Advertisement

Applied Microbiology and Biotechnology

, Volume 98, Issue 2, pp 979–986 | Cite as

Comparison of CO-dependent H2 production with strong promoters in Thermococcus onnurineus NA1

  • Seong Hyuk Lee
  • Min-Sik Kim
  • Seung Seob Bae
  • Ae Ran Choi
  • Jin-Won Lee
  • Tae Wan Kim
  • Jung-Hyun Lee
  • Hyun Sook LeeEmail author
  • Sung Gyun KangEmail author
Bioenergy and biofuels

Abstract

To overproduce biotechnologically valuable products, the expression level of target genes has been modulated by using strong promoters. In a hyperthermophilic archaeon Thermococcus onnurineus NA1, two promoters, P TN0413 and P TN0157 , which drive expression of the genes encoding the S-layer protein and glutamate dehydrogenase were inserted in front of a gene cluster encoding a carbon monoxide dehydrogenase, a hydrogenase and a Na+/H+ antiporter. Two promoters exhibited strong activity by increasing the transcription and translation levels of the gene cluster in the mutant strains by 2.5- to 49-folds and 1.4- to 3.3-folds, respectively, than the native promoter in the wild-type strain. While KS0413 with P TN0413 promoter exhibited 2.7 to 4.7 times higher transcript level than KS0157 with P TN0157 promoter, the levels of proteins were a little different between them. The biomass concentrations and H2 production rates of two mutants were 2- to 3-fold higher than those of the wild-type strain in a bioreactor where CO was supplied at a flow rate of 120 ml min−1. Two mutants showed differential response to the higher CO flow rate, 240 ml min−1, in terms of growth pattern and product formation, indicating two promoters were regulated by culture conditions. The results demonstrate that not only promoter strength but also product-forming conditions should be considered in promoter engineering.

Keywords

Strong promoter Hyperthermophilic archaeon Carbon monoxide H2 production 

Notes

Acknowledgments

This work was supported by the KIOST in-house program, the Marine and Extreme Genome Research Centre, and the Development of Biohydrogen Production Technology Using the Hyperthermophilic Archaea Program of the Ministry of Oceans and Fisheries in the Republic of South Korea.

