Development of safe food using lactic acid bacteria

• Project/Area Number: 14560221
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Zootechnical science/Grassland science
• Research Institution: Hirosaki University
• Principal Investigator: TOBA Takahiro Hirosaki University, Faculty of Agriculture and Life Science, Professor, 農学生命科学部, 教授 (10108483)
• Project Period (FY): 2002 – 2003
• Project Status: Completed (Fiscal Year 2003)
• Budget Amount: ¥3,100,000 (Direct Cost: ¥3,100,000); Fiscal Year 2003: ¥1,400,000 (Direct Cost: ¥1,400,000); Fiscal Year 2002: ¥1,700,000 (Direct Cost: ¥1,700,000)
• Keywords: Lactobacillus crispatus / hydrogen peroxide / bifidobacteria / PCR / Lactobacillus gasseri / S-layer protein

Research Abstract

Adhesion assay was performed using Lactobacillus and Bifidobacterium strains isolated from chicken and human faeces to select the strains which strongly adhered to chicken crop epithelium, human intestinal Caco-2 cells and HT-29 cells, and immobilized extracellular matrix proteins. For rapid identification of Lactobacillus crispatus we newly developed PCR primers targeting S-layer protein gene(Horie et al.2002). Adhesive lactobaciller and bifidobacterial strains inhibited the adhesion of pathogens such as Salmonella to Caco-2 cells Adhesion of L.crispatus and L.gasseri strains tested were induced on plate cultures under anaerobic conditions. A 46.5kDa S-layer protein(named as LbsA) was identified as an adhesin of highly adhesive L.crispatus MH 3 1 5 strain. The DNA region coding for LbsA was cloned and sequenced. Alignment comparisons of the predicted amino acid sequence with S-layer proteins of other Lactobacillus strains suggested the presence of laminin-binding site in LbsA (Mukai et al.2003). An 58.7kDa surface protein was identified an putative adhesin of L.gasseri MH 411 by Western bolting assay.
We also selected hydrogen peroxide(H_2O_2)-producing Lactococcus lactis subsp.lactis and L.plantarum strains from foods. They produced >300ppm H_2O_2 under the condition tested. One of these strains, L.lactis subsp.lactis AI62 inhibited pathogens such as Listeria ivanovii, Yersinia enterocolitica and Aeromonas hydrophila efficiently(Ito et al.2003).

Research Products

• [Publications] Horie M, Kajikawa HS, Toba T: "Identification of Lactobacillus crispatus by polymerase chain reaction targeting S-layer protein gene."Letters in Applied Microbiology. 35. 57-61 (2002)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Ito A, Sato Y, Kudo S, Sato S, Nakajima H, Toba T: "The screening of hydrogen peroxide-producing lactic acid bacteria and their application to inactivating psychrotrophic food-borne pathogens."Current Microbiology. 47. 231-236 (2003)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] 向井孝夫, 戸羽隆宏, 大堀均: "乳酸菌における細胞接着の分子機構"ミルクサイエンス. 52. 167-174 (2003)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Horie M, Kajikawa HS, Toba T: "Identification of Lactobacillus crispatus by polymerase chain reaction targeting S-layer protein gene."Letters in Applied Microbiology. 38. 57-61 (2002)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Ito A, Sato Y, Kudo S, Sato S, Nakajima H, Toba T: "The screening of hydrogen peroxide-producing lactic acid bacteria and their application to inactivating psychrotrophic food-borne pathogens."Current Microbiology. 47. 231-236 (2003)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Mukai T, Toba T, Ohori H: "Molecular characterization of adhesion of lactic acid bacteria to mucosal surface of intestine."Milk Science. 52. 167-174 (2003)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Horie M, Kajikawa HS, Toba T: "Identification of Lactobacillus crispatus by polymerase chain reaction targeting S-layer protein gene"Letters in Applied Microbiology. 35. 57-61 (2002)
• [Publications] Ito A, Sato Y, Kudo S, Sato S, Nakajima H, Toba T: "The screening of hydrogen peroxide-producing lactic acid bacteria and their application to inactivating psychrotrophic food-borne pathogens"Current Microbiology. 47. 231-236 (2003)
• [Publications] 向井孝夫, 戸羽隆宏, 大堀均: "乳酸菌における細胞接着の分子機構"ミルクサイエンス. 52. 167-174 (2003)
• [Publications] Horie M, Kajikawa HS, Toba T: "Identification of Lactobacillus crispatus by polymerase chain reaction targeting S-layer protein gene"Letters in Applied Microbiology. 35. 57-61 (2002)
• [Publications] Ito A, Sato Y, Kudo S, Sato S, Nakajima H, Toba T: "The screening of hydrogen peroxide-producing lactic acid bacteria and their application to inactivating psychrotrophic food-borne pathogens"Current Microbiology. 46. (2003)