[Effects of different methanol feeding strategy on hirudin production in high-density fermentation by recombinant Pichia pastoris].
Four different methanol feeding modes were evaluated in the hirudin production in high-density fermentation by Pichia pastoris. It was difficult to avoid methanol excessive in the broth with the feeding strategy only based on DO level. On the other hand, the fluctuation in methanol concentration was observed with methanol feeding strategy by off-line gas chromatography. However, the stable methanol concentration was perfectly achieved by the on-line monitoring with methanol sensor. The supply of energy was improved by feeding glycerol at a limited rate as well as methanol in the induction phase. Therefore, the high cell dry weight (162 g/L) and high hirudin activity (2.4 x 10(4) ATU/mL or 1.7 g/L) was obtained in the fed-batch fermentation of recombinant Pichia pastoris by methanol-glycerol mixed feeding.
State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China.
Xiang-Shan
XS
Zhou
Wei-Min
WM
Fan
Yuan-Xing
YX
Zhang
D005285:Fermentation; D005990:Glycerol / Q000494:pharmacology; D006629:Hirudins / Q000096:biosynthesis*; D000432:Methanol / Q000494:pharmacology*; D010843:Pichia / Q000235:genetics / Q000378:metabolism*; D011995:Recombination, Genetic
D006629:Hirudins; D005990:Glycerol; D000432:Methanol