The goal of this study was screening and molecular identification of Lactic Acid Bacteria (LAB) producing β-glucan from different species isolated from boza and cider compared to a standard strain for Lactobacillus rhamnosus NRRL 1937 (LGG). From 48 unknown isolates, four LAB strains were selected. Based on the NCBI database, their nomenclature was A3, B6, and C9 for Limosilactobacillus fermentum SH1, SH2, and SH3 along with D6 for Leuconostoc mesenteroides SH4. Also, their similarity values were 100%, 99.8%, 100%, and 100%, respectively. The potential of Exopolysaccharide (EPS) (as β-glucan) production for selected LAB strains by gtf gene, conventional PCR and gene expression using both LGG as a control and LAB 16S rRNA gene as a house-keeping gene was investigated. In addition, EPS (mg/100 mL), cell mass (mg/100 mL), pH, total carbohydrate%, total protein% and β-glucan% by the HPLC for all selected LAB isolates were studied. All results of genetic and chemical tests proved the superiority of B6 treatment for L. fermentum SH2. The results showed the superiority of B6 treatment in gtf gene expression (14.7230 ± 0.070-fold) followed by C9 and A3 treatments, which were 10.1730 ± 0.231-fold and 8.6139 ± 0.320-fold, respectively. while D6 treatment recorded the lowest value of gene expression (0.8566 ± 0.040-fold) compared to the control LGG (one-fold). The results also demonstrated that B6 treatment was superior to the other treatments in terms of EPS formation, with a value of 481 ± 1.00 mg/100 mL, followed by the C9 treatment at 440 ± 2.00 mg/100 mL, compared to the LGG (control) reaching 199.7 ± 3.51 mg/100 mL. Also, the highest % of quantitative and qualitative β-glucan in EPS was observed in B6 followed by C9, D6 and A3 which were 5.56 ± 0.01%, 4.46 ± 0.01%, 0.25 ± 0.008% and 0.12 ± 0.008%, respectively compared to control (0.31 ± 0.01%). Finally, the presented results indicate the importance of screening the local LAB isolates to obtain a superior strain for β-glucan production which will be introduced in a subsequent study under optimum conditions.

Background: This study aims to identify the types of lactic acid bacteria (LAB) in raw cow milk obtained from different locations in Babylon, Iraq using genetic analysis and study the phylogenetic relationship among them. Methods: The study was based on 74 isolates collected from different milk samples. LAB was isolated on selective media from MRS agar after conducting a series of decimal dilutions. LAB was diagnosed depending on the morphological characteristics and nitrogenous base sequences for the 16S rRNA gene, which was 1500 bp. the whole genome was extracted from all isolates. Result: This study identified 74 isolated bacteria as gram-positive lactic acid bacteria (LAB), with most being cocci and the rest bacilli. Six LAB species were identified through genetic analysis (16S rRNA sequencing): Lactobacillus rhamnosus, L. kefiri, L. curvatus, Lactococcus lactis, Pediococcus pentosaceus and Pediococcus acidilactici. Phylogenetic analysis revealed relationships between these species and the study highlighted how environmental factors can lead to genetic variations within LAB. The research emphasizes the importance of molecular methods for identifying and understanding the genetic diversity of LAB in food and beverage samples.