Rare-Mating and Protoplast Fusion for the Improvement of Ethanol Producibility and Cell-Viability of Yeast

효모의 에탄올 생산능 및 세포 생존능의 증진을 위한 Rare-mating과 원형질체 융합

  • Kang, Tae-Young (Department of Genetic Engineering, The University of Suwon) ;
  • Kim, Keun (Department of Genetic Engineering, The University of Suwon)
  • 강태영 (수원대학교 유전공학과) ;
  • 김근 (수원대학교 유전공학과)
  • Published : 2001.12.01

Abstract

To improve the ethanol fermentability, four Saccharomyces yeast strains with efficient ethanol fermentability were subjected to rare-mating and protoplast fusion. Using these 4 strains, 5 different combinations of mating-pair or fusion-pair were constructed and their hybrids or fusants were obtained. From the statistical analysis of the results of the ethanol fermentation by the hybrids of the different mating-pair or fusion-pair, no difference was found in ethanol production, but [S. kluveri $khl{\times}S$ cerevisiae cp3] pair was shown to be the best combination which can produce high cell-viability. In fact, the clone No. 3 of the [S. kluveri $khl{\times}S$ cerevisiae cp3] pair was selected as the best strain which produced ethanol of 10.11% (w/v) or 12.81% (v/v) from 25% (w/v) glucose at $33^{\circ}C$ for 3 days with the residual sugar of 3.53% (w/v), viability of 62.65%, fermentation efficiency of 92.2%.

에탄올 발효능이 우수한 Saccharomyces에 속하는 4 균주를 가지고 여러 조합의 mating-pair 또는 fusion-pair를 만들고 이들 pair들로부터 만들어진 hybrid주들의 에탄올 생성능과 생존능을 통계적으로 분석한 결과, 에탄올 생성능에서는 차이가 없었으나, 생존능의 경우는 [S. kluveri $khl{\times}S.$ cerevisiae cp3]의 균주조합이 가장 우수한 hybrid를 낼 수 있는 것으로 나타났다. 실제로 에탄올 생성능과 잔당, 효율, 생존능에서 두루 우수한 균주는 [S. kluveri $khl{\times}S$ cerevisiae cp3] 조합에서 얻어진 융합주 clone No. 3가 에탄올 생성능 10.11%(w/v) 또는 12.81%(v/v), 잔당 3.53%(w/v), 생존능 62.65%, 발효 효율 92.2%로서 가장 발효능과 생존능이 우수한 균주로 선정되었다.

Keywords

References

  1. Method of enzymatic analysis v.3 Ethanol determination with alcohol dehydrogenase and β-NAD Bernet, T.;I. Gutman;H. U. Bergmeyer(ed.)
  2. Methods Enzymol. v.1 Amylases α and β Bernfeld, P.
  3. Biotechnol. Bioeng. Symp. v.8 Alternatives for energy savings at plant level for the production of alcohol for use as automotive fuel Espinosa, R.;V. Cojulun;F. Marroguim
  4. J. Bacteriol. v.135 Ployethylene Glycol-induceed fusion of heat-inactivated and living protoplast of Bacillus megaterium Fordor, K.;E. Dimiri
  5. Arch. Microbiol. v.115 Recombination after protoplast fusion in the yeast Candidia tropicalis Fouriner, P.;A. Provost;C. Bourguignon;H. Heslot
  6. Curr. Genet. v.12 Genetic analysis of highly ethanol-tolerant wine yeast Jimenez J.;T. Benitez
  7. Kor. J. Appl. Microbiol. Biotechnol. v.28 Isolation and identification of yeast strains producing high concentraion of ethanol with high viability Kang, T.Y.;G.H. Oh;K. Kim
  8. J. Microbiol. Biotechnol. v.8 Strain improvement of yeast for ethanol production using a combined treatment of electric field and chemical mutagen N-methyl-N'-nitro-N-nitrosoguanidine Kim, K.;J.Y. Lee.
  9. Kor. J. Appl. Microbial. Biotechnol. v.28 Protoplast fusion of Saccharomyces and Kluyveromyces to develop thermotolerant ethanol-producing yeast strains Kim, M-S;K. Kim
  10. Appl. Biochem. Biotechnol. v.59 The construction of a stable starch-fermenting yeast strain using genetic engineering and raremating Kim, T-G.;K. Kim
  11. Agricultural experimentation design and analysis Little, T.M.;F.J. Hills
  12. Biotechnol & Bioengin. Symp. v.11 Rapid determination of yeast viability Lee, S.S.;F.M. Robinson;H.Y. Wang
  13. Kor. J. Appl. Microbiol. Biotechnol. v.24 Improvement of starchfermentability of recombinant haploid yeast strain by hybridization Park, S-Y.;K. Kim;C-H. Lee.
  14. J. Am. Soc. Brew. Chem. v.43 Valuable techniques in the genetic manipulation of industrial strains Russell, L.;G.G. Stewart
  15. SAS/STAT guide for personal computers(ver 6.03) SAS institute Inc.
  16. Biotechnol. Lett. v.5 Genetic construction of yeast strains for high ethanol production Seki, T.;S. Myoga;S. Limtong;S. Uedono;J. Kumnuanta;H. Taguchi
  17. Method in genetics;laboratory course manual Sherman, F.;G. Fink;J.B. Hicks
  18. Kor. J. Appl. Microbiol. biotechnol. v.22 Improvement of ethanol-tolerance of haploid Saccharomyces diastaticus Song, S.H.;K. Kim;M.W. Lee
  19. Curr. Genet. v.9 The use of mitochondrial mutant in hybridization of industrial yeast strains Spencer, J.F.T.;C. Bizeau;N. Reynolds;D. M. Spencer
  20. Yeast;A practical approach Yeast genetics Spencer, J.F.T.;D.M. Spencer;A.R.W. Smith;I. Campbell(ed.);J.H.Duffus(ed.)