Cultivation and Induction Process Exploration and Optimization of HPV 18 L1 Protein Expressing Pichia Pastorius in 30 L Sartotious Bioreactor
Author(s): XU Dao-jun, MA Bo, QIAN Wen, ZENG Xian-fang, WALVAX Biotechnology Co., Ltd., Shanghai Zerun Biotech Co., Ltd.
Pages: 38-44
Year: 2015 Issue: 1
Journal: Pharmaceutical Biotechnology
Keyword: Late HPV18L1 protein; Recombinant Pichia pastoris; Bioreactor; Glycerol feeding; Starvation; Methanol feeding;
Abstract: Using improved low-salt medium,the recombinant HPV18L1 late protein was expressed by Pichia pastorius at 30 L Sartorious bioreactor( C30-3). Various parameters affecting expression of HPV18L1 protein in accumulation and induction phases,such as medium components,p H value,temperature,stirring speed,dissolved oxygen,addition of glycerol and methanol,were evaluated.The results showed that the optimized low-salt medium is more suitable for the growth of the yeast Pichia cells. During accumulation stage,p H of the culture medium 5. 0,temperature 31℃,DO content of 20% to 25% and stirring speed 500 r / min were maintained,3 000 m L of 40% glycerol solution was added at a constant feed rate for the first 20 h until the Pichia wet weight ≥190 mg / m L and then feeding of glycerol was stopped and Pichia was cultivated for additional 4 h to exhaust glycerol. After entering the recombinant Pichia expression HPV18L1 protein induced late stage,to maintain the culture p H5. 2,temperature 29 ℃,DO content of 45%,stirring speed 600 r / min,methanol concentration control culture medium supplemented with 4% methanol online and induction 60 h.Late HPV18L1 protein expression reached 311. 43 mg / L. 30 L Stable expression of recombinant HPV18L1 protein by Pichia at 30 L Sartorious bioreactor( C30-3) was established.
Pages: 38-44
Year: 2015 Issue: 1
Journal: Pharmaceutical Biotechnology
Keyword: Late HPV18L1 protein; Recombinant Pichia pastoris; Bioreactor; Glycerol feeding; Starvation; Methanol feeding;
Abstract: Using improved low-salt medium,the recombinant HPV18L1 late protein was expressed by Pichia pastorius at 30 L Sartorious bioreactor( C30-3). Various parameters affecting expression of HPV18L1 protein in accumulation and induction phases,such as medium components,p H value,temperature,stirring speed,dissolved oxygen,addition of glycerol and methanol,were evaluated.The results showed that the optimized low-salt medium is more suitable for the growth of the yeast Pichia cells. During accumulation stage,p H of the culture medium 5. 0,temperature 31℃,DO content of 20% to 25% and stirring speed 500 r / min were maintained,3 000 m L of 40% glycerol solution was added at a constant feed rate for the first 20 h until the Pichia wet weight ≥190 mg / m L and then feeding of glycerol was stopped and Pichia was cultivated for additional 4 h to exhaust glycerol. After entering the recombinant Pichia expression HPV18L1 protein induced late stage,to maintain the culture p H5. 2,temperature 29 ℃,DO content of 45%,stirring speed 600 r / min,methanol concentration control culture medium supplemented with 4% methanol online and induction 60 h.Late HPV18L1 protein expression reached 311. 43 mg / L. 30 L Stable expression of recombinant HPV18L1 protein by Pichia at 30 L Sartorious bioreactor( C30-3) was established.
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