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Food Bioscience
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A R T I C LE I N FO A B S T R A C T
Keywords: The influences of fermentation temperature on metabolic changes in Kimchi using a gas chromatography–mass
Kimchi spectrometry (GC-MS) method were studied. Kimchi mixtures fermented at 4, 12, and 20 °C were taken on the
Chinese cabbage 1st, 5th, 10th, and 50th day of fermentation to determine overall changes in metabolites of Kimchi during
Fermentation fermentation. Based on the principal component analysis (PCA) score plot, Kimchi samples were confirmed to
Metabolomics
have separation trends by the first principal component 1 (PC1). One group included all samples on fermentation
GC-MS
day 0 and day 1, and samples fermented at 4 °C for up to 10 days after fermentation, demonstrating that me-
tabolites of Kimchi fermented at 4 °C changed slowly. The partial least squares discriminant analysis (PLS-DA)
score plot also showed clear differences in metabolites among Kimchi samples with fermentation temperature.
The changing metabolites were identified to be alanine, propylene glycol, fumaric acid, malic acid, citric acid,
and galactaric acid. These results highlight that a GC-MS-based metabolomics approach can be used to monitor
distinct metabolite changes in Kimchi with fermentation temperature.
1. Introduction Chang and Kim (2000) have reported that the edible period of Kimchi
depended on storage temperature. In addition, it has been reported that
Kimchi is Korea's representative ethnic food. It is a unique food low storage temperatures are better for the stability and quality of
made by fermenting vegetables (such as Chinese cabbage and radish), Kimchi than high storage temperatures (Choi et al., 1990; Park et al.,
fish sauce, and spices (such as garlic and ginger) (Jang, Chung, Yang, 2008). Therefore, differences in Kimchi quality might be simply due to
Kim, & Kwon, 2015). According to the Korean Nutrition survey, Korean difference in fermentation speed depending on fermentation tempera-
adults consume 50–200 g/day/person, accounting for 12.5% of their ture. However, recent studies have shown that differences in fermen-
total daily food intake (Mheen & Kwon, 1984). tation microbial community also depended on temperature (Park et al.,
Kimchi is usually manufactured by spontaneous fermentation 2003; Tabatabaei Yazdi, Alizadeh Behbahani, Mohebbi, Mortazavi &
without using a starter culture. Lactic acid bacteria (LAB) such as Ghaitaranpour, 2013). For example, Lee, Kim, Cho, and Kim (2008)
Lactobacillus, Leuconostoc, Weissella, Lactococcus, and Pediococcus are have reported that Lactobacillus sakei was predominant in Kimchi due to
involved in Kimchi fermentation (Park et al., 2003). Recently, many proper fermentation temperature (5–9 °C) and storage temperature
studies have determined the dynamics of microbial communities in (−2 °C). However, few studies have shown the overall fermentation
Kimchi using a metagenomics approach (Jeong, Lee, Jung, Choi, & pattern or metabolite differences of Kimchi according to fermentation
Jeon, 2013; Jung et al., 2012, 2013; Park et al., 2003). Microbial temperature.
communities in Kimchi during its fermentation are affected by several Metabolomics refers to systematic identification and quantification
factors, including raw material (Chinese cabbage), temperature, pH, of metabolites present within an organism, cell, or tissue. Many meta-
salt concentration, and fermentation period (Kim et al., 2017; Lee, bolomics studies have been done for component analysis (Farag, Porzel,
Song, Jung, Lee, & Chang, 2017). Among these, fermentation tem- & Wessjohann, 2012), quality evaluation (Vikram, Lui, Hossain, &
perature is a critical factor for determining its final quality. To maintain Kushalappa, 2006), safety evaluation (Castro-Puyana and Herrero,
the quality of Kimchi and store it for a long time, controlling fermen- 2013), or microbial monitoring (MacKenzie et al., 2008). A previous
tation rate through temperature regulation is well-known. metabolomics study using GC-MS with multivariate statistical analysis
Many previous studies have reported the relationship between fer- has shown the metabolic characteristics of Kimchi according to starter
mentation temperature and Kimchi quality. Mheen and Kwon (1984) cultures (Park et al., 2016). Kimchi metabolites are important factors
have reported that Kimchi samples stored at high temperatures will that determine the taste of Kimchi. However, little is known about
reach the ripening stage faster than those stored at low temperatures. changes of Kimchi metabolites with temperature during the
⁎
Corresponding author.
