Jae Seon Kang

Isolation and characterization of surfactin produced by Bacillus polyfermenticus KJS-2

Bacillus polyfermenticus KJS-2 (BP-KJS-2) was used to produce a lipopeptide-type surfactin. To accomplish this, a surfactin-producing BP-KJS-2 was fermented by soybeans. The surfactin was then purified by a procedure including ethanol treatment and preparative chromatography. Next, the biochemical structure of the purified surfactin was analyzed by electrospray ionization mass spectrometry (ESI-MS) and high-resolution ESI Q-Tof mass spectrometry (Q-Tof MS). In addition, the masses of the four peaks were determined to be 1007, 1021, 1035, and 1049 m/z revealing that the compound was mixture with quasi-molecular ions. Taken together, these findings indicated that the lipopeptide had a cyclic structure and amino acid composition of Gln-Leu-Leu-Leu-Val-Asp-Leu-Leu, and that the major lipopeptide product of BP-KJS-2 is the surfactin isoform. In addition, this lipopeptide showed strong antimicrobial activity against bacteria at the level of 0.05 mg/mL.

Antibacterial activities of macrolactin a and 7-O-succinyl macrolactin a from Bacillus polyfermenticus KJS-2 against vancomycin-resistant enterococci and methicillin-resistant Staphylococcus aureus

The principal objective of this study was to evaluate the antibacterial activities of macrolactin A (MA) and 7-O-succinyl macrolactin A (SMA) generated from Bacillus polyfermenticus KJS-2 against vancomycin-resistant enterococci (VREs) and methicillin-resistant Staphylococcus aureus. The minimal inhibitory concentrations (MICs) of MA and SMA against VREs were 16 and 2∼1 μg/mL, respectively, and the MICs of MA and SMA against methicillin-resistant Staphylococcus aureus were 2 and < 0.25 μg/mL, respectively. Their MIC values were comparable or superior to those of teicoplanin. In evaluating the inhibitory effects of intestinal VRE colonization in mice, the oral MA and SMA effected a rapid inhibition of intestinal VRE colonization in mice, and the intraperitoneal SMA also inhibited VRE colonization, whereas intraperitoneal MA did not.

Preparation and characterization of salmon calcitonin-sodium triphosphate ionic complex for oral delivery.

Even though salmon calcitonin (sCT) has been known as a potent hypocalcemic agent, only injection or nasal spray products are available on the market. In order to develop oral delivery system of the agent, a novel sCT-sodium tripolyphosphate (STPP) ionic complex was fabricated and also characterized. For the optimization of the ionic complexation, the effect of incubation time and molar ratio between sCT and STPP was evaluated. Particle size of the ionic complex in aqueous media, SEM images, DSC, FT-IR, in vitro release test, stability within the simulated intestinal fluid, and hypocalcemic effect were evaluated. The optimal molar complexation ratio of sCT to STPP was ranged from 1:5 to 1:10 and the complexation efficiency was about 95%. The SEM image has shown that the freeze dried ionic complex has rough morphology in their surface and the particle size in PBS (pH 7.4) was about 220nm. The DSC and FT-IR results provided evidences for ionic interaction between -NH(2) groups and -P horizontal lineO groups of sCT and STPP, respectively. The sCT ionic complex has shown sustained sCT releasing characteristics for 3weeks. The sCT-STPP ionic complex was protective to enzymatic attack and in vivo animal data revealed that the present ionic complex would show continuous hypocalcemic effect. Conclusively, the present sCT-STPP ionic complex formulation thought to be a novel oral delivery candidate for the treatment of osteoporosis.

Characterization of Bacillus mojavensis KJS-3 for industrial applications

One of the strains of the Bacillus mojavensis group, Bacillus mojavensis KJS-3 (B. mojavensis KJS-3), which has been demonstrated to play a role in protecting plants against diseases as a bacterial endophyte and in reducing the accumulation of mycotoxins generated by an endophytic fungus, was recently discovered in food waste. In this study, the identification and characterization of B. mojavensis KJS-3 was performed via TEM analysis, API-zym test, API 50 CHB test, assays of catalase and oxidase activity, lactic acid production, stability under various conditions, antibiotic susceptibility, and cellular fatty acid composition. The overall results of this study demonstrate that B. mojavensis KJS-3 may have great potential as a probiotic product, as this bacterium is quite stable in somewhat harsh environments. B. mojavensis KJS-3 was positive on oxidase and catalase tests, and the conversion rate of glucose to lactic acid was 58.9%. Finally, anteiso-C15:0 (43.10%) was identified as the major fatty acid.

