Comparison of Isoflavone Contents and Antioxidant Effect in Cheonggukjang with Black Soybean Cultivars by Bacillus subtilis CSY191

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  • ABSTRACT

    BACKGROUND:

    Soybeans are the rich sources of isoflavones. To date, the changes of isoflavone contents in various black soybeans cheonggukjang during fermentation by Bacillus subtilis CSY191 has not been investigated.

    METHODS AND RESULTS:

    This study investigated the changes of total phenolic and isoflavone contents and antioxidant effects during cheonggukjang fermentation made with four black soybean (BS) cultivars including Cheongja, Cheongja#3, Geomjeong#5, and Ilpumgeomjeong with a potential probiotic Bacillus subtilis CSY191. The total phenolic contents, isoflavone-malonylglycoside and -aglycone contents, and antioxidant activity were increased in cheonggukjang at 48 h fermentation, while the content of isoflavone-glycosides was decreased during cheonggukjang fermentation. In particular, the Cheongja#3 soybean fermented at 37℃ for 48 h displayed the highest antioxidant activities, compared to those of the other BS cultivars tested. Also, the highest levels of total phenolic, daidzein, glycitein, and genistein were present at concentrations of 17.28 mg/g, 283.7 g/g, 39.9 g/g, and 13.2 g/g at the end of Cheongja#3 soybean fermentation.

    CONCLUSION:

    The results from this study suggested that the enhanced antioxidant activity of cheonggukjang of BS might be related to increased levels of total phenolic, isoflavon-aglycone, and malonyl-glycoside contents achieved during fermentation. Furthermore, fermented Cheongja#3 soybean showed the highest levels of enhanced antioxidant activities than the other BS cultivars.


  • KEYWORD

    Antioxidant , Bacillus subtilis CSY191 , Black soybean , Cheonggukjang , Isoflavone