E-mail address: hsson@dsu.ac.kr (H.-S. Son).
https://doi.org/10.1016/j.fbio.2018.03.009
Received 3 November 2017; Received in revised form 19 March 2018; Accepted 24 March 2018
Available online 27 March 2018
2212-4292/ © 2018 Elsevier Ltd. All rights reserved.
S.-E. Park et al. Food Bioscience 23 (2018) 100–106
fermentation process. Thus, the objective of this study was to monitor (Agilent). The mass spectrometer was programmed using electron im-
overall changes in metabolites of Kimchi with fermentation tempera- pact in a full scan mode at m/z 50–550 with a scanning rate of 2 scans/
ture using a GC-MS-based metabolomics approach. sec.
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Fig. 2. PCA score plot derived from GC-MS data of Kimchi during ripening at different temperatures (4, 12, and 20 °C). Each symbol (point) in the score plot
represents a Kimchi sample. Symbols with different colors and shapes denote Kimchi samples taken at different temperatures on different days of fermentation.
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Fig. 3. PCA score plots derived from GC-MS data of Kimchi during ripening at 4 °C (A), 12 °C (B), and 20 °C (C), showing different metabolic changes (fermentation
rates) depending on different temperature. The color and shape of each point are the same as shown in Fig. 2.
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Fig. 4. PCA (A) and PLS-DA (B) score plots derived from GC-MS data of Kimchi after 50 days of fermentation at different temperatures. A permutation test was
carried out with 200 random permutations in the PLS-DA model (C). The color and shape of each point are the same as shown in Fig. 2.
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Fig. 5. Changes in identified metabolites of Kimchi during ripening at different temperatures. Metabolites contributing to differentiation in the PLS-DA model (VIP
score > 1.5, p < 0.05) were selected. Values are presented as mean ± standard deviations (n = 5).
and R2 values were found to be higher than their original values, Park, Hong, & Lee, 2009). When lactic acid is produced in Kimchi, the
proving the suitability and validity of this model (Fig. 4C). To de- growth of aerobic bacteria is suppressed (Jang & Kim, 2013). Park et al.
termine which metabolites caused segregation, the VIP score of > 1.5 (2012) have reported changes in bacterial community structure of
and p < 0.01 were used. A total of six metabolites were identified Kimchi after a long period of incubation at room temperature (22 °C).
based on fragmentation patterns of GC–MS library in NIST 14 and other These changes in microbial communities might cause differences in
researchers’ experimental data. Metabolites responsible for metabolic metabolites, thus affecting the quality of Kimchi. Levels of propylene
differences among these groups included some organic acids (fumaric glycol and galactaric acid in Kimchi stored at 4 °C were increased
acid, malic acid, citric acid, and galactaric acid), propylene glycol, and sharply between the 10th and 50th day of fermentation. Interestingly,
alanine. these metabolites showed very little changes during the entire fer-
Changes of these six metabolites according to fermentation tem- mentation period in Kimchi samples stored at 12 or 20 °C. In this study,
perature are shown in Fig. 5. Levels of fumaric acid, malic acid, and different contents of metabolites in Kimchi with fermentation tem-
citric acid in Kimchi samples fermented at 20 °C decreased rapidly peratures indicated that fermentation temperature can affect microbial
during the early fermentation period. However, levels of malic acid and community and fermentation rate.
citric acid in Kimchi samples fermented at 4 °C decreased after 10 days Kimchi generally undergoes fermentation at low temperature (about
of fermentation. These results were consistent with large changes in 4–6 °C). However, it will take weeks to finish the process. Commercially
metabolites of Kimchi fermented at 4 °C between 10 and 50 days of produced Kimchi is usually fermented for a short period (1–2 days) at
fermentation as shown in Fig. 3A. Reduction in malic acid and citric 20 °C. It is then stored at low temperature (5–8 °C) to improve orga-
acid levels indicated the presence of malolactic fermentation since LAB noleptic quality (Hong et al., 1994; Lee et al., 2017). It is difficult to
could convert malic acid and citric acid into lactic acid (Son, Hwang, clearly determine the optimum duration of ripening time according to
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the fermentation temperature of Kimchi. Cheigh, Park, and Lee (1994) Cheigh, H. S., Park, K. Y., & Lee, C. (1994). Biochemical, microbiological, and nutritional
have reported that optimized time for Kimchi fermented at 5 and 20 °C aspects of Kimchi (Korean fermented vegetable products). Critical Reviews in Food
Science and Nutrition, 34, 175–203.
were 35–180 days and 2–3 days, respectively. According to a study of Choi, S. Y., Kim, Y. B., Yoo, J. Y., Lee, I. S., Chung, K. S., & Koo, Y. J. (1990). Effect of
Chang and Kim (2000), optimized time for Kimchi ripening at 15, 10, temperature and salts concentration of kimchi manufacturing on storage. Korean
and 5 °C were 4, 10, and 18 days, respectively. Similar to these studies, Journal of Food Science and Technology, 22, 707–710.
