Lactic acid bacteria (LAB) were isolated from Kung-Som at various fermentation periods. Only ten strains, named D2SM22, D6SM3, D6SM24, D6SM26, D8SM21, D10SM5, D10SM11, D10SM16, D10SM20 and D16SM26 showed a survival rate of more than 50% under the simulated gastric juice. After being subjected to simulated gastric juice, four strains (D6SM3, D8SM21, D10SM16 and D10SM20) showed a survival rate of more than 50% in simulated small intestinal juices. Growth of strain D6SM3, D8SM21 and D10SM16 under micro-aerobic and anaerobic conditions was not different. Tested pathogenic strains (Escherichia coli, Staphylococcus aureus, Bacillus cereus, Vibrio parahaemolyticus and Salmonella sp.) were inhibited by probiotic LAB. However, none of strains could produce bacteriocins. All strains were identified as Lactobacillus plantarum. No differences in pH, acidity, LAB count and liking scores between Kung-Som produced with starter culture and conventional method were observed (P > 0.01).

In Lactococcus lactis and some other lactic acid bacteria, respiratory metabolism has been reported upon supplementation with only heme, leading to enhanced biomass formation, reduced acidification, resistance to oxygen, and improved long-term storage. Genes encoding a complete respiratory chain with all components were found in genomes of L. lactis and Leuconostoc mesenteroides, but menaquinone biosynthesis was found to be incomplete in Lactobacillaceae (except L. mesenteroides). Lactiplantibacillus plantarum has only two genes (menA, menG) encoding enzymes in the biosynthetic pathway (out of eight), and Lentilactobacillus buchneri has only four (menA, menB, menE, and menG). We constructed knock-out strains of L. lactis defective in menA, menB, menE, and menG (encoding the last steps in the pathway) and complemented these by expression of the extant genes from Lactipl. plantarum and Lent. buchneri to verify their functionality. Three of the Lactipl. plantarum biosynthesis genes, lpmenA1, lpmenG1, and lpmenG2, as well as lbmenB and lbmenG from Lent. buchneri, reconstituted menaquinone production and respiratory growth in the deficient L. lactis strains when supplemented with heme. We then reconstituted the incomplete menaquinone biosynthesis pathway in Lactipl. plantarum by expressing six genes from L. lactis homologous to the missing genes in a synthetic operon with two inducible promoters. Higher biomass formation was observed in Lactipl. plantarum carrying this operon, with an OD600 increase from 3.0 to 5.0 upon induction.

No abstract