Expression and fermentation optimization of oxidized polyvinyl alcohol hydrolase in E. coli |
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Authors: | Yu Yang Dongxu Zhang Song Liu Dongxu Jia Guocheng Du Jian Chen |
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Institution: | (1) The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Lihu Ave. 1800, Wuxi, 214122, Jiangsu, China; |
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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. |
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