A total of sixty-six samples of Nuruk, a fermention starter used to make the Korean traditional rice wine, Makgeolli, were collected from central and southern regions of Korea in 2013 and 2014. We classified two groups of the Nuruk samples, "commercial" and "home-made", according to the manufacturing procedure and purpose of use. Commercial Nuruks were made in a controlled environment where the temperature and humidity are fixed and the final product is supplied to Makgeolli manufacturers. Home-made Nuruks were made under uncontrolled conditions in the naturally opened environment and were intended for use in the production of small amounts of home-brewed Makgeolli. We obtained more than five hundred isolates including filamentous fungi and yeasts from the Nuruk samples followed by identification of fungal species. Also we stored glycerol stocks of each single isolate at $-70^{\circ}C$. We identified the species of each isolate based on the sequences of ITS regions amplified with two different universal primer pairs. We also performed morphological characterization of the filamentous fungi and yeast species through observations under the microscope. We investigated the major fungal species of commercial and home-made Nuruks by counting the colony forming units (CFU) and analyzing the occurrence tendency of fungal species. While commercial Nuruks contained mostly high CFU of yeasts, home-made Nuruks showed relatively high occurrence of filamentous fungi. One of the representative Nuruk manufacturers used both domestic wheat bran and imported ones, mainly from US, as raw material. Depending on the source of ingredient, the fungal diversity was somewhat different. Another commercial Nuruk sample was collected twice, once in 2013 and again in 2014, and showed different diversity of fungal species in each year. Nuruks obtained from the southern regions of Korea and Jeju island showed high frequency of yeast such as Saccharomycopsis fibuligera and Pichia species as well as unique filamentous fungus, Monascus species. S. fibuligera was easily found in many Nuruk samples with high CFU. The major filamentous fungi were Aspergillus, Lichtheimia, Mucor and Penicillium species. In order to further our understanding of the isolates and their potential industrial applications, we assayed three enzymes, alpha amylase, glucoamylase and acid protease from 140 isolates out of about five hundred isolates and selected about 10 excellent strains with high enzyme activities. With these fungal isolates, we will perform omics analyses including genomics, transcriptomics, metabolic pathway analyses, and metabolomics followed by whole genome sequencing of unique isolates associated with the basic research of Nuruk and that also has applications in the Makgeolli making process.
Reliable data on the exposure of capsaicin, which evokes hot sensation in hot red pepper, are important prerequisites for studying biological functions of capsaicin in human body since its roles are controversial according to animal and epidemiological studies. To get accurate data, the use of urinary biomarkers was considered as a measure of internal exposure of capsaicin. After 2-day-capsaicin depletion five to six women in their 20's were instructed to eat 60 or 80 g/day hot red pepper soybean paste as a capsaicin source with meal which did not contain capsaicinoid, and to collect their urine separately. HPLC conditions were set to detect capsaicinoid and urinary metabolites at the same time. Most of capsaicinoid were excreted in the form of 4-hydroxy-3-methoxybenzoic acid in three subjects at the highest dose. This result suggests the oxidation of the 4-hydroxy3-methoxybenzaldehyde, the hydrolysis product of capsaicin, is the major metabolic pathway in the human body, whereas the reduction of the aldehyde is the major route in rats. However, neither any metabolite nor the parent compound was ever shown in two of the subjects reflecting large individual differences of capsaicin absorption and/or biotransformation.
The cultural and physiological conditions for the T-2 toxin [4,15-diacetoxy-8-(3-mety1butyloxy)-12,13- epoxy-trichothec-9-en-3-01, $C_{24}H_{30}O_9$] production by Fusarium spp. were studied. Thin layer chromatography (TLC) assay and the microbiological assay uslng Rhodotomla rubra were used to quantitate tbe T- 2 toxin. Among the four strains of Fusarium spp., F tn'cinctum NRRL 3299 was best for T-2 toxin production. In solid culture, white com grit medium was best for T-2 toxm production. Temperature played a critical role in the production of T-2 toxin. T-2 toxin production was favored by long duration of low-temperature incubation. The growth and toxin production were relatively high on galactose, fructose, glucose, and sucrose media, when each was used as a sole carbon source, and relatively low on sorbitol, glycerol, and lactose media. For nitrogen sources, $NH_4^(+) and NO_3^{-}were used well as a sole nitrogen source, but $NO_2^-$ was not used. Initial pH and speed of shaker also affected the production of T-2 toxin. From temperature shifting experiment, it is clear that T-2 toxin metabolic pathway is regulated by temperature-dependent enzyme depression or enzyme induction system.
