Biotechnology and Bioprocess Engineering

, Volume 18, Issue 1, pp 1–7 | Cite as

High throughput screening methods for ω-transaminases

  • Sam Mathew
  • Giyoung Shin
  • Minsu Shon
  • Hyungdon Yun
Review Paper
First Online:
Received:
Revised:
Accepted:

Abstract

Recently, ω-transaminases have been increasingly used to synthesize amine compounds by reductive amination of prochiral ketones which are of high pharmacological significance. However, the conventional methods for evaluating these enzymes are time consuming and have often been regarded as a bottle neck in developing these enzymes as industrial biocatalysts. In the past few years, several high throughput screening methods have been developed for fast evaluation and identification of ω-transaminase. This review summarizes the various methodologies developed for rapidly screening ω-transaminases.

Keywords

transaminase high throughput screening techniques biocatalysis chiral amines 
This is a preview of subscription content, log in to check access.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Cassimjee, K. E., C. Branneby, V. Abedi, A. Wells, and P. Berglund (2010) Transaminations with isopropyl amine: Equilibrium displacement with yeast alcohol dehydrogenase coupled to in situ cofactor regeneration. Chem. Commun. 46: 5569–5571.CrossRefGoogle Scholar
  2. 2.
    Bea, H. S., H. J. Park, S. H. Lee, and H. Yun (2011) Kinetic resolution of aromatic β-amino acids by ω-transaminase. Chem. Commun. 47: 5894–5896.CrossRefGoogle Scholar
  3. 3.
    Abrahamson, M. J., E. Vazquez-Figueroa, N. B. Woodall, J. C. Moore, and A. S. Bommarius (2012) Development of an amine dehydrogenase for synthesis of chiral amines. Angew. Chem. Int. Ed. Engl. 51: 3969–3972.CrossRefGoogle Scholar
  4. 4.
    Mutti, F. G., C. S. Fuchs, D. Pressnitz, J. H. Sattler, and W. Kroutil (2011) Stereoselectivity of four (R)-selective transaminases for the asymmetric amination of ketones. Adv. Synth. Catal. 353: 3227–3233.CrossRefGoogle Scholar
  5. 5.
    Hwang, B. Y., B. K. Cho, H. Yun, and B. G. Kim (2005) Revisit of aminotransferase in the genomic era and its application to biocatalysis. J. Mol. Catal. B: Enzym. 37: 47–55.CrossRefGoogle Scholar
  6. 6.
    Shin, J. S. and B. G. Kim (2002) Exploring the active site of amine: Pyruvate aminotransferase on the basis of the substrate structure-reactivity relationship: How the enzyme controls substrate specificity and stereoselectivity. J. Org. Chem. 67: 2848–2853.CrossRefGoogle Scholar
  7. 7.
    Shin, J. S. and B. G. Kim (1997) Kinetic resolution of α-methylbenzylamine with ω-transaminase screened from soil microorganisms: Application of a biphasic system to overcome product inhibition. Biotechnol. Bioeng. 55: 348–358.CrossRefGoogle Scholar
  8. 8.
    Bea, H. S., Y. M. Seo, M. N. Cha, B. G. Kim, and H. Yun (2010) Kinetic REsolution of α-methylbenzylamine by recombinant Pichia pastoris expressing ω-transaminase. Biotechnol. Bioeng. 15: 429–434.Google Scholar
  9. 9.
    Yun, H. and B. G. Kim (2008) Asymmetric synthesis of (S)-α-methylbenzylamine by recombinant Escherichia coli co-expressing omega-transaminase and acetolactate synthase. Biosci. Biotechnol. Biochem. 72: 3030–3033.CrossRefGoogle Scholar
  10. 10.
    Malik, M. S., E. S. Park, and J. S. Shin (2012) ω-Transaminasecatalyzed kinetic resolution of chiral amines using l-threonine as an amino acceptor precursor. Green Chem. 14: 2137–2140.CrossRefGoogle Scholar
  11. 11.
    Seo, Y. M., S. Mathew, H. S. Bea, Y. H. Khang, S. H. Lee, B. G. Kim, and H. Yun (2012) Deracemization of unnatural amino acid: homoalanine using D-amino acid oxidase and ω-transaminase. Org. Biomol. Chem. 10: 2482–2485.CrossRefGoogle Scholar
  12. 12.
    Hohne, M. and U. T. Bornscheuer (2009) Biocatalytic routes to optically active amines. Chemcatchem. 1: 42–51.CrossRefGoogle Scholar
  13. 13.
