Expression and non-chromatographic purification of 1,3-propanediol oxidoreductase in Escherichia coli

The gene dhaT from Klebsiella pneumoniae 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 w...

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Autores principales:	Li,Wei, Ng,I-Son, Fang,Baishan, Zhang,Guangya
Lenguaje:	English
Publicado:	Pontificia Universidad Católica de Valparaíso 2011
Materias:	1,3-propanediol oxidoreductase elastin-like polypeptides Escherichia coli fusion protein non-chromatographic purification
Acceso en línea:	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582011000600010
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spelling	oai:scielo:S0717-345820110006000102012-06-06Expression and non-chromatographic purification of 1,3-propanediol oxidoreductase in Escherichia coliLi,WeiNg,I-SonFang,BaishanZhang,Guangya 1,3-propanediol oxidoreductase elastin-like polypeptides Escherichia coli fusion protein non-chromatographic purification The gene dhaT from Klebsiella pneumoniae 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.info:eu-repo/semantics/openAccessPontificia Universidad Católica de ValparaísoElectronic Journal of Biotechnology v.14 n.6 20112011-11-01text/htmlhttp://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582011000600010en
institution	Scielo Chile
collection	Scielo Chile
language	English
topic	1,3-propanediol oxidoreductase elastin-like polypeptides Escherichia coli fusion protein non-chromatographic purification
spellingShingle	1,3-propanediol oxidoreductase elastin-like polypeptides Escherichia coli fusion protein non-chromatographic purification Li,Wei Ng,I-Son Fang,Baishan Zhang,Guangya Expression and non-chromatographic purification of 1,3-propanediol oxidoreductase in Escherichia coli
description	The gene dhaT from Klebsiella pneumoniae 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.
author	Li,Wei Ng,I-Son Fang,Baishan Zhang,Guangya
author_facet	Li,Wei Ng,I-Son Fang,Baishan Zhang,Guangya
author_sort	Li,Wei
title	Expression and non-chromatographic purification of 1,3-propanediol oxidoreductase in Escherichia coli
title_short	Expression and non-chromatographic purification of 1,3-propanediol oxidoreductase in Escherichia coli
title_full	Expression and non-chromatographic purification of 1,3-propanediol oxidoreductase in Escherichia coli
title_fullStr	Expression and non-chromatographic purification of 1,3-propanediol oxidoreductase in Escherichia coli
title_full_unstemmed	Expression and non-chromatographic purification of 1,3-propanediol oxidoreductase in Escherichia coli
title_sort	expression and non-chromatographic purification of 1,3-propanediol oxidoreductase in escherichia coli
publisher	Pontificia Universidad Católica de Valparaíso
publishDate	2011
url	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582011000600010
work_keys_str_mv	AT liwei expressionandnonchromatographicpurificationof13propanedioloxidoreductaseinescherichiacoli AT ngison expressionandnonchromatographicpurificationof13propanedioloxidoreductaseinescherichiacoli AT fangbaishan expressionandnonchromatographicpurificationof13propanedioloxidoreductaseinescherichiacoli AT zhangguangya expressionandnonchromatographicpurificationof13propanedioloxidoreductaseinescherichiacoli
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Expression and non-chromatographic purification of 1,3-propanediol oxidoreductase in Escherichia coli

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