• Search Research Projects
  • Search Researchers
  • How to Use
  1. Back to previous page

Development of probiotic bacteria which have an ability of degrading cholesterol and starch

Research Project

Project/Area Number 10556019
Research Category

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

Allocation TypeSingle-year Grants
Section展開研究
Research Field 応用微生物学・応用生物化学
Research InstitutionOsaka University

Principal Investigator

MUROOKA Yoshikatsu  Osaka Univ.Eng. Professor, 大学院・工学研究科, 教授 (60029882)

Co-Investigator(Kenkyū-buntansha) ONO Hisayo  Osaka Univ.Eng. Assis.Prof., 遺伝情報実験施設, 助手 (40224274)
YAMASHITA Mitsuo  Osaka Univ.Eng. Assoc.Prof, 大学院・工学研究科, 助教授 (40220347)
KANEKO Yoshinobu  Osaka Univ.Eng. Assoc.Prof., 大学院・工学研究科, 助教授 (90161182)
ASHIKARI Toshihiko  Suntory Senior Res., 基礎研究所, 主席研究員
Project Period (FY) 1998 – 2000
Project Status Completed (Fiscal Year 2000)
Budget Amount *help
¥12,500,000 (Direct Cost: ¥12,500,000)
Fiscal Year 2000: ¥2,300,000 (Direct Cost: ¥2,300,000)
Fiscal Year 1999: ¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 1998: ¥8,000,000 (Direct Cost: ¥8,000,000)
Keywordscholesterol / cholesterol oxidase / probiotics / lactic acid bacteria / propionic acid bacteria / dairy food
Research Abstract

Probiotic bacteria, such as lactic acid and propionic acid bacteria and bifidobacteria, are believed to contribute to promoting human health in different ways. Consequently, genetic engineering to improve dairy lactobacilli has been actively pursued, promising numerous benefits in food production and health promotion.
In this project, host-vector systems for lactic acid and propionic acid bacteria have been developed. The shuttle vectors pRN14 and pPK705 were constructed from indigenous plasmids derived from Lactobacillus plantarum L137 and Propionibacter acidipropionici, respectively. Successful development of transformation systems in various species of Lactobacillus and Propionibacterium using shuttle vectors pRN14 and pPK705 prompted to develop as expression system for heterologous genes in lactic acid and propionic acid bacteria. Our choA gene from Streptomyces has been widely used as a diagnostic tool or for degradation of cholesterol in several species of lactic acid bacteria. In … More these studies, promoters from E.coli and Streptomyces were used for the expression of choA.In contrast, we constructed a plasmid, pTACO-1, that consisted of the acetyl coenzyme A carboxylase (acc) promoter and the aminoterminal region of the first open reading frame of the acc operon, which is essential for fatty acid biosynthesis, plus choA in the promoter probe vector pCVE1. Furthermore, we constructed an expression vector for lactic acid bacteria using the acc promoter-choA cassette and a shuttle vector, pRN14. For propionibacteria, native promoters (P138, P8, P116, and P4) were isolated from P.freudenrechii subsp. freudenreichii IFO 12424 using the promoter probe vector pCVE1. The Propionibacterium promoters allow as strong expression of cholesterol oxidase in E.coli were used to construct an expression vector for propionibacteria with the shuttle vector pPK705.
Thus, the extension of the functional diversity in probiotic bacteria will increase their potential utility as a "health care microbe" for bringing about desirable compositional changes in food, such as reduction of cholesterol content in dairy products. Less

Report

(4 results)
  • 2000 Annual Research Report   Final Research Report Summary
  • 1999 Annual Research Report
  • 1998 Annual Research Report
  • Research Products

    (49 results)

All Other

All Publications (49 results)

