Functional expression of recombinant human trefoil factor 1 by Escherichia coli and Brevibacillus choshinensis

BMC Biotechnology

Yueh-Mei Cheng²  Meng-Ting Lu²  Chuan Mei Yeh¹ 

[1] Agricultural Biotechnology Center, National Chung-Hsing University, Taichung, Taiwan, Republic of China

[2] Department of Food Science and Biotechnology, National Chung-Hsing University, Taichung, Taiwan, Republic of China

关键词: Secretion;    Brevibacillus choshinensis;    Escherichia coli;    Glycosylated recombinant trefoil factor 1;   

Others  :  1210258
DOI  :  10.1186/s12896-015-0149-5

 received in 2014-12-15, accepted in 2015-04-22,  发布年份 2015

【 摘 要 】

Background

Trefoil factor 1 (TFF1) mediates mucosal repair and belongs to a highly conserved trefoil factor family proteins which are secreted by epithelial cells in the stomach or colon mucous membrane. TFF1 forms a homodimer via a disulphide linkage that affects wound healing activity. Previous recombinant expressions of TFF1 were too low yield for industrial application. This study aims to improve the expression level of bioactive recombinant TFF1 (rTFF1) and facilitate application potency.

Methods

The rTFF1 gene rtff1 was synthesized, expressed by Escherichia coli and secreted by Brevibacillus choshinensis. The rTFF1s were purified. The polymeric patterns and wound healing capacities of purified rTFF1s were checked.

Results

In Escherichia coli, 21.08 mg/L rTFF1 was stably expressed as monomer, dimer and oligomer in soluble fraction. In Brevebacillus choshinensis, the rTFF1 was secreted extracellularly at high level (35.73 mg/L) and formed monomer, dimer and oligomer forms. Both proteins from different sources were purified by Ni-NTA chromatography and exhibited the wound healing activities. The rTFF1 produced by B. choshinensis had better wound healing capability than the rTFF1 produced by E. coli. After pH 2.4 buffer treatments, the purified rTFF1 formed more oligomeric forms as well as better wound healing capability. Glycosylation assay and LC-MS/MS spectrometry experiments showed that the rTFF1 produced by B. choshinensis was unexpectedly glycosylated at N-terminal Ser residue. The glycosylation may contribute to the better wound healing capacity.

Conclusions

This study provides a potent tool of rTFF1 production to be applied in gastric damage protection and wound healing. The protein sources from B. choshinensis were more efficient than rTFF1 produced by E. coli.

【 授权许可】

   
2015 Cheng et al.; licensee BioMed Central.

【 图 表 】

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