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Enzymatic synthesis of nylon precursors by 4-aminobutyrate aminotransferase and 6-oxohexanoate dehydrogenase

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Abstract

6-Aminocaproic acid and adipic acid are the key value-added chemical precursors in the pharmaceutical, solvent and polyamide industry, including nylon-6, and nylon-6, 6. An enzymatic interconversion of the two precursors can provide a convenient and eco-friendly biosynthetic route to each of the precursors and thus, require analysis of the reaction process. Herein, an in vitro enzymatic method was employed to convert the two precursors while most studies so far have focused on the whole cell bioconversion to investigate the process. 4-Aminobutyrate aminotransferase was utilized to mediate the reactions between 6-aminocaproic acid and the intermediate 6-oxohexanoic acid with the aid of pyridoxal 5’-phosphate and amine donor/acceptor. 6-Oxohexanoate dehydrogenase was utilized for the reaction from 6-oxohexanoic acid to adipic acid with NADP+. A range of reaction conditions were investigated including the type of amine donor, pH conditions, the concentrations of enzyme and amine donor/acceptor. The optimum condition resulted in 78% yield for the reaction from 6-oxohexanoic acid to 6-aminocaproic acid. The yield for the one-pot, two-step enzymatic cascade from 6-aminocaproic acid via 6-oxohexanoate intermediate to adipic acid was 88%, which was higher than the yield for each individual step in the cascade, with 31% and 32%, for the first and second step, respectively. Furthermore, structural analysis on the active site of the 4-aminobutyrate aminotransferase docked with a range of amine donors implicates the optimal donor is glutamate in accordance with the experimental data and suggests enzyme engineering possibilities for more readily available donors to facilitate the industrial application of the process.

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References

  1. Schaffer S, Haas T (2014) Biocatalytic and fermentative production of α, ω-bifunctional polymer precursors. Org Process Res Dev 18:752–766. https://doi.org/10.1021/op5000418

    Article  CAS  Google Scholar 

  2. Mares F, Sheehan D (1978) Kinetics of caprolactam formation from 6-aminocaproic acid, ester, and amide. Ind Eng Chem Process Des Dev 17:9–16. https://doi.org/10.1021/i260065a003

    Article  CAS  Google Scholar 

  3. Polen T, Spelberg M, Bott M (2013) Toward biotechnological production of adipic acid and precursors from biorenewables. J Biotechnol 167:75–84. https://doi.org/10.1016/j.jbiotec.2012.07.008

    Article  CAS  PubMed  Google Scholar 

  4. Zuidhof KT, de Croon MHJM, Schouten JC (2010) Beckmann rearrangement of cyclohexanone oxime to ε-caprolactam in microreactors. AIChE J 56:1297–1304. https://doi.org/10.1002/aic.12051

    Article  ADS  CAS  Google Scholar 

  5. Srinivasamurthy VST, Böttcher D, Bornscheuer UT (2019) A multi-enzyme cascade reaction for the production of 6-hydroxyhexanoic acid. Z Naturforsch C J Biosci 74:71–76. https://doi.org/10.1515/znc-2018-0216

    Article  CAS  PubMed  Google Scholar 

  6. Schmidt S, Scherkus C, Muschiol J et al (2015) An enzyme cascade synthesis of ε-caprolactone and its oligomers. Angew Chem Int Ed Engl 54:2784–2787. https://doi.org/10.1002/anie.201410633

    Article  CAS  PubMed  Google Scholar 

  7. Yin DL, Bernhardt P, Morley KL et al (2010) Switching catalysis from hydrolysis to perhydrolysis in pseudomonas fluorescens esterase. Biochemistry 49:1931–1942. https://doi.org/10.1021/bi9021268

    Article  CAS  PubMed  Google Scholar 

  8. Rudat J, Brucher BR, Syldatk C (2012) Transaminases for the synthesis of enantiopure beta-amino acids. AMB Express 2:11. https://doi.org/10.1186/2191-0855-2-11

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Liu W, Peterson PE, Langston JA et al (2005) Kinetic and crystallographic analysis of active site mutants of Escherichia coli gamma-aminobutyrate aminotransferase. Biochemistry 44:2982–2992. https://doi.org/10.1021/bi048657a

    Article  CAS  PubMed  Google Scholar 

  10. Priestman MA, Shell TA, Sun L et al (2012) Merging of confocal and caging technologies: selective three-color communication with profluorescent reporters. Angew Chem Int Ed Engl 51:7684–7687. https://doi.org/10.1002/anie.201202820

