RESUMO

Microbial exopolysaccharides (EPSs) have attracted extensive attention for their biological functions in antioxidant activities. In this study, we characterized a novel EPS produced by Bifidobacterium pseudocatenulatum Bi-OTA128 which exhibited the highest antioxidant capacity compared to nine other ropy bacterial strains, achieving 76.50 % and 93.84 % in DPPH· and ABTS·+ scavenging activity, and ferric reducing power of 134.34 µM Fe2+. Complete genomic analysis identified an eps gene cluster involved in the EPS biosynthesis of Bi-OTA128 strain, which might be responsible for its ropy phenotype. The EPS was then isolated and purified by a DEAE-Sepharose Fast Flow column. A single elution part EPS128 was obtained with a recovery rate of 43.5 ± 1.78 % and a total carbohydrate content of 93.6 ± 0.76 %. Structural characterization showed that EPS128 comprised glucose, galactose, and rhamnose (molar ratio 4.0:1.2:1.1), featuring a putative complex backbone structure with four branched chains and an unusual acetyl group at O-2 of terminal rhamnose. Antioxidant assay in vitro indicated that EPS128 exhibited antioxidant potential with 50.52 % DPPH· and 65.40 % ABTS·+ scavenging activities, reaching 54.3 % and 70.44 % of the efficacy of standard Vitamin C at 2.0 mg/L. Furthermore, EPS128 showed protective effects against H2O2-induced oxidative stress in HepG2 cells by reducing cellular reactive oxygen species (ROS) and increasing cell viability. These findings present the first comprehensive report of an antioxidant EPS from B. pseudocatenulatum, highlighting its potential as a natural antioxidant for applications in the food industry and clinical settings.

RESUMO

Glucagon-like peptide-1(GLP-1) is an incretin hormone secreted primarily from the intestinal L-cells in response to meals. GLP-1 is a key regulator of energy metabolism and food intake. It has been proven that P9 protein from A. muciniphila could increase GLP-1 release and improve glucose homeostasis in HFD-induced mice. To obtain an engineered Lactococcus lactis which produced P9 protein, mature polypeptide chain of P9 was codon-optimized, fused with N-terminal signal peptide Usp45, and expressed in L. lactis NZ9000. Heterologous secretion of P9 by recombinant L. lactis NZP9 were successfully detected by SDS-PAGE and western blotting. Notably, the supernatant of L. lactis NZP9 stimulated GLP-1 production of NCI-H716 cells. The relative expression level of GLP-1 biosynthesis gene GCG and PCSK1 were upregulated by 1.63 and 1.53 folds, respectively. To our knowledge, this is the first report on the secretory expression of carboxyl-terminal processing protease P9 from A. muciniphila in L. lactis. Our results suggest that genetically engineered L. lactis which expressed P9 may have therapeutic potential for the treatment of diabetes, obesity and other metabolic disorders.

Assuntos

Akkermansia , Peptídeo 1 Semelhante ao Glucagon , Lactococcus lactis , Peptídeo 1 Semelhante ao Glucagon/metabolismo , Peptídeo 1 Semelhante ao Glucagon/genética , Akkermansia/genética , Akkermansia/metabolismo , Lactococcus lactis/genética , Lactococcus lactis/metabolismo , Humanos , Células L , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Animais , Camundongos , Linhagem Celular , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo