Applied Microbiology and Biotechnology

, Volume 99, Issue 12, pp 5217–5225 | Cite as

Metabolic engineering of Klebsiella pneumoniae for the production of cis,cis-muconic acid

Applied microbial and cell physiology
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Abstract

cis,cis-Muconic acid (ccMA), a metabolic intermediate of Klebsiella pneumoniae, can be converted to adipic acid and terephthalic acid, which are important monomers of synthetic polymers. However, wild-type K. pneumoniae does not produce ccMA because intracellular carbon flow does not favor ccMA biosynthesis. In this study, several metabolic engineering strategies were used in an attempt to modify the wild-type strain to induce it to produce ccMA. First, by blocking the synthesis of aromatic amino acids, 343 mg/L of catechol, a precursor of ccMA, was produced. Then, the native catechol 1,2-dioxygenasegene (catA) was overexpressed, which caused the strain to convert the catechol to ccMA. The production of ccMA was further improved by deletion of the muconate cycloisomerase gene (catB) and by deleting a feedback inhibitor of the aromatic amino acid pathway. Further improvement was achieved by adjusting the pH of the culture broth. The developed strain produced 2.1 g/L of ccMA in flask cultivation. The results showed the potential of K. pneumoniae as a ccMA producer.

Keywords

cis,cis-Muconic acid Klebsiella pneumoniae Aromatic amino acid pathway Benzoate degradation pathway 
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Notes

Acknowledgments

This work was supported by grants from the National Research Foundation of Korea funded by the Korean Government (2012M1A2A2026560 and 2014R1A2A2A03007094).

Supplementary material

253_2015_6442_MOESM1_ESM.pdf (67 kb)
ESM 1 (PDF 66 kb)

