Characteristics of yeast with low temperature adaptation for Yakju brewed

약주 제조를 위한 저온 적응성 효모의 특성

  • Seo, Dong-Jun (Fermented Food Division, Department of Agro-food Resource, NAAS, RDA) ;
  • Yeo, Soo-Hwan (Fermented Food Division, Department of Agro-food Resource, NAAS, RDA) ;
  • Mun, Ji-Young (Fermented Food Division, Department of Agro-food Resource, NAAS, RDA) ;
  • Jung, Woo-Jin (Division of Applied Bioscience and Biotechnology, Institute of Environmentally-Friendly Agriculture, Chonnam National University) ;
  • Cho, Yong Sik (Fermented Food Division, Department of Agro-food Resource, NAAS, RDA) ;
  • Baek, Seong Yeol (Fermented Food Division, Department of Agro-food Resource, NAAS, RDA)
  • 서동준 (농촌진흥청 국립농업과학원 농식품자원부 발효식품과) ;
  • 여수환 (농촌진흥청 국립농업과학원 농식품자원부 발효식품과) ;
  • 문지영 (농촌진흥청 국립농업과학원 농식품자원부 발효식품과) ;
  • 정우진 (전남대학교 농업생명대학 친환경농업센터, 농식품생명화학부) ;
  • 조용식 (농촌진흥청 국립농업과학원 농식품자원부 발효식품과) ;
  • 백성열 (농촌진흥청 국립농업과학원 농식품자원부 발효식품과)
  • Received : 2015.02.02
  • Accepted : 2015.04.10
  • Published : 2015.12.30


The objectives of this study were to isolate and characterize low temperature adaptation yeast and to obtain suitable yeasts strains for manufacturing Yakju. In this study, we isolated 482 wild yeasts from fermented foods. Out of these, 5 yeast strains were selected based on increased growth at low temperature ($15^{\circ}C$) and high ${\beta}$-glucosidase activity. To screen the aromatic level of isolates, media containing cerulenin and 5,5,5-trifluor-DL-leucine (TFL) were used. Y297 strain demonstrated tolerance against TFL and produced more than 13% alcohol. Y297 strain was identified a Saccharomyces cerevisiae based on the 26S rDNA gene sequences. Maximum cell growth was observed after 19 hr and 38 hr of incubation at $25^{\circ}C$ and $15^{\circ}C$, respectively. The exponential phase was followed by a lengthy stationary phase, at $15^{\circ}C$, when the cells remained high viable. Y297 strain demonstrated tolerance against alcohol (10%), glucose (60%) and salt(NaCl, 8%). ${\beta}$-glucosidase and esterase activity in Y297 were higher than those of controls at $15^{\circ}C$. Overall, these results indicated that using wild yeast strain, isolated from fermented food, affects the chemical characteristics of the brewed Yakju.