Jo, Mi Na;Jung, Ji En;Yoon, Hyun Joo;Chang, Kyung Hoon;Jee, Hee Sook;Kim, Kee-Tae;Paik, Hyun-Dong
Microbiology and Biotechnology Letters
/
v.42
no.4
/
pp.347-353
/
2014
${\beta}$-Glucosidase from Aspergillus usamii KCTC 6954 was used to convert ginsenoside Rb1 to compound K, which has a high bio-functional activity. The enzymatic activities during culturing for 15 days were determined using ${\rho}$-nitrophenyl-${\beta}$-glucopyranoside. The growth rate of the strain and the enzymatic activity were maximized after 6 days (IU; $175.93{\mu}M\;ml^{-1}\;min^{-1}$). The activities were maximized at $60^{\circ}C$ in pH 6.0. During culturing, Rb1 was converted to Rd after 9 d and then finally converted to compound K at 15 d. In the enzymatic reaction, Rb1 was converted to the ginsenoside Rd within 1 h of reaction time and compound K could be detected after 8 h. As a result, this study demonstrates that $Rb1{\rightarrow}Rd{\rightarrow}F2{\rightarrow}$compound K is the main metabolic pathway catalyzed by ${\beta}$-glucosidase and that ${\beta}$-glucosidase is a feasible option for the development of specific bioconversion processes to obtain minor ginsenosides such as Rd and compound K.
Oxidized low-density lipoprotein (oxLDL) has been shown to modulate transactivations by the peroxisome proliferator activated receptor (PPAR)$\gamma$ and nuclear factor-kappa B (NF$\kappa$B). In this study, the oxLDL signaling pathways involved with the NF$\kappa$B transactivation were investigated by utilizing a reporter construct driven by three upstream NF$\kappa$B binding sites, and various pharmacological inhibitors. OxLDL and its constituent lysophophatidylcholine (lysoPC) induced a rapid and transient increase of intracellular calcium and stimulated the NF-KB transactivation in resting RAW264.7 macrophage cells in an oxidation-dependent manner. The NF$\kappa$B activation by oxLDL or lysoPC was inhibited by protein kinase C inhibitors or an intracellular calcium chelator. Tyrosine kinase or PI3 kinase inhibitors did not block the NF$\kappa$B transactivation. Furthermore, the oxLDL-induced NF$\kappa$B activity was abolished by the PPAR$\gamma$ ligands. When the endocytosis of oxLDL was blocked by cytochalasin B, the NF$\kappa$B transactivation by oxLDL was synergistically increased, while PPAR transactivation was blocked. These results suggest that oxLDL activates NF-$\kappa$B in resting macrophages via protein kinase C- and/or calcium-dependent pathways, which does not involve the endocytic processing of oxLDL. The endocytosis-dependent PPAR$\gamma$ activation by oxLDL may function as an inactivation route of the oxLDL induced NF$\kappa$B signal. Short heterodimer partner (SHP), specifically expressed in liver and a limited number of other tissues, is an unusual orphan nuclear receptor that lacks the conventional DNA-binding domain. In this work, we found that SHP expression is abundant in murine macrophage cell line RAW 264.7 but suppressed by oxLDL and its constituent I3-HODE, a ligand for peroxisome proliferator-activated receptor y. Furthermore, SHP acted as a transcription coactivator of nuclear factor-$\kappa$B (NF$\kappa$B) and was essential for the previously described NF$\kappa$B transactivation by lysoPC, one of the oxLDL constituents. Accordingly, NF$\kappa$B, transcriptionally active in the beginning, became progressively inert in oxLDL-treated RAW 264.7 cells, as oxLDL decreased the SHP expression. Thus, SHP appears to be an important modulatory component to regulate the transcriptional activities of NF$\kappa$B in oxLDL-treated, resting macrophage cells.