References

  1. Adachi O, Ano Y, Toyama H, Matsushita K (2007) Biooxidation with PQQ-and FAD-dependent dehydrogenases. In: Schmid RD, Urlacher V (eds) Modern biooxidation: enzymes, reactions and applications. Wiley-VCH, Weinheim, pp 1–41CrossRefGoogle Scholar
  2. Bang SG, Choi CY (1995) DO-stat fed-batch production of cis, cis-muconic acid from benzoic acid by Pseudomonas putida BM014. J Ferment Bioeng 79(4):381–383. doi: 10.1016/0922-338x(95)94001-8 CrossRefGoogle Scholar
  3. Basu A, Apte SK, Phale PS (2006) Preferential utilization of aromatic compounds over glucose by Pseudomonas putida CSV86. Appl Environ Microbiol 72(3):2226–2230. doi: 10.1128/AEM.72.3.2226-2230.2006 CrossRefPubMedCentralPubMedGoogle Scholar
  4. Baumeister C, Peersman G (2013) Time-varying effects of oil supply shocks on the US economy. Am Econ J Macroecon 5(4):1–28CrossRefGoogle Scholar
  5. Bongaerts J, Kramer M, Muller U, Raeven L, Wubbolts M (2001) Metabolic engineering for microbial production of aromatic amino acids and derived compounds. Metab Eng 3(4):289–300. doi: 10.1006/mben.2001.0196 CrossRefPubMedGoogle Scholar
  6. Burnham A, Han J, Clark CE, Wang M, Dunn JB, Palou-Rivera I (2012) Life-cycle greenhouse gas emissions of shale gas, natural gas, coal, and petroleum. Environ Sci Technol 46(2):619–627. doi: 10.1021/es201942m CrossRefPubMedGoogle Scholar
  7. Cotter PD, Hill C (2003) Surviving the acid test: responses of gram-positive bacteria to low pH. Microbiol Mol Biol Rev 67(3):429–453. doi: 10.1128/MMBR.67.3.429-453.2003 CrossRefPubMedCentralPubMedGoogle Scholar
  8. Curran KA, Leavitt JM, Karim AS, Alper HS (2013) Metabolic engineering of muconic acid production in Saccharomyces cerevisiae. Metab Eng 15:55–66. doi: 10.1016/j.ymben.2012.10.003 CrossRefPubMedGoogle Scholar
  9. Datsenko KA, Wanner BL (2000) One-step inactivation of chromosomal genes in Escherichia coli K-12 using PCR products. Proc Natl Acad Sci USA 97(12):6640–6645. doi: 10.1073/pnas.120163297
  10. Dresselhaus MS, Thomas IL (2001) Alternative energy technologies. Nature 414(6861):332–337. doi: 10.1038/35104599 CrossRefPubMedGoogle Scholar
  11. Gorke B, Stulke J (2008) Carbon catabolite repression in bacteria: many ways to make the most out of nutrients. Nat Rev Microbiol 6(8):613–624. doi: 10.1038/nrmicro1932 CrossRefPubMedGoogle Scholar
  12. Hommes RWJ, Postma PW, Tempest DW, Neijssel OM (1989) The Influence of the culture pHvalue on the direct glucose oxidative pathway in Klebsiella pneumoniae NCTC418. Arch Microbiol 151(3):261–267. doi: 10.1007/Bf00413140 CrossRefPubMedGoogle Scholar
  13. Im SW, Davidson H, Pittard J (1971) Phenylalanine and tyrosine biosynthesis in Escherichia coli K-12: mutants derepressed for 3-deoxy-D-arabinoheptulosonic acid 7-phosphate synthetase (phe), 3-deoxy-D-arabinoheptulosonic acid 7-phosphate synthetase (tyr), chorismate mutase T-prephenate dehydrogenase, and transaminase A. J Bacteriol 108(1):400–409PubMedCentralPubMedGoogle Scholar
  14. Ji XJ, Huang H, Ouyang PK (2011) Microbial 2,3-butanediol production: a state-of-the-art review. Biotechnol Adv 29(3):351–364. doi: 10.1016/j.biotechadv.2011.01.007 CrossRefPubMedGoogle Scholar
  15. Jimenez JI, Minambres B, Garcia JL, Diaz E (2002) Genomic analysis of the aromatic catabolic pathways from Pseudomonas putida KT2440. Environ Microbiol 4(12):824–841. doi: 10.1046/j.1462-2920.2002.00370.x CrossRefPubMedGoogle Scholar
  16. Jung MY, Ng CY, Song H, Lee J, Oh MK (2012) Deletion of lactate dehydrogenase in Enterobacter aerogenes to enhance 2,3-butanediol production. Appl Microbiol Biotechnol 95:461–469. doi: 10.1007/s00253-012-3883-9 CrossRefPubMedGoogle Scholar
