Expression difference of heterologous recombinant protein in Pichia pastoris with different specific growth rate (mu) was observed. The expression conditions of recombinant human interferon omega (rhIFN(omega)) in logarithm phrase Pichia pastoris with higher mu were optimized by shake flask tests under various initial pH, methanol concentration, duration of the induction, cell density, and medium volume. The results showed that there were prominent influences of mu on expression of rhlFN(omega). The maximum yield of rhIFN(omega) in the Pichia pastoris with mu of 0.1612 h(-1) was 558 mg/L. However, these in the Pichia pastoris with mu of 0.1321, 0.0505 and 0.0052 h(-1) were reduced by 50%, 68% and 99%, respectively. In 250 mL shake flask, the optimal medium volume, cell density, initial pH, methanol concentration, frequency and duration of methanol induction were 30 mL, 200 - 300 g/L (WCW), natural, 15 g/L, 1 time in every 24h and 4d, respectively. Under the optimal expression condition, the maximum yield of rhIFN(omega) in logarithm phrase Pichia pastoris was 1070 mg/L which was increased by 149% more than that under the initial condition.
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J Agric Food Chem
January 2025
State Key Laboratory of Materials-Oriented Chemical Engineering, College of Biotechnology and Pharmaceutical Engineering, Nanjing Tech University, Nanjing 211800, P.R. China.
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January 2025
School of Biotechnology and Key Laboratory of Industrial Biotechnology of Education, School of Biotechnology, Jiangnan University, Wuxi 214122 China. Electronic address:
Achieving enzyme catalysis at high substrate concentrations is a substantial challenge in industrial biocatalysis, and the role of glycosylation in post-translational modifications that modulate enzyme substrate inhibition remains poorly understood. This study provides insights into the role of N-glycosylation in substrate inhibition by comparing the catalytic properties of d-lactonohydrolase (d-Lac) derived from Fusarium moniliforme expressed in prokaryotic and eukaryotic hosts. Experimental evidence indicates that recombinant d-Lac expressed in Pichia pastoris (PpLac-WT) exhibits higher hydrolysis rates at a substrate concentration of 400 g/L, with reduced substrate inhibition and enhanced stability compared to the recombinant d-Lac expressed in Escherichia coli (EcLac-WT).
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College of Life Science and Technology, Wuhan Polytechnic University, Wuhan, 430023, China.
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Laboratory of Molecular Studies and Experimental Therapy-LEMTE, Department of Genetics, Federal University of Pernambuco, Recife 50670-901, Brazil.
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