Shin, Bo Su;Lee, Seul Ah;Moon, Sung Min;Han, Seul Hee;Hwang, Eun Ju;Kim, Su-Gwan;Kim, Do Kyung;Kim, Jin-Soo;Park, Bo-Ram;Kim, Chun Sung
International Journal of Oral Biology
/
v.42
no.4
/
pp.183-190
/
2017
Ficus carica L. (common fig), one of the first plants cultivated by humans, originated in the Mediterranean basin and currently grows worldwide, including southwest Asia and South Korea. It has been used as a traditional medicine for treatment of metabolic, cardiovascular, and respiratory diseases as well as hemorrhoids and skin infections. Its pharmacological properties have recently been studied in detail, but research on the anti-cancer effect of its latex has been only been studied on a limited basis on several cell lines, such prostate cancer, breast cancer, and leukemia. In this study, we investigated the anti-cancer activity of the latex of Ficus carica L.and its underlying mechanism in FaDu human hypopharynx squamous carcinoma cells. (See Ed. note above) We confirmed through SDS-PAGE analysis and gelatinolytic activity analysis that the latex of Ficus carica contains cysteine protease ficin. Our data showed that the latex inhibited cell growth in a dose-dependent manner. In addition, the latex treatment markedly induced apoptosis in FaDu cells as determined by FACS analysis, elevated expression level of cleaved caspase-9, -3 and PARP (poly (ADP-ribose) polymerase), and. increased the expression of Bax (pro-apoptotic factor) while decreasing the expression of Bcl-2 (anti-apoptotic factor). Taken together, these results suggested that latex containing the ficin inhibited cell growth and induced apoptosis by caspase and the Bcl-2 family signaling pathway in FaDu human hypopharynx squamous carcinoma cells. These findings point to the potential of latex of Ficus carica to provide a novel chemotherapeutic drug due to its growth inhibition effects and induction of apoptosis in human oral cancer cells.
Panax ginseng seedlings were grown at various temperature regimes from 15 to $30^{\circ}C$ for 19 days under dark and the linear relationship between various regrowth efficiencies and thermal stress indices, cumulative superoptimum temperatures corrected with factors. Gross growth efficiency(shoot weight l root weight loss) was 37.5 % at the optimum temperature $15^{\circ}C$/$15^{\circ}C$, and 12.3% at the highest temperature, $30^{\circ}C$/$30^{\circ}C$ while net growth efficiency (shoot weight + Sm)l(root loss-Rm), which corrected by maintenance respiration for shoot(Sm) and root(Rm) was 39.6% and 16.7 at optimum and highest temperature respectively. All growth efficiencies showed negative correlations (p = 0.001) with all thermal stress indices and negative(p = 0.001) with shoot growth(St). When growth temperature difference in a day was nil or above $15^{\circ}C$ growth efficiency decreased greatly. Thermal stress indices showed negative correlation with root dry matter loss(RDL) but positive with Rm. St showed positive correlation with RDL. Thermal stress appeared to inhibit substrate supply for shoot growth resulting in the extremely low growth efficiency comparing with other crops that seems to be main rate limiting factor of slow growth, Thus it is necessary that growth efficiency and thermal stress must be elucidated in terms of metabolic pathway.
Yi, Sang Ah;Lee, Jieun;Park, Sun Kyu;Kim, Jeom Yong;Park, Jong Woo;Lee, Min Gyu;Nam, Ki Hong;Park, Jee Hun;Oh, Hwamok;Kim, Saetbyul;Han, Jihoon;Kim, Bo Kyung;Jo, Dong-Gyu;Han, Jeung-Whan
Journal of Ginseng Research
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v.44
no.1
/
pp.58-66
/
2020
Background: The biological and pharmacological effects of BST204, a fermented ginseng extract, have been reported in various disease conditions. However, its molecular action in metabolic disease remains poorly understood. In this study, we identified the antiadipogenic activity of BST204 resulting from its inhibition of the S6 kinase 1 (S6K1) signaling pathway. Methods: The inhibitory effects of BST204 on S6K1 signaling were investigated by immunoblot, nuclear fractionation, immunoprecipitation analyses. The antiadipogenic effect of BST204 was evaluated by measuring mRNA levels of adipogenic genes and by chromatin immunoprecipitation and quantitative real-time polymerase chain reaction analysis. Results: Treatment with BST204 inhibited activation and nuclear translocation of S6K1, further decreasing the interaction between S6K1 and histone H2B in 10T1/2 mesenchymal stem cells. Subsequently, phosphorylation of H2B at serine 36 (H2BS36p) by S6K1 was reduced by BST204, inducing an increase in the mRNA expression of Wnt6, Wnt10a, and Wnt10b, which disturbed adipogenic differentiation and promoted myogenic and early osteogenic gene expression. Consistently, BST204 treatment during adipogenic commitment suppressed the expression of adipogenic marker genes and lipid drop formation. Conclusion: Our results indicate that BST204 blocks adipogenesis of mesenchymal stem cells through the inhibition of S6K1-mediated histone phosphorylation. This study suggests the potential therapeutic strategy using BST204 to combat obesity and musculoskeletal diseases.