References

  1. Amend JP, Shock EL (2001) Energetics of overall metabolic reactions of thermophilic and hyperthermophilic archaea and bacteria. FEMS Microbiol Rev 25:175–243PubMedCrossRefGoogle Scholar
  2. Alper H, Fischer C, Nevoigt E, Stephanopoulos G (2005) Tuning genetic control through promoter engineering. Proc Natl Acad Sci USA 102:12678–12683PubMedCrossRefGoogle Scholar
  3. Bae SS, Kim YJ, Yang SH, Lim JK, Jeon JH, Lee HS, Kang SG, Kim SJ, Lee JH (2006) Thermococcus onnurineus sp. nov., a hyperthermophilic archaeon isolated from a deep-sea hydrothermal vent area at the PACMANUS field. J Microbial Biotechnol 16:1826–1831Google Scholar
  4. Bae SS, Kim TW, Lee HS, Kwon KK, Kim YJ, Kim MS, Lee JH, Kang SG (2012) H2 production from CO, formate or starch using the hyperthermophilic archaeon, Thermococcus onnurineus. Biotechnol Lett 34:75–79PubMedCrossRefGoogle Scholar
  5. Balch WE, Wolfe RS (1976) New approach to the cultivation of methanogenic bacteria: 2-mercaptoethanesulfonic acid (HS-CoM)-dependent growth of Methanobacterium ruminantium in a pressurized atmosphere. Appl Environ Microbiol 32:781–791PubMedCentralPubMedGoogle Scholar
  6. Balzer S, Kucharova V, Megerle J, Lale R, Brautaset T, Valla S (2013) A comparative analysis of the properties of regulated promoter systems commonly used for recombinant gene expression in Escherichia coli. Microb Cell Fact 12:26PubMedCentralPubMedCrossRefGoogle Scholar
  7. Cardoza RE, Moralejo FJ, Gutiérrez S, Casqueiro J, Fierro F, Martín JF (1998) Characterization and nitrogen-source regulation at the transcriptional level of the gdhA gene of Aspergillus awamori encoding an NADP-dependent glutamate dehydrogenase. Curr Genet 34:50–59PubMedCrossRefGoogle Scholar
  8. Cartwright CP, Li Y, Zhut YS (1994) Use of β-lactamase as a secreted reporter of promoter function in yeast. Yeast 10:497–508PubMedCrossRefGoogle Scholar
  9. Chandrayan SK, McTernan PM, Hopkins RC, Sun JS, Jenney FE, Adams MWW (2011) Engineering hyperthermophilic archaeon Pyrococcus furiosus to overproduce its cytoplasmic [NiFe]-Hydrogenase. J Biol Chem 287:3257–3264PubMedCrossRefGoogle Scholar
  10. Díez B, Mellado E, Rodríguez M, Bernasconi E, Barredo JL (1999) The NADP-dependent glutamate dehydrogenase gene from Penicillium chrysogenum and the construction of expression vectors for filamentous fungi. Appl Microbiol Biotechnol 52:196–207PubMedCrossRefGoogle Scholar
  11. de Smit MH, van Duin J (1994) Translational initiation on structured messengers. Another role for the Shine-Dalgarno interaction. J Mol Biol 235:173–184PubMedCrossRefGoogle Scholar
  12. Hammer K, Mijakovic I, Jensen PR (2006) Synthetic promoter libraries-tuning of gene expression. Trends Biotechnol 24:53–55PubMedCrossRefGoogle Scholar
  13. Holden JF, Takai K, Summit M, Bolton S, Zyskowski J, Baross JA (2001) Diversity among three novel groups of hyperthermophilic deep-sea Thermococcus species from three sites in the northeastern Pacific Ocean. FEMS Microbiol Ecol 36:51–60PubMedCrossRefGoogle Scholar
  14. Jeong JY, Yim HS, Ryu JY, Lee SH, Lee JH, Seen DS, Kang SG (2012) One-step sequence- and ligation-independent cloning as a rapid and versatile cloning method for functional genomics studies. Appl Environ Microbiol 78:5440–5443PubMedCentralPubMedCrossRefGoogle Scholar
  15. Kengen SWM, Stams AJM (1994) Formation of L-alanine as a reduced end-product in carbohydrate fermentation by the hyperthermophilic archaeon Pyrococcus furiosus. Arch Microbiol 161:168–175CrossRefGoogle Scholar
  16. Kim MS, Bae SS, Kim YJ, Kim TW, Lim JK, Lee SH, Choi AR, Jeon JH, Lee JH, Lee HS, Kang SG (2013) CO-dependent H2 production by a genetically engineered Thermococcus onnurineus NA1. Appl Environ Microbiol 79:2048–2053PubMedCentralPubMedCrossRefGoogle Scholar
  17. Kim YJ, Lee HS, Kim ES, Bae SS, Lim JK, Matsumi R, Lebedinsky AV, Sokolova TG, Kozhevnikova DA, Cha SS, Kim SJ, Kwon KK, Imanaka T, Atomi H, Bonch-Osmolovskaya EA, Lee JH, Kang SG (2010) Formate-driven growth coupled with H2 production. Nature 467:352–355PubMedCrossRefGoogle Scholar
  18. Kuen B, Lubitz W (1996) Analysis of S-layer proteins and genes. In: Sleytr UB, Messner P, Pum D, Sára M (eds) Crystalline bacterial cell surface proteins. R.G. Landes Comp. and Academic, Austin, pp 77–102Google Scholar
  19. Lee HS, Kang SG, Bae SS, Lim JK, Cho Y, Kim YJ, Jeon JH, Cha SS, Kwon KK, Kim HT, Park CJ, Lee HW, Kim SI, Chun J, Colwell RR, Kim SJ, Lee JH (2008) The complete genome sequence of Thermococcus onnurineus NA1 reveals a mixed heterotrophic and carboxydotrophic metabolism. J Bacteriol 190:7491–7499PubMedCentralPubMedCrossRefGoogle Scholar