Farag, M. A., Porzel, A., & Wessjohann, L. A. (2012). Comparative metabolite profiling
Lee, Cho, and Pyun (1991) have reported that Kimchi's shelf life 4 and and fingerprinting of medicinal licorice roots using a multiplex approach of GC–MS,
28 °C were 33 and 3 days, respectively, based on kinetic modeling of LC–MS and 1D NMR techniques. Phytochemistry, 76, 60–72.
total acidity for quality prediction. Hong, Lee, Kim, and Ahn (2016) Hong, S. P., Lee, E. J., Kim, Y. H., & Ahn, D. U. (2016). Effect of fermentation temperature
on the volatile composition of Kimchi. Journal of Food Science, 81, C2623–C2629.
have reported that the optimum ripening time for Kimchi fermented at Hongu, N., Kim, A. S., Suzukim, A., Wilson, H., Tsui, K. C., & Park, S. (2017). Korean
4 and 20 °C was 35 and 2 days, respectively, after analyzing volatile Kimchi: Promoting healthy meals through cultural tradition. Journal of Ethnic Foods,
compositions of Kimchi. Nowadays, it is common to store homemade or 4, 172–180.
Hotelling, H. (1931). The generalization of Student's ratio. The Annals of Mathematical
commercially produced Kimchi products in a Kimchi refrigerator in
Statistics, 2, 360–378.
Korea to reduce microbial growth and avoid over-fermenting or souring Jang, D. J., Chung, K. R., Yang, H. J., Kim, K. S., & Kwon, D. Y. (2015). Discussion on the
by rapid shifting to subzero temperatures (Hongu et al., 2017; Lee et al., origin of Kimchi, representative of Korean unique fermented vegetables. Journal of
2008). Although the optimum ripening time for Kimchi is different, it Ethnic Foods, 2, 126–136.
Jang, J. Y., & Kim, T. W. (2013). Lactic acid bacteria in kimchi and their im-
obviously depends on storage temperature. Storage temperature of munomodulatory activities. Current Topic in Lactic Acid Bacteria and Probiotics, 1,
Kimchi is directly related to the growth of microorganisms which will 28–37.
affect Kimchi metabolites. Therefore, it is important to know the me- Jeong, S. H., Lee, S. H., Jung, J. Y., Choi, E. J., & Jeon, C. O. (2013). Microbial succession
and metabolite changes during long-term storage of Kimchi. Journal of Food Science,
tabolic difference in Kimchi because it affects the quality of Kimchi. 78, M763–M769.
These results suggest that the optimum duration for Kimchi fermenta- Jung, J. Y., Lee, S. H., Jin, H. M., Hahn, Y., Madsen, E. L., & Jeon, C. O. (2013).
tion can be determined using a metabolomics approach to simulta- Metatranscriptomic analysis of lactic acid bacterial gene expression during Kimchi
fermentation. International Journal of Food Microbiology, 163, 171–179.
neously observe metabolites during fermentation. Jung, J. Y., Lee, S. H., Lee, H. J., Seo, H. Y., Park, W. S., & Jeon, C. O. (2012). Effects of
Leuconostoc mesenteroides starter cultures on microbial communities and metabolites
4. Conclusion during Kimchi fermentation. International Journal of Food Microbiology, 153, 378–387.
Kim, D. W., Kim, B. M., Lee, H. J., Jang, G. J., Song, S. H., Lee, J. I., ... Kim, H. J. (2017).
Effects of different salt treatments on the fermentation metabolites and bacterial
A GC-MS based metabolomics approach coupled with multivariate profiles of Kimchi. Journal of Food Science, 82, 1124–1131.
statistics were applied in this study to understand relationships between Lee, D., Kim, S., Cho, J., & Kim, J. (2008). Microbial population dynamics and tem-
perature changes during fermentation of kimjang Kimchi. The Journal of Microbiology,
fermentation temperature and metabolite differences of Kimchi.
