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Biotechnology and Bioprocess Engineering

, Volume 24, Issue 1, pp 176–182 | Cite as

Activity Improvements of an Engineered ω-transaminase for Ketones Are Positively Correlated with Those for Cognate Amines

  • Sang-Woo Han
  • Jong-Shik ShinEmail author
Research Paper
  • 2 Downloads

Abstract

Chiral amines are broadly used as essential building blocks of diverse pharmaceutical drugs. ω-Transaminase (ω-TA) is one of the promising enzymes for biocatalytic preparation of chiral amines owing to its capability for reductive amination of ketones. However, industrial application of ω-TA to asymmetric synthesis of chiral amines is often limited by marginal activities for ketones. Here, we explore whether activity improvements of ω-TA for ketones caused by active site engineering are correlated with those for cognate amines. We examined amino donor reactivities of 20 amines with ω-TA from Ochrobactrum anthropi (OATA) and its engineered variant carrying a W58L mutation (OATAW58L). Consistent with the previously observed activity increases for ketones, OATAW58L showed 3 to 79-fold activity improvements for amines. Docking simulations suggested that the activity improvements resulted from increased proximity of a bound amine to an internal aldimine owing to relocation of a small binding pocket. Activity comparison indicated that the W58L mutation induced activity increases for ketones better than it did those for amines. This result motivated us to carry out Pearson correlation analysis among the activity improvements for amines and ketones, revealing a strong positive correlation (r = 0.84). This suggests that active site engineering beneficial for activity of ω-TA toward a target amine is likely to evoke an activity improvement for its cognate ketone. Our finding is expected to provide a clue to design a robust high-throughput screening method required for creation of an engineered mutant displaying a better activity toward a target ketone.

Keywords

chiral amine ω-transaminase asymmetric synthesis protein engineering docking simulation 

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References

  1. 1.
    Ghislieri, D. and N. J. Turner (2014) Biocatalytic approaches to the synthesis of enantiomerically pure chiral amines. Top. Catal. 57: 284–300.CrossRefGoogle Scholar
  2. 2.
    Nugent, T. C. and M. El-Shazly (2010) Chiral amine synthesis-Recent developments and trends for enamide reduction, reductive amination, and imine reduction. Adv. Synth. Catal. 352: 753–819.CrossRefGoogle Scholar
  3. 3.
    Boumehira, A. Z., H. A. El-Enshasy, H. Hacène, E. A. Elsayed, R. Aziz, and E. Y. Park (2016) Recent progress on the development of antibiotics from the genus Micromonospora. Biotechnol. Bioprocess Eng. 21: 199–223.CrossRefGoogle Scholar
  4. 4.
    Liu, S. P., R. X. Liu, J. Mao, L. Zhang, Z. Y. Ding, Z. H. Gu, and G. Y. Shi (2016) Structural-based screening of L-phenylglycine aminotransferase using L-phenylalanine as the optimal amino donor: Recycling of L-phenylalanine to produce L-phenylglycine. Biotechnol. Bioprocess Eng. 21: 153–159.CrossRefGoogle Scholar
  5. 5.
    Sheldon, R. A. (2017) The E factor 25 years on: the rise of green chemistry and sustainability. Green Chem. 19: 18–43.CrossRefGoogle Scholar
  6. 6.
    Malik, M. S., E. S. Park, and J. S. Shin (2012) Features and technical applications of omega-transaminases. Appl. Microbiol. Biotechnol. 94: 1163–1171.CrossRefGoogle Scholar
  7. 7.
    Koszelewski, D., K. Tauber, K. Faber, and W. Kroutil (2010) ω-Transaminases for the synthesis of non-racemic α-chiral primary amines. Trends Biotechnol. 28: 324–332.CrossRefGoogle Scholar
  8. 8.
    Bornscheuer, U. T., G. W. Huisman, R. J. Kazlauskas, S. Lutz, J. C. Moore, and K. Robins (2012) Engineering the third wave of biocatalysis. Nature 484: 185–194.CrossRefGoogle Scholar
  9. 9.
    Savile, C. K., et al. (2010) Biocatalytic asymmetric synthesis of chiral amines from ketones applied to sitagliptin manufacture. Science 329: 305–309.CrossRefGoogle Scholar
  10. 10.
    Fuchs, M., J. E. Farnberger, and W. Kroutil (2015) The industrial age of biocatalytic transamination. Eur. J. Org. Chem. 2015: 6965–6982.Google Scholar
  11. 11.
    Kelly, S. A., S. Pohle, S. Wharry, S. Mix, C. C. R. Allen, T. S. Moody, and B. F. Gilmore (2018) Application of ω-transaminases in the pharmaceutical industry. Chem. Rev. 118: 349–367.CrossRefGoogle Scholar
  12. 12.
    Reetz, M. T. (2013) Biocatalysis in organic chemistry and biotechnology: Past, present, and future. J. Am. Chem. Soc. 135: 12480–12496.CrossRefGoogle Scholar
  13. 13.
    Hammer, S. C., A. M. Knight, and F. H. Arnold (2017) Design and evolution of enzymes for non-natural chemistry. Curr. Opin. Green Sustain. Chem. 7: 23–30.CrossRefGoogle Scholar
  14. 14.
    Guo, F. and P. Berglund (2017) Transaminase biocatalysis: optimization and application. Green Chem. 19: 333–360.CrossRefGoogle Scholar
  15. 15.
    Han, S.-W., J. Kim, H.-S. Cho, and J.-S. Shin (2017) Active site engineering of ω-transaminase guided by docking orientation analysis and virtual activity screening. ACS Catal. 7: 3752–3762.CrossRefGoogle Scholar
  16. 16.
    Han, S. W., E. S. Park, J. Y. Dong, and J. S. Shin (2015) Mechanism-guided engineering of omega-transaminase to accelerate reductive amination of ketones. Adv. Synth. Catal. 357: 1732–1740.CrossRefGoogle Scholar
  17. 17.
    Rausch, C., A. Lerchner, A. Schiefner, and A. Skerra (2013) Crystal structure of the omega-aminotransferase from Paracoccus denitrificans and its phylogenetic relationship with other class III aminotransferases that have biotechnological potential. Proteins 81: 774–787.CrossRefGoogle Scholar
  18. 18.
    Han, S. W., E. S. Park, J. Y. Dong, and J. S. Shin (2015) Expanding substrate specificity of w-transaminase by rational remodeling of a large substrate?binding pocket. Adv. Synth. Catal. 357: 2712–2720.CrossRefGoogle Scholar
  19. 19.
    Shin, J. S. and B. G. Kim (1999) Asymmetric synthesis of chiral amines with ω-transaminase. Biotechnol. Bioeng. 65: 206–211.CrossRefGoogle Scholar
  20. 20.
    Seo, J.-H., W.-K. Min, S.-G. Lee, H. Yun, and B.-G. Kim (2018) To the final goal: can we predict and suggest mutations for protein to develop desired phenotype? Biotechnol. Bioprocess Eng. 23: 134–143.CrossRefGoogle Scholar

Copyright information

© The Korean Society for Biotechnology and Bioengineering and Springer 2019

Authors and Affiliations

  1. 1.Department of BiotechnologyYonsei UniversitySeoulKorea

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