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