  • 1. Cho K. M., Lee J. H., Yun H. D., Ahn B. Y., Kim H., Seo W. T. (2011) Changes of phytochemical constituents (isoflavones, flavanols, and phenolic acids) during cheonggukjang soybeans fermentation using potential probiotics Bacillus subtilis CS90. [Journal of Food Composition and Analysis] Vol.24 P.402-410 google doi
  • 2. Cho K. M., Hong S. Y., Math R. K., Lee J. H., Kambiranda D. M., Kim J. M., Islam S. M. A., Yun M. G., Cho J. J., Lim W. J., Yun H. D. (2009) Biotransformation of phenolics (isoflavones, flavanols and phenolic acids) during the fermentation of cheonggukjang by Bacillus pumilus HY1. [Food Chemistry] Vol.114 P.413-419 google doi
  • 3. Choi J. S., Kim H. Y., Seo W. T., Lee J. H., Cho K. M. (2012) Roasting enhances antioxidant effect of bitter melon (Momordica charantia L.) increasing in flavan-3-ol and phenolic acid contents. [Food Science and Biotechnology] Vol.21 P.19-26 google doi
  • 4. Coward L., Smith M., Kirk M., Barnes S. (1998) Chemical modification of isoflavones in soyfoods during cooking and processing. [American Journal of Clinical Nutrition] Vol.68 P.1486-1491 google
  • 5. Da Silva L. H., Celeghini R. M. S., Chang Y. K. (2011) Effect of the fermentation of whole soybean flour on the conversion of isolfavones from glycosides to aglycones. [Food Chemistry] Vol.128 P.640-644 google doi
  • 6. Hu Y., Ge C., Yuan W., Zhu R., Zhang W., Du L., Xue J. (2010) Characterization of fermented black soybean natto inoculated with Bacillus natto during fermentation. [Journal of the Science of Food and Agriculture] Vol.90 P.1194-1202 google doi
  • 7. Hwang C. E., An M. J., Lee H. Y., Lee B. W., Kim H. T., Ko J. M., Baek I. Y., Seo W. T., Cho K. M. (2014) Potential probiotic Lactobacillus plantarum P1201 to produce soy-yougurt with enhanced antioxidant activity. [Korean Journal of Food Science and Technology] Vol.46 P.556-565 google doi
  • 8. Hwang C. E., Seo W. T., Cho K. M. (2013) Enhanced antioxidant effect of black soybean by cheonggukjang with potential probiotic Bacillus subtilis CSY191. [Korean Journal of Microbiology] Vol.49 P.391-397 google doi
  • 9. Jang C. H., Lim J. K., Kim J. H., Park C. S., Kwon D. Y., Kim Y. S., Shin D. H., Kim J. S. (2006) Change of isoflavone content during manufacturing of cheonggukjang, a traditional Korean fermented soy food. [Food Science and Biotechnology] Vol.15 P.643-646 google
  • 10. Juan M. Y., Chou C. C. (2010) Enhancement of antioxidant activity, total phenolic and flavonoid content of black soybeans by solid state fermentation with Bacillus subtilis BCRC 14715. [Food Microbiology] Vol.27 P.586-591 google doi
  • 11. Kim H. G., Kim G. W., Oh H., Yoo S. Y., Kim Y. O., Oh M. S. (2011) Influence of roasting on the antioxidant activity of small black soybean (Glycine max L. Merrill). [LWT-Food Science and Technology] Vol.44 P.992-998 google doi
  • 12. Kim J. S., Kang O. J., Gweon O. C. (2013) Comparison of phenolic acids and flavonoids in black garlic at different thermal processing steps. [Journal of Functional Foods] Vol.5 P.80-86 google doi
  • 13. Kim J. H., Hwan C. E., Lee C. K., Lee J. H., Kim G. M., Jeong S. H., Shin J. H., Kim J. S., Cho K. M. (2014) Characteristics and antioxidant effect of garlic in the fermentation of cheonggukjang by Bacillus amyloliquefaciens MJ1-4. [Journal of Microbiology and Biotechnology] Vol.24 P.959-968 google
  • 14. Kim N. Y., Song E. J., Kwon D. Y., Kim H. P., Heo M. Y. (2008) Antioxidant and antigenotoxic activities of Korean fermented soybean. [Food and Chemical Toxicology] Vol.46 P.1184-1189 google doi
  • 15. Kwak C. S., Kim S. A., Lee M. S. (2005) The correlation of antioxidative effects of 5 Korean common edible seaweeds and total polyphenol content. [Journal of the Korean Society of Food Science and Nutrition] Vol.34 P.1143-1150 google doi
  • 16. Kwak C. S., Lee M. S., Park S. C. (2007) Higher antioxidant of chungkookjang, a fermented soybean paste, may be due to increased aglycone and malonylgycoside isoflavone during fermentation. [Nutrition Research] Vol.27 P.719-727 google doi
  • 17. Lee J. H., Cho K. M. (2012) Changes occurring in compositional components of black soybeans maintained at room temperature for different storage periods. [Food Chemistry] Vol.131 P.161-169 google doi
  • 18. Lee J. H., Choung M. G. (2011) Determination of optimal acid hydrolysis time of soybean isoflavones using drying oven and microwave assisted methods. [Food Chemistry] Vol.129 P.577-582 google doi
