Overexpression of cyclodextrin glycosyltransferase gene from Brevibacillus brevis in Escherichia coli by control of temperature and mannitol concentration

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Title
Overexpression of cyclodextrin glycosyltransferase gene from Brevibacillus brevis in Escherichia coli by control of temperature and mannitol concentration
Author(s)
Myung Hee Kim; Jung Kee Lee; Hyung Kwoun Kim; Cheon Bae Sohn; Tae Kwang Oh
Bibliographic Citation
Biotechnology Techniques, vol. 13, pp. 765-770
Publication Year
1999
Abstract
Expression of Brevibacillus brevis CD 162 cyclodextrin glycosyltransferase (CGTase) gene using pET22b(+) vector in Escherichia coli BL21 (DE3) resulted in the formation of inactive inclusion bodies under the usual induction conditions. However, by lowering the induction temperature to 30°C and/or adding 0.5 M mannitol as an osmolyte, the formation of insoluble aggregates was prevented and about a 34-fold increase (8.51 U ml-1) in biologically active soluble form was achieved after 6 h induction. The active CGTase enzyme was estimated to comprise as much as 24% of the total soluble proteins. In addition, other polyols such as glycerol, erythritol, xylitol, sorbitol, and arabitol showed similar effects with mannitol on the production of active CGTase enzyme.
Keyword
Brevibacilluscyclodextringene expressionglycosyltransferasemannitolpolyol
ISSN
0951-208X
Publisher
Springer
DOI
http://dx.doi.org/10.1023/A:1008909105229
Type
Article
Appears in Collections:
Division of Biomedical Research > Metabolic Regulation Research Center > 1. Journal Articles
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