| Title : Biochemical characterization and high-level production of oxidized polyvinyl alcohol hydrolase from Sphingopyxis sp. 113P3 expressed in methylotrophic Pichia pastoris - Yang_2014_Bioprocess.Biosyst.Eng_37_777 |
| Author(s) : Yang Y , Liu L , Li J , Du G , Chen J |
| Ref : Bioprocess Biosyst Eng , 37 :777 , 2014 |
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Abstract :
The Sphingopyxis sp. 113P3 gene oph, encoding oxidized polyvinyl alcohol hydrolase (OPH), was optimized with the preferred codons of Pichia pastoris and ligated into the pPIC9K vector behind the alpha-factor signal sequence. The vector was then transfected into P. pastoris GS115 and genomic integration was confirmed. Large-scale production of recombinant protein was performed by induction with 14.4 g/L methanol at 22 degrees C in a 3-L bioreactor. The maximal OPH activity obtained was 68.4 U/mL, which is the highest activity reported. The optimal pH and temperature of recombinant OPH were 8.0 and 45 degrees C, respectively. OPH activity was stable over a pH range of 5.0-8.5, and at a maximal temperature of 45 degrees C. The K cat /K m of recombinant OPH was 598 mM(-1) s(-1), which was 4.27-fold higher than that of recombinant OPH derived from Escherichia coli. The improved catalytic efficiency of OPH expressed in recombinant P. pastoris makes it favorable for industrial applications. |
| PubMedSearch : Yang_2014_Bioprocess.Biosyst.Eng_37_777 |
| PubMedID: 24068496 |
Yang Y, Liu L, Li J, Du G, Chen J (2014)
Biochemical characterization and high-level production of oxidized polyvinyl alcohol hydrolase from Sphingopyxis sp. 113P3 expressed in methylotrophic Pichia pastoris
Bioprocess Biosyst Eng
37 :777
Yang Y, Liu L, Li J, Du G, Chen J (2014)
Bioprocess Biosyst Eng
37 :777