Recombinant bioprocess optimization for heterologous protein production using two-stage, cyclic fed-batch culture

Applied Microbiology and Biotechnology
C C ChangD M Ogrydziak

Abstract

A two-stage, cyclic fed-batch bioprocess was designed, and its performance evaluated to improve rice alpha-amylase productivity by the yeast Yarrowia lipolytica SMY2 (MatA, ade1, ura3, xpr2), ATCC 201847, containing a replicative plasmid coding for a rice alpha-amlyase. Transcription of the recombinant gene is controlled by the XPR2 promoter. The first stage (or growth stage) was operated in the fed-batch mode, and the growth medium, designed to maintain a constant high cell density (i.e., 60 g/l), was fed according to a predetermined and preprogrammed optimal feed rate which, in turn, maintained the specific cell growth rate at an optimal value (i.e., 0.1 h-1). Typically, when the volume in the first stage reached a preset value, a portion of culture broth (i.e., 55%) was transferred to the second stage (or production stage). The remaining cells in the growth stage were then fed with fresh growth medium according to the bioprocess control strategy developed, while induction of alpha-amylase expression and its production was taking place in the second stage. The second stage was also operated in the fed-batch mode, and the production medium designed to maintain a constant high cell density and high productivity of heterologous ...Continue Reading

Citations

Mar 12, 2008·Applied Microbiology and Biotechnology·Dheepak MaharajhSantosh Ramchuran
Oct 1, 2005·Journal of Industrial Microbiology & Biotechnology·Daniel MandersonYusuf Chisti
Aug 5, 2000·Applied and Environmental Microbiology·G PignèdeJ M Nicaud
Sep 2, 2009·Progress in Lipid Research·Athanasios BeopoulosJean-Marc Nicaud
Apr 8, 2011·Biotechnology Journal·Jens FrickeReiner Luttmann
May 2, 2018·Microorganisms·Antonio Milton Vieira GomesNádia Skorupa Parachin
Jul 2, 2021·Microbial Cell Factories·Aravind MadhavanParameswaran Binod

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