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Lam S (1990) High performance liquid chromatographic assay of Amicar, epsilon-aminocaproic acid, in plasma and urine after pre-column derivatization with o-phthalaldehyde for fluorescence detection. Biomed Chromatogr 4:175–177. https://doi.org/10.1002/bmc.1130040414

    Article  CAS  PubMed  Google Scholar 

  12. Lee HS, Na JG, Lee J et al (2019) Structure-based mutational studies of D-3-hydroxybutyrate dehydrogenase for substrate recognition of aliphatic hydroxy acids with a variable length of carbon chain. Biotechnol Bioprocess Eng 24:605–612. https://doi.org/10.1007/s12257-019-0135-1

    Article  CAS  Google Scholar 

  13. Shin JS, Kim BG (2001) Comparison of the omega-transaminases from different microorganisms and application to production of chiral amines. Biosci Biotechnol Biochem 65:1782–1788. https://doi.org/10.1271/bbb.65.1782

    Article  CAS  PubMed  Google Scholar 

  14. Der Garabedian PA, Lotti AM, Vermeersch JJ (1986) 4-Aminobutyrate: 2-Oxoglutarate aminotransferase from Candida. Purification and properties Eur J Biochem 156:589–596. https://doi.org/10.1111/j.1432-1033.1986.tb09618.x

    Article  Google Scholar 

  15. Iijima K, Kojima N (1986) 4-Aminobutyrate: 2-Oxoglutarate transaminase-containing neurons in the perinuclear zone of the rat supraoptic nucleus. Acta Histochem 79:211–221. https://doi.org/10.1016/S0065-1281(86)80085-X

    Article  CAS  PubMed  Google Scholar 

  16. White HL (1979) 4-Aminobutyrate: 2-Oxoglutarate aminotransferase in blood platelets. Science 205:696–698. https://doi.org/10.1126/science.462176

    Article  ADS  CAS  PubMed  Google Scholar 

  17. Hwang BY, Cha M, Park HY et al (2011) Aminotransferase-catalyzed asymmetric synthesis of benazepril intermediate. Biotechnol Bioprocess Eng 16:625–630. https://doi.org/10.1007/s12257-011-0066-y

    Article  CAS  Google Scholar 

  18. Bergmeyer HU, Scheibe P, Wahlefeld AW (1978) Optimization of methods for aspartate aminotransferase and alanine aminotransferase. Clin Chem 24:58–73. https://doi.org/10.1093/clinchem/24.1.58

    Article  CAS  PubMed  Google Scholar 

  19. Maître M, Ciesielski L, Cash C et al (1975) Purification and studies on some properties of the 4-aminobutyrate: 2-oxoglutarate transaminase from rat brain. Eur J Biochem 52:157–169. https://doi.org/10.1111/j.1432-1033.1975.tb03983.x

    Article  PubMed  Google Scholar 

  20. Ramírez-Palacios C, Wijma HJ, Thallmair S et al (2021) Computational prediction of ω-transaminase specificity by a combination of docking and molecular dynamics simulations. J Chem Inf Model 61:5569–5580. https://doi.org/10.1021/acs.jcim.1c00617

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Acknowledgements

This research was supported by National Research Foundation of Korea (NRF) grant funded by the Ministry of Science and ICT (2022M3J4A1091452) and the Ministry of Education (2021R1I1A3060595).

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Authors and Affiliations

  1. Department of Biochemical Engineering, Gangneung-Wonju National University, Gangneung, 25457, Korea

    Hoe-Suk Lee & Young Joo Yeon

  2. Department of Biological Engineering, Konkuk University, Seoul, 05029, Korea

    Yung-Hun Yang

  3. Department of Biotechnology, Duksung Women’s University, Seoul, 01369, Korea

    Hyun June Park

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  1. Hoe-Suk Lee
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  2. Yung-Hun Yang
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Correspondence to Young Joo Yeon or Hyun June Park.

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Lee, HS., Yang, YH., Yeon, Y.J. et al. Enzymatic synthesis of nylon precursors by 4-aminobutyrate aminotransferase and 6-oxohexanoate dehydrogenase. Biotechnol Bioproc E 29, 211–218 (2024). https://doi.org/10.1007/s12257-024-00011-x

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  • DOI: https://doi.org/10.1007/s12257-024-00011-x

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