Cherl-Ho Lee

Metabolomic studies on geographical grapes and their wines using 1H NMR analysis coupled with multivariate statistics.

Environmental vineyard conditions can affect the chemical composition or metabolites of grapes and their wines. Grapes grown in three different regions of South Korea were collected and separated into pulp, skin, and seed. The grapes were also vinified after crushing. (1)H NMR spectroscopy with pattern recognition (PR) methods was used to investigate the metabolic differences in pulp, skin, seed, and wines from the different regions. Discriminatory compounds among the grapes were Na, Ca, K, malate, citrate, threonine, alanine, proline, and trigonelline according to PR methods of principal component analysis (PCA) or partial least-squares discriminant analysis (PLS-DA). Grapes grown in regions with high sun exposure and low rainfall showed higher levels of sugar, proline, Na, and Ca together with lower levels of malate, citrate, alanine, threonine, and trigonelline than those grown in regions with relatively low sun exposure and high rainfall. Environmental effects were also observed in the complementary wines. This study demonstrates that (1)H NMR-based metabolomics coupled with multivariate statistical data sets can be useful for determining grape and wine quality.

Lactic acid fermented foods and their benefits in Asia

Abstract This paper reviews many types of the worlds lactic acid fermented foods and discusses the beneficial effects of lactic acid fermentation of food by focusing on two examples taken from Korean cuisine, kimchi and sikhae. Sikhae is the generic name for a class of Korean lactic acid fermented fish products that contain 6–8% salt and generally are at pH 4–5. Koreans are able to preserve fish for 1–2 months at ambient temperatures by this method. Due to the low salt content, sikhae contributes much-needed protein to the Korean rice-based diet. Kimchi is the generic name for a class of Korean lactic acid fermented vegetables that contain 3–4% salt and generally are at pH 4.0–4.5. Kimchi is an important source of vitamins and minerals especially during the wintertime. It is a popular side-dish and provides a source of intestinal lactic acid bacteria. The physiological effects of kimchi have been studied widely in Korea and recent results are summarized in this paper.

Importance of lactic acid bacteria in Asian fermented foods

Lactic acid bacteria play important roles in various fermented foods in Asia. Besides being the main component in kimchi and other fermented foods, they are used to preserve edible food materials through fermentation of other raw-materials such as rice wine/beer, rice cakes, and fish by producing organic acids to control putrefactive microorganisms and pathogens. These bacteria also provide a selective environment favoring fermentative microorganisms and produce desirable flavors in various fermented foods. This paper discusses the role of lactic acid bacteria in various non-dairy fermented food products in Asia and their nutritional and physiological functions in the Asian diet.

1H Nuclear Magnetic Resonance-Based Metabolomic Characterization of Wines by Grape Varieties and Production Areas

(1)H NMR spectroscopy was used to investigate the metabolic differences in wines produced from different grape varieties and different regions. A significant separation among wines from Campbell Early, Cabernet Sauvignon, and Shiraz grapes was observed using principal component analysis (PCA) and partial least squares-discriminant analysis (PLS-DA). The metabolites contributing to the separation were assigned to be 2,3-butanediol, lactate, acetate, proline, succinate, malate, glycerol, tartarate, glucose, and phenolic compounds by PCA and PLS-DA loading plots. Wines produced from Cabernet Sauvignon grapes harvested in the continental areas of Australia, France, and California were also separated. PLS-DA loading plots revealed that the level of proline in Californian Cabernet Sauvignon wines was higher than that in Australian and French Cabernet Sauvignon, Australian Shiraz, and Korean Campbell Early wines, showing that the chemical composition of the grape berries varies with the variety and growing area. This study highlights the applicability of NMR-based metabolomics with multivariate statistical data sets in determining wine quality and product origin.

