Development of an industrial medium and a novel fed-batch strategy for high-level expression of recombinant β-mananase by Pichia pastoris.
An industrial medium, Corn Steep Liquor Powder Dextrose (CSD medium) was developed for constitutive expression of recombinant β-mananase by Pichia pastoris. The β-mananase activity (513 U/mL) with CSD medium was 1.64- and 2.5-fold higher than with YPD and BSM in shaken flasks. The β-mananase productivity with CSD medium was 61.0 U/mL h, which was 1.7- and 2.5-fold higher than with YPD and BSM in a 5-L fermenter based on a novel fed-batch strategy combining the real-time exponential feed mode with the DO-stat feed mode. The β-mananase activity, dry cell weight and the recombinant enzyme reached up to 5132 U/mL, 110.0 g/L and 4.50 g/L after 50 h cultivation in a 50-L fermenter. The high efficient expression of recombinant β-mananase by P. pastoris indicated that CSD medium and the novel fed-batch strategy have great potential for the production of recombinant β-mananase in industrial fermentation.
School of Minerals Processing and Bioengineering, Central South University, Changsha 410083, China.
Jia
J
Zheng
Wei
W
Zhao
Ning
N
Guo
Fulai
F
Lin
Jian
J
Tian
Lishuang
L
Wu
Hongbo
H
Zhou
D061249:Batch Cell Culture Techniques / Q000191:economics / Q000379:methods*; D018533:Biomass; D019149:Bioreactors / Q000191:economics / Q000382:microbiology; D003470:Culture Media / Q000138:chemical synthesis*; D005285:Fermentation; D006863:Hydrogen-Ion Concentration; D007218:Industrial Microbiology / Q000191:economics / Q000379:methods*; D010843:Pichia / Q000166:cytology / Q000201:enzymology* / Q000254:growth & development; D011994:Recombinant Proteins / Q000191:economics / Q000378:metabolism*; D013997:Time Factors; D044902:beta-Mannosidase / Q000191:economics / Q000378:metabolism*
D003470:Culture Media; D011994:Recombinant Proteins; D044902:beta-Mannosidase