|
KMID : 1007519980070010041
|
|
Food Science and Biotechnology
1998 Volume.7 No. 1 p.41 ~ p.45
|
|
|
Purification and Characterization of Kluyveromyces marxianus Inulinase from Recombinant Saccharomyces cerevisiae
|
|
Kim, Byung Woo
Kim, Youn Hee/NAM, SOO WAN/Kim, Han Woo
|
|
|
Abstract
|
|
|
|
The inulinase gene of Kluyveromyces marxianus was overexpressed by using GAL1 promoter in Saccharomyces cerevisiae. The recombinant inulinase produced in the extracellular medium was partially purified by ammonium sulfate precipitation and polyethylene glycol treatment. The specific activity of purified recombinant inulinase was 15.3 unit/㎎-protein toward inulin and 88.7 unit/㎎-protein toward sucrose, resulting in the S/I ratio of 5.8. The recombinant inulinase migrated as a disperse band at 94 to 171 kDa on SDS-polyacrylamide gel. The deglycosylated forms of recombinant and wild-type inulinases showed a distinct band at 62 kDa. The carbohydrate contents of recombinant and wild-type enzymes were estimated to be about 50% and 40% (w/w), respectively. The enzyme activity was stably maintained above 80% in the range of pH 4.0 to pH 8.0. The half life of the recombinant enzyme activity at 60℃ was about 50 min, whereas the wild-type was about 15 min. The enhanced thermal stability seems to be due to the higher content of carbohydrate moeity attached to the recombinant inulinase than that of the wild-type inulinase.
|
|
|
KEYWORD
|
|
|
|
|
|
FullTexts / Linksout information
|
|
|
|
|
|
Listed journal information
|
|
 
|