  17. Jung MY, Park BS, Lee J, Oh MK (2013a) Engineered Enterobacter aerogenes for efficient utilization of sugarcane molasses in 2,3-butanediol production. Bioresour Technol 139:21–27. doi: 10.1016/j.biortech.2013.04.003 CrossRefPubMedGoogle Scholar
  18. Jung SG, Jang JH, Kim AY, Lim MC, Kim B, Lee J, Kim YR (2013b) Removal of pathogenic factors from 2,3-butanediol-producing Klebsiella species by inactivating virulence-related wabG gene. Appl Microbiol Biotechnol 97(5):1997–2007. doi: 10.1007/s00253-012-4284-9 CrossRefPubMedGoogle Scholar
  19. Jung MY, Mazumdar S, Shin SH, Yang KS, Lee J, Oh MK (2014) Improvement of 2,3-butanediol yield in Klebsiella pneumoniae by deleting pyruvate formate-lyase gene. Appl Environ Microbiol 80(19):6195–6203. doi: 10.1128/AEM.02069-14 CrossRefPubMedCentralPubMedGoogle Scholar
  20. Kim B, Lee S, Park J, Lu M, Oh M, Kim Y, Lee J (2012) Enhanced 2,3-butanediol production in recombinant Klebsiella pneumoniae via overexpression of synthesis-related genes. J Microbiol Biotechnol 22:1258–1263. doi: 10.4014/jmb.1201.01044 CrossRefPubMedGoogle Scholar
  21. Liao JC, Hou SH, Chao YP (1996) Pathway analysis, engineering, and physiological considerations for redirecting central metabolism. Biotechnol Bioeng 52(1):129–140CrossRefPubMedGoogle Scholar
  22. Lin YH, Sun XX, Yuan QP, Yan YJ (2014) Extending shikimate pathway for the production of muconic acid and its precursor salicylic acid in Escherichia coli. Metab Eng 23:62–69. doi: 10.1016/j.ymben.2014.02.009 CrossRefPubMedGoogle Scholar
  23. Markovic M, Markov S, Pejin D, Mojovic L, Vukasinovic M, Pejin J, Jokovic N (2011) The possibility of lactic acid fermentation in the triticale stillage. Chem Ind Chem Eng Q 17(2):153–162. doi: 10.2298/Ciceq100916065m CrossRefGoogle Scholar
  24. Mazumdar S, Clomburg JM, Gonzalez R (2010) Escherichia coli strains engineered for homofermentative production of D-lactic acid from glycerol. Appl Environ Microbiol 76(13):4327–4336. doi: 10.1128/AEM.00664-10 CrossRefPubMedCentralPubMedGoogle Scholar
  25. Mazumdar S, Lee J, Oh MK (2013) Microbial production of 2,3 butanediol from seaweed hydrolysate using metabolically engineered Escherichia coli. Bioresour Technol 136:329–336. doi: 10.1016/j.biortech.2013.03.013 CrossRefPubMedGoogle Scholar
  26. Mizuno S, Yoshikawa N, Seki M, Mikawa T, Imada Y (1988) Microbial production of cis, cis-muconic acid from benzoic acid. Appl Microbiol Biotechnol 28(1):20–25Google Scholar
  27. Niu W, Draths KM, Frost JW (2002) Benzene-free synthesis of adipic acid. Biotechnol Prog 18(2):201–211. doi: 10.1021/bp010179x CrossRefPubMedGoogle Scholar
  28. Pittard J, Camakaris H, Yang J (2005) The TyrR regulon. Mol Microbiol 55(1):16–26. doi: 10.1111/j.1365-2958.2004.04385.x CrossRefPubMedGoogle Scholar
  29. Polen T, Spelberg M, Bott M (2013) Toward biotechnological production of adipic acid and precursors from biorenewables. J Biotechnol 167(2):75–84. doi: 10.1016/j.jbiotec.2012.07.008 CrossRefPubMedGoogle Scholar
  30. Prior JE, Lynch MD, Gill RT (2010) Broad-host-range vectors for protein expression across gram negative hosts. Biotechnol Bioeng 106(2):326–332. doi: 10.1002/bit.22695 PubMedGoogle Scholar
  31. Sambrook J, Russell DW (2001) Molecular cloning: a laboratory manual, 3rd edn. Cold Spring Harbor Laboratory Press, Cold Spring HarborGoogle Scholar
  32. Saxena RK, Anand P, Saran S, Isar J (2009) Microbial production of 1,3-propanediol: recent developments and emerging opportunities. Biotechnol Adv 27(6):895–913. doi: 10.1016/j.biotechadv.2009.07.003 CrossRefPubMedGoogle Scholar