  20. Li J, Wang J, Wang S, Xing M, Yu S, Liu G (2012) Achieving efficient protein expression in Trichoderma reesei by using strong constitutive promoters. Microbial Cell Fact 11:84CrossRefGoogle Scholar
  21. Lim JK, Kang SG, Lebedinsky AV, Lee JH, Lee HS (2010) Identification of a novel class of membrane-bound [NiFe]-hydrogenases in Thermococcus onnurineus NA1 by in silico analysis. Appl Environ Microbiol 76:6286–6289PubMedCentralPubMedCrossRefGoogle Scholar
  22. Matsumi R, Manabe K, Fukui T, Atomi H, Imanaka T (2007) Disruption of a sugar transporter gene cluster in a hyperthermophilic archaeon using a host-marker system based on antibiotic resistance. J Bacteriol 189:2683–2691PubMedCentralPubMedCrossRefGoogle Scholar
  23. Mueller M, Takemasa R, Schwarz A, Atomi H, Nidetzky B (2009) “Short-chain” α-1,4-glucan phosphorylase having a truncated N-terminal domain: functional expression and characterization of the enzyme from Sulfolobus solfataricus. Biochim Biophys Acta 1794:1709–1714PubMedCrossRefGoogle Scholar
  24. Pouwels P, Kolen CPAM, Boot HJ (1997) S-layer protein genes in Lactobacillus. FEMS Microbiol Rev 20:78–82Google Scholar
  25. Qi F, Yao L, Tan X, Lu X (2013) Construction, characterization and application of molecular tools for metabolic engineering of Synechocystis sp. Biotechnol Lett 35:1655–1661PubMedCrossRefGoogle Scholar
  26. Ragsdale SW (2004) Life with carbon monoxide. Crit Rev Biochem Mol Biol 39:165–195PubMedCrossRefGoogle Scholar
  27. Rud I, Jensen PR, Naterstad K, Axelsson L (2006) A synthetic promoter library for constitutive gene expression in Lactobacillus plantarum. Microbiology 152:1011–1019PubMedCrossRefGoogle Scholar
  28. Seghezzi N, Amar P, Koebmann B, Jensen PR, Virolle MJ (2011) The construction of a library of synthetic promoters revealed some specific features of strong Streptomyces promoters. Appl Microbiol Biotechnol 90:615–623PubMedCrossRefGoogle Scholar
  29. Sleytr UB, Messner P, Pum D, Sára M (1999) Crystalline bacterial cell surface layers (S layers): from supramolecular cell structure to biomimetics and nanotechnology. Angew Chem 38:1034–1054CrossRefGoogle Scholar
  30. Sokolova TG, Jeanthon C, Kostrikina NA, Chernyh NA, Lebedinsky AV, Stackebrandt E, Bonch-Osmolovskaya EA (2004) The first evidence of anaerobic CO oxidation coupled with H2 production by a hyperthermophilic archaeon isolated from a deep-sea hydrothermal vent. Extremophiles 8:317–323PubMedCrossRefGoogle Scholar
  31. Sun J, Shao Z, Nair N, Wen F, Xu J, Zhao H (2012) Cloning and characterization of a panel of constitutive promoters for applications in pathway engineering in Saccharomyces cerevisiae. Biotechnol Bioeng 109:2082–2092PubMedCrossRefGoogle Scholar
  32. Tangney M, Mitchell WJ, Dolberg-Rasmussen M (1999) Development of a shuttle vector for screening of strong promoter sequences. Biotechnol Tech 13:141–144CrossRefGoogle Scholar
  33. Takemasa R, Yokooji Y, Yamatsu A, Atomi H, Imanaka T (2011) Thermococcus kodakarensis as a host for gene expression and protein secretion. Appl Environ Microbiol 77:2392–2398PubMedCentralPubMedCrossRefGoogle Scholar
  34. van Gorcom RFM, Pouwels PH, Goosen T, Visser J, van den Broek HWJ, Hamer JE, Timberlake WE, van den Hondel CAMJJ (1985) Expression of an Escherichia coli beta-galactosidase fusion gene in Aspergillus nidulans. Gene 40:99–106PubMedCrossRefGoogle Scholar
  35. van Vliet AHM (2010) Next generation sequencing of microbial transcriptomes: challenges and opportunities. FEMS Microbiol Lett 302:1–7PubMedCrossRefGoogle Scholar
  36. Vellanoweth RL, Rabinowitz JC (1992) The influence of ribosome-binding-site elements on translational efficiency in Bacillus subtilis and Escherichia coli in vivo. Mol Microbiol 6:1105–1114PubMedCrossRefGoogle Scholar
  37. Wang T, Ma X, Du G, Chen J (2012) Overview of regulatory strategies and molecular elements in metabolic engineering of bacteria. Mol Biotechnol 52:300–308PubMedCrossRefGoogle Scholar
  38. Yokooji Y, Tomita H, Atomi H, Imanaka T (2009) Pantoate kinase and phosphopantothenate synthetase, two novel enzymes necessary for CoA biosynthesis in the archaea. J Biol Chem 284:28137–28145PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2013

Authors and Affiliations

  • Seong Hyuk Lee
    • 1
    • 2
  • Min-Sik Kim
    • 1
  • Seung Seob Bae
    • 1
    • 2
  • Ae Ran Choi
    • 1
  • Jin-Won Lee
    • 3
  • Tae Wan Kim
    • 1
    • 2
  • Jung-Hyun Lee
    • 1
    • 2
  • Hyun Sook Lee
    • 1
    • 2
    Email author
  • Sung Gyun Kang
    • 1
    • 2
    Email author
  1. 1.Korea Institute of Ocean Science and TechnologyAnsanSouth Korea
  2. 2.Department of Marine BiotechnologyUniversity of Science and TechnologyDaejeonSouth Korea
  3. 3.Department of Integrated BiotechnologySogang UniversitySeoulRepublic of Korea

Personalised recommendations