  1. Baek SY, Lee YJ, Kim JH, Yeo SH (2015) Isolation and characterization of wild yeasts for improving liquor flavors style and quality. Microbiol Biotechnol Lett, 43, 1-9
  2. Lee SR (1986) Korean Fermentation Foods. Eeha women's university press, Seoul, Korea, p 224-294
  3. Shin KR, Kim BC, Wang JY (1999) Characterization of Yakju prepared with yeasts from fruits. J Korean Soc Food Sci Nutr, 28, 794-800
  4. Beltran G, Rozes N, Mas A, Guillamon JM (2007) Effect of low-temperature fermentation on yeast nitrogen metabolism. World J Microbiol Biotechnol, 23, 809-815
  5. Chiva R, Lopez-malo M, Salvado Z, Mas A, Guillamon JM (2012) Analysis of low temperature-induced gene (LTIG) in wine yeast during alcoholic fermentation. FEMS Yeast Res, 831-843
  6. Torija MJ, Rozes N, poblet M, Guillamon JM, Mas A (2003) Effects of fermentation temperature on the strain population of Saccharomyces cerevisiae. Int J Food Microbiol, 80, 47-53
  7. Engan S, Aubert O (1977) Relations between fermentation temperature and the formation of some favour components. In: 16th European brewery convention congress, Amsterdam, Netherlands, p 591-607
  8. Schade B, Jansen G, Whiteway M, Entian KD, Thomas DY (2004) Clod adaption in budding yeast. Mol Biol Cell, 15, 5492-5502
  9. Aguilera J, Randez-Gil F, Prieto JA (2007) Cold response in Saccharomyces cerevisiae : new functions for old mechanisms. FEMS Microbiol Rev, 31, 327-341
  10. Lilly M, Lambrechts MG, Pretorius IS (2000) Effect of increased yeast alcohol acetyltransferase activity on flavor profiles of wine and distillates. Appl Environ Microbiol, 66, 744-753
  11. Mason AB, Dufour JP (2000) Alcohol acetyltransferase and the significance of ester synthesis in yeast. Yeast, 16, 1287-1298<1287::AID-YEA613>3.0.CO;2-I
  12. Cauet G, Degryse E, Ledoux C, Spagnol R, Achstetter T (1999) Pregnenolone esterification in Sacchromyces cerevisiae. A potential detoxification mechanism. Eur J Microchem, 261, 317-324
  13. Fujii T, Nagasawa N, Iwamatsu A, Bogaki T, Tamai Y, Hamachi M (1994) Molecular cloning, sequence analysis, and expression of the yeast alcohol acetyltransferase gene. Appl Environ Microbiol, 60, 2786-2792
  14. Fukuda K, Kiyokawa Y, Yanagiuchi T, Wakai Y, Kitamoto K, Inoue Y (2000) Purification and characterization of isoamyl acetate-hydrolyzing esterase encoded by the IAH1 gene of Saccharomyces cerevisiae from a recombinant Escherichia coli. Appl Microbiol Biothchnol, 53, 596-600
  15. Saerens SMG, Verstrepen KJ, Van laere SDM, Voet ARD, Van Dijck P, Delvaux FR (2006) The Saccharomyces cerevisiae EHT1 and EEB1 genes encode novel enzymes with medium-chain fatty acid ethyl ester synthesis and hydrolysis capacity. J Biologic Chem, 281, 4446-4456
  16. Verstrepen KJ, Van Laere SDM, Vanderhaegen BMP, Derdelinckx G, Dufour JP, Pretorius IS (2003) Expression levels of the yeast alcohol acetyltransferase genes ATF1, Lg-ATF, and ATF2 control the formation of a broad range of volatile esters. Appl Environ Microbiol, 69, 5228-5237
  17. Ichikawa E, Hosokawa N, Hata Y, Abe Y, Suginami K, Imayasu S (1991) Breeding of sake yeast with improved ethyl caproate productivity. Agric Biol Chem, 55, 2153-2154
  18. Bussry H, Umbarger HE (1970) Biosynthesis of branched chain amino acids in yeast: a trifluoroleucine-sesistant mutant with altered regulation of leucine uptake. J Bacteriol, 103, 286-294
  19. Comitini F, Gobbi M, Domizio P, Romani C, Lencioni L Mannazzu I, Ciani M (2011) Selected non-Saccharomyces cerevisiae wine yeasts in controlled multistarter fermentations with Saccharomyces cerevisiae. Food Microbiol, 28, 873-882
  20. Grover AK, Macmurche DD, Cushley RJ (1977) Studies on almond emulsion $\beta$-D-glucosidase. I. Isolation and characterization of a bifunctional isozyme. Biochem Biophys Acta, 482, 98-108
  21. Frank B, Bjorn D, Sabrina R, Christian Z, Helmut S, Manfred JS (2006) Cell surface expression of bacterial esterase A by Saccharomyces cerevisiae and its enhancement by constitutive activation of the cellular unfolded protein response. Appl Environ Microbiol, 72, 7140-7147
  22. Jung HK, Park CD, Park HH, Lee GD, Lee IS, Hong JH (2006) Manufacturing and characteristic of Korea traditional liquor, Hahyangju prepared by Saccharomyces cerevisiae HA3 isolated from traditional Nuruk. Korean J Food Sci Technol, 38, 659-667
  23. Hernandez LF, Espinosa JC, Fernandez-Gonzalez M, Briones A (2003) $\beta$-Glucosidase activity in a Saccharomyces cerevisiae wine strain. Int J Food Microbiol, 80, 171-176
  24. Strauss MLA, Jolly NP, Lambrechts MG van Rensburg P (2001) Screening for the production of extracellular hydrolytic enzymes by non-Saccharomyces cerevisiae wine yeasts. Food Microbiol, 28, 873-882
  25. Yamaoka C, Kurita O, Kubo T (2014) Improved ethanol tolerance of Sacchromyces cerevisiae in mixed cultures with Kluyveromyces lactis on high-sugar fermentation. Microbiol Res, 169, 907-914
  26. Wu G, Wang G, Ji J, Li Y, Gao H, Wu J, Guan W (2015) A chimeric vacuolar Na+/H+ antiporter gene evolved by DNA family shuffling confers increased salt tolerance in yeast. J Biotechnol, 203, 1-8