The Korean Journal of Mycology (한국균학회지)

The Korean Society of Mycology (한국균학회)
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Intergeneric Hybrid Constructed by Nuclear Transfer of Saccharomycopsis into Saccharomyces

핵전이를 이용한 Saccharomycopsis 속과 Saccharomyces 속간의 잡종형성

  • Yang, Young-Ki (Department of Biological Sciences, College of Natural Sciences, Chosun University) ;
  • Lim, Chae-Young (Department of Biological Sciences, College of Natural Sciences, Chosun University) ;
  • Kang, Hee-Kyoung (Department of Biological Sciences, College of Natural Sciences, Chosun University) ;
  • Moon, Myeng-Nim (Department of Biological Sciences, College of Natural Sciences, Chosun University) ;
  • Rhee, Young-Ha (Department of Microbiology, College of Natural Sciences, Chungnam National University)
  • 양영기 (조선대학교 자연과학대학 생물과학부) ;
  • 임채영 (조선대학교 자연과학대학 생물과학부) ;
  • 강희경 (조선대학교 자연과학대학 생물과학부) ;
  • 문명님 (조선대학교 자연과학대학 생물과학부) ;
  • 이영하 (충남대학교 자연과학대학 미생물학과)
  • Published : 1999.12.30
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Abstract

Intergeneric hybrids between Saccharomyccopsis fiburigera KCTC 7393 and Saccharomyces cerevisiae KCTC 7049 (tyr-, ura-) were obtained by nuclear transfer technique. Nuclei isolated from the wild type S. fiburigera strain were transfered into auxotrophic S. cerevisiae mutants and new strains showing an increased starch degrading capability were selected. Maximum production of protoplasts was obtained from the treatment with 0.1 % Novozym 234 at $30^{\circ}C$ for 90 min, and most effective osmotic stabilizer for the isolation of protoplasts was 0.6 M KCl at pH 5.8. The frequency of protoplast regeneration was 14.64% under the conditions. Genectic stability, conidial size, DNA content, and nuclear stain suggested that the fusants were aneuploidy. The specific activity of ${\alpha}-amylase$ was observed to increase about $1.2{\sim}1.9$ folds.

${\alpha}-amylase$ 생산력이 우수한 야생형인 S. fiburigera의 핵을 순수분리하여 영양요구성 변이주인 S. cerevisiae(tyr-, ura-)의 원형질체에 전달하는 핵전이를 이용하여 종간잡종을 형성시킨 후 이들 형질전환체로부터 ${\alpha}-amylase$ 고생산 균주를 획득하였다. 핵전이를 위한 원형질체 형성 및 재생조건에서는 Novozym 234의 농도 1%, 삼투안정제는 0.6M KC1, 효소처리 시간은 90분, 그리고 최적 pH는 5.8로 나타났다. 핵전이에 의한 형질전환율은 0.25%로 비교적 낮은 편이었으며, 유전적 안정성, 세포크기의 체적, DNA함량의 측정 그리고 핵염색의 결과 형질전환체의 핵형은 aneuploid로 추정되었다. 형질전환 체의 ${\alpha}-amylase$ 활성은 모균주와 비교하여 약 $1.2{\sim}1.9$배 증가하였다.

Keywords

References

  1. Protoplast fusion in the yeast, Candida pseudotropicalis CBS 607 Bai, S.
  2. General Foods World v.21 Enzyme in starch processing Barfoed, H.C.
  3. Biotechnology Dictionary Biotechnology Dictionary Compilation Comimittee
  4. Hakkokogaku v.60 Production of cellulolytic enzymes by Penicillium verruculosum Chung, K.C.;Kawai, K.;Yoshima, S.;Eguchi, Y.
  5. Curr. Microbiol. v.7 Transfer of nuclei into protoplasts of Saccharomyces cerevisiae Ferenczy, L.;Pesti, M.
  6. 6th Intern. Protoplast Sumposium Current guestion of gene transfer via protoplast fusion in microorganism Ferenczy, L.
  7. Nature v.206 An improved diphenylamine method for estimation of deoxyribonucleic acid Giles, K.W.;Myers, W.
  8. J. Appl. Chem. Biotech. v.25 Hernandez, E.;Pilt, D.J.
  9. Trichoderma koningii. Kor. J. Microbiol. v.22 The conidial protoplast fusion of cellulolytic fungus Hong, S.W.;Hah, Y.C.;Park, H.M.
  10. The protoplast fusion from conidiospore of Penicillium verruculosum Kim, J.H.
  11. J. Ferment. Technol. v.64 Interspecific protoplast fusion of citric acid producing strain of Aspergilus niger Kirimura, K.;Yanguchi, T.;Usami, S.
  12. Nature v.227 Cleveage of structural proteins during the assembly of the head of bcteriophage T4 Laemmli, U.K.
  13. Kor. J. Mycol. v.17 Synergistic effect of substrate on the biosynthesis of cellulase and xylanase complexes from Aspergillus nidulans Lee, J.A.;Maeng, J.S.;Maeng, P.J.;Rhee, Y.H.
  14. Protoplast fusion Aspergillus oryzae and Saccharomyces cerevisiae Lee, S.Y.
  15. Charaterization of lipase producing hybrid constructed by intergeneric nuclear transfer between Aspergillus oryzae var. oryzae and Penicilllium verruculosum F-3 Lim, C.Y.
  16. J. Biol. Chem. v.193 Protein measuremant with the folin phenol reagent Lowry, O.H.;Rosevroung, N.J.;Farr, A.L.;Randall, R.J.
  17. J. Appl. Bacteriol. v.50 Characterization of thermostable ${\alpha}$-amylase from Bacillus licheniformis NCIB 6346 Morgan, F.J.;Priest, F.G.
  18. Can. J. Microbiol. v.27 Production of protoplast in different yeast by mutanase Nasim, A.;Stephen, O.E.R.
  19. J. Biochem. v.71 Production of extracellular enzymes in mutants isolated from Trichoderma viride Nevalaninen, K.M.H.;Palva, F.T.
  20. Br. Mycol. Soc. v.67 Factors affecting protoplast release in some filamentous fungi Trans Peberdy, J.F.;Buckley, C.E.;Daltrey, D.C.;Moore, P.M.
  21. Study on interspecific hybrid formation by nuclear transfer in Aspergillus spp Rho, H.S.
  22. Enzyme Handbook Vol. 4 Schomburg, D.;Salzmann, M.
  23. Appl. Environm. Microbiol. v.56 Transfer of isolated nuclei into protoplast of Trichoderma harzianum Sivan, A.;Harman, G.E.;Stasz, T.E.
  24. Microbios v.26 Protoplast from Aspergillus nidulans Van den Borek, H.W.;Stunninberg, J.G.;Wennekes, L.M.J.
  25. Agric. Biol. Chem. v.47 Molecular cloning of a glucoamylase proceeding gene in the yeast Saccharomyces Yamashita, I.;Fukui, S.
  26. Handbook of amylase and related enzymes, their sources isolation methods, properties and applications Yammamoto, T.
  27. Kor. J. Mycol. v.17 Construction of intraspecific hybrids by nuclear transfer in A. nidulans Yang, Y.K.;Park, Y.;Rhee, Y.H.;Maeng, P.J.