Decursin and decursinol angelate affect spermatogenesis in the adult rat at oral administration

The aim of this study was to investigate the positive effects of decursin and decursinol angelate (D/DA) on the reproductive system of male rats as well as to examine whether D/DA is able to ameliorate the effects of bisphenol A (BPA). Adult rats were divided into the following four groups: (1) control, (2) BPA, (3) D/DA, and (4) BPA+D/DA. Organ weight ratio of vital organs decreased in the kidney (9.5%) and increased in the prostate (24.07%) and epididymis (22.14%) in the BPA+D/DA group compared to the values in the BPA group. A significant decrease (P<0.05) in sperm count was found in the BPA+D/DA (18.91%) and BPA(54.05%) groups compared to the count in the control group. In only D/DA (20.14%) group, sperm count was significantly increased. D/DA ameliorated the histopathological changes induced by BPA in the testis, consistent with these data. D/DA was also identified as promoting the activities of antioxidant enzymes such as catalase (CAT) and glutathione peroxidase (GPx), the levels of which were significantly increased compared to the corresponding levels in the BPA group. Fas/FasL (FAS) and caspase-3 (CAP) levels were significantly decreased in rats. The results suggest that D/DA reduces apoptosis of Leydig and germ cells in the mouse testis through the Fas-signaling pathway.

Testicular antioxidant mechanism of cultivated wild ginseng extracts

Recent studies have reported the relationship between reduced sperm counts and male infertility. The effects of cultivated wild ginseng extracts, which include ginsenosides, on cellular antioxidant activities were studied in testicular cells and animal models. In a study of rats, the weight of testis-right (15.38%, P<0.05) and testis-left (16.98%, P<0.05) in group D (cultivated wild ginseng extracts with bisphenol A) increased in comparison with group A (only bisphenol A). Reactive oxygen species of TM3 Leydig and TM4 Sertoli cells treated with bisphenol A significantly increased, by 36-41%, compared to the controls. Cultivated wild ginseng extracts at 10 and 25 μg/mL significantly increased the expression of catalase in TM3 cells, and catalase, superoxide dismutase 1, and glutathione peroxidase enzymes in TM4 cells. The induction of expression of catalase and superoxide dismutase 1 by cultivated wild ginseng extracts in rats occurs via the ERK and p38 pathways. Cultivated wild ginseng extracts also ameliorated the histopathological changes induced by bisphenol A in the testis.

Simultaneous Analysis Method for Polar and Non-polar Ginsenosides in Cultivated Wild Ginseng by Reversed-phase HPLC-CAD

Cultivated wild ginseng is a widely used dietary supplement and medicinal herb. The aim of this study was to optimize the ginseng using high performance liquid chromatography (HPLC)- charged aerosol detection (CAD) for ginsenoside analysis. CAD measures the physical property of an analyte and responds to almost all non-volatile species, independent of their nature, spectral properties, or particle size. It has become widely employed in pharmaceutical analysis. The cultivated wild ginseng extracts were analyzed for compositions of ginsenosides Rb1, Rd, Rg1, Rf, Re, and Rh1. The optimal analysis condition was set up from an experiment using a gradient. Ten grams of cultivated wild ginseng were extracted with 95% EtOH 100 ml for 24 hr at 80℃. The contents of the 6six major ginsenosides in the cultivated wild ginseng extract were Rb1 (5.48±0.12 mg/g), Rd (5.33±0.14 mg/g), Rg1 (12.80± 0.05 mg/g), Rf (19.08±0.68 mg/g), Re (19.87±0.05 mg/g), and Rh1 (16.47±0.16 mg/g), respectively. HPLC showed that the protopanaxatriol group (Rg1, Rf, Re, Rh1) had more content than the protopanaxadiol group (Rb1, Rd) in cultivated wild ginseng extract. In summary, the ginsenosides were identified with HPLC-CAD analysis, and their presence and quantity imply the importance of quality control, as well as the pharmacological activity of the ginseng root.

