Applied Microbiology and Biotechnology

, Volume 100, Issue 21, pp 9103–9110 | Cite as

A novel cell autolysis system for cost-competitive downstream processing

  • Ivan Hajnal
  • Xiangbin Chen
  • Guo-Qiang ChenEmail author
Biotechnological products and process engineering


The industrial production of low value-added biological products poses significant challenges due to cost pressures. In recent years, it has been argued that synthetic biology approaches will lead to breakthroughs that eliminate price bottlenecks for the production of a wide range of biological products including bioplastics and biofuels. One significant bottleneck lies in the necessity to break the tough cell walls of microbes in order to release intracellular products. We here report the implementation of the first synthetic biology standard part based on the lambda phage SRRz genes and a synthetic ribosome binding site (RBS) that works in Escherichia coli and Halomonas campaniensis, which enables the producer strains to induce lysis after the addition of small amounts (1–5 %) of solvents or to spontaneously lyse during the stresses of downstream processing, and thus has the potential to eliminate the mechanical cell disruption step as both an efficiency bottleneck and a significant capex barrier when implementing downstream bioprocesses.


Autolysis PHB PHA Bioplastics Halomonas Synthetic biology 



Plasmid pSEVA311 was kindly donated by Prof. Victor de Lorenzo of Spanish National Centre of Biotechnology (CSIC, CNB, Syst Biol Program). This research was financially supported by 973 Basic Research Fund (Grant No. 2012CB725201) and a grant from National Natural Science Foundation of China (Grant No. 31430003).

Author contributions

Ivan Hajnal devised the experiments. Ivan Hajnal and Xiangbin Chen conducted the experiments. Guo-Qiang Chen supervised the studies and revised the manuscript.

Compliance with ethical standards

Conflict of interest

All authors declare that they have no conflict of interest.

Ethical approval

This article does not contain any studies with human participants or animals performed by any of the authors.

Supplementary material

253_2016_7669_MOESM1_ESM.pdf (928 kb)
ESM 1 (PDF 928 kb)


