Mucin Degradation Test of Probiotics

Mucin degradation is an important aspect to assessing the safety of probiotics. Creative Enzymes offers a wide range of tests to help clients perform a comprehensive analysis of the mucin-degrading activity of probiotics.

Mucin Degradation Activity of Probiotics

Mucins are highly glycosylated proteins released by the intestinal goblet cells that provide structure and viscosity to the mucus layer, covering and protecting the intestinal epithelial surface from adverse external conditions and infection by pathogenic microorganisms. Therefore, any change in mucus content and structure can impair the mucosal defense barrier function of the mucus layer. This may lead to increased displacement of normal intestinal bacteria. Creative Enzymes uses our science to test for possible harmful effects of probiotics on intestinal tissues.

Fig. 1 In vitro mucin degradation assay of B. thetaiotaomicron. (Nakjang, 2017)

Mucin Degradation Testing Services at Creative Enzymes

Creative Enzymes is a well-known probiotic safety testing services provider. We are committed to applying our advanced technology and experience in probiotic safety testing to provide clients with comprehensive evaluation solutions to ensure the safety of products. In Creative Enzymes' assay lab, we determine the mucoadhesive protein degrading activity of probiotics by a combination of in vitro culture of strains, transcriptomic and untargeted metabolomics approaches.

Testing Methods of Creative Enzymes for Mucin Degradation Activity

• In Vitro Culture Method
  In vitro culture methods have been applied to research the response of probiotics to different levels of mucins. We tested the mucin degradation activity of probiotics in three experiments. In all the mucin degradation tests, the relevant tests were performed using porcine gastric mucin (HGM).
  • Growth in liquid medium

  Probiotic cultures were incubated in liquid media containing different carbon sources (no carbon source, mucin as the only carbon source, glucose as the only carbon source, mucus and glucose as the carbon source) and their absorbance was measured.

  • SDS-PAGE analysis of degraded mucin residues

  To detect whether the composition of mucin changed after incubation in different liquid media. We analyzed the electrophoretic profiles of mucin samples by SDS-PAGE. Any new band with a smaller molecular weight compared to the control band was defined as positive for mucin degradation.

  • Degradation test in Petri dish

  Probiotic cultures were inoculated on agar plates containing mucin. Staining was performed after incubation under suitable conditions, and the mucin degradation ability of the strains was analyzed by observing whether mucolytic zones (discolored halos) appeared around the colonies.

• Transcriptome Analysis
  Probiotics may undergo morphological and molecular changes when culture conditions are altered. Therefore, we analyzed the mucin degradation ability of probiotics by performing whole genome sequencing to establish a set of reference sequences for transcriptome analysis.
• Metabolomics Analysis
  To explore the metabolomic profile of probiotics when different amounts of mucins were added to the medium, we performed non-targeted metabolic analysis of cultures under different culture conditions.

Creative Enzymes is a professional and experienced probiotic supplier and service provider. Our professional scientific team works closely with you to provide a full range of assays for the research of probiotic activity on mucin degradation. To get more information, please contact us and we will be happy to serve your research demands.

Reference

1. Nakjang, S.; et al. A novel extracellular metallopeptidase domain shared by animal host-associated mutualistic and pathogenic microbes. PloS one. 2012, 7(1): e30287.