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



Título del documento: Expression and non-chromatographic purification of 1,3-propanediol oxidoreductase in Escherichia coli
Revista: Electronic journal of biotechnology
Base de datos: PERIÓDICA
Número de sistema: 000359629
ISSN: 0717-3458
Autores: 1
2
2
3
Instituciones: 1Huaqiao University, The Key Laboratory for Industrial Biotechnology of Fujian Higher Education, Xiamen, Fujian. China
2Xiamen University, College of Chemistry and Chemical Engineering, Xiamen, Fujian. China
3Xiamen University, The Key Laboratory for Chemical Biology of Fujian Province, Xiamen, Fujian. China
Año:
Volumen: 14
Número: 6
Paginación: 10-10
País: Chile
Idioma: Inglés
Tipo de documento: Artículo
Enfoque: Experimental
Resumen en inglés 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
Disciplinas: Biología,
Química
Palabras clave: Bacterias,
Genética,
Bioquímica,
1,3-propanodiol oxidorreductasa,
Elastina-como Polipéptidos,
Escherichia coli,
Proteína de fusión,
Purificación no cromatográfica
Keyword: Biology,
Chemistry,
Bacteria,
Genetics,
Biochemistry,
1,3-propanediol oxidoreductase,
Elastin-like polypeptides,
Escherichia coli,
Fusion protein,
Non-chromatographic purification
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