Geographical and Climatic Dependencies of Green Tea (Camellia sinensis) Metabolites: A 1H NMR-Based Metabolomics Study

The effects of climatic conditions on green tea metabolites in three different growing areas of Jeju Island, South Korea, were investigated through global metabolite profiling by (1)H nuclear magnetic resonance (NMR) spectroscopy. Pattern recognition methods, such as principal component analysis (PCA) and orthogonal projection on latent structure-discriminant analysis (OPLS-DA), revealed clear discriminations of green teas from the three different growing areas. Variations of theanine, isoleucine, leucine, valine, alanine, threonine, glutamine, quinic acid, glucose, epicatechin (EC), epigallocatechin (EGC), epigallocatechin-3-gallate (EGCG), and caffeine levels were responsible for the discriminations. Green teas grown in an area with high temperature, long sun exposure time, and high rainfall had higher levels of theanine but lower levels of isoleucine, leucine, valine, alanine, EC, EGC, EGCG, and caffeine than those grown in areas with relatively low temperature, short sun exposure time, and low rainfall. These results indicate that high temperature, long sun exposure, and high preciptation stimulate theanine synthesis in green tea during the spring season. This study highlights how metabolomics coupled with multivariate statistical analysis can illuminate the metabolic characteristics of green tea associated with climatic variables, thereby allowing for the assessment of quality strategy in green tea production.

Estimation of Excess Cancer Risk on Time-Weighted Lifetime Average Daily Intake of PAHs from Food Ingestion

ABSTRACT The purposes of this study were to quantify the time-weighted, lifetime average, daily intake (LADI) of polycyclic aromatic hydrocarbons (PAHs) through food ingestion and to estimate the excess cancer risk based on lifetime dietary PAH intake. Twenty-seven different food commodities were selected from the 2001 Korean National Health and Nutrition survey based on their frequent consumption and high PAH level. The foods were analyzed for the profile of 14 PAH congeners using high performance liquid chromatography (HPLC) and fluorescence detector. Considering the toxic equivalent (TEQ) level converted with the toxic equivalent factors (TEFs), the highest total TEQ level of PAHs in foods was detected from roasted laver at 1.2 ug TEQ/kg. For the PAH exposure assessment according to ingested foods, the average body weight was separated according to the following age groups, 1–6, 7–19, 20–64 and over 64 years, and the daily food ingestion rates from the National Health and Nutrition survey were used. The estimated Lifetime Average Daily Intake (LADI) of PAHs was 3.22 × 10–3 ug/kg/day for carcinogenic effects and was higher in the younger age groups under 20 years old than in the older groups. The dietary excess cancer risk estimated using the cancer potency of benzo(a)pyrene (7.3(mg/kg/day)−1) was 2.3 × 10−5, which is equivalent to a probability of tumor eruption in the upper gastrointestinal tract of two per hundred thousand persons.

1H NMR-based metabolomic approach for understanding the fermentation behaviors of wine yeast strains.

(1)H NMR spectroscopy coupled with multivariate statistical analysis was used for the first time to investigate metabolic changes in musts during alcoholic fermentation and wines during aging. Three Saccharomyces cerevisiae yeast strains (RC-212, KIV-1116, and KUBY-501) were also evaluated for their impacts on the metabolic changes in must and wine. Pattern recognition (PR) methods, including PCA, PLS-DA, and OPLS-DA scores plots, showed clear differences for metabolites among musts or wines for each fermentation stage up to 6 months. Metabolites responsible for the differentiation were identified as valine, 2,3-butanediol (2,3-BD), pyruvate, succinate, proline, citrate, glycerol, malate, tartarate, glucose, N-methylnicotinic acid (NMNA), and polyphenol compounds. PCA scores plots showed continuous movements away from days 1 to 8 in all musts for all yeast strains, indicating continuous and active fermentation. During alcoholic fermentation, the highest levels of 2,3-BD, succinate, and glycerol were found in musts with the KIV-1116 strain, which showed the fastest fermentation or highest fermentative activity of the three strains, whereas the KUBY-501 strain showed the slowest fermentative activity. This study highlights the applicability of NMR-based metabolomics for monitoring wine fermentation and evaluating the fermentative characteristics of yeast strains.