  • [Publications] Y.Nishiya: "Effect of non-ionic detergents on apparent enzyme mechanism : V121A mutant of Streptomyces cholesterol oxidase endowed with enhanced sensitivity against detergents."Protein Engineering. 11(8). 609-611 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] M.Yamashita: "Separation of the two reactions, oxidation and isomerization, catalyzed by Streptomyces cholesterol oxidase"Protein Engineering. 11(11). 1075-1081 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] H.Ono: "Characterization of biosynthetic enzymes for ectoine as a compatible solute in a moderately halophilic eubacteria, Halomonas elongata."Journal of Bacteriology. 181(1). 91-99 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] S.Nishikawa: "Rhodobacter sphaeroides mutants which accumulate 5-aminolevulinic acid under aerobic and dark conditions."Journal of Bioscience and Bioengneering. 87(6). 798-804 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Y.Kaneko: "Development of a host-vector system for Lactobacillus plantarum L137 isolated from a traditional fermented food produced in the Philippines."Journal of Bioscience and Bioengneering. 89(1). 62-67 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] M.Yamashita: "Isolation, characterization, and molecular cloning of a thermostable xylitol oxidase from Streptomyces sp.IKD472."Journal of Bioscience and Bioengneering. 89(4). 350-360 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] S.-H.Hong: "Molecular design of novel metal-binding oligomeric human metallothioneins."Applied Microbiology and Biotechnology. 54. 84-89 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] H.Nkayama: "Ectoine, the compatible solute of Halomonas elongata, confers hyperosmotic tolerance in cultured tobacco cells."Plant Physiology. 122. 1239-1247 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] P.Kiatpapan: "Characterization of pRGO1, a plasmid from Prpionibacterium acidipropionici, and its use for development of a host-vector system in Propionibacteria."Applied and Environmental Microbiology. 66(11). 4688-4695 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] P.Kiatpapan: "Molecular characterization of Lactobacillus plantarum genes for β-ketoacyl-acyl carrier protein synthase III(fabH) and acetyl coenzyme A carboxylase (accBCDA) which are essntial for fatty acid biosynthesis."Applied and Environmental Microbiology. 67(1). 426-433 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] S.-H.Hong: "High yield expression and single step purification of human thionein/metallothionnein."Protein Expression and Purification. 21(1). 243-250 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] P.Kiatpapan: "P.Kiatpapan, Construction of an expression vector for propionibacteria and its use in production of aminolevulinic acid (ALA) by Propionibacterium freudenreichii."Applied Microbiology and Biotechnology. (印刷中). (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Y.Murooka: "Genetic and protein engineering of diagnostic enzymes, cholesterol oxidase and xylitol oxidase."Journal of Bioscience and Bioengneering. (印刷中). (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Yoshikatsu Murooka: "Progress in Biotechnology Vol.15, Stability and Stabilization of Biocatalysts"eds.A.Ballesteros, et al.,Elsevier, Amsterdam. 757 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] 鈴木孝仁 ら編集: "微生物の資材化:研究の最前線"ソフトサイエンス社. 364 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] S.Nishikawa: "Biotechnology & Genetic Engineering Reviews."eds.A.Ballesteros, et al., Elsevier, Amsterdam(印刷中). (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Y.Nishiya: "Effect of non-ionic detergents on apparent enzyme mechanism : V121A mutant of Streptomyces cholesterol oxidase endowed with enhanced sensitivity towards detergents."Protein Eng.. 11(8). 609-611 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] M.Yamashita: "Separation of the two reactions, oxidation and isomerization, catalyzed by Streptomyces cholesterol oxidase."Protein Eng.. 11(11). 1075-1081 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] H.Ono: "Characterization of biosynthetic enzymes for ectoine as a compatible solute in a moderately halophilic eubacteria, Halomonas elongata."J.Bacteriol.. 181(1). 91-99 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] S.Nishikawa: "Rhodobacter sphaeroides mutants which accumulate 5-aminolevulinic acid under aerobic and dark conditions."J.Biosci.Bioeng.. 87(6). 798-804 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Y.Kaneko: "Development of a host-vector system for Lactobacillus plantarum L137 isolated from a traditional fermented food produced in the Philippines."J.Biosci.Bioeng.. 89(1). 62-67 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] M.Yamashita: "Isolation, characterization, and molecular cloning of a thermostable xylitol oxidase from Streptomyces sp. IKD472."J.Biosci.Bioeng.. 89(4). 350-360 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] P.Kiatpapan: "Characterization of pRGO1, a plasmid from Propionibacterium acidipropionici, and its use for development of a host-vector system in Propionibacteria."Appl.Environ.Microbiol.. 66(11). 4688-4695 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] P.Kiatpapan: "Molecular characterization of Lactobacillus plantarum genes for β-ketoacyl-acyl carrier protein synthase III (fabH) and acetyl coenzyme A carboxylase (accBCDA) which are essential for fatty acid biosynthesis."Appl.Environ.Microbiol.. 67(1). 426-433 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] K.Nanda: "Characterizations of acetic acid bacteria in traditional acetic acid fermentation of rice vinegar (komesu) and unpolished rice vinegar (kurosu) produced in Japan."Appl.Envirol.Microbiol.. 67(2). 986-990 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] P.Kiatpapan: "Construction of an expression vector for propionibacteria and its use in production of aminolevulinic acid (ALA) by Propionibacterium freudenreichii."Appl.Microbiol.Biotechnol.. (in press).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Y.Murooka: "Genetic and protein engineering of diagnostic enzymes, cholesterol oxidase and xylitol oxidase."J.Biosci.Bioeng.. (in press).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] S.Nishikawa: "5-aminolevulinic acid (ALA) : the production by fermentation and agricultural and biomedical applications."Biotechnology & Genetic Engineering Reviews. Vol.18 (in press).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] S.-H.Hong: "Molecular design of novel metal-binding oligomeric human metallothioneins."Applied Microbiology and Biotechnology. 54. 84-89 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] H.Nakayama: "Ectoine, the compatible solute of Halomonas elongata, confers hyperosmotic tolerance in cultured tobacco cells."Plant Physiology. 122. 1239-1247 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] P.Kiatpapan: "Characterization of pRGO1, a plasmid from Propionibacterium acidipropionici, and its use for development of a host-vector system in Propionibacteria."Applied and Environmental Microbiology. 66(11). 4688-4695 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] P.Kiatpapan: "Molecular characterization of Lactobacillus plantarum genes for β-ketoacyl-acyl carrier protein synthase III (fabH) and acetyl coenzyme A carboxylase (accBCDA) which are essential for fatty acid biosynthesis."Applied and Environmental Microbiology. 67(1). 426-433 (2001)

