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Production of acrylic acid and propionic acid by constructing a portion of the 3-hydroxypropionate/4-hydroxybutyrate cycle from Metallosphaera sedula in Escherichia coli

  • Metabolic Engineering and Synthetic Biology - Original Paper
  • Published:
Journal of Industrial Microbiology & Biotechnology

Abstract

Acrylic acid and propionic acid are important chemicals requiring affordable, renewable production solutions. Here, we metabolically engineered Escherichia coli with genes encoding components of the 3-hydroxypropionate/4-hydroxybutyrate cycle from Metallosphaera sedula for conversion of glucose to acrylic and propionic acids. To construct an acrylic acid-producing pathway in E. coli, heterologous expression of malonyl-CoA reductase (MCR), malonate semialdehyde reductase (MSR), 3-hydroxypropionyl-CoA synthetase (3HPCS), and 3-hydroxypropionyl-CoA dehydratase (3HPCD) from M. sedula was accompanied by overexpression of succinyl-CoA synthetase (SCS) from E. coli. The engineered strain produced 13.28 ± 0.12 mg/L of acrylic acid. To construct a propionic acid-producing pathway, the same five genes were expressed, with the addition of M. sedula acryloyl-CoA reductase (ACR). The engineered strain produced 1430 ± 30 mg/L of propionic acid. This approach can be expanded to synthesize many important organic chemicals, creating new opportunities for the production of chemicals by carbon dioxide fixation.

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References

  1. Akawi L, Srirangan K, Liu X, Moo-Young M, Chou CP (2015) Engineering Escherichia coli for high-level production of propionate. J Ind Microbiol Biotechnol 42:1–16

    Article  CAS  Google Scholar 

  2. Alber B, Olinger M, Rieder A, Kockelkorn D, Jobst B, Hügler M, Fuchs G (2006) Malonyl-coenzyme a reductase in the modified 3-hydroxypropionate cycle for autotrophic carbon fixation in archaeal Metallosphaera and Sulfolobus spp. J Bacteriol 188:8551–8559

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Beerthuis R, Rothenberg G, Shiju NR (2015) Catalytic routes towards acrylic acid, adipic acid and ε-caprolactam starting from biorenewables. Green Chem 17:1341–1361

    Article  CAS  Google Scholar 

  4. Bennett BD, Kimball EH, Gao M, Osterhout R, Van Dien SJ, Rabinowitz JD (2009) Absolute metabolite concentrations and implied enzyme active site occupancy in Escherichia coli. Nat Chem Biol 5:593–599

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Berg IA, Kockelkorn D, Buckel W, Fuchs G (2007) A 3-hydroxypropionate/4-hydroxybutyrate autotrophic carbon dioxide assimilation pathway in Archaea. Science 318:1782–1786

    Article  CAS  PubMed  Google Scholar 

  6. Bogorad IW, Lin T-S, Liao JC (2013) Synthetic non-oxidative glycolysis enables complete carbon conservation. Nature 502:693–697

    Article  CAS  PubMed  Google Scholar 

  7. Chen X, Zhou L, Kangming T, Kumar A, Singh S, Prior BA, Wang Z (2013) Metabolic engineering of Escherichia coli: a sustainable industrial platform for bio-based chemical production. Biotechnol Adv 31:1200–1223

    Article  CAS  PubMed  Google Scholar 

  8. Chu HS, Ahn J-H, Yun J, Choi IS, Nam T-W, Cho KM (2015) Direct fermentation route for the production of acrylic acid. Metab Eng 32:23–29

    Article  CAS  PubMed  Google Scholar 

  9. Corma A, Iborra S, Velty A (2007) Chemical routes for the transformation of biomass into chemicals. Chem Rev 107:2411–2502

    Article  CAS  PubMed  Google Scholar 

  10. Datsenko KA, Wanner BL (2000) One-step inactivation of chromosomal genes in Escherichia coli K-12 using PCR products. Proc Natl Acad Sci 97:6640–6645

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Davis MS, Solbiati J, Cronan JE (2000) Overproduction of acetyl-CoA carboxylase activity increases the rate of fatty acid biosynthesis in Escherichia coli. J Biol Chem 275:28593–28598

    Article  CAS  PubMed  Google Scholar 

  12. Fuchs G (2011) Alternative pathways of carbon dioxide fixation: insights into the early evolution of life? Ann Rev Microbiol 65:631–658

    Article  CAS  Google Scholar 

  13. Guo D, Zhu J, Deng Z, Liu T (2014) Metabolic engineering of Escherichia coli for production of fatty acid short-chain esters through combination of the fatty acid and 2-keto acid pathways. Metab Eng 22:69–75

    Article  CAS  PubMed  Google Scholar 

  14. Jiang X, Meng X, Xian M (2009) Biosynthetic pathways for 3-hydroxypropionic acid production. Appl Microbiol Biotechnol 82:995–1003