  • 19. Lee L. S., Choi E. J., Kim C. H., Kim Y.B., Kum J. S., Park J. D. (2014) Quality characteristics and antioxidant properties of black and yellow soybeans. [Korean Journal of Food Science and Technology] Vol.46 P.757-761 google doi
  • 20. Nam Y. D., Yi S. H., Lim S. I. (2012) Bacterial diversity of cheonggukjang, a traditional Korean fermented food, analyzed by barcoded pyrosequencing. [Food Control] Vol.28 P.135-142 google doi
  • 21. Prabhakaran M. P., Perera C. O., Valiyaveettil S. (2006) Effect of different coagulants on the isoflavones levels and physical properties of prepared firm tofu. [Food Chemistry] Vol.99 P.492-499 google doi
  • 22. Pratt D. E. (1980) Natural antioxidants of soybeans and other oil seeds. In: Simic, M. G., Karel M, editors. Autoxidation in food and biological systems P.283-293 google
  • 23. Shin E. C., Lee J. H., Hwang C. E., Lee B. W., Kim H. T., Ko J. M., Baek I. Y., Shin J. H., Nam S. H., Seo W. T., Cho K. M. (2014) Enhancement of total phenolic and isoflavone-aglycone contents and antioxidant activities during cheonggukjang fermentation of brown soybeans by the potential probiotic Bacillus subtilis CSY191. [Food Science and Biotechnology] Vol.23 P.531-538 google doi
  • 24. Shon M. Y., Lee J., Choi S. Y., Nam S. H., Seo K. I., Lee S. W., Sung N. J., Park S. K. (2007) Antioxidant and free radical scavenging activity of methanol extract of chungkukjang. [Journal of Food Composition and Analysis] Vol.20 P.113-118 google doi
  • 25. Slavin M., Cheng Z., Luther M., Kenworthy W., Yu L. (2009) Antioxidant properties and phenolic, isoflavone, tocopherol and carotenoid composition of Maryland-grown soybean lines with altered fatty acid profiles. [Food Chemistry] Vol.114 P.20-27 google doi
  • 26. Velioglu Y. S., Mazza G., Gao L., Oomah B. D. (1998) Antioxidant activity and total phenolics in selected fruits, vegetables and green products. [Journal of Agricultural and Food Chemistry] Vol.46 P.4113-4117 google doi
  • 27. Wang L., Yin L., Li D., Zou L., Saito M., Tatsumi E., Li L. (2007) Influecnes of processing and NaCl supplementation on isoflavone contents and composition during douche manufacturing. [Food Chemistry] Vol.101 P.1247-1253 google doi
  • 28. Yang S. O., Chang P. S., Lee J. H. (2006) Isoflavone distribution and -glycosidase activity in cheonggukjang, a traditional Korean whole soybean-fermented food. [Food Science and Biotechnology] Vol.15 P.96-101 google
  • [Table 1.] Change of viable cell, pH, and β-glucosidase activity during four black soybean cultivars with cheonggukjang fermentation by B. subtilis CSY191
    Change of viable cell, pH, and β-glucosidase activity during four black soybean cultivars with cheonggukjang fermentation by B. subtilis CSY191
  • [Fig. 1.] Change of total phenolic contents during cheonggukjang fermentation with four black soybean cultivars by B. subtilis CSY191. UFBS, unfermented black soybeans; and FBS, fermented black soybeans at 37℃ for 48 h. All values are means of determinations in three independent experiments. Means with different lowercase letters (a, b, c, d, and e) indicate significant differences of fermentation times by Tukey’s multiple range test (p< 0.05).
    Change of total phenolic contents during cheonggukjang fermentation with four black soybean cultivars by B. subtilis CSY191. UFBS, unfermented black soybeans; and FBS, fermented black soybeans at 37℃ for 48 h. All values are means of determinations in three independent experiments. Means with different lowercase letters (a, b, c, d, and e) indicate significant differences of fermentation times by Tukey’s multiple range test (p< 0.05).
  • [Fig. 2.] Change of isoflavone β-glucoside, -malonyl- β-glucoside, and -aglycones and total isoflavones contents during cheonggukjang fermentation with four black soybean cultivars by B. subtilis CSY191. UFBS, unfermented black soybeans; and FBS, fermented black soybeans at 37℃ for 48 h.
    Change of isoflavone β-glucoside, -malonyl- β-glucoside, and -aglycones and total isoflavones contents during cheonggukjang fermentation with four black soybean cultivars by B. subtilis CSY191. UFBS, unfermented black soybeans; and FBS, fermented black soybeans at 37℃ for 48 h.
