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2003 Fiscal Year Final Research Report Summary

Studies on lactic acid producing bacteria in fermented seafoods

Research Project

Project/Area Number 13460090
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Fisheries chemistry
Research Institution東京水産大学

Principal Investigator

FUJII Tateo  Tokyo University of Marine Science and Technology, Department of Marine Science, Professor, 海洋科学部, 教授 (30093305)

Project Period (FY) 2001 – 2003
Keywordslactic acid bacteria / fish sauce / kusaya / funazushi / Tetragenococcus / histamine / histidine decarboxylase
Research Abstract

(1)Chemical composition of fish sauce produced in Tobishima Island are NaCl 20.7%, VBN 181mg/100g, lactic acid 1,620mg/100g, glutamic acid 1,322mg/100g, histamine 163mg/100mg and halophilic histamine-producing bacterium of Tetragenococus muriaticus appeared in the early stage of fermentation, while T.halophilus appeared throughout the period.
(2)Histamine formation in cultures of Tetragenococcus muriaticus was observed in low acidity (pH 5.8), O_2 limiting conditions with optimal NaCl and glucose concentrations of 5-7% (w/v) and above 1%, respectively. Histamine formation could not be prevented even at 20% (w/v) NaCl.
(3)A histidine decarboxylase from Tetragenococcus muriaticus was purified to homogeneity, for the first time. The pure enzyme consisted of two polypeptide chains with molecular mass of 28.8 and 13.4kDa. The N-terminal amino acid sequences of these polypeptides highly correlated with those of the alpha-and beta-chains of other Gram-positive bacterial histidine decarboxylases … More . The optimum and stable pH for the enzyme was 4.5-7.0 and 4.0-7.0, respectively. The enzyme activity decreased with the addition of NaCl.
(4)The gene co d ing histidine decarboxylase (HDC) has been cloned by EcoRI fragment of chromosomal DNA from Tetragenococcus muriaticus. Sequence analysis of this fragment identified two open reading frames (ORFs). The first ORF (hdcA) encoded 316 amino acid residue s and it was identical to N-terminal amino acid sequence of HDC from T.muriaticus. The deduced amino acid sequence was 98% identity with HDC of Oenococcus oeni. Enzyme activity was reconstituted by treatment with 8M urea solution. The specific activity of renatured HdcA was 3.6×10^<-4> unit/mg and pH optimum at 4.8. The transcript analysis revealed that the hdcA was expressed in stationary-phase cell under O_2-limiting and low pH condition.
(5)DGGE analysis of 16S rDNA fragments amplified directly from kusaya gravy revealed the existence of Bacteroides, Fusobacterium, Eggerthela lenta, and Clostridium, suggesting that unculturable organisms prevail in the gravy.
(6)Dominant microflora of funazushi crucian carp fermented with rice, were Lactobacillus kefir in the late stage of fermentation, and L.alimentarius in the early stage. Less

  • Research Products

    (12 results)

All Other

All Publications (12 results)

  • [Publications] Y.Konagaya: "Purification and properties of a histidine decarboxylase from Tetragenococcus muriaticus, a halophilic lactic acid bacterium"J.Appl.Microbiol.. 92・. 1136-1142 (2002)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] H.Takahashi: "Analysis of bacterial communieties in kusya gravy by denaturing gradient gel electrophoresis of PCR-amplified DNA fragments"Japan.J.Food Microbiol.. 19・4. 179-185 (2002)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] H.Takahashi: "Cloning arid sequencing of the histidine decarboxylase gene of gram-negative, histamine-producing bacteria and their application in detection and identification of these organisms in fish"Appl.Environ.Microbiol.. 69・5. 2568-2579 (2003)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] M.Satomi: "Marinospilillum insulare sp.nov., a novel halophilic helical bacterium isolated from kusaya gravy"Int.J.Syst.Evol.Microbiol.. 54・1. 163-167 (2004)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] B.Kimura: "Histamine formation by Tetragenococcus muriaticus, a halophilic lactic acid bacterium isolated from fish sauce"Int.J.Food Mierobiol.. 70・1-2. 71-77 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 藤井建夫: "増補 塩辛・くさや・かつお節-水産発酵食品の製法と旨味"恒星社厚生閣. 121 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 藤井建夫(編著): "食品の保全と微生物"幸書房. 262 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] B.Kimura: "Histamine formation by Tetragenococcus muriaticus, a halophilic lactic acid bacterium"Int.J.food Microbiol.. 92. 1136-1142 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Y.Konagaya: "Purification and properties of a histidine decarboxylase from Tetragenococcus muriaticus, a halophilic lactic acid bacterium."J.Appl.Microbiol.. 92. 1136-1142 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] H.Takahashi: "Analysis of bacterial communieties in kusya gravy by denaturing gradient gel electrophoresis of PCR-amplified DNA fragments."Japan.J.Food Microbiol.. 19. 179-185 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] H.Takahashi: "Cloning and sequencing of the histidine decarboxylase gene of gram-negative, histamine-producing bacteria and their application in detection and identification of these organisms in fish."Appl.Environ.Microbiol.. 69. 2568-2579 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] M.Satomi: "Marinospilillum insulare sp.nov., a novel halophilic helical bacterium isolated from kusaya gravy."Int.J.Syst.Evol.Microbiol.. 54. 163-167 (2004)

    • Description
      「研究成果報告書概要(欧文)」より

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Published: 2005-04-19  

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