|
KMID : 0380020040190050368
|
|
Korean Journal of Biotechnology and Bioengineering
2004 Volume.19 No. 5 p.368 ~ p.373
|
|
|
Purification and Characterization of Glucosyltransferase and Fructosyltransferase in Leuconostoc mesenteroides NRRL B-1149
|
|
Lee Jin-Ha
Park Jun-Sung
Lee Hee-Sun
Kim Do-Man
|
|
|
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 yeast extract, peptone, K2HP04 and 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 polyacrylamide gel was isolated and it synthesized soluble fructan. The 1149GTase showed a band of 180 kDa on 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.
키워드
|
|
|
KEYWORD
|
|
|
Purification, glucosyltransferase, fructosyltransferase, response surface methodology
|
|
|
FullTexts / Linksout information
|
|
|
|
|
|
Listed journal information
|
|
|