Optimal expression and characterization of a fusion enzyme having dextransucrase and dextranase activities

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Title
Optimal expression and characterization of a fusion enzyme having dextransucrase and dextranase activities
Author(s)
H J Ryu; X Jin; J H Lee; H J Woo; Young Min Kim; G J Kim; E S Seo; H K Kang; J Kim; D L Cho; A Kimura; D Kim
Bibliographic Citation
Enzyme and Microbial Technology, vol. 47, no. 5, pp. 212-215
Publication Year
2010
Abstract
Long isomalto-oligosaccharides (IMOs) were generated via an engineered fusion enzyme of dextransucrase and dextranase (DSXR). To increase the expression level, response surface methodology (RSM) was utilized for optimization of protein expression conditions for enhancement of protein production by the effects of three-level-three-factors and their mutual interaction in Escherichia coli. Seventeen experiments were designed and conducted for investigation of cell density to start induction, induction temperature, and induction time. Optimal induction conditions included a cell density to start induction (A600) of 0.76 at 12.16°C for 18h for dextransucrase activity and a cell density to start induction (A600) of 0.75 at 10.5°C for 20.9h for dextranase activity. The produced dextransucrase or dextranase activity was obtained at 120.1±7.2 or 871±58U, respectively, from 1L cultures.
Keyword
DextransucraseFusion enzymeIsomalto-oligosaccharideDextranaseResponse surface methodology
ISSN
0141-0229
Publisher
Elsevier
DOI
http://dx.doi.org/10.1016/j.enzmictec.2010.07.006
Type
Article
Appears in Collections:
1. Journal Articles > Journal Articles
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