    • Related Report
      2000 Annual Research Report
  • [Publications] S.-H.Hong: "High yield expression and single step purification of human thionein/metallothionein."Protein Expression and Purification. 21(1). 243-250 (2001)

    • Related Report
      2000 Annual Research Report
  • [Publications] Y.Murooka: "Genetic design of stable metal-binding biomolecules, oligomeric metallothioneins"Biocatalysis and Biotransformation. 9(3)(印刷中). (2001)

    • Related Report
      2000 Annual Research Report
  • [Publications] 鈴木孝仁 ら 編集: "微生物の資材化:研究の最前線"ソフトサイエンス社. 364 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] S. Nishikawa: "Rhodobacter sphaeroides mutants which accumulate 5-aminolevulinic acid under aerobic and dark conditions"Journal of Bioscience and Bioengineering. 87(6). 798-804 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] 西矢 芳昭: "機能改変コレステロールオキシダーゼの開発と応用"生物工学会誌. 77(10). 429-432 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Yoshinobu Kaneko: "Development of a host-vector system for Lactobacillus plantarum L137 isolated from a traditional fermented food produced in the Philippines"Journal of Bioscience and Bioengineering. 89(1)(印刷中). (2000)

    • Related Report
      1999 Annual Research Report
  • [Publications] S.-H. Hong: "Molecular design of novel metal-binding oligomeric human metallothioneins"Applied Microbiology and Biotechnology. (印刷中). (2000)

    • Related Report
      1999 Annual Research Report
  • [Publications] H. Nakayama: "Ectoine,the compatible solute of Halomonas,elongata,confer hyperosmotic tolerance in cultured tobacco cells"Plant Physioligy. (印刷中). (2000)

    • Related Report
      1999 Annual Research Report
  • [Publications] M. Yamashita: "Isolation,characterization,and molecular cloning of a thermostable xylitol oxidase from Streptomyces sp. IKD472"Journal of Bioscience and Bioengineering. 89(1)(印刷中). (2000)

    • Related Report
      1999 Annual Research Report
  • [Publications] S.-L.Ryu: "Reciprocal translocation at duplicated RPL2 loci might cause speciation of Saccharomyces bayanus and Saccharomyces cerevisiae." Current Genetics. 33. 345-351 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Y.Nishiya: "Effect of non-ionic detergents on apparent enzyme mechanism:V121A mutant of Streptomyces cholesterol oxidase endowed with enhanced sensitivity against detergents." Protein Engineering. 11(8). 609-611 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] S.-L.Ryu: "A simple PCR method for distinguishing Saccharomyces cerevisiae from its sibling species by amplification of the RPL2 region." Journal of Fermentation and Bioengineering. 86(3). 249-252 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Mitsuo Yamashita: "Separation of the two reactions,oxidation and isomerization,catalyzed by Streptomyces cholesterol oxidase" Protein Engineering. 11(11). 1075-1081 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] H.-J.Cho: "Agrobacterium rhizogenes-mediated transformation and regeneration of the legume Astragalus sinicus(Chinese milk vetch)." Plant Science. 138(1). 53-65 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Hisayo Ono: "Characterization of biosynthetic enzymes for ectoine as a compatible solute in a moderately halophilic eubacteria,Halomonas elongata." Journal of Bacteriology. 181(1). 91-99 (1999)

    • Related Report
      1998 Annual Research Report
  • [Publications] Yoshikatsu Murooka: "Biological Nitrogen Fixation for the 21^<st> Century" eds.C.Elmerich,et al.,Kluwer Academic Publishers.London, 708 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Yoshikatsu Murooka: "Progress in Biotechnology Vol.15,Stability and Stabilization of Biocatalysts" eds.A.Ballesteros.et al.,Elsevier.Amsterdam, 757 (1998)

    • Related Report
      1998 Annual Research Report

URL: 

Published: 1998-04-01   Modified: 2016-04-21  

Information User Guide FAQ News Terms of Use Attribution of KAKENHI

Powered by NII kakenhi