  1. Bläsi U, Chang CY, Zagotta MT, Nam KB, Young R (1990) The lethal lambda S gene encodes its own inhibitor. EMBO J 9:981PubMedPubMedCentralGoogle Scholar
  2. Cai Z, Xu W, Xue R, Lin Z (2008) Facile, reagentless and in situ release of Escherichia coli intracellular enzymes by heat-inducible autolytic vector for high-throughput screening. Protein Eng Des Sel 21:681–687CrossRefPubMedGoogle Scholar
  3. Durante-Rodríguez G, de Lorenzo V, Martínez-García E (2014) The Standard European Vector Architecture (SEVA) plasmid toolkit. In: Pseudomonas methods and protocols. Springer, New York, pp. 469–478CrossRefGoogle Scholar
  4. Fu XZ, Tan D, Aibaidula G, Wu Q, Chen JC, Chen GQ (2014) Development of Halomonas TD01 as a host for open production of chemicals. Metab Eng 23:78–91CrossRefPubMedGoogle Scholar
  5. Gao Y, Feng X, Xian M, Wang Q, Zhao G (2013) Inducible cell lysis systems in microbial production of bio-based chemicals. Appl Microbiol Biotechnol 97:7121–7129CrossRefPubMedGoogle Scholar
  6. García-Fruitós E, Vázquez E, Díez-Gil C, Corchero JL, Seras-Franzoso J, Ratera I, Veciana J, Villaverde A (2012) Bacterial inclusion bodies: making gold from waste. Trends Biotechnol 30:65–70CrossRefPubMedGoogle Scholar
  7. Gefen O, Fridman O, Ronin I, Balaban NQ (2014) Direct observation of single stationary-phase bacteria reveals a surprisingly long period of constant protein production activity. Proc Natl Acad Sci U S A 111:556–561CrossRefPubMedGoogle Scholar
  8. Grima EM, Belarbi EH, Fernández FA, Medina AR, Chisti Y (2003) Recovery of microalgal biomass and metabolites: process options and economics. Biotechnol Adv 20:491–515CrossRefGoogle Scholar
  9. Gründling A, Manson MD, Young R (2001) Holins kill without warning. Proc Natl Acad Sci U S A 98:9348–9352CrossRefPubMedPubMedCentralGoogle Scholar
  10. Jacquel N, Lo CW, Wei YH, Wu HS, Wang SS (2008) Isolation and purification of bacterial poly (3-hydroxyalkanoates). Biochem Eng J 39:15–27CrossRefGoogle Scholar
  11. Li R, Zhang HX, Qi QS (2007) The production of polyhydroxyalkanoates in recombinant Escherichia coli. Appl Microbiol Biotechnol 98:2313–2320Google Scholar
  12. Li T, Chen XB, Chen JC, Wu Q, Chen GQ (2014) Open and continuous fermentation: products, conditions and bioprocess economy. Biotechnol J 9:1503–1511CrossRefPubMedGoogle Scholar
  13. Liu X, Curtiss R (2009) Nickel-inducible lysis system in Synechocystis sp PCC 6803. Proc Natl Acad Sci U S A 106:21550–21554CrossRefPubMedPubMedCentralGoogle Scholar
  14. Minamino T, Imae Y, Oosawa F, Kobayashi Y, Oosawa K (2003) Effect of intracellular pH on rotational speed of bacterial flagellar motors. J Bacteriol 185:1190–1194CrossRefPubMedPubMedCentralGoogle Scholar
  15. Park T, Struck DK, Deaton JF, Young R (2006) Topological dynamics of holins in programmed bacterial lysis. Proc Natl Acad Sci U S A 103:19713–19718CrossRefPubMedPubMedCentralGoogle Scholar
  16. Pédelacq JD, Cabantous S, Tran T, Terwilliger TC, Waldo GS (2006) Engineering and characterization of a superfolder green fluorescent protein. Nat Biotechnol 24:79–88CrossRefPubMedGoogle Scholar
  17. Pylypiw HM, Grether MT (2000) Rapid high-performance liquid chromatography method for the analysis of sodium benzoate and potassium sorbate in foods. J Chromatogr A 883:299–304CrossRefPubMedGoogle Scholar
  18. Resch S, Gruber K, Wanner G, Slater S, Dennis D, Lubitz W (1998) Aqueous release and purification of poly (β-hydroxybutyrate) from Escherichia coli. J Biotechnol 65:173–182CrossRefPubMedGoogle Scholar
  19. Romano I, Giordano A, Lama L, Nicolaus B, Gambacorta A (2005) Halomonas campaniensis sp nov, a haloalkaliphilic bacterium isolated from a mineral pool of Campania region, Italy. Syst Appl Microbiol 28:610–618CrossRefPubMedGoogle Scholar
  20. Salmond CV, Kroll RG, Booth IR (1984) The effect of food preservatives on pH homeostasis in Escherichia coli. J Gen Microbiol 130:2845–2850PubMedGoogle Scholar
  21. Silva-Rocha R, Martínez-García E, Calles B, Chavarría M, Arce-Rodríguez A, de las Heras A, Platero R, de Lorenzo V (2013) The standard European vector architecture (SEVA): a coherent platform for the analysis and deployment of complex prokaryotic phenotypes. Nucleic Acids Res 41:666–675CrossRefGoogle Scholar
  22. Simon R, Priefer U, Pühler A (1983) A broad host range mobilization system for in vivo genetic engineering: transposon mutagenesis in gram negative bacteria. Nat Biotechnol 1:784–791CrossRefGoogle Scholar
  23. Wang Y, Yin J, Chen GQ (2014) Polyhydroxyalkanoates, challenges and opportunities. Curr Opin Biotechnol 30:59–65CrossRefPubMedGoogle Scholar
  24. White R, Tran TA, Dankenbring CA, Deaton J, Young R (2010) The N-terminal transmembrane domain of λ S is required for holin but not antiholin function. J Bacteriol 192:725–733CrossRefPubMedGoogle Scholar
  25. Xu L, Li S, Ren C, Cai Z, Lin Z (2006) Heat-inducible autolytic vector for high-throughput screening. Biotechniques 41:319CrossRefPubMedGoogle Scholar
  26. Young RY (1992) Bacteriophage lysis: mechanism and regulation. Microbiol Rev 56:430PubMedPubMedCentralGoogle Scholar
  27. Young RY (2002) Bacteriophage holins: deadly diversity. J Mol Microbiol 4:21–36Google Scholar
  28. Young RY, Wang N, Roof WD (2000) Phages will out: strategies of host cell lysis. Trends Microbiol 8:120–128CrossRefPubMedGoogle Scholar
  29. Yue H, Ling C, Yang T, Chen X, Chen Y, Deng H, Chen GQ (2014) A seawater-based open and continuous process for polyhydroxyalkanoates production by recombinant Halomonas campaniensis LS21 grown in mixed substrates. Biotechnol Biofuels 7:108–119CrossRefGoogle Scholar
  30. Zhang X, Pan Z, Fang Q, Zheng J, Hu M, Jiao X (2009) An auto-inducible Escherichia coli lysis system controlled by magnesium. J Microbiol Methods 79:199–204CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2016

Authors and Affiliations

  1. 1.Peking-Tsinghua Center for Life Sciences, School of Life ScienceTsinghua UniversityBeijingChina
  2. 2.Center for Synthetic and Systems BiologyTsinghua UniversityBeijingChina
  3. 3.MOE Key Lab of Industrial BiocatalysisTsinghua UniversityBeijingChina

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