    Article  CAS  PubMed  Google Scholar 

  15. Kandasamy V, Vaidyanathan H, Djurdjevic I, Jayamani E, Ramachandran K, Buckel W, Jayaraman G, Ramalingam S (2013) Engineering Escherichia coli with acrylate pathway genes for propionic acid synthesis and its impact on mixed-acid fermentation. Appl Microbiol Biotechnol 97:1191–1200

    Article  CAS  PubMed  Google Scholar 

  16. Leonard E, Ajikumar PK, Thayer K, Xiao W-H, Mo JD, Tidor B, Stephanopoulos G, Prather KL (2010) Combining metabolic and protein engineering of a terpenoid biosynthetic pathway for overproduction and selectivity control. Proc Natl Acad Sci 107:13654–13659

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Liu C, Ding Y, Zhang R, Liu H, Xian M, Zhao G (2016) Functional balance between enzymes in malonyl-CoA pathway for 3-hydroxypropionate biosynthesis. Metab Eng 34:1034–1111

    Article  CAS  Google Scholar 

  18. Liu H, Lu T (2015) Autonomous production of 1, 4-butanediol via a de novo biosynthesis pathway in engineered Escherichia coli. Metab Eng 29:135–141

    Article  CAS  PubMed  Google Scholar 

  19. Liu L, Guan N, Zhu G, Li J, Shin H-d DuG, Chen J (2016) Pathway engineering of Propionibacterium jensenii for improved production of propionic acid. Sci Rep 6:19963

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Liu L, Zhu Y, Li J, Wang M, Lee P, Du G, Chen J (2012) Microbial production of propionic acid from propionibacteria: current state, challenges and perspectives. Crit Rev Biotechnol 32:374–381

    Article  CAS  PubMed  Google Scholar 

  21. Liu L, Zhuge X, Shin H-d, Chen RR, Li J, Du G, Chen J (2015) Improved production of propionic acid via combinational overexpression of glycerol dehydrogenase and malate dehydrogenase from Klebsiella pneumoniae in Propionibacterium jensenii. Appl Environ Microbiol AEM 81:2256–2264

    Article  CAS  Google Scholar 

  22. Liu Q, Wu K, Cheng Y, Lu L, Xiao E, Zhang Y, Deng Z, Liu T (2015) Engineering an iterative polyketide pathway in Escherichia coli results in single-form alkene and alkane overproduction. Metab Eng 28:82–90

    Article  CAS  PubMed  Google Scholar 

  23. Liu T, Vora H, Khosla C (2010) Quantitative analysis and engineering of fatty acid biosynthesis in E. coli. Metab Eng 12:378–386

    Article  CAS  PubMed  Google Scholar 

  24. Liu Z, Gao Y, Chen J, Imanaka T, Bao J, Hua Q (2013) Analysis of metabolic fluxes for better understanding of mechanisms related to lipid accumulation in oleaginous yeast Trichosporon cutaneum. Bioresour Technol 130:144–151

    Article  CAS  PubMed  Google Scholar 

  25. Lv L, Ren Y-L, Chen J-C, Wu Q, Chen G-Q (2015) Application of CRISPRi for prokaryotic metabolic engineering involving multiple genes, a case study: controllable P (3HB-co-4HB) biosynthesis. Metab Eng 29:160–168

    Article  CAS  PubMed  Google Scholar 

  26. McMahon MD, Prather KL (2014) Functional screening and in vitro analysis reveal thioesterases with enhanced substrate specificity profiles that improve short-chain fatty acid production in Escherichia coli. Appl Environ Microbiol 80:1042–1050

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Nanchen A, Fuhrer T, Sauer U (2007) Determination of metabolic flux ratios from 13C-experiments and gas chromatography-mass spectrometry data. Metabol Methods Protoc 358:177–197

    CAS  Google Scholar 

  28. Rathnasingh C, Raj SM, Lee Y, Catherine C, Ashok S, Park S (2012) Production of 3-hydroxypropionic acid via malonyl-CoA pathway using recombinant Escherichia coli strains. J Biotechnol 157:633–640

    Article  CAS  PubMed  Google Scholar 

  29. Rodriguez BA, Stowers CC, Pham V, Cox BM (2014) The production of propionic acid, propanol and propylene via sugar fermentation: an industrial perspective on the progress, technical challenges and future outlook. Green Chem 16:1066–1076

    Article  CAS  Google Scholar 

  30. Rogers JK, Church GM (2016) Genetically encoded sensors enable real-time observation of metabolite production. Proc Natl Acad Sci 113:2388–2393

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Wang Z, Ammar EM, Zhang A, Wang L, Lin M, Yang S-T (2015) Engineering Propionibacterium freudenreichii subsp. shermanii for enhanced propionic acid fermentation: effects of overexpressing propionyl-CoA: succinate CoA transferase. Metab Eng 27:46–56