Evaluation of genotoxicity of Bacillus mojavensis KJS-3 on culture supernatant for use as a probiotic

The genotoxicity of crude substances cultured in Bacillus mojavensis KJS-3 (B. mojavensis KJS-3) was investigated by using Ames tester strains of Salmonella typhimurium (S. typhimurium) with or without metabolic activation (S9 mix). No increase in the number of revertants was observed in response to any of the doses of the culture supernatant fermented by B. mojavensis KJS-3 (5, 10, 50, 100, and 500 μg/plate) in S. typhimurium TA 98, 100, 1535 and 1537 both in the absence and in the presence of S-9 metabolic activation system. The half-maximal inhibitory concentration (IC50) value of the culture supernatant was >83.33 μg/mL. Crude substances had no effect on the DNA repair system for mutagenesis. Furthermore, the cell number of S. typhimurium strains was observed to have decreased compared to the solvent control. These results showed dose-dependent activity on the mutagenicity of crude substances in Ames test. These results strongly indicate that antimutagenic compounds are produced during fermentation by B. mojavensis KJS-3. On using the Chinese hamster lung cell line in mammalian cell system, no clastogenicity of the culture supernatant fermented by B. mojavensis KJS-3 was observed in the absence or presence of the S9 mix at the concentrations of 1.66, 8.33, 16.67, and 83.33 μg/mL. No significant increase in chromosome aberrations was observed in response to treatment with any of these concentrations, regardless of activation of the metabolic system. Consequently, the mutagenic potential of the culture supernatant was not induced with respect to in vitro bacterial reverse mutation and clastogenicity in the range of concentrations evaluated in these experiments in our study.

Cultivated wild ginseng extracts upregulate the anti-apoptosis systems in cells and mice induced by bisphenol A

Cultivated wild ginseng has a variety of pharmacological effects. The aim of this study was to investigate the effects of cultivated wild ginseng extracts (CWGE) on apoptosis. CWGE showed the ability to protect hormone production (testosterone and progesterone from in vitro results) in TM3 Leydig and TM4 Sertoli cells damaged by Bisphenol A (BPA). CWGE also showed the ability to protect production of testosterone (7.24%, P<0.05), plasma luteinizing hormone (LH) (32.67%, P<0.05), and follicle-stimulating hormone (FSH) (37.34%, P<0.05) compared to the control and BPA-only groups. Reduction of apoptotic protein expression and induction of anti-apoptotic protein expression was observed in cells and rats pretreated with CWGE. These proteins are expressed through the ERK and p38 signalling pathways. In addition, CWGE might be a useful herbal medicine of cellular defense agents as our results in cells and animals.

Recovery from the Two-generation Reproductive Toxicity in Sprague-Dawley Rats by Treatment with Decursin and Decursinol Angelate

The aim of this study was to determine the effect of decursin (D) and decursinol angelate (DA) against bisphenol A (BPA) toxicity in a rat two-generation study. Adult rats were divided into the following three groups: (1) control, (2) BPA, and (3) BPA+D/DA. The D and DA treatment of F0 parents increased the terminal body weight and relative adult organ weights (testes, kidneys, spleen, and liver) when compared with the BPA group. A significant decrease in sperm count was found in the BPA+D/DA (7.69%) and BPA (64.70%, p<0.01) groups, when compared with the sperm count in the control group. No offspring were obtained in the F1 generation of the BPA (50 mg/kg/day) group, but the addition of D/DA in the BPA+D/DA group significantly restored fertility (55.78%) and gestation indices (98.87%) in the F1 generation. No significant differences were found in the fertility index between the control (75.02%) and the BPA+D/DA (78.11%) groups in the two-generation study, when compared with the one-generation study. The viability ratio during lactation in the D/DA group was also similar to that of the control group. These data indicate that D/DA (50 mg/kg/day) administered over two generations causes significant positive changes in reproductive or developmental parameters.