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Electronic Journal of Biotechnology
Universidad Católica de Valparaíso
ISSN: 0717-3458
Vol. 14, No. 6, 2011
Bioline Code: ej11070
Full paper language: English
Document type: Short Communication
Document available free of charge

Electronic Journal of Biotechnology, Vol. 14, No. 6, 2011

 en Expression and non-chromatographic purification of 1,3-propanediol oxidoreductase in Escherichia coli check for this species in other resources
Li, Wei; Ng, I-Son; Fang, Baishan & Zhang, Guangya

Abstract

The gene dhaT from Klebsiella pneumoniae check for this species in other resources encodes 1,3-propanediol oxidoreductase (PDOR). Thermally responsive elastin-like polypeptides (ELPs) was used as a fusion tag to purify the proteins (PDOR). The ELP gene was attached to dhaT and ligated into the pET-22b vector. Different NaCl concentrations were employed to decrease the transition temperature (Tt) which was diminished as salt concentration increased. The optimal final concentration of NaCl was 1 M and the corresponding Tt was 39.5ºC. Enzymatic assays were determined via every step for purification of fusion PDOR. PDOR showed good stability during the purification process, the specific activity in the first and second round of inverse transition cycling (ITC) was 276.1 ± 13.3 and 213.3 ± 10.8 U/mg, respectively. The ELPs fusion PDOR was superior to histidine tagged PDOR in both yield and activity after the purification.

Keywords
1,3-propanediol oxidoreductase, elastin-like polypeptides, Escherichia coli, fusion protein, non-chromatographic purification

 
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