Brazilian artisanal ripened cheeses as sources of proteolytic lactic acid bacteria capable of reducing cow milk allergy

J Appl Microbiol. 2018 Aug;125(2):564-574. doi: 10.1111/jam.13779. Epub 2018 May 4.

Abstract

Aim: The objective was to obtain lactic acid bacteria (LAB) capable of hydrolysing immunoreactive proteins in milk, to optimize the hydrolysis, to determine the proteolysis kinetics and to test the safety of the best hydrolytic strain.

Methods and results: Brazilian cheese was used as source of LAB capable of hydrolysing main milk allergens. Proteolytic isolates were submitted to RAPD-PCR for the characterization of clonal diversity. Optimized hydrolysis was strain and protein fraction dependent. 16S rDNA sequencing identified three proteolytic strains: Enterococcus faecalis VB43, that hydrolysed αS1 -, αS2 - and β-caseins, α-lactalbumin and β-lactoglobulin (partial hydrolysis), and Pediococcus acidilactici VB90 and Weissella viridescens VB111, that caused partial hydrolysis of αS1 - and αS2 -caseins. Enterococcus faecalis VB43 tested negative for virulence genes asa1, agg, efaA, hyl, esp, cylLL and cylLS but positive for genes ace and gelE. Ethylenediamine tetra-acetic acid inhibited the proteolysis, indicating that the main proteases of E. faecalis VB43 are metalloproteases.

Conclusion: Brazilian artisanal cheese is a good source of LAB capable of hydrolysing allergenic proteins in milk. One isolate (E. faecalis VB43) presented outstanding activity against these proteins and lacked most of the tested virulence genes.

Significance and impact of the study: Enterococcus faecalis VB43 presents good potential for the manufacture of hypoallergenic dairy products.

Keywords: Enterococcus faecalis; cheese; immunoreactive proteins; lactic acid bacteria; milk allergy; proteolysis.

MeSH terms

  • Animals
  • Brazil
  • Cattle
  • Cheese / microbiology*
  • Lactobacillales* / isolation & purification
  • Lactobacillales* / metabolism
  • Milk / microbiology
  • Milk Hypersensitivity*
  • Milk Proteins / chemistry
  • Milk Proteins / metabolism

Substances

  • Milk Proteins

Associated data

  • GENBANK/UR1268
  • GENBANK/KM497509
  • GENBANK/KJ779095.1
  • GENBANK/KJ580423.1