Proteome analysis of Lactobacillus helveticus H9 during growth in skim milk

J Dairy Sci. 2014 Dec;97(12):7413-25. doi: 10.3168/jds.2014-8520. Epub 2014 Oct 11.

Abstract

Lactobacillus helveticus H9 was isolated from traditionally fermented yak milk in Tibet (China) with the ability to produce the antihypertensive peptides Val-Pro-Pro (VPP) and Ile-Pro-Pro (IPP) during milk fermentation. To understand the changes in the protein expression of L. helveticus H9, proteome analysis was performed at 3 different growth stages, lag phase (pH 6.1), log phase (pH 5.1), and stationary phase (pH 4.5) using 2-dimensional electrophoresis (2-DE). Further analysis showed that 257 differential protein spots were found and 214 protein spots were identified using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF/MS). The cellular functions of the differentially expressed proteins were complex. Interestingly, the proteolytic system-related proteins aminopeptidase N (PepN), aminopeptidase E (PepE), endopeptidase O2 (PepO2), and oligopeptide transport system permease protein (OppC) were observed only on the maps of pH 5.1 and pH 4.5, which was consistent with the presence of angiotensin I-converting enzyme (ACE)-inhibitory peptides VPP and IPP during these 2 growth stages (log phase and stationary phase). These results, combined with a previous study of gene expression of the proteolytic system, led us to conclude that the Opp transport system, pepE, and pepO2 are likely related to the production of ACE-inhibitory peptides.

Keywords: Lactobacillus helveticus H9; proteolytic system; proteomics.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bacterial Proteins / metabolism
  • Cattle
  • Electrophoresis, Gel, Two-Dimensional
  • Endopeptidases / metabolism
  • Female
  • Fermentation
  • Hypertension / therapy
  • Lactobacillus helveticus / chemistry
  • Lactobacillus helveticus / growth & development
  • Lactobacillus helveticus / metabolism*
  • Membrane Transport Proteins / metabolism
  • Milk / microbiology*
  • Oligopeptides / analysis
  • Oligopeptides / metabolism
  • Peptides / analysis
  • Peptidyl-Dipeptidase A / analysis
  • Peptidyl-Dipeptidase A / metabolism
  • Proteolysis
  • Proteome*
  • Proteomics*
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
  • Tibet

Substances

  • Bacterial Proteins
  • Membrane Transport Proteins
  • Oligopeptides
  • Peptides
  • Proteome
  • isoleucyl-prolyl-proline
  • valyl-prolyl-proline
  • oligopeptide permease, Bacteria
  • Endopeptidases
  • Peptidyl-Dipeptidase A