Improving RNA content of salt-tolerant Zygosaccharomyces rouxii by atmospheric and room temperature plasma (ARTP) mutagenesis and its application in soy sauce brewing

  • Jian Guo
  • Wen Luo
  • Xue-ming Wu
  • Jun Fan
  • Wen-xue ZhangEmail author
  • Taikei Suyama
Original Paper


Derived from RNA, 5ʹ-ribonucleotides, especially Inosine-5ʹ-monophosphate (IMP) and guanosine-5ʹ-monophosphate (GMP), can enhance the umami taste of soy sauce. In this study, the RNA content of three different salt-tolerant yeasts was examined. The most valuable strain was subjected to atmospheric and room-temperature plasma (ARTP) mutagenesis, which improved its RNA content by 160.54%. Regular fermentation with RNA-enhanced strain failed to increase the amount of 5ʹ-ribonucleotides in the soy sauce due to hydrolysis by phosphatase. A two-stage fermentation strategy was then carried out. Aroma compounds were mainly synthesized in the first stage, and RNA-enriched biomass was massively produced in the second stage followed by heat treatment to inactivate phosphatase. After the proposed strategy was applied, IMP and GMP in the soy sauce reached 68.54 and 89.37 mg/L, respectively. Moreover, the amounts of key aroma compounds and organic acids significantly increased. Results may provide new insights for improving the quality of soy sauce through microorganism breeding and fermentation control.


5ʹ-Ribonucleotides Organic acids RNA Zygosaccharomyces rouxii 



This study was financially supported by the Natural Science Foundation of China (Grant No. 31871788).

Supplementary material

11274_2019_2743_MOESM1_ESM.doc (470 kb)
Electronic supplementary material 1 (DOC 470 kb)