Alcohol consumption increases the risk of type 2 diabetes. However, its effects on prediabetes or early diabetes have not been studied. We investigated endoplasmic reticulum (ER) stress in the pancreas and liver resulting from chronic alcohol consumption in the prediabetes and early stages of diabetes. We separated Otsuka Long-Evans Tokushima Fatty (OLETF) rats, a type-2 diabetic animal model, into two groups based on diabetic stage: prediabetes and early diabetes were defined as occurrence between the ages of 11 to 16 weeks and 17 to 22 weeks, respectively. The experimental group received an ethanol-containing liquid diet for 6 weeks. An intraperitoneal glucose tolerance test was conducted after 16 and 22 weeks for the prediabetic and early diabetes groups, respectively. There were no significant differences in body weight between the control and ethanol groups. Fasting and 120-min glucose levels were lower and higher, respectively, in the ethanol group than in the control group. In prediabetes rats, alcohol induced significant expression of ER stress markers in the pancreas; however, alcohol did not affect the liver. In early diabetes rats, alcohol significantly increased most ER stress-marker levels in both the pancreas and liver. These results indicate that chronic alcohol consumption increased the risk of diabetes in prediabetic and early diabetic OLETF rats; the pancreas was more susceptible to damage than was the liver in the early diabetic stages, and the adaptive and proapoptotic pathway of ER stress may play key roles in the development and progression of diabetes affected by chronic alcohol ingestion.
Lee, Jun Hee;Lee, Sang Hun;Lee, Hyang Seon;Ji, Seung Taek;Jung, Seok Yun;Kim, Jae Ho;Bae, Sun Sik;Kwon, Sang-Mo
The Korean Journal of Physiology and Pharmacology
/
v.20
no.5
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pp.459-466
/
2016
Adipogenic differentiation of mesenchymal stem cells (MSCs) is critical for metabolic homeostasis and nutrient signaling during development. However, limited information is available on the pivotal modulators of adipogenic differentiation of MSCs. Adaptor protein Lnk (Src homology 2B3 [SH2B3]), which belongs to a family of SH2-containing proteins, modulates the bioactivities of different stem cells, including hematopoietic stem cells and endothelial progenitor cells. In this study, we investigated whether an interaction between insulin-like growth factor-1 receptor (IGF-1R) and Lnk regulated IGF-1-induced adipogenic differentiation of MSCs. We found that wild-type MSCs showed greater adipogenic differentiation potential than $Lnk^{-/-}$ MSCs. An ex vivo adipogenic differentiation assay showed that $Lnk^{-/-}$ MSCs had decreased adipogenic differentiation potential compared with wild-type MSCs. Interestingly, we found that Lnk formed a complex with IGF-1R and that IGF-1 induced the dissociation of this complex. In addition, we observed that IGF-1-induced increase in the phosphorylation of Akt and mammalian target of rapamycin was triggered by the dissociation of the IGF-1R-Lnk complex. Expression levels of a pivotal transcription factor peroxisome proliferator-activated receptor gamma ($PPAR-{\gamma}$) and its adipogenic target genes (LPL and FABP4) significantly decreased in $Lnk^{-/-}$ MSCs. These results suggested that Lnk adaptor protein regulated the adipogenesis of MSCs through the $IGF-1/Akt/PPAR-{\gamma}$ pathway.
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