Skip to main content
SpringerLink
Log in

Deletion of putative oxidoreductases from Klebsiella pneumoniae J2B could reduce 1,3-propanediol during the production of 3-hydroxypropionic acid from glycerol

  • Research Paper
  • Published:
Biotechnology and Bioprocess Engineering Aims and scope Submit manuscript

Abstract

Recombinant Klebsiella pneumoniae over-expressing 3-hydroxypropionaldehyde (3-HPA) dehydrogenase can produce 3-hydroxypropionic acid (3-HP), an important platform chemical, from glycerol. However, K. pneumoniae co-produces 1,3-propanediol (1,3-PDO) due to the presence of 1,3-propanediol oxidoreductases, which decreases the titer and yield of 3-HP. Previously, two major oxidoreductases, dhaT and yqhD, were removed from K. pneumoniae; however the mutant still produced a significant amount of 1,3-PDO, indicating the probable existence of other oxidoreductase(s). Genome analysis of K. pneumoniae revealed the presence of five putative oxidoreductases having high amino acid similarities to both DhaT (primary 1,3-propanediol oxidoreductase) and YqhD (aldehyde dehydrogenase). Among them, adhE was highly expressed in the absence of DhaT and YqhD. Additionally, an alkyl hydroperoxide oxidoreductase (ahpF), albeit dissimilar to both DhaT and YqhD, was highly expressed in the absence of DhaT and YqhD. To examine the role of adhE and ahpF in 1,3-PDO production, mutant strains devoid of dhaT, yqhD, ahpF and/or adhE genes were developed. However, these mutants neither reduced the production of 1,3-PDO nor improved the production of 3-HP when engineered to over-express an aldehyde dehydrogenase (KGSADH). These results indicate that, apart from DhaT, YqhD, AhpF and AdhE, K. pneumoniae has other, unknown oxidoreductases that are involved in 1,3-PDO production. It is concluded that complete elimination of 1,3-PDO during 3-HP production from glycerol by K. pneumoniae is highly challenging.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Wang, Z., Z. Wu, and T. Tan (2013) Studies on purification of 1, 3-propanediol by molecular distillation. Biotechnol. Bioproc. Eng., 18: 697–702.

    Article  CAS  Google Scholar 

  2. Gokarn, R. R., O. V. Selifonova, H. J. Jessen, S. J. Gort, T. Selmer, and W. Buckel (2007) 3-Hydroxypropionic acid and other organic compounds. US Patent 7,186,541B2.

    Google Scholar 

  3. Werpy, T. and G. Petersen (2004) Top value added chemicals from biomass, volume 1: Results of screening for potential candidates from sugars and synthesis gas. US Department of Energy. [http://www.osti.gov/bridge].

    Google Scholar 

  4. Sankaranarayanan, M., S. Ashok, and S. Park (2014) Production of 3-hydroxypropionic acid from glycerol by acid tolerant Escherichia coli. J. Ind. Microbiol. Biotechnol., 41: 1039–1050.

    Article  CAS  Google Scholar 

  5. Rathnasingh, C., S. M. Raj, J. E. Jo, and S. Park (2009) Development and evaluation of efficient recombinant Escherichia coli strains for the production of 3-hydroxypropionic acid from glycerol. Biotechnol. Bioeng., 104: 729–739.

    CAS  Google Scholar 

  6. Arasu, M. V., R. Sarkar, B. S. Sekar, V. Kumar, C. Rathnasingh, H. Song, D. Seung, and S. Park (2013) Isolation of a novel Pseudomonas species SP2 producing vitamin B12 under aerobic condition. Biotechnol. Bioproc. Eng., 18: 43–51.

    Article  CAS  Google Scholar 

  7. Ashok, S., S. Mohan Raj, Y. Ko, M. Sankaranarayanan, S. Zhou, V. Kumar, and S. Park (2013) Effect of puuC overexpression and nitrate addition on glycerol metabolism and anaerobic 3-hydroxypropionic acid production in recombinant Klebsiella pneumoniae ΔglpKΔdhaT. Metab. Eng., 15: 10–24.

    Article  CAS  Google Scholar 

  8. Huang, Y., Z. Li, K. Shimizu, and Q. Ye (2013) Co-production of 3-hydroxypropionic acid and 1,3-propanediol by Klebseilla pneumoniae expressing aldH under microaerobic conditions. Bioresour. Technol., 128: 505–512.

    Article  CAS  Google Scholar 

  9. Ashok, S., S. M. Raj, C. Rathnasingh, and S. Park (2011) Development of recombinant Klebsiella pneumoniae ΔdhaT strain for the co-production of 3-hydroxypropionic acid and 1,3-propanediol from glycerol. Appl. Microbiol. Biotechnol., 90: 1253–1265.

    Article  CAS  Google Scholar 

  10. Huang, Y., Z. Li, K. Shimizu, and Q. Ye (2012) Simultaneous production of 3-hydroxypropionic acid and 1,3-propanediol from glycerol by a recombinant strain of Klebsiella pneumoniae. Bioresour. Technol., 103: 351–359.

