Archives of Microbiology

, Volume 201, Issue 2, pp 209–214 | Cite as

Fatty acid biosynthesis is enhanced in Escherichia coli strains with deletion in genes encoding the PII signaling proteins

  • Thiago Estefano Rodrigues
  • Guilherme Lanzi Sassaki
  • Gláucio Valdameri
  • Fábio Oliveira Pedrosa
  • Emanuel Maltempi Souza
  • Luciano Fernandes HuergoEmail author
Original Paper


The committed and rate-limiting step in fatty acid biosynthesis is catalyzed by acetyl-CoA carboxylase (ACC). In previous studies we showed that ACC activity is inhibited through interactions with the PII signaling proteins in vitro. Here we provide in vivo support for that model; we noted that PII proteins are able to reduce malonyl-CoA levels in vivo in Escherichia coli. Furthermore, we show that fatty acid biosynthesis is strongly enhanced in E. coli strains carrying deletions in PII coding genes. Given that PII proteins act as conserved negative regulators of ACC in Bacteria, our findings may be explored to engineer other prokaryotes to improve fatty acid yields, thereby turning microbial biofuel production economically competitive in the future.


Acetyl-CoA carboxylase PII proteins Free fatty acids Microbial biofuels Metabolic engineering 



This work was supported by CNPq and CAPES. We are grateful to Prof. Mike Merrick (John Innes Centre), Prof. Brian F. Pfleger (University of Wisconsin-Madison) and Prof. Hugo Gramajo (University of Rosario) for providing strains and plasmids used in this work.


