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Process conditions affecting proteolysis of recombinant proteins in Escherichia coli
Authors:Shaojun Yang  Sven-Olof Enfors
Institution:(1) Department of Biochemistry and Biotechnology, Royal Institute of Technology, S-10044 Stockholm, Sweden;(2) Present address: Department of Biology, Hong Kong University of Science and Technology, Kowloon, Hong Kong
Abstract:The effects of process conditions on proteolysis of three recombinant IgG-binding proteins ZT3, ZZT3 and protein A in E. coli were studied. SDS/PAGE shows that the relative amount of degradation intermediates of ZT3 in a shake flask cultivation is much higher than that in a bioreactor culture. The rate of proteolysis of ZZT3 and protein A was also higher in shake flask cultures than in bioreactor cultures. The proteolysis rate constant of ZZT3 was 12/h in a shake flask but only 2.1/h in a bioreactor. Corresponding values for protein A were 2.4/h and 1.0/h, respectively. High proteolysis rate constants correlated with lower product yields.
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