Won-Mok Park

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.

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.

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.

New Method Development for Nanoparticle Extraction of Water-Soluble β-(1→3)-d-Glucan from Edible Mushrooms, Sparassis crispa and Phellinus linteus

Sparassis crispa and Phellinus linteus are edible/medicinal mushrooms that have remarkably high contents of beta-(1-->3)-D-glucan, which acts as a biological response modifier, but difficulty in cultivating the fruiting bodies and extraction of beta-D-glucan have restricted detailed studies. Therefore, a novel process for nanoparticle extraction of Sparan, the beta-D-glucan from Sparassis crispa, and Phellin, the beta-D-glucan from Phellinus linteus, has been investigated using insoluble tungsten carbide as a model for nanoknife technology. This is the first report showing that the nanoknife method results in high yields of Sparan (70.2%) and Phellin (65.2%) with an average particle size of 150 and 390 nm, respectively. The extracted Sparan with beta-(1-->3) linkages showed a remarkably high water solubility of 90% even after 10 min of incubation at room temperature. Therefore, it is likely that this nanoknife method could be used to produce beta-D-glucan for food, cosmetic, and pharmaceutical industries.

Metabolomic Characterization of Malolactic Fermentation and Fermentative Behaviors of Wine Yeasts in Grape Wine

Wine contains a number of metabolites that are produced during alcoholic and malolactic fermentations (MLF) or aging, which are important compounds for determining wine quality. This study investigated changes in metabolites in wines to characterize malolactic fermentation (MLF) and to assess fermentative behaviors of wine yeast strains using (1)H nuclear magnetic resonance (NMR) spectroscopy coupled with multivariate statistics. Principal component analysis (PCA) showed clear differentiation between non- and induced-malolactic fermented wines by wine lactic acid bacteria (LAB) and between wines fermented with various wine yeast strains. Metabolites such as glycerol, lactate, 2,3-butanediol, succinate, leucine, isoleucine, alanine, valine, proline, choline, gamma-aminobutyric acid (GABA), and polyphenols contributed to the differentiations. Decreased levels of malate and citrate along with increased levels of lactate were the metabolites most responsible for the differentiation of induced-MLF wines from non-MLF wines. In particular, high succinate levels provided evidence of an inhibitory effect of Saccharomyces bayanus against spontaneous MLF. Furthermore, dependence of metabolites on wine yeast strains was observed, demonstrating their different fermentative behaviors. This study demonstrates that wine fermentation by yeast and LAB can be characterized through global and multivariate statistical analysis of (1)H NMR spectral data.

Mycelial Growth and Fruiting Body Formation of Hericium erinaceum in Sawdust and Agricultural By-product Substrates

This study was carried out to investigate the suitability of various agricultural by-products as basal substrates for the mycelial growth and fruiting body formation of Hericium erinaceum. For this aim, oak sawdust, cotton waste, sugarcane bagasse, Jobs tears, rice hull, Chinese cabbage, and coconut waste were used as sole or mixed substrate(s). Corn waste and rice bran were used as nutrient supplements. The growth and density of mycelium, yield of fruiting body, and biological efficiency were compared among tested substrates colonized by Hericium erinaceum. The best measurement of mycelial growth and density, yield of fruiting body, and biological efficiency in a laboratory test was found in a spawn substrate composed with oak sawdust 80% and rice bran 20%. The suitability of this spawn substrate composition for Hericium fruiting body production was testified through practical tests in plastic bottles (850 ml) in a mushroom farm which had bottle cultivation facility. However, test in a mushroom farm which had plastic bag cultivation facility, best production of Hericium fruiting body (520 g per one bag) was observed in a spawn substrate composed of cotton waste 40%, saw dust 40%, corn waste 10%, and rice bran 10%.