escherichia coli,its purificat on and Molecular structure" Journal of Fermentation and Bioengineering. 74. 12-16 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] 大嶋 孝之: "λ-ファージP_RP_Lプロモーターを利用した組換え大腸菌の高濃度培養による効率的な遺伝子産物の生産" 化学工学論文集. 18. 684-692 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] Yong Soo Park: "Enhanced β-galacosidase production by high cell-density culture of recombinant Bacillus subtilis with glucose concentration control" Biotechnology and Bioengineering. 40. 686-696 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] 大嶋 孝之: "二種類の炭素源を用いる酵母SUC2プロモーターからの遺伝子発現制御" 化学工学論文集. 18. 693-700 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] Tomoyasu Kawabe: "Inducible production of recombinant xylose isomerase by Escherichia coli in fed-batch culture" Journal of Chemical Engineering of Japan. 25. 702-708 (1992)

    • Related Report
      1992 Annual Research Report

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Published: 1992-04-01   Modified: 2016-04-21  

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