  33. Schweigert N, Zehnder AJB, Eggen RIL (2001) Chemical properties of catechols and their molecular modes of toxic action in cells, from microorganisms to mammals. Environ Microbiol 3(2):81–91. doi: 10.1046/j.1462-2920.2001.00176.x CrossRefPubMedGoogle Scholar
  34. Silveira MM, Berbert-Molina M, Prata AMR, Schmidell W (1998) Production of 2,3-butanediol from sucrose by Klebsiella pneumoniae NRRL B199 in batch and fed-batch reactors. Braz Arch Biol Technol 41(3):329–334CrossRefGoogle Scholar
  35. Spanning A, Neujahr HY (1990) The effect of glucose on enzyme activities and phenol utilization in Trichosporon cutaneum grown in continuous culture. J Gen Microbiol 136:1491–1495CrossRefGoogle Scholar
  36. Sun XX, Lin YH, Huang Q, Yuan QP, Yan YJ (2013) A novel muconic acid biosynthesis approach by shunting tryptophan biosynthesis via anthranilate. Appl Environ Microbiol 79(13):4024–4030. doi: 10.1128/Aem.00859-13 CrossRefPubMedCentralPubMedGoogle Scholar
  37. Umbarger HE (1978) Amino acid biosynthesis and its regulation. Annu Rev Biochem 47:532–606. doi: 10.1146/annurev.bi.47.070178.002533 CrossRefPubMedGoogle Scholar
  38. Vesely M, Knoppova M, Nesvera J, Patek M (2007) Analysis of catRABC operon for catechol degradation from phenol-degrading Rhodococcus erythropolis. Appl Microbiol Biotechnol 76(1):159–168. doi: 10.1007/s00253-007-0997-6 CrossRefPubMedGoogle Scholar
  39. Weber C, Bruckner C, Weinreb S, Lehr C, Essl C, Boles E (2012) Biosynthesis of cis, cis-muconic acid and its aromatic precursors, catechol and protocatechuic acid, from renewable feedstocks by Saccharomyces cerevisiae. Appl Environ Microbiol 78(23):8421–8430. doi: 10.1128/AEM.01983-12 CrossRefPubMedCentralPubMedGoogle Scholar
  40. Wei D, Xu JQ, Sun JS, Shi JP, Hao J (2013) 2-Ketogluconic acid production by Klebsiella pneumoniae CGMCC 1.6366. J Ind Microbiol Biotechnol 40(6):561–570. doi: 10.1007/s10295-013-1261-y CrossRefPubMedGoogle Scholar
  41. Will MA, Clark NA, Swain JE (2011) Biological pH buffers in IVF: help or hindrance to success. J Assist Reprod Genet 28(8):711–724. doi: 10.1007/s10815-011-9582-0 CrossRefPubMedCentralPubMedGoogle Scholar
  42. Wu CM, Wu CC, Su CC, Lee SN, Lee YA, Wu JY (2006) Microbial synthesis of cis, cis-muconic acid from benzoate by Sphingobacterium sp mutants. Biochem Eng J 29(1–2):35–40. doi: 10.1016/j.bej.2005.02.034 CrossRefGoogle Scholar
  43. Xie NZ, Wang QY, Zhu QX, Qin Y, Tao F, Huang RB, Xu P (2014) Optimization of medium composition for cis, cis-muconic acid production by a Pseudomonas sp. mutant using statistical methods. Prep Biochem Biotechnol 44(4):342–354. doi: 10.1080/10826068.2013.829497 CrossRefPubMedGoogle Scholar
  44. Yu EKC, Saddler JN (1983) Fed-batch approach to production of 2,3-butanediol by Klebsiella pneumoniae grown on high substrate concentrations. Appl Environ Microbiol 46(3):630–635PubMedCentralPubMedGoogle Scholar
  45. Zelle RM, de Hulster E, Kloezen W, Pronk JT, van Maris AJ (2010) Key process conditions for production of C(4) dicarboxylic acids in bioreactor batch cultures of an engineered Saccharomyces cerevisiae strain. Appl Environ Microbiol 76(3):744–750. doi: 10.1128/AEM.02396-09 CrossRefPubMedCentralPubMedGoogle Scholar
  46. Zhang QR, Xiu ZL (2009) Metabolic pathway analysis of glycerol metabolism in Klebsiella pneumoniae incorporating oxygen regulatory system. Biotechnol Prog 25(1):103–115. doi: 10.1021/Bp.70 CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2015

Authors and Affiliations

  1. 1.Department of Chemical and Biological EngineeringKorea UniversitySeoulKorea

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