  1. Beksan E, Schieberle P, Robert F, Blank I, Fay LB, Schlichtherle-Cerny H, Hofmann T (2003) Synthesis and sensory characterization of novel umami-tasting glutamate glycoconjugates. J Agric Food Chem 51:5428–5436CrossRefGoogle Scholar
  2. Cao S, Zhou X, Jin W, Wang F, Tu R, Han S, Ma F (2017) Improving of lipid productivity of the oleaginous microalgae Chlorella pyrenoidosa via atmospheric and room temperature plasma (ARTP). Bioresour Technol 244:1400–1406CrossRefGoogle Scholar
  3. Feng Y, Cui C, Zhao H, Gao X, Zhao M, Sun W (2013) Effect of koji fermentation on generation of volatile compounds in soy sauce production. Int J Food Sci Technol 48:609–619CrossRefGoogle Scholar
  4. Feng Y, Cai Y, Su G, Zhao H, Wang C, Zhao M (2014) Evaluation of aroma differences between high-salt liquid-state fermentation and low-salt solid-state fermentation soy sauces from China. Food Chem 145:126–134CrossRefGoogle Scholar
  5. Frewer LJ, Scholderer J, Bredahl L (2003) Communicating about the risks and benefits of genetically modified foods: The mediating role of trust. Risk Anal 23:1117–1133CrossRefGoogle Scholar
  6. Gao XL, Cui C, Zhao HF, Zhao MM, Yang L, Ren JY (2010) Changes in volatile aroma compounds of traditional Chinese-type soy sauce during moromi fermentation and heat treatment. Food Sci Biotechnol 19:889–898CrossRefGoogle Scholar
  7. Grüner W, Giegerich R, Strothmann D, Reidys C, Weber J, Hofacker IL, Schuster P (1996) Analysis of RNA sequence structure maps by exhaustive enumeration I. Neutral Netw 127:355–374Google Scholar
  8. Hayashida Y, Kuriyama H, Nishimura K, Slaughter JC (1999) Production of 4-hydroxyfuranones in simple media by fermentation. Biotechnol Lett 21:505–509CrossRefGoogle Scholar
  9. Kaneko S, Kumazawa K, Nishimura O (2012) Comparison of key aroma compounds in five different types of Japanese soy sauces by aroma extract dilution analysis (AEDA). J Agric Food Chem 60:3831–3836CrossRefGoogle Scholar
  10. Kaneko S, Kumazawa K, Nishimura O (2013) Studies on the key aroma compounds in raw (unheated) and heated Japanese soy sauce. J Agric Food Chem 61:3396–3402CrossRefGoogle Scholar
  11. Kawai M, Okiyama A, Ueda Y (2002) Taste enhancements between various amino acids and IMP. Chem Senses 27:739–745CrossRefGoogle Scholar
  12. Kim JS, Lee YS (2008) A study of chemical characteristics of soy sauce and mixed soy sauce: chemical characteristics of soy sauce. Eur Food Res Technol 227:933–944CrossRefGoogle Scholar
  13. Kong Y, Zhang LL, Zhang YY, Sun BG, Sun Y, Zhao J, Chen HT (2018) Evaluation of non-volatile taste components in commercial soy sauces. Int J Food Prop 21:1854–1866CrossRefGoogle Scholar
  14. Li X, Staszewski L, Xu H, Durick K, Zoller M, Adler E (2002) Human receptors for sweet and umami taste. Proc Natl Acad Sci USA 99:4692–4696CrossRefGoogle Scholar
  15. Lin Z, Luo Z, Dou J, Wang H, Qiu W, Cao J, Zhou C (2011) High RNA accumulation in Candida tropicalis is affected by specific growth rate and different medium composition. J China Pharm Univ 42:169–175Google Scholar
  16. Lioe HN, Wada K, Aoki T, Yasuda M (2007) Chemical and sensory characteristics of low molecular weight fractions obtained from three types of Japanese soy sauce (shoyu)–Koikuchi, tamari and shiro shoyu. Food Chem 100:1669–1677CrossRefGoogle Scholar
  17. Lioe HN, Selamat J, Yasuda M (2010) Soy sauce and its umami taste: a link from the past to current situation. J Food Sci 75:71–76CrossRefGoogle Scholar
  18. Marques AF, Teixeira NA, Gambaretto C, Sillero A, Günther Sillero MA (1998) IMP-GMP 5ʹ-nucleotidase from rat brain: activation by polyphosphates. J Neurochem 71:1241–1250CrossRefGoogle Scholar
  19. Olmedo Iturbe F, Gomez-Hernández J, Lopez-Munguia A (1994) Continuous production of 5ʹ-ribonucleotides from yeast RNA by hydrolysis with immobilized 5ʹ-phosphodiesterase and 5ʹ-adenylate deaminase. World J Microbiol Biotechnol 10:36–40CrossRefGoogle Scholar
  20. Qi W, Fan ZC, Wang CL, Hou LH, Wang XH, Liu JF, Cao XH (2014a) Comparative study of physiological adaptation to salt stress in the genome shuffled Candida versatilis and a wild-type salt-tolerant yeast strain. Eur Food Res Technol 238:675–682CrossRefGoogle Scholar
  21. Qi W, Hou LH, Guo HL, Wang CL, Fan ZC, Liu JF, Cao XH (2014b) Effect of salt-tolerant yeast of Candida versatilis and Zygosaccharomyces rouxii on the production of biogenic amines during soy sauce fermentation. J Sci Food Agric 94:1537–1542CrossRefGoogle Scholar
  22. Schmitt ME, Brown TA, Trumpower BL (1990) A rapid and simple method for preparation of RNA from Saccharomyces cerevisiae. Nucleic Acids Res 18:3091–3096CrossRefGoogle Scholar
  23. Soga T, Ross GA (1999) Simultaneous determination of inorganic anions, organic acids, amino acids and carbohydrates by capillary electrophoresis. J Chromatogr A 837:231–323CrossRefGoogle Scholar
  24. Steinhaus P, Schieberle P (2007) Characterization of the key aroma compounds in soy sauce using approaches of molecular sensory science. J Agric Food Chem 55:6262–6269CrossRefGoogle Scholar
  25. Su NW, Wang ML, Kwok KF, Lee MH (2005) Effects of temperature and sodium chloride concentration on the activities of proteases and amylases in soy sauce koji. J Agric Food Chem 53:1521–1525CrossRefGoogle Scholar
  26. Sukkwai S, Kijroongrojana K, Chonpracha P, Pujols KD, Alonso-Marenco José R, Ardoin R (2018) Effects of colorant concentration and ‘natural colour’ or ‘sodium content’ claim on saltiness perception, consumer liking and emotion, and purchase intent of dipping sauces. Int J Food Sci Technol 53:1246–1254CrossRefGoogle Scholar
  27. Sun SY, Jiang WG, Zhao YP (2010) Profile of volatile compounds in 12 Chinese soy sauces produced by a high-salt-diluted state fermentation. J Inst Brew 116:316–328CrossRefGoogle Scholar
  28. Taing O, Taing K (2007) Production of malic and succinic acids by sugar-tolerant yeast Zygosaccharomyces rouxii. Eur Food Res Technol 224:343–347CrossRefGoogle Scholar
  29. Tanaka Y, Watanabe J, Mogi Y (2012) Monitoring of the microbial communities involved in the soy sauce manufacturing process by PCR-denaturing gradient gel electrophoresis. Food Microbiol 31:100–106CrossRefGoogle Scholar
  30. Vandamme EJ, Soetaert W (2002) Bioflavours and fragrances via fermentation and biocatalysis. J Chem Technol Biotechnol 77:1323–1332CrossRefGoogle Scholar
  31. Yamaguchi S, Yoshikawa T (1971) Measurement of the relative taste intensity of some l-α-amino acids and 5ʹ-nucleotides. J Food Sci 36:846–849CrossRefGoogle Scholar
  32. Yongmei L, Xiaohong C, Mei J, Xin L, Rahman N, Mingsheng D, Yan G (2009) Biogenic amines in Chinese soy sauce. Food Control 20:593–597CrossRefGoogle Scholar
  33. Yoshikawa S, Kurihara H, Kawai Y, Yamazaki K, Tanaka A, Nishikiori T, Ohta T (2010) Effect of halotolerant starter microorganisms on chemical characteristics of fermented chum salmon (Oncorhynchus keta) sauce. J Agric Food Chem 58:6410–6417CrossRefGoogle Scholar
  34. Zhang D, Song H, Cheng H, Hao D, Wang H, Kan G, Yu D (2014) The acid phosphatase-encoding gene GmACP1 contributes to soybean tolerance to low-phosphorus stress. PLoS Genet 10:1004061–1004069CrossRefGoogle Scholar

Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  • Jian Guo
    • 1
    • 2
  • Wen Luo
    • 2
  • Xue-ming Wu
    • 2
  • Jun Fan
    • 2
  • Wen-xue Zhang
    • 1
    • 3
    Email author
  • Taikei Suyama
    • 4
  1. 1.College of Biomass Science and EngineeringSichuan UniversityChengduChina
  2. 2.Qianhe Condiment and Food Co., LtdMeishanChina
  3. 3.School of Liquor-Making EngineeringSichuan University Jinjiang CollegeMeishanChina
  4. 4.Akashi National College of TechnologyAkashiJapan

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