title: Expression and fermentation optimization of oxidized polyvinyl alcohol hydrolase in E. coli.
abstract: Oxidized polyvinyl alcohol (PVA) hydrolase (OPH) is a key enzyme in the degradation of PVA, suggesting that OPH has a great potential for application in textile desizing processes. In this study, the OPH gene from Sphingopyxis sp. 113P3 was modified, by artificial synthesis, for overexpression in Escherichia coli. The OPH gene, lacking the sequence encoding the original signal peptide, was inserted into pET-20b (+) expression vector, which was then used to transform E. coli BL21 (DE3). OPH expression was detected in culture medium in which the transformed E. coli BL21 (DE3) was grown. Nutritional and environmental conditions were investigated for improved production of OPH protein by the recombinant strain. The highest OPH activity measured was 47.54 U/mL and was reached after 84 h under optimal fermentation conditions; this level is 2.64-fold higher that obtained under sub-optimal conditions. The productivity of recombinant OPH reached 565.95 U/L/h. The effect of glycine on the secretion of recombinant OPH was examined by adding glycine to the culture medium to a final concentration of 200 mM. This concentration of glycine reduced the fermentation time by 24 h and increased the productivity of recombinant OPH to 733.17 U/L/h. Our results suggest that the recombinant strain reported here has great potential for use in industrial applications.
authors.affiliation: The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Lihu Ave. 1800, Wuxi, 214122, Jiangsu, China.
authors.forename: Yu
authors.initials: Y
authors.lastname: Yang
authors.forename: Dongxu
authors.initials: D
authors.lastname: Zhang
authors.forename: Song
authors.initials: S
authors.lastname: Liu
authors.forename: Dongxu
authors.initials: D
authors.lastname: Jia
authors.forename: Guocheng
authors.initials: G
authors.lastname: Du
authors.forename: Jian
authors.initials: J
authors.lastname: Chen
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000096:biosynthesis* / Q000235:genetics; D001483:Base Sequence; D002265:Carboxylic Ester Hydrolases / Q000096:biosynthesis* / Q000235:genetics; D003470:Culture Media; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D005285:Fermentation; D005998:Glycine / Q000494:pharmacology; D008969:Molecular Sequence Data; D042301:Sphingomonadaceae / Q000201:enzymology* / Q000235:genetics
chemical_list: D001426:Bacterial Proteins; D003470:Culture Media; D002265:Carboxylic Ester Hydrolases; D005998:Glycine