Open AccessHuman G‐CSF synthesis using stress‐responsive bacterial proteins
Author(s) -
Song JongAm,
Han KyungYeon,
Park JinSeung,
Seo HyukSeong,
Ahn KeumYoung,
Lee Jeewon
Publication year - 2009
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.2009.01616.x
Subject(s) - escherichia coli , dihydrofolate reductase , inclusion bodies , biology , biochemistry , protein disulfide isomerase , fusion protein , cytoplasm , circular dichroism , population , microbiology and biotechnology , recombinant dna , enzyme , demography , sociology , gene
We previously reported that under the stress condition caused by the addition of 2‐hydroxyethyl disulfide, a thiol‐specific oxidant, to growing cultures of Escherichia coli BL21(DE3), a population of stress‐responsive proteins [peptidyl‐prolyl cis – trans isomerase B (PpiB), bacterioferritin (Bfr), putative HTH‐type transcriptional regulator yjdC (YjdC), dihydrofolate reductase (FolA), chemotaxis protein cheZ (CheZ), and glutathione synthetase (GshB)] were significantly upregulated when compared with the nonstress condition. When those stress‐responsive proteins were used as fusion partners for the expression of human granulocyte colony‐stimulating factor (hG‐CSF), the solubility of hG‐CSF was dramatically enhanced in E. coli cytoplasm, whereas almost all of the directly expressed hG‐CSF were aggregated to inclusion bodies. In addition, the spectra of circular dichroism measured with the purified hG‐CSF were identical to that of standard hG‐CSF, implying that the synthesized hG‐CSF has native conformation. These results indicate that the bacterial stress‐responsive proteins could be potent fusion expression partners for aggregation‐prone heterologous proteins in E. coli cytoplasm.