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Comparative reaction engineering studies for succinic acid production from sucrose by metabolically engineered E s cherichia coli in fed‐batch‐operated stirred tank bioreactors
Author(s) -
Hoefel Torben,
Faust Georg,
Reinecke Liv,
Rudinger Nicolas,
WeusterBotz Dirk
Publication year - 2012
Publication title -
biotechnology journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.144
H-Index - 84
eISSN - 1860-7314
pISSN - 1860-6768
DOI - 10.1002/biot.201200046
Subject(s) - bioreactor , succinic acid , corynebacterium glutamicum , metabolic engineering , biochemistry , sucrose , escherichia coli , industrial microbiology , biology , fed batch culture , fermentation , chemistry , enzyme , botany , gene
This study presents a comparative reaction engineering analysis of metabolically engineered sucrose‐utilizing Escherichia coli derived from E. coli K12 MG1655 for the anaerobic production of succinic acid. Production capacities of 16 different recombinant strains were evaluated in 48 parallel fed‐batch‐operated milliliter‐scale stirred tank bioreactors (10 mL) with continuous CO 2 sparging. The effects of recombinant sucrose‐utilization systems ( csc ‐operon or scr ‐operon), enhancements of anaplerotic reactions ( pck , ppc , maeA , maeB or heterologous pyc ) and gene deletions ( ldhA , adhE , ack‐pta and ptsG ) were studied with respect to the overall process performances of the respective recombinant strains. Both sucrose‐utilization systems enabled the production of succinic acid from sucrose in E. coli K12 MG1655. Maximum succinate production was observed by overexpressing the pyruvate carboxylase from Corynebacterium glutamicum resulting in a succinate concentration of 26.8 g L –1 after 48 h and a cell‐specific productivity of 0.14 g g –1 h –1 . Further experiments in a fed‐batch‐operated laboratory‐scale stirred tank bioreactor (2 L) showed that micro‐aerobic conditions preceding the anaerobic phase enhance succinic acid production of E. coli K12 MG1655‐derived strains. The work demonstrates the importance of parallel approaches within the scope of applied metabolic engineering studies.