Shengyu Li

Antioxidant activity of Lactobacillus plantarum strains isolated from traditional Chinese fermented foods

Eleven Lactobacillus plantarum strains isolated from traditional Chinese fermented foods were investigated for their in vitro scavenging activity against hydroxyl and 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radicals, and their resistance to hydrogen peroxide. L. plantarum C88 at a dose of 10(10) CFU/ml showed the highest hydroxyl radical and DPPH scavenging activities, with inhibition rates of 44.31% and 53.05%, respectively. Resistance of intact cells to hydrogen peroxide was also found in all strains. L. plantarum C88 was the most resistant strain against hydrogen peroxide. When L. plantarum C88 was administered to senescent mice suffering oxidative stress induced by d-galactose, the serum superoxide dismutase activity, the glutathione peroxidase activity and the total antioxidant capacity in liver increased significantly, while the level of malondialdehyde in liver decreased significantly. L. plantarum C88 isolated from traditional Chinese fermented dairy tofu could be considered as a potential antioxidant to be applied in functional foods.

Capsular and slime-polysaccharide production by Lactobacillus rhamnosus JAAS8 isolated from Chinese sauerkraut: Potential application in fermented milk products

Exopolysaccharides (EPSs) produced by lactic acid bacteria (LAB) play an important role in the improvement of the physical properties of fermented dairy products. To find EPS-producing LAB strains with potential industrial applications, a Lactobacillus rhamnosus strain, L. rhamnosus JAAS8, that is capable of producing two forms of EPS when grown in MRS broth or semi-defined medium with glucose as a carbon source was isolated and identified from Chinese sauerkraut. The capsular-polysaccharide (CPS) present surrounding the bacterial surface of L. rhamnosus JAAS8 was observed by both optical microscopy and transmission electron microscopy. The slime-polysaccharide (SPS) present in the growth medium was produced mainly during the exponential growth phase, while the CPS was produced only in fermentation. Monosaccharide analysis of the purified polysaccharide samples showed that the CPS was composed of galactose and N-acetylglucosamine in a molar ratio of 5:1, and the SPS was composed of galactose, glucose and N-acetylglucosamine in a molar ratio of 4:1:1. The use of L. rhamnosus JAAS8 could be considered for potential applications in the dairy industry to improve the rheological properties of fermented milk products by increasing their water-holding capacity and viscosity.

Reduction of Aflatoxin B1 Toxicity by Lactobacillus plantarum C88: A Potential Probiotic Strain Isolated from Chinese Traditional Fermented Food “Tofu”

In this study, we investigated the potential of Lactobacillus plantarum isolated from Chinese traditional fermented foods to reduce the toxicity of aflatoxin B1 (AFB1), and its subsequent detoxification mechanism. Among all the investigated L. plantarum strains, L. plantarum C88 showed the strongest AFB1 binding capacity in vitro, and was orally administered to mice with liver oxidative damage induced by AFB1. In the therapy groups, the mice that received L. plantarum C88, especially heat-killed L. plantarum C88, after a single dose of AFB1 exposure, showed an increase in unabsorbed AFB1 in the feces. Moreover, the effects of L. plantarum C88 on the enzymes and non-enzymes antioxidant abilities in serum and liver, histological alterations of liver were assayed. The results indicated that compared to the control group, L. plantarum C88 alone administration induced significant increase of antioxidant capacity, but did not induce any significant changes in the histological picture. Compared to the mice that received AFB1 only, L. plantarum C88 treatment could weaken oxidative stress by enhancing the activity of antioxidant enzymes and elevating the expression of Glutathione S-transferase (GST) A3 through Nuclear factor erythroid (derived factor 2) related factor 2 (Nrf2) pathway. Furthermore, cytochrome P450 (CYP 450) 1A2 and CYP 3A4 expression was inhibited by L. plantarum C88, and urinary aflatoxin B1-N7-guanine (AFB-N7-guanine), a AFB1 metabolite formed by CYP 1A2 and CYP 3A4, was significantly reduced by the presence of viable L. plantarum C88. Meanwhile, the significant improvements were showed in histological pictures of the liver tissues in mice orally administered with viable L. plantarum C88. Collectively, L. plantarum C88 may alleviate AFB1 toxicity by increasing fecal AFB1 excretion, reversing deficits in antioxidant defense systems and regulating the metabolism of AFB1.

Genome shuffling of Lactobacillus plantarum C88 improves adhesion

Genome shuffling is an important method for rapid improvement in microbial strains for desired phenotypes. In this study, ultraviolet irradiation and nitrosoguanidine were used as mutagens to enhance the adhesion of the wild-type Lactobacillus plantarum C88. Four strains with better property were screened after mutagenesis to develop a library of parent strains for three rounds of genome shuffling. Fusants F3-1, F3-2, F3-3, and F3-4 were screened as the improved strains. The in vivo and in vitro tests results indicated that the population after three rounds of genome shuffling exhibited improved adhesive property. Random Amplified Polymorphic DNA results showed significant differences between the parent strain and recombinant strains at DNA level. These results suggest that the adhesive property of L. plantarum C88 can be significantly improved by genome shuffling. Improvement in the adhesive property of bacterial cells by genome shuffling enhances the colonization of probiotic strains which further benefits to exist probiotic function. Graphical abstract RAPD analysis of L. plantarum C88 and fusants F3-1, F3-2, F3-3, and F3-4. Primers Z3, Z4, Z5, Z6, Z7, Z8, Z9, Coc, and OPA-3 were used in this test.

Involvement of Lysosome Membrane Permeabilization and Reactive Oxygen Species Production in the Necrosis Induced by Chlamydia muridarum Infection in L929 Cells.

Chlamydiae, obligate intracellular bacteria, are associated with a variety of human diseases. The chlamydial life cycle undergoes a biphasic development: replicative reticulate bodies (RBs) phase and infectious elementary bodies (EBs) phase. At the end of the chlamydial intracellular life cycle, EBs have to be released to the surrounded cells. Therefore, the interactions between Chlamydiae and cell death pathways could greatly influence the outcomes of Chlamydia infection. However, the underlying molecular mechanisms remain elusive. Here, we investigated host cell death after Chlamydia infection in vitro, in L929 cells, and showed that Chlamydia infection induces cell necrosis, as detected by the propidium iodide (PI)-Annexin V double-staining flow-cytometric assay and Lactate dehydrogenase (LDH) release assay. The production of reactive oxygen species (ROS), an important factor in induction of necrosis, was increased after Chlamydia infection, and inhibition of ROS with specific pharmacological inhibitors, diphenylene iodonium (DPI) or butylated hydroxyanisole (BHA), led to significant suppression of necrosis. Interestingly, live-cell imaging revealed that Chlamydia infection induced lysosome membrane permeabilization (LMP). When an inhibitor upstream of LMP, CA-074-Me, was added to cells, the production of ROS was reduced with concomitant inhibition of necrosis. Taken together, our results indicate that Chlamydia infection elicits the production of ROS, which is dependent on LMP at least partially, followed by induction of host-cell necrosis. To our best knowledge, this is the first live-cell-imaging observation of LMP post Chlamydia infection and report on the link of LMP to ROS to necrosis during Chlamydia infection.