    Mathew, S. and H. Yun (2012) ω-Transaminases for the production of optically pure amines and unnatural amino acids. ACS Catal. 2: 993–1001.CrossRefGoogle Scholar
  14. 14.
    Weingart, U., Y. Lavi, and D. Horn (2009) Data mining of enzymes using specific peptides. BMC Bioinformatics 10: 446–455.CrossRefGoogle Scholar
  15. 15.
    Sharan, R., I. Ulitsky, and R. Shamir (2007) Network-based prediction of protein function. Mol. Syst. Biol. 3: 88.CrossRefGoogle Scholar
  16. 16.
    Eisenberg, D., E. M. Marcotte, I. Xenarios, and T. O. Yeates (2000) Protein function in the post genomic era. Nature 405: 823–826.CrossRefGoogle Scholar
  17. 17.
    Kazlauskas, R. J. and U. T. Bornscheuer (2009) Finding better protein engineering strategies. Nat. Chem. Biol. 5: 526–529.CrossRefGoogle Scholar
  18. 18.
    Savile, C. K., J. M. Janey, E. C. Mundorff, J. C. Moore, S. Tam, W. R. Jarvis, J. C. Colbeck, A. Krebber, F. J. Fleitz, J. Brands, P. N. Devine, G. W. Huisman, and G. J. Hughes (2010) Biocatalytic asymmetric synthesis of chiral amines from ketones applied to sitagliptin manufacture. Science 329: 305–309.CrossRefGoogle Scholar
  19. 19.
    Desai, A. A. (2011) Sitagliptin manufacture: A compelling tale of green chemistry, process intensification, and industrial asymmetric catalysis. Angew. Chem. Int. Ed. Engl. 50: 1974–1976.CrossRefGoogle Scholar
  20. 20.
    Koszelewski, D., K. Tauber, K. Faber, and W. Kroutil (2010) ω-Transaminases for the synthesis of non-racemic a-chiral primary amines. Trends Biotechnol. 28: 324–332.CrossRefGoogle Scholar
  21. 21.
    Malik, M. S., E. S. Park, and J. S. Shin (2012) Features and technical applications of ω-transaminases. Appl. Microbiol. Biotechnol. 94: 1163–1171.CrossRefGoogle Scholar
  22. 22.
    Tufvesson, P., J. Lima-Ramos, J. S. Jensen, N. Al-Haque, W. Neto, and J. M. Woodley (2011) Process considerations for the asymmetric synthesis of chiral amines using transaminases. Biotechnol. Bioeng. 108: 1479–1493.CrossRefGoogle Scholar
  23. 23.
    Hwang, B. Y. and B. G. Kim (2004) High-throughput screening method for the identification of active and enantioselective ω-transaminases. Enz. Microb. Technol. 34: 429–436.CrossRefGoogle Scholar
  24. 24.
    Shin, J. S. and B. G. Kim (2001) Comparison of the w-transaminases from different microorganisms and application to production of chiral Amines. Biosci. Biotechnol. Biochem. 65: 1782–1788.CrossRefGoogle Scholar
  25. 25.
    Yun, H., S. Lim, B. K. Cho, and B. G. Kim (2004) ω-amino acid: Pyruvate transaminase from alcaligenes denitrificans Y2k-2: A new catalyst for kinetic resolution of ω-amino acids and amines Appl. Environ. Microbiol. 70: 2529–2534.CrossRefGoogle Scholar
  26. 26.
    Hanson, R. L., B. L. Davis, Y. Chen, S. L. Goldberg, W. L. Parker, T. P. Tully, M. A. Montana, and R. N. Patel (2008) Preparation of (R)-amines from racemic amines with an (S)-amine transaminase from Bacillus megaterium Adv. Synth. Catal. 350: 1367–1375.CrossRefGoogle Scholar
  27. 27.
    Kim, J., D. Kyung, H. Yun, B. K. Cho, and B. G. Kim (2006) Screening and purification of a novel transaminase catalyzing the transamination of Aryl β-Amino Acid from Mesorhizobium sp. LUK. J. Microbiol. Biotechnol. 16: 1832–1836.Google Scholar
  28. 28.
    Iwasaki, A., Y. Yamada, N. Kizaki, Y. Ikenaka, and J. Hasegawa (2006) Microbial synthesis of chiral amines by (R)-specific transamination with Arthrobacter sp. KNK168 Appl. Microbiol. Biotechnol. 69: 499–505.CrossRefGoogle Scholar
  29. 29.
    Liszka, M. J., M. E. Clark, E. Schneider, and D. S. Clark (2012) Nature versus nurture: Developing enzymes that function under extreme conditions. Annu. Rev. Chem. Biomol. Eng. 3: 77–102.CrossRefGoogle Scholar
  30. 30.
    Bornscheuer, U. T. and M. Pohl (2001) Improved biocatalysts by directed evolution and rational protein design. Curr. Opin. Chem. Biol. 5: 137–143.CrossRefGoogle Scholar
  31. 31.