Evidence of vintage effects on grape wines using 1H NMR-based metabolomic study.

The chemical composition of grape wines varies with grape variety, environmental factors of climate and soil, and bacterial strains, which can each affect the wine quality. Using (1)H NMR analysis coupled with multivariate statistical data sets, we investigated the effects of grape vintage on metabolic profiles of wine and the relationship between wine metabolites and meteorological data. Principal component analysis (PCA) showed a clear differentiation between Meoru wines that were vinified with the same yeast strain and Meoru grapes harvested from the same vineyard but with a different vintage. The metabolites contributing to the differentiation were identified as 2,3-butandiol, lactic acid, alanine, proline, gamma-aminobutyric acid (GABA), choline, and polyphenols, by complementary PCA loading plot. Markedly higher levels of proline, lactic acid and polyphenols were observed in the 2006 vintage wines compared to those of 2007 vintage, showing excellent agreement with the meteorological data that the sun-exposed time and rainfall in 2006 were approximately two times more and four times less, respectively, than those in 2007. These results revealed the important role of climate during ripening period in the chemical compositions of the grape. This study highlights the reliability of NMR-based metabolomic data by integration with meteorological data in characterizing wine or grape.

Identification of LDL-receptor transcription stimulating peptides from soybean hydrolysate in human hepatocytes.

Soybean protein and its hydrolysate have been reported to have cholesterol-lowering property, but the responsible components are still largely unknown. In previous study, we found that soybean protein hydrolysate (SPH) prepared with the protease from Bacillus amyloliquefaciens FSE-68, strongly stimulates transcription of low density lipoprotein receptor (LDL-R). To identify LDL-R transcription stimulating peptides in human hepatocytes, the SPH was fractionated with gel permeation chromatograpy and the active fraction was further separated by using reverse-phase chromatography. Several peptides in the most active fraction were identified by LC/MS and MS/MS analysis. LDL-R transcription stimulating peptides were synthesized on the basis of identified sequences, and their effect on LDL-R transcription was tested in vitro. Among the synthesized peptides, Phe-Val-Val-Asn-Ala-Thr-Ser-Asn (FVVNATSN) showed the strongest activity, and LDL-R transcription of hepatic cells was increased to 248.8% (compared to 100% of untreated control) by FVVNATSN at a concentration of 100 microM. This study provides direct evidence that peptides derived from soybean protein can influence LDL-R transcription in hepatocytes.

Characterization of Fermentative Behaviors of Lactic Acid Bacteria in Grape Wines through 1H NMR- and GC-Based Metabolic Profiling

The effects of five commercial Oenococcus oeni strains (MCW, Enoferm alpha, Wyeast, Vinibacti111, and Vinibacti222) on fermentative behaviors, and variations of metabolites in Meoru ( Vitis coigneties ) wines during malolactic fermentation (MLF) were investigated by metabolomic analysis of (1)H NMR and GC data sets. In the development of MLF with various O. oeni strains, the fastest conversions of malic acid to lactic acid occurred in wines fermented with Enoferm alpha and Vinibacti111 strains. Seventeen primary metabolites and 65 secondary metabolites of volatile compounds in the wines were identified by (1)H NMR spectroscopy and GC-MS, respectively. In pattern recognition models of principal component analysis (PCA) and orthogonal projection to latent structures discriminant analysis (OPLS-DA), significant differentiations between wines with O. oeni strains were identified by the secondary metabolites rather than by the primary metabolites, showing the effects of O. oeni strains only on the secondary metabolites. Twelve volatile compounds, 2-phenylethanol, isoamyl alcohol, 2-butanol, ethyl octanoate, ethyl hexanoate, hexadecanoic acid, diethyl succinate, butyl butyrate, octanoic acid, 9-hexadecanoic acid, isobutyric acid, and 2-ethyl-1-hexanol, contributed to the differentiation of wines according to O. oeni strain, including spontaneous MLF. This study demonstrates that O. oeni strains affect the secondary metabolites, which are easily identified through multivariate statistical analysis of GC-MS data set.