Detection of riboflavin-producing probiotic strains isolated from dairy products by ۱۶SrRNA sequencing

Background and Aim : The probiotic bacteria has the main effect in boosting the immunity, reducing the duration of certain diarrheal diseases, treatment of gastro intestinal disorders. Probiotic bacteria are able to synthesize B-group vitamins particularly riboflavin to obtain fermented bio-enriched food. The aim of our study was detection of riboflavin-producing probiotic strains isolated from dairy products by ۱۶ S rRNA sequencing.Methods : A riboflavin producing lactic acid bacterium was isolated from raw milk and traditional dairy products in Tehran. Samples were cultured on MRS.broth) for isolation of lactic acid bacteria, MRI broth (for isolation of Bifidobacterium), MRI broth (for isolation of Lactobacillus acidophilus), a broth neutrino (for yeast isolation). Diagnostic tests to identify lactic acid bacteria and the probiotic properties of this strain were examined. After DNA extraction accordance with the kit protocol, PCR amplification, and DNA sequencing of the ۱۶S rRNA gene were done by Lactobacillus ۱۶s rRNA Sequence Specific Primers included: ۵-CTCAAAACTAAACAAAGTTTC-۳ and ۵-CTTGTACACACCGCCCGTCA -۳. The sequence of the PCR product was compared with known ۱۶S rRNA gene sequences in GenBank by multiple sequence alignment with the CLUSTAL W program. The riboflavin quantification was performed by High Performance Liquid Chromatography (HPLC) analysis.Results : In the molecular study, the results of blast strains at EZ Taxon identified five isolates: L. salivarius, L. johnsonii, L. paracasei, L. xiangfangensis, L. helveticus. Also, the HPLC result of an over-the-counter solution for isolating cultures showed that one of the isolates, L. paracasei, was able to produce vitamin B۲.Conclusion : The results of the present study showed that vitamin B۲-producing isolates are present in traditional milk and dairy samples, and by purifying and optimizing the amount of vitamin production by these isolates, they can be supplemented as He used a variety of foods.

کلیدواژه ها:

riboflavin-producing ، ۱۶SrRNA sequencing ، probiotic ، dairy products