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Bioprocess Engineering

, Volume 21, Issue 4, pp 355–361 | Cite as

Studies on growth kinetics and plasmid stability of a recombinant Escherichia coli expressing a Schistosoma mansoni antigen

  • A. C. Chaves
  • F. G. C. Abath
  • J. L. Lima-Filho
  • J. M. S. Cabral
  • N. Lucena-Silva
Article

Abstract

A 13 kDa Schistosoma mansoni tegumental antigen (Sm13) was cloned and expressed in Escherichia coli. The fermentation product of 55 kDa corresponds to the maltose binding protein-recombinant Sm13 fusion protein (MBP-rSm13). The growth conditions for maximum IPTG inducible MBP-rSm13 production was investigated by examining the following parameters: innoculum size, agitation, temperature of induction and concentration of the inducer, IPTG. The maximum MBP-rSm13 production was achieved by two steps-fermentation. First, the recombinant strain was cultivated in a 37 °C orbital shaker, at 160 rpm, for 3 hours. The MBP-rSm13 was then induced by adding 0.3 mM IPTG and placing the culture in a 28 °C orbital shaker for 2 hours. Under these conditions the MBP-rSm13 production yield was approximately 50% of total intracellular proteins and the degradation by bacterial intracellular proteases seemed to be minimized. The plasmid stability was also studied during the fermentation of the Sm13-expressing E. coli in non induced and induced conditions. In both conditions there was a slightly plasmid loss before induction, however after the addition of IPTG the loss was increased. The rate of plasmid loss was 5.5 and 13.5 before and after IPTG induction respectively.

Abbreviations LB Lurian-Bertani medium; PBS phosphate-buffered saline pH 7.4; IPTG isopropil-6-D-thiogalactoside; MBP-rSm13 maltose binding protein-recombinant Sm13 fusion protein; SDS-PAGE sodium dodecyl sulphate-polyacrylamide gel electrophoresis. 

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Copyright information

© Springer-Verlag Berlin Heidelberg 1999

Authors and Affiliations

  • A. C. Chaves
    • 1
  • F. G. C. Abath
    • 2
  • J. L. Lima-Filho
    • 3
  • J. M. S. Cabral
    • 4
  • N. Lucena-Silva
    • 2
  1. 1.Departamento de Patologia-ICB e de Ciências Exatas e Naturais-FFPNM da Universidade Estadual de Pernambuco e Fundação de Ensino Superior de Olinda
  2. 2.Departamento de Imunologia do Centro de Pesquisas Aggeu Magalhães/Fundação Oswaldo Cruz e-mail: norma@cpqam.fiocruz.br
  3. 3.Departamento de Bioquímica e Laboratório de Imunologia Keizo Asami-UFPE, Brasil
  4. 4.Laboratório de Engenharia Bioquímica, Centro de Engenharia Biológica e Química do Instituto Superior Técnico de Lisboa, PortugalPT

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