Biotechnology Bulletin ›› 2022, Vol. 38 ›› Issue (4): 269-277.doi: 10.13560/j.cnki.biotech.bull.1985.2021-0148
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WANG Yue(), GAO Qing-hua, DONG Cong, LUO Tong-yang, WANG Qing-qing
Received:
2021-02-04
Online:
2022-04-26
Published:
2022-05-06
WANG Yue, GAO Qing-hua, DONG Cong, LUO Tong-yang, WANG Qing-qing. Expression of Pyranose Oxidase with Optimized Codon in Pichia pastoris[J]. Biotechnology Bulletin, 2022, 38(4): 269-277.
Fig. 1 Gene sequence after the optimization of codons The boxed sequences indicate restriction sites EcoR Iand Not I. The underlined sequences show 6×His-tag
Fig. 7 Production of P2O by high-density fermentation in 10 L bioreactor A:Growth curve of P2O activity. B:SDS-PAGE result. M:Protein marker. 1-12:Accumulation of recombinant protein after 0,12,24,36,48,60,72,84,96,108,120 and 132 h
Fig. 8 Optimal pH and pH stability A:Relative activity at different pH conditions at 30℃. B:The relative enzyme activity of P2O kept in the same temperature for 24 h at different pH conditions at 30℃. The used buffers were 100 mmol/L sodium citrate(■,black line),potassium phosphate(●,red line),Tris-HCl(▲,blue line)and glycine-NaOH(▼,pink line)
Fig. 9 Optimal temperature and temperature stability A:Relative activity of P2O at different temperatures at pH 6.5. B:The relative enzyme activity of P2O kept in the different temperatures for 30 min at pH 6.5
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