46, 590–593.
Metabolites of Kimchi fermented at 4 °C changed more slowly during Lee, K. H., Cho, H. Y., & Pyun, Y. R. (1991). Kinetic modelling for the prediction of shelf-
early fermentation than those of Kimchi fermented at higher tempera- life of kimchi based on total acidity as a quility index. Korean Journal of Food Science
tures (4 and 20 °C). After fermentation, different concentrations of and Technology, 23, 306–310.
Lee, M., Song, J. H., Jung, M. Y., Lee, S. H., & Chang, J. Y. (2017). Large-scale targeted
metabolites were observed in Kimchi samples depending on fermenta- metagenomics analysis of bacterial ecological changes in 88 Kimchi samples during
tion temperature. It is possible to determine the optimum duration of fermentation. Food Microbiology, 66, 173–183.
Kimchi fermentation by metabolomics approach which can simulta- MacKenzie, D. A., Defernez, M., Dunn, W. B., Brown, M., Fuller, L. J., de Herrera, S. R., ...
Philo, M. (2008). Relatedness of medically important strains of Saccharomyces cere-
neously observe metabolites during fermentation. However, more de- visiae as revealed by phylogenetics and metabolomics. Yeast, 25, 501–512.
tailed studies are needed to better understand relationships of sensory Mheen, T. I., & Kwon, T. W. (1984). Effect of temperature and salt concentration on
characteristics, microbial communities, and metabolites of Kimchi with kimchi fermentation. Korean Journal of Food Science and Technology, 16, 443–450.
Park, E. J., Chun, J., Cha, C. J., Park, W. S., Jeon, C. O., & Bae, J. W. (2012). Bacterial
fermentation temperature. community analysis during fermentation of ten representative kinds of Kimchi with
barcoded pyrosequencing. Food Microbiology, 30, 197–204.
Acknowledgements Park, J., Heo, G., Oh, Y., Kim, B., Miheen, T., Kim, C., ... Lee, J. (2003). Change of mi-
crobial communities in kimchi fermentation at low temperature. Korean Journal of
Microbiology, 39, 45–50.
This work was supported by a grant (NRF- 2014R1A1A1002817) Park, J. G., Kim, J. H., Park, J. N., Kim, Y. D., Kim, W. G., Lee, J. W., ... Byun, M. W.
from the National Research Foundation funded by the Korean (2008). The effect of irradiation temperature on the quality improvement of Kimchi,
Korean fermented vegetables, for its shelf stability. Radiation Physics and Chemistry,
Government.
77, 497–502.
Park, S. E., Yoo, S. A., Seo, S. H., Lee, K. I., Na, C. S., & Son, H. S. (2016). GC–MS based
Conflict of interest metabolomics approach of Kimchi for the understanding of Lactobacillus plantarum
fermentation characteristics. LWT-Food Science and Technology, 68, 313–321.
Son, H. S., Hwang, G. S., Park, W. M., Hong, Y. S., & Lee, C. H. (2009). Metabolomic
The authors declare that there are no conflicts of interest. characterization of malolactic fermentation and fermentative behaviors of wine
yeasts in grape wine. Journal of Agricultural and Food Chemistry, 57, 4801–4809.
References Tabatabaei Yazdi, F., Alizadeh Behbahani, B., Mohebbi, M., Mortazavi, A., &
Ghaitaranpour, A. (2013). Effect of temperature on microbial changes during kimchi
fermentation. Scientific Journal of Microbiology, 2, 9–14.
Castro-Puyana, M., & Herrero, M. (2013). Metabolomics approaches based on mass Tautenhahn, R., Boettcher, C., & Neumann, S. (2008). Highly sensitive feature detection
spectrometry for food safety, quality and traceability. TrAC Trends in Analytical for high resolution LC/MS. BMC Bioinformatics, 9, 504.
Chemistry, 52, 74–87. Vikram, A., Lui, L., Hossain, A., & Kushalappa, A. (2006). Metabolic fingerprinting to
Chang, M. J., & Kim, M. H. (2000). Fermentation property of Chinese cabbage kimchi by discriminate diseases of stored carrots. Annals of Applied Biology, 148, 17–26.
fermentation temperature and salt concentration. Applied Biological Chemistry, 43,
7–11.
106