Metabolic engineering of itaconate production in Escherichia coli

Kiira S. Vuoristo (Corresponding Author), Astrid E. Mars, Jose Vidal Sangra, Jan Springer, Gerrit Eggink, Johan P.M. Sanders, Ruud A. Weusthuis

Research output: Contribution to journalArticleScientificpeer-review

35 Citations (Scopus)

Abstract

Interest in sustainable development has led to efforts to replace petrochemical-based monomers with biomass-based ones. Itaconic acid, a C5-dicarboxylic acid, is a potential monomer for the chemical industry with many prospective applications. cis-aconitate decarboxylase (CadA) is the key enzyme of itaconate production, converting the citric acid cycle intermediate cis-aconitate into itaconate. Heterologous expression of cadA from Aspergillus terreus in Escherichia coli resulted in low CadA activities and production of trace amounts of itaconate on Luria-Bertani (LB) medium (<10 mg/L). CadA was primarily present as inclusion bodies, explaining the low activity. The activity was significantly improved by using lower cultivation temperatures and mineral medium, and this resulted in enhanced itaconate titres (240 mg/L). The itaconate titre was further increased by introducing citrate synthase and aconitase from Corynebacterium glutamicum and by deleting the genes encoding phosphate acetyltransferase and lactate dehydrogenase. These deletions in E. coli’s central metabolism resulted in the accumulation of pyruvate, which is a precursor for itaconate biosynthesis. As a result, itaconate production in aerobic bioreactor cultures was increased up to 690 mg/L. The maximum yield obtained was 0.09 mol itaconate/mol glucose. Strategies for a further improvement of itaconate production are discussed.

Original languageEnglish
Pages (from-to)221-228
Number of pages8
JournalApplied Microbiology and Biotechnology
Volume99
Issue number1
DOIs
Publication statusPublished - 2015
MoE publication typeA1 Journal article-refereed

Keywords

  • Aconitase
  • cis-aconitate decarboxylase
  • Citrate synthase
  • Escherichia coli
  • Itaconic acid
  • Metabolic engineering

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