  • [Fig. 3.] Typical HPLC chromatograms of isoflavones. HPLC chromatogram of isoflavone extract in cheonggukjang made with black soybean of (A) Cheongja cultivar according to fermentation period (0 h), (B) Cheongja cultivar according to fermentation period (48 h), (C) Cheongja#3 cultivar according to fermentation period (0 h), (D) Cheongja#3 cultivar according to fermentation period (48 h), (E) Geomjeong#5 cultivar according to fermentation period (0 h), (F) Geomjeong#5 cultivar according to fermentation period (48 h), (G) Ilpumgeomjeong cultivar according to fermentation period (0 h), and (H) Ilpumgeomjeong cultivar according to fermentation period (48 h). 1, diadzin; 2, glycitin; 3, genistin; 4, malonyldaidzin; 5, malonyl glycitin; 6, malonyl genistin; 7, daidzein; 8, glycitein; and 9, genistein.
    Typical HPLC chromatograms of isoflavones. HPLC chromatogram of isoflavone extract in cheonggukjang made with black soybean of (A) Cheongja cultivar according to fermentation period (0 h), (B) Cheongja cultivar according to fermentation period (48 h), (C) Cheongja#3 cultivar according to fermentation period (0 h), (D) Cheongja#3 cultivar according to fermentation period (48 h), (E) Geomjeong#5 cultivar according to fermentation period (0 h), (F) Geomjeong#5 cultivar according to fermentation period (48 h), (G) Ilpumgeomjeong cultivar according to fermentation period (0 h), and (H) Ilpumgeomjeong cultivar according to fermentation period (48 h). 1, diadzin; 2, glycitin; 3, genistin; 4, malonyldaidzin; 5, malonyl glycitin; 6, malonyl genistin; 7, daidzein; 8, glycitein; and 9, genistein.
  • [Table 2.] Distributions of isoflavone contents in cheonggukjang made of four black soybean cultivars by the B. subtilis CSY191
    Distributions of isoflavone contents in cheonggukjang made of four black soybean cultivars by the B. subtilis CSY191
  • [Fig. 4.] Change of diphenyl picrylhydrazyl (DPPH) radical scavenging activity during cheonggukjang fermentation with four black soybean cultivars by B. subtilis CSY191. UFBS, unfermented black soybeans; and FBS, fermented black soybeans at 37℃ for 48 h. All values are means of determinations in three independent experiments. Means with different lowercase letters (a, b, c, and d) indicate significant differences of fermentation times by Tukey’s multiple range test (p<0.05).
    Change of diphenyl picrylhydrazyl (DPPH) radical scavenging activity during cheonggukjang fermentation with four black soybean cultivars by B. subtilis CSY191. UFBS, unfermented black soybeans; and FBS, fermented black soybeans at 37℃ for 48 h. All values are means of determinations in three independent experiments. Means with different lowercase letters (a, b, c, and d) indicate significant differences of fermentation times by Tukey’s multiple range test (p<0.05).
  • [Fig. 5.] Change of ABTS radical scavenging activity during cheonggukjang fermentation with black soybean cultivars by B. subtilis CSY191. UFBS, unfermented black soybeans; and FBS, fermented black soybeans at 37℃ for 48 h. All values are means of determinations in three independent experiments. Means with different lowercase letters (a, b, c, and d) indicate significant differences of fermentation times by Tukey’s multiple range test (p< 0.05).
    Change of ABTS radical scavenging activity during cheonggukjang fermentation with black soybean cultivars by B. subtilis CSY191. UFBS, unfermented black soybeans; and FBS, fermented black soybeans at 37℃ for 48 h. All values are means of determinations in three independent experiments. Means with different lowercase letters (a, b, c, and d) indicate significant differences of fermentation times by Tukey’s multiple range test (p< 0.05).
  • [Fig. 6.] Change of ferric reducing/antioxidant power value during cheonggukjang fermentation with black soybean cultivars by B. subtilis CSY191. UFBS, unfermented black soybeans; and FBS, fermented black soybeans at 37℃ for 48 h. All values are means of determinations in three independent experiments. Means with different lowercase letters (a, b, c, and d) indicate significant differences of fermentation times by Tukey’s multiple range test (p< 0.05).
    Change of ferric reducing/antioxidant power value during cheonggukjang fermentation with black soybean cultivars by B. subtilis CSY191. UFBS, unfermented black soybeans; and FBS, fermented black soybeans at 37℃ for 48 h. All values are means of determinations in three independent experiments. Means with different lowercase letters (a, b, c, and d) indicate significant differences of fermentation times by Tukey’s multiple range test (p< 0.05).