Cyclodextrin enhanced the soluble expression of Bacillus clarkii ?-CGTase in Escherichia coli.

ABSTRACT: BACKGROUND:Cyclodextrin glycosyltransferases (CGTases) catalyze the synthesis of cyclodextrins, which are circular ?-(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 ?-CGTase, requires their efficient production. In this study, the effects of chemical chaperones, temperature and inducers on cell growth and the production of soluble ?-CGTase by Escherichia coli were investigated. RESULTS:The yield of soluble ?-CGTase in shake-flask culture approximately doubled when ?-cyclodextrin was added to the culture medium as a chemical chaperone. When a modified two-stage feeding strategy incorporating 7.5 mM ?-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 ?-CGTase activity (50.29 U·mL-?1) was 1.71-fold greater than that observed in the absence of ?-cyclodextrin (29.33 U·mL-?1). CONCLUSIONS:Since ?-cyclodextrin is inexpensive and nontoxic to microbes, these results suggest its universal application during recombinant protein production. The higher expression of soluble ?-CGTase in a semi-synthetic medium showed the potential of the proposed process for the economical production of many enzymes on an industrial scale.

SUBMITTER: Wang L 

PROVIDER: S-EPMC6233531 | biostudies-literature | 2018 Nov

REPOSITORIES: biostudies-literature