    Article  CAS  Google Scholar 

  11. Ko, Y., S. Ashok, S. Zhou, V. Kumar, and S. Park (2012) Aldehyde dehydrogenase activity is important to the production of 3-hydroxypropionic acid from glycerol by recombinant Klebsiella pneumoniae. Proc. Biochem., 47: 1135–1143.

    Article  CAS  Google Scholar 

  12. Jarboe, L. R. (2011) YqhD: A broad-substrate range aldehyde reductase with various applications in production of biorenewable fuels and chemicals. Appl. Microbiol. Biotechnol., 89: 249–257.

    Article  CAS  Google Scholar 

  13. Ashok, S., M. Sankaranarayanan, Y. Ko, K. E. Jae, S. K. Ainala, V. Kumar, and S. Park (2013) Production of 3-hydroxypropionic acid from glycerol by recombinant Klebsiella pneumoniae ΔdhaTΔyqhD which can produce vitamin B12 naturally. Biotechnol. Bioeng., 110: 511–524.

    Article  CAS  Google Scholar 

  14. Hall, R. H. and E. S. Stern (1950) Acid-catalysed hydration of acrylaldehyde: Kinetics of the reaction and isolation of b-hydroxypropaldehyde. J. Chem. Soc. 490–498.]

    Google Scholar 

  15. Laemmli, U. K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. JNature, 227: 680–685.

    CAS  Google Scholar 

  16. Yim, S. H., T. M. Kim, H. J. Hu, J. H. Kim, B. J. Kim, J. Y. Lee, B. G. Han, S. H. Shin, S. H. Jung, and Y. J. Chung (2010) Copy number variations in East-Asian population and their evolutionary and functional implications. Hum. Mol. Genet., 19: 1001–1008.

    Article  CAS  Google Scholar 

  17. Link, A. J., D. Phillips, and G. M. Church (1997) Methods for generating precise deletions and insertions in the genome of wild-type Escherichia coli: Application to open reading frame characterization. J. Bacteriol., 179: 6228–6237.

    CAS  Google Scholar 

  18. Bradford, M. M. (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem., 72: 248–254.

    Article  CAS  Google Scholar 

  19. Raj, S. M., C. Rathnasingh, J.-E. Jo, and S. Park (2008) Production of 3-hydroxypropionic acid from glycerol by a novel recombinant Escherichia coli BL21 strain. Proc. Biochem., 43: 1440–1446.

    Article  CAS  Google Scholar 

  20. Johnson, E. A. and E. C. Lin (1987) Klebsiella pneumoniae 1,3-propanediol:NAD+ oxidoreductase. J. Bacteriol., 169: 2050–2054.

    CAS  Google Scholar 

  21. Wang, W., J. Sun, M. Hartlep, W. D. Deckwer, and A. P. Zeng (2003) Combined use of proteomic analysis and enzyme activity assays for metabolic pathway analysis of glycerol fermentation by Klebsiella pneumoniae. Biotechnol. Bioeng., 83: 525–536.

    Article  CAS  Google Scholar 

  22. Li Calzi, and L. B. Poole (1997) Requirement for the two AhpF cystine disulfide centers in catalysis of peroxide reduction by alkyl hydroperoxide reductase. Biochem., 36: 13357–13364.

    Article  Google Scholar 

  23. Kim, H. J., B. K. Hou, S. G. Lee, J. S. Kim, D. W. Lee, and S. J. Lee (2013) Genome-wide analysis of redox reactions reveals metabolic engineering targets for D-lactate overproduction in Escherichia coli. Metab. Eng., 18: 44–52.

    Article  CAS  Google Scholar 

  24. Arasu, M., V. Kumar, S. Ashok, H. Song, C. Rathnasingh, H. Lee, D. Seung, and S. Park (2011) Isolation and characterization of the new Klebsiella pneumoniae J2B strain showing improved growth characteristics with reduced lipopolysaccharide formation. Biotechnol. Bioproc. Eng., 16: 1134–1143.

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. School of Chemical and Biomolecular Engineering, Pusan National University, Busan, 609-735, Korea

    Yeounjoo Ko, Somasundar Ashok, Eunhee Seol, Satish Kumar Ainala & Sunghoon Park

Authors
  1. Yeounjoo Ko
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Somasundar Ashok
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Eunhee Seol
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Satish Kumar Ainala
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Sunghoon Park
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Sunghoon Park.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Ko, Y., Ashok, S., Seol, E. et al. Deletion of putative oxidoreductases from Klebsiella pneumoniae J2B could reduce 1,3-propanediol during the production of 3-hydroxypropionic acid from glycerol. Biotechnol Bioproc E 20, 834–843 (2015). https://doi.org/10.1007/s12257-015-0166-1

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12257-015-0166-1

Keywords

Search

Navigation