Applied Microbiology and Biotechnology

, Volume 85, Issue 3, pp 553–561 | Cite as

Delayed supplementation of glycine enhances extracellular secretion of the recombinant α-cyclodextrin glycosyltransferase in Escherichia coli

  • Zhaofeng Li
  • Zhengbiao Gu
  • Miao Wang
  • Guocheng Du
  • Jing WuEmail author
  • Jian ChenEmail author
Biotechnological Products and Process Engineering


The targeting of recombinant proteins for secretion to the culture medium of Escherichia coli presents significant advantages over cytoplasmic or periplasmic expression. However, a major barrier is inadequate secretion across two cell membranes. In the present study, we attempted to circumvent this secretion problem of the recombinant α-cyclodextrin glycosyltransferase (α-CGTase) from Paenibacillus macerans strain JFB05-01. It was found that glycine could promote extracellular secretion of the recombinant α-CGTase for which one potential mechanism might be the increase in membrane permeability. However, further analysis indicated that glycine supplementation resulted in impaired cell growth, which adversely affected overall recombinant protein production. Significantly, delayed supplementation of glycine could control cell growth impairment exerted by glycine. As a result, if the supplementation of 1% glycine was optimally carried out at the middle of the exponential growth phase, the α-CGTase activity in the culture medium reached 28.5 U/ml at 44 h of culture, which was 11-fold higher than that of the culture in regular terrific broth medium and 1.2-fold higher than that of the culture supplemented with 1% glycine at the beginning of culture.


Cyclodextrin glycosyltransferase Paenibacillus macerans Extracellular secretion Glycine Escherichia coli Delayed supplementation 



This work was supported financially by the Natural Science Foundation of Jiangsu Province (BK2007019), the National Outstanding Youth Foundation of China (20625619), the National High-tech Research and Development Program of China (863 Program; 2006AA10Z335), Research Program of State Key Laboratory of Food Science and Technology (SKLF-MB-200802), Program of Innovation Team of Jiangnan University (2008CXTD01), and the Graduate Student Creative Research Program of Jiangsu Province in 2008 (CX08B_127Z).


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Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  1. 1.State Key Laboratory of Food Science and TechnologyJiangnan UniversityWuxiPeople’s Republic of China
  2. 2.School of Food Science and TechnologyJiangnan UniversityWuxiPeople’s Republic of China
  3. 3.School of Biotechnology and the Key Laboratory of Industrial Biotechnology, Ministry of EducationJiangnan UniversityWuxiPeople’s Republic of China

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