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Cyclodextrin enhanced the soluble expression of Bacillus clarkii γ-CGTase in Escherichia coli
- Source :
- BMC Biotechnology, Vol 18, Iss 1, Pp 1-9 (2018)
- Publication Year :
- 2018
- Publisher :
- BMC, 2018.
-
Abstract
- Abstract Background Cyclodextrin glycosyltransferases (CGTases) catalyze the synthesis of cyclodextrins, which are circular α-(1,4)-linked glucans used in many applications in the industries related to food, pharmaceuticals, cosmetics, chemicals, and agriculture, among others. Economic use of these CGTases, particularly γ-CGTase, requires their efficient production. In this study, the effects of chemical chaperones, temperature and inducers on cell growth and the production of soluble γ-CGTase by Escherichia coli were investigated. Results The yield of soluble γ-CGTase in shake-flask culture approximately doubled when β-cyclodextrin was added to the culture medium as a chemical chaperone. When a modified two-stage feeding strategy incorporating 7.5 mM β-cyclodextrin was used in a 3-L fermenter, a dry cell weight of 70.3 g·L− 1 was achieved. Using this cultivation approach, the total yield of γ-CGTase activity (50.29 U·mL− 1) was 1.71-fold greater than that observed in the absence of β-cyclodextrin (29.33 U·mL− 1). Conclusions Since β-cyclodextrin is inexpensive and nontoxic to microbes, these results suggest its universal application during recombinant protein production. The higher expression of soluble γ-CGTase in a semi-synthetic medium showed the potential of the proposed process for the economical production of many enzymes on an industrial scale.
Details
- Language :
- English
- ISSN :
- 14726750
- Volume :
- 18
- Issue :
- 1
- Database :
- Directory of Open Access Journals
- Journal :
- BMC Biotechnology
- Publication Type :
- Academic Journal
- Accession number :
- edsdoj.58eeea9ea8f4d8db4f1df8658f474d2
- Document Type :
- article
- Full Text :
- https://doi.org/10.1186/s12896-018-0480-8