Gye-Won Kim

Composition of Organic Acids and Physiological Functionality of Commercial Makgeolli

Makgeolli is Korean traditional alcoholic beverage that has historically been brewed. In this study, we analyzed the profile of organic acids in makgeolli and also evaluated its physiological characteristics. Makgeolli contained excess lactic acid, which is produced by lactic acid bacteria (LAB). Anti-obesity effects of makgeolli were investigated in 3T3-L1 preadipocytes. Compared to the negative control, makgeolli inhibited the differentiation of preadipocyte as quantified by Oil red O dye. In particular, 100 μg/mL makgeolli reduced 40 to 70% of differentiation. To evaluate the anti-angiogenic and anti-inflammatory effects of makgeolli, we performed chorioallantoic membrane assay and measured nitric oxide production from lipopolysaccharide-induced RAW264.7 cells. Most makgeolli interrupted the formation of neo-vasculature and significantly inhibited NO production in a dose-dependent manner. Taken together, these findings suggest that commercial makgeolli has inhibitory activities against adipogenesis, neo-vascularization, and inflammation, and also they are influenced by second metabolites from nuruk microflora containing fungi and LAB.

Bioactive 3,4-seco-Triterpenoids from the Fruits of Acanthopanax sessiliflorus.

Eight new 3,4-seco-lupane triterpenes and glycosides, acanthosessiligenins I and II (1, 3) and acanthosessiliosides A-F (2, 4-8), as well as six known 3,4-seco-lupane triterpenes (9-14) were isolated from an ethanolic extract of Acanthopanax sessiliflorus fruits. The chemical structures of 1-8 were determined by spectroscopic data interpretation. All isolated compounds were tested for their cytotoxicity against six human cancer cell lines and their ability to inhibit LPS-induced nitric oxide production in RAW 264.7 macrophages.

Anti-inflammatory lignans from the fruits of Acanthopanax sessiliflorus.

A new lignan, named acanthosessilin A (1), as well as eight known lignan and lignan glycosides 2−9 were isolated from an ethanolic extract of Acanthopanax sessiliflorus fruits. The chemical structures were determined by spectroscopic methods, including HR-EIMS, 1D NMR (1H, 13C, DEPT), 2D NMR (gCOSY, gHSQC, gHMBC, NOESY), and IR spectroscopy. All isolated compounds were tested for the ability to inhibit LPS-induced nitric oxide production in RAW264.7 macrophages.

Cyanidin-3-O-Sambubioside from Acanthopanax sessiliflorus Fruit Inhibits Metastasis by Downregulating MMP-9 in Breast Cancer Cells MDA-MB-231

Anthocyanins play an important role in physiological functions related to human health. This study aimed to investigate the inhibitory effect of cyanidin-3-O-sambubioside isolated from Acanthopanax sessiliflorus fruit on cancer cell metastasis. The inhibitory effect of cyanidin-3-O-sambubioside on angiogenesis was assessed by chorioallantoic membrane and tube formation assays. Cyanidin-3-O-sambubioside interrupted the formation of neovasculature and tube-like structures in a dose-dependent manner. It inhibited the wound healing migration and invasion of MDA-MB-231 cells through reconstituted extracellular matrix (Matrigel). Gelatin zymography revealed that cyanidin-3-O-sambubioside significantly decreased the secretion of matrix metalloproteinase-9 and inhibited matrix metalloproteinase-9 expression. In additional, cyanidin-3-O-sambubioside exerted an inhibitory effect on Akt phosphorylation. These results suggest that cyanidin-3-O-sambubioside inhibits metastasis processes, such as angiogenesis and invasion, in breast cancer cells through regulation of matrix metalloproteinase-9 activity.

Melanin Biosynthesis Inhibition Effects of Ginsenoside Rb2 Isolated from Panax ginseng Berry.

Ginsenoside Rb2 (Gin-Rb2) was purified from the fruit extract of Panax ginseng. Its chemical structure was measured by spectroscopic analysis, including HR-FAB-MS, (1)H-NMR, and IR spectroscopy. Gin-Rb2 decreased potent melanogenesis in melan-a cells, with 23.4% at 80 μM without cytotoxicity. Gin-Rb2 also decreased tyrosinase and MITF protein expression in melan-a cells. Furthermore, Gin-Rb2 presented inhibition of the body pigmentation in the zebrafish in vivo system and reduced melanin contents and tyrosinase activity. These results show that Gin-Rb2 isolated from P. ginseng may be an effective skin-whitening agent via the in vitro and in vivo systems.

Differentiation of lactic acid bacteria based on RFLP analysis of the tuf gene

The tuf gene, which encodes the elongation factor Tu, is universally distributed in Gram-positive bacteria. A small region of the tuf gene was evaluated for the differentiation of Lactobacillus strains using restriction fragment length polymorphism (RFLP) analysis. The 803 bp portion of tuf genes from 9 closely-related Lactobacillus reference strains were successfully amplified by polymerase chain reaction (PCR), and restriction patterns of each were evaluated with a set of restriction enzymes, which included AluI, HaeIII, and Sau3AI. The results implied that restriction patterns obtained from 2 of these restriction enzymes (AluI and HaeIII) could effectively differentiate closely-related Lactobacillus species.

Diversity analysis of Saccharomyces cerevisiae isolated from natural sources by multilocus sequence typing (MLST)

We used multilocus sequence typing (MLST) to analyze the diversity of natural isolates of Saccharomyces cerevisiae, the most important microorganism in alcoholic fermentation. Six loci, ADP1, RPN2, GLN4, ACC1, MET4, and NUP116, in S. cerevisiae genome were selected as MLST markers. To investigate genetic diversity within S. cerevisiae, 42 S. cerevisiae isolated from natural sources in Korea as well as six S. cerevisiae obtained from Genbank and four industrial S. cerevisiae were examined using MLST. Twenty-six polymorphic sites were found in the six loci. Among them, ACC1 had the most genetic variation with eight polymorphic sites. MLST differentiated the 52 strains into three clades. Alcohol fermentation results revealed that S. cerevisiae in Clade III produced less alcohol than those in Clades I and II. These results suggested that MLST is a powerful tool to differentiate S. cerevisiae and can potentially be used to select S. cerevisiae suitable for industrial use.

Extract of Makgeolli suldeot downregulates NF-κB-mediated MMP-9 expression, thereby inhibiting invasive activity in a human breast cancer cell line, MDA-MB-231

Makgeolli is a Korean traditional alcoholic beverage prepared by fermenting rice starch using nuruk. It is known for its health benefits, but the mechanisms underlying its inhibitory effects on cancer remain unclear. Here a 70% ethanol extract of Makgeolli suldeot (70EMS) was tested for its inhibitory activities against cancer metastasis. This extract interrupted the formation of the neovasculature and reduced the migratory and invasive activity of breast cancer cells. Furthermore, it dose-dependently suppressed the activity of matrix metalloproteinase (MMP)-9 and phosphorylation of ERK1/2, JNK, and Akt. Moreover, 70EMS strongly inhibited the translocation of nuclear factor-kappa B (NF-κB) into the nucleus and expression of activator protein-1 (AP-1). These data indicate that the anti-invasive effect of 70EMS may be related to the inhibition of ERK1/2, phosphorylation of JNK, and reduced expression of NF-κB and AP-1, thereby attenuating MMP-9 in human breast cancer cells.