  1. Atkinson MR, Ninfa AJ (1998) Role of the GlnK signal transduction protein in the regulation of nitrogen assimilation in Escherichia coli. Mol Microbiol 29:431–447CrossRefGoogle Scholar
  2. Atkinson MR, Blauwkamp TA, Ninfa AJ (2002) Context-dependent functions of the PII and GlnK signal transduction proteins in Escherichia coli. J Bacteriol 184:5364–5375CrossRefGoogle Scholar
  3. Baud S, Feria Bourrellier AB, Azzopardi M, Berger A, Dechorgnat J, Daniel-Vedele F et al (2010) PII is induced by WRINKLED1 and fine-tunes fatty acid composition in seeds of Arabidopsis thaliana. Plant J 64:291–303CrossRefGoogle Scholar
  4. Cho H, Cronan JE Jr (1995) Defective export of a periplasmic enzyme disrupts regulation of fatty acid synthesis. J Biol Chem 270:4216–4219CrossRefGoogle Scholar
  5. Climate Change Synthesis Report (2014) Intergovernamental panel on climate change (IPCC report, 2014). Accessed Mar 2018
  6. Cronan JE Jr, Waldrop GL (2002) Multi-subunit acetyl-CoA carboxylases. Prog Lipid Res 41:407–435CrossRefGoogle Scholar
  7. Davis MS, Cronan JE Jr (2001) Inhibition of Escherichia coli acetyl coenzyme A carboxylase by acyl-acyl carrier protein. J Bacteriol 183:1499–1503CrossRefGoogle Scholar
  8. Davis MS, Solbiati J, Cronan JE Jr (2000) Overproduction of acetyl-CoA carboxylase activity increases the rate of fatty acid biosynthesis in Escherichia coli. J Biol Chem 275:28593–28598CrossRefGoogle Scholar
  9. Evans A, Ribble W, Schexnaydre E, Waldrop GL (2017) Acetyl-CoA carboxylase from Escherichia coli exhibits a pronounced hysteresis when inhibited by palmitoyl-acyl carrier protein. Arch Biochem Biophys 636:100–109CrossRefGoogle Scholar
  10. Feria Bourrellier AB, Valot B, Guillot A, Ambard-Bretteville F, Vidal J, Hodges M (2010) Chloroplast acetyl-CoA carboxylase activity is 2-oxoglutarate-regulated by interaction of PII with the biotin carboxyl carrier subunit. Proc Natl Acad Sci USA 107:502–507CrossRefGoogle Scholar
  11. Forchhammer K (2008) P-II signal transducers: novel functional and structural insights. Trends Microbiol 16:65–72CrossRefGoogle Scholar
  12. Gerhardt EC, Rodrigues TE, Muller-Santos M, Pedrosa FO, Souza EM, Forchhammer K, Huergo LF (2015) The Bacterial signal transduction protein GlnB regulates the committed step in fatty acid biosynthesis by acting as a dissociable regulatory subunit of acetyl-CoA carboxylase. Mol Microbiol 95:1025–1035CrossRefGoogle Scholar
  13. Hauf W, Schmid K, Gerhardt EC, Huergo LF, Forchhammer K (2016) Interaction of the nitrogen regulatory protein GlnB (PII) with biotin carboxyl carrier protein (BCCP) controls acetyl-CoA levels in the cyanobacterium Synechocystis sp. PCC 6803. Front Microbiol 7:1700CrossRefGoogle Scholar
  14. Huergo LF, Dixon R (2015) The emergence of 2-oxoglutarate as a master regulator metabolite. Microbiol Mol Biol Rev 79:419–435CrossRefGoogle Scholar
  15. Huergo LF, Filipaki A, Chubatsu LS, Yates MG, Steffens MB, Pedrosa FO, Souza EM (2005) Effect of the over-expression of PII and PZ proteins on the nitrogenase activity of Azospirillum brasilense. FEMS Microbiol Lett 253:47–54CrossRefGoogle Scholar
  16. Huergo LF, Chandra G, Merrick M (2013) P(II) signal transduction proteins: nitrogen regulation and beyond. FEMS Microbiol Rev 37:251–283CrossRefGoogle Scholar
  17. Jiang P, Peliska JA, Ninfa AJ (1998) The regulation of Escherichia coli glutamine synthetase revisited: role of 2-ketoglutarate in the regulation of glutamine synthetase adenylylation state. Biochemistry 37:12802–12810CrossRefGoogle Scholar
  18. Kuwayama Y, Olmstead SM, Krupnick AJ (2013) Water resources and unconventional fossil fuel development: linking physical impacts to social costs. Accessed Mar 2018
  19. Lennen RM, Pfleger BF (2012) Engineering Escherichia coli to synthesize free fatty acids. Trends Biotechnol 30:659–667CrossRefGoogle Scholar
  20. Lennen RM, Braden DJ, West RA, Dumesic JA, Pfleger BF (2010) A process for microbial hydrocarbon synthesis: overproduction of fatty acids in Escherichia coli and catalytic conversion to alkanes. Biotechnol Bioeng 106:193–202CrossRefGoogle Scholar
  21. Liu T, Vora H, Khosla C (2010) Quantitative analysis and engineering of fatty acid biosynthesis in E. coli. Metab Eng 12:378–386CrossRefGoogle Scholar
  22. Liu H, Yu C, Feng D, Cheng T, Meng X, Liu W et al (2012) Production of extracellular fatty acid using engineered Escherichia coli. Microb Cell Fact 11:41CrossRefGoogle Scholar
  23. Macneil D, Zhu J, Brill WJ (1981) Regulation of nitrogen fixation in Klebsiella pneumoniae: isolation and characterization of strains with nif–lac fusions. J Bacteriol 145:348–357Google Scholar
  24. Marella ER, Holkenbrink C, Siewers V, Borodina I (2018) Engineering microbial fatty acid metabolism for biofuels and biochemicals. Curr Opin Biotechnol 50:39–46CrossRefGoogle Scholar
  25. Merrick M (2014) Post-translational modification of PII signal transduction proteins. Front Microbiol 5:763Google Scholar
  26. Reyes-Ramirez F, Little R, Dixon R (2001) Role of Escherichia coli nitrogen regulatory genes in the nitrogen response of the Azotobacter vinelandii NifL–NifA complex. J Bacteriol 183:3076–3082CrossRefGoogle Scholar
  27. Rodrigues TE, Gerhardt EC, Oliveira MA, Chubatsu LS, Pedrosa FO, Souza EM et al (2014) Search for novel targets of the PII signal transduction protein in bacteria identifies the BCCP component of acetyl-CoA carboxylase as a PII binding partner. Mol Microbiol 91:751–761CrossRefGoogle Scholar
  28. Shin KS, Lee SK (2017) Introduction of an acetyl-CoA carboxylation bypass into Escherichia coli for enhanced free fatty acid production. Bioresour Technol 245:1627–1633CrossRefGoogle Scholar
  29. Tong L (2013) Structure and function of biotin-dependent carboxylases. Cell Mol Life Sci 70:863–891CrossRefGoogle Scholar
  30. Zalutskaya Z, Kharatyan N, Forchhammer K, Ermilova E (2015) Reduction of PII signaling protein enhances lipid body production in Chlamydomonas reinhardtii. Plant Sci 240:1–9CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Department of Biochemistry and Molecular BiologyUFPRCuritibaBrazil
  2. 2.Setor Litoral, UFPRMatinhosBrazil

Personalised recommendations