    Article  CAS  PubMed  Google Scholar 

  32. Wang Z, Jin Y, Yang ST (2015) High cell density propionic acid fermentation with an acid tolerant strain of Propionibacterium acidipropionici. Biotechnol Bioeng 112:502–511

    Article  CAS  PubMed  Google Scholar 

  33. Wang Z, Lin M, Wang L, Ammar EM, Yang S-T (2015) Metabolic engineering of Propionibacterium freudenreichii subsp. shermanii for enhanced propionic acid fermentation: effects of overexpressing three biotin-dependent carboxylases. Process Biochem 50:194–204

    Article  CAS  Google Scholar 

  34. Wang Z, Yang S-T (2013) Propionic acid production in glycerol/glucose co-fermentation by Propionibacterium freudenreichii subsp. shermanii. Bioresour Technol 137:116–123

    Article  CAS  PubMed  Google Scholar 

  35. Yang JE, Choi YJ, Lee SJ, Kang K-H, Lee H, Oh YH, Lee SH, Park SJ, Lee SY (2014) Metabolic engineering of Escherichia coli for biosynthesis of poly (3-hydroxybutyrate-co-3-hydroxyvalerate) from glucose. Appl Microbiol Biotechnol 98:95–104

    Article  CAS  PubMed  Google Scholar 

  36. Yang Y, Lin Y, Li L, Linhardt RJ, Yan Y (2015) Regulating malonyl-CoA metabolism via synthetic antisense RNAs for enhanced biosynthesis of natural products. Metab Eng 29:217–226

    Article  CAS  PubMed  Google Scholar 

  37. Ye Z, Li X, Cheng Y, Liu Z, Tan G, Zhu F, Fu S, Deng Z, Liu T (2016) Evaluation of 3-hydroxypropionate biosynthesis in vitro by partial introduction of the 3-hydroxypropionate/4-hydroxybutyrate cycle from Metallosphaera sedula. J Ind Microbiol Biotechnol 43:1313–1321

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  38. Yin J, Wang H, Fu X-Z, Gao X, Wu Q, Chen G-Q (2015) Effects of chromosomal gene copy number and locations on polyhydroxyalkanoate synthesis by Escherichia coli and Halomonas sp. Appl Microbiol Biotechnol 99:5523–5534

    Article  CAS  PubMed  Google Scholar 

  39. Yu X, Liu T, Zhu F, Khosla C (2011) In vitro reconstitution and steady-state analysis of the fatty acid synthase from Escherichia coli. Proc Natl Acad Sci 108:18643–18648

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  40. Yuzawa S, Chiba N, Katz L, Keasling JD (2012) Construction of a part of a 3-hydroxypropionate cycle for heterologous polyketide biosynthesis in Escherichia coli. Biochemistry 51:9779–9781

    Article  CAS  PubMed  Google Scholar 

  41. Zhang H, Stephanopoulos G (2013) Engineering E. coli for caffeic acid biosynthesis from renewable sugars. Appl Microbiol Biotechnol 97:3333–3341

    Article  CAS  PubMed  Google Scholar 

  42. Zhu F, Zhong X, Hu M, Lu L, Deng Z, Liu T (2014) In vitro reconstitution of mevalonate pathway and targeted engineering of farnesene overproduction in Escherichia coli. Biotechnol Bioeng 111:1396–1405

    Article  CAS  PubMed  Google Scholar 

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Acknowledgments

This work was supported by grants from the 973 (2012CB721000, 2011CBA00800) and 863 programs (2012AA02A701) of the Ministry of Science and Technology of China, and by a grant from the National Natural Science Foundation of China (31222002). The authors are grateful to Prof. Kristala Prather of Massachusetts Institute of Technology for providing the E. coli MG1655 ΔrecAΔendA (DE3) strain.

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Authors and Affiliations

  1. Key Laboratory of Combinatorial Biosynthesis and Drug Discovery (Wuhan University), Ministry of Education, Wuhan University School of Pharmaceutical Sciences, 185 Donghu Road, Wuhan, 430071, People’s Republic of China

    Zhijie Liu & Tiangang Liu

  2. Hubei Engineering Laboratory for Synthetic Microbiology, Wuhan Institute of Biotechnology, Wuhan, 430075, People’s Republic of China

    Tiangang Liu

  3. Hubei Provincial Cooperative Innovation Center of Industrial Fermentation, Wuhan, People’s Republic of China

    Tiangang Liu

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  1. Zhijie Liu
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Correspondence to Tiangang Liu.

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Liu, Z., Liu, T. Production of acrylic acid and propionic acid by constructing a portion of the 3-hydroxypropionate/4-hydroxybutyrate cycle from Metallosphaera sedula in Escherichia coli . J Ind Microbiol Biotechnol 43, 1659–1670 (2016). https://doi.org/10.1007/s10295-016-1843-6

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  • DOI: https://doi.org/10.1007/s10295-016-1843-6

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