Genetic and proteomic analysis of factors affecting serum cholesterol reduction by Lactobacillus acidophilus A4

Jin Lee, Younghoon Kim, Hyun Sun Yun, Jong Gun Kim, Sejong Oh, Sae Hun Kim

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

This article identifies novel factors involved in cholesterol reduction by probiotic bacteria, which were identified using genetic and proteomic approaches. Approximately 600 Lactobacillus acidophilus A4 mutants were created by random mutagenesis. The cholesterol-reducing ability of each mutant was determined and verified using two different methods: the o-phthalaldehyde assay and gas chromatographic analysis (GC). Among screened mutants, strain BA9 showed a dramatically diminished ability to reduce cholesterol, as demonstrated by a 7.7% reduction rate, while the parent strain had a more than 50% reduction rate. The transposon insertion site was mapped using inverse PCR (I-PCR), and it was determined using bioinformatic methods that the deleted region contained the Streptococcus thermophilus catabolite control protein A gene (ccpA). In addition, we have shown using two-dimensional gel electrophoresis (2-DE) that several proteins, including a transcription regulator, FMN-binding protein, major facilitator superfamily permease, glycogen Phosphorylase, the YknV protein, and fructose/tagatose bisphosphate aldolase, were strongly regulated by the ccpA gene. In addition, in vivo experiments investigating ccpA function were conducted with rats. Rats fed wild-type L. acidophilus A4 showed a greater than 20% reduction in total serum cholesterol, but rats fed BA9 mutant L. acidophilus showed only an approximately 10% reduction in cholesterol. These results provide important insights into the mechanism by which these lactic acid bacteria reduce cholesterol.

Original languageEnglish
Pages (from-to)4829-4835
Number of pages7
JournalApplied and Environmental Microbiology
Volume76
Issue number14
DOIs
Publication statusPublished - 2010 Jul 1

Fingerprint

Lactobacillus acidophilus
proteomics
Proteomics
Statistical Factor Analysis
serum
Cholesterol
cholesterol
protein
Staphylococcal Protein A
Serum
mutants
gene
metabolites
proteins
Genes
rats
genes
o-Phthalaldehyde
Streptococcus thermophilus
Bacteria

ASJC Scopus subject areas

  • Applied Microbiology and Biotechnology
  • Food Science
  • Biotechnology
  • Ecology

Cite this

Genetic and proteomic analysis of factors affecting serum cholesterol reduction by Lactobacillus acidophilus A4. / Lee, Jin; Kim, Younghoon; Sun Yun, Hyun; Kim, Jong Gun; Oh, Sejong; Kim, Sae Hun.

In: Applied and Environmental Microbiology, Vol. 76, No. 14, 01.07.2010, p. 4829-4835.

Research output: Contribution to journalArticle

Lee, Jin ; Kim, Younghoon ; Sun Yun, Hyun ; Kim, Jong Gun ; Oh, Sejong ; Kim, Sae Hun. / Genetic and proteomic analysis of factors affecting serum cholesterol reduction by Lactobacillus acidophilus A4. In: Applied and Environmental Microbiology. 2010 ; Vol. 76, No. 14. pp. 4829-4835.
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