Research Article: A two-stage temperature control strategy enhances extracellular secretion of recombinant α-cyclodextrin glucosyltransferase in Escherichia coli

Date Published: August 23, 2017

Publisher: Springer Berlin Heidelberg

Author(s): Yang Li, Jia Liu, Yinglan Wang, Bingjie Liu, Xiaofang Xie, Rui Jia, Caiming Li, Zhaofeng Li.


The effects of temperature on extracellular secretion of the α-cyclodextrin glucosyltransferase (α-CGTase) from Paenibacillus macerans JFB05-01 by Escherichia coli were investigated. When protein expression was induced at constant temperature, the greatest amount of extracellular recombinant α-CGTase was produced at 25 °C. Higher or lower induction temperatures were not conducive to extracellular secretion of recombinant α-CGTase. To enhance extracellular secretion of α-CGTase by E. coli, a two-stage temperature control strategy was adopted. When expression was induced at 25 °C for 32 h, and then the temperature was shifted to 30 °C, the extracellular α-CGTase activity at 90 h was 45% higher than that observed when induction was performed at a constant temperature of 25 °C. Further experiments suggested that raising the induction temperature can benefit the transport of recombinant enzyme and compensate for the decreased rate of recombinant enzyme synthesis during the later stage of expression. This report provides a new method of optimizing the secretory expression of recombinant enzymes by E. coli.

Partial Text

The cyclic oligosaccharides α-, β-, and γ-cyclodextrin consist of 6, 7, and 8 glucose units, respectively, linked by α-1, 4-glycosidic bonds. Cyclodextrins form inclusion complexes with many different small, hydrophobic guest molecules, improving their solubility and stability in aqueous environments. This property makes it have many applications in scientific, medical and industrial fields (Roy et al. 2017). The industrial use of α-cyclodextrin is in its infancy, yet is still expanding because of its small internal cavity, high water solubility, and resistance to enzymatic hydrolysis. Previous reports have shown that α-cyclodextrin can be used as a carrier of active ingredients, a solubilizer of lipids, a stabilizer of oils, a modifier of flavors or aromas, and a natural soluble dietary fiber (Aytac and Uyar 2016; Li et al. 2010b, 2014a).




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