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The Korean Society for Biotechnology and Bioengineering (한국생물공학회)
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Purification and Characterization of Glucosyltransferase and Fructosyltransferase in Leuconostoc mesenteroides NRRL B-1149

Leuconostoc mesenteroides NRRL B-1149의 Glucosyltransferase와 Fructosyltransferase의 분리와 특성 연구

  • Lee Jin Ha (The Engineering Research Institute, Chonnam National University) ;
  • Park Jun Seong (Department of Materials and Biochemical Engineering, Chonnam National University) ;
  • Lee Hee Sun (Department of Molecular Biotechnology, Chonnam National University) ;
  • Kim Do Man (School of Biological Sciences and Technology and Research Institue for Catalysis, Chonnam National University)
  • 이진하 (전남대학교 공업기술연구소) ;
  • 박준성 (전남대학교 물질생물화학공학과) ;
  • 이희선 (전남대학교 분자생물공학협동과정) ;
  • 김도만 (전남대학교 생명과학기술학부, 촉매연구소)
  • Published : 2004.10.01
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Abstract

The optimal condition for the production of a glucan and a fructan synthesizing enzymes from Leuconostoc mesenteroides NRRL B-1149 were studied based on the different medium compositions. Response surface methodology was applied to find the optimistic condition showing the relationship between the fermentation response (enzyme activities) and the fermentation variable concentrations of yeast extract, peptone concentration, K2HP04 concentration and sucrose. Optimum medium composition for both enzymes production was $0.75\%$ yeast extract, $0.72\%$ peptone, $1\%$ K2HP04 and $2.17\%$ sucrose. Using this medium, the activities produced in culture was 0.90 U/m~ for glucosyltransferase (GTase) and 0.96 U/ml for fructosyltransferase (FTase). After purification of 1149FTase by consecutive chromatographies using Sephadex G-150 and DEAE-Sepharose, a 1149FTase of 210 kDa on $7\%$ polyacrylamide gel was isolated and it synthesized soluble fructan. The 1149GTase showed a band of 180 kDa on $8\%$ polyacrylamide gel after purification using Bio-Gel P-100 gel chromatography and DEAE-Sepharose ion exchange chromatography and it synthesized insoluble glucan. The linkages of polymers were determined by methylation using Hakomori reagent and following NMR analysis. The glucan was composed of a(1~6) and a(1~3) linkages and the fructan was levan.

Keywords

References

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