Anti-Inflammatory and Immunostimulatory Effects of Extract from Culture of Effective Microorganisms (ECEM) Revealed by Functional Genomics and Metabolome Analyses

The extract from culture of effective microorganisms (ECEM) is a secondary metabolite produced in symbiotic culture of a mixture of yeasts and lactic bacteria followed by photosynthetic bacteria. Metabolomics studies have demonstrated that ECEM contained several active ingredients associated with anti-inflammation through interactions with nuclear factor-κB. Functional genomics using DNA microarrays revealed that ECEM had distinctive anti-inflammatory and immunostimulatory actions in RAW264.7 cells. ECEM inhibited the LPS-upregulated expression of one group of pro-inflammatory genes (iNos, Il6 and Cox2) but not another group genes (Tnf, Ccl7 and Dusp2);it also upregulated the expression of Cd1d1, which participates in antigen presentation on lipids, resulting in activation of natural killer T (NKT) cells related to infection defense and cancer. Bacterial glycosylated lipid and other related complex lipids in ECEM are thought to trigger this upregulation of Cd1d1 expression. These results suggest that ECEM functions in the immune response in both inflammation and antigen presentation and that it contains components important for these functions. These novel effects of ECEM could be invaluable for developing functional drinks or health beverages.

Harnessing a T1 Phage-Derived Spanin for Developing Phage-Based Antimicrobial Development

The global increase in the prevalence of drug-resistant bacteria has necessitated the development of alternative treatments that do not rely on conventional antimicrobial agents.Using bacteriophage-derived lytic enzymes in antibacterial therapy shows promise;however,a thorough comparison and evaluation of their bactericidal efficacy are lacking.This study aimed to compare and investigate the bactericidal activity and spectrum of such lytic enzymes,with the goal of harnessing them for antibacterial therapy.First,we examined the bactericidal activity of spanins,endolysins,and holins derived from 2 Escherichia coli model phages,T1 and T7.Among these,T1-spanin exhibited the highest bactericidal activity against E.coli.Subsequently,we expressed T1-spanin within bacterial cells and assessed its bactericidal activity.T1-spanin showed potent bactericidal activity against all clinical isolates tested,including bacterial strains of 111 E.coli,2 Acinetobacter spp.,3 Klebsiella spp.,and 3 Pseudomonas aeruginosa.In contrast,T1 phage-derived endolysin showed bactericidal activity against E.coli and P.aeruginosa,yet its efficacy against other bacteria was inferior to that of T1-spanin.Finally,we developed a phage-based technology to introduce the T1-spanin gene into target bacteria.The synthesized non-proliferative phage exhibited strong antibacterial activity against the targeted bacteria.The potent bactericidal activity exhibited by spanins,combined with the novel phage synthetic technology,holds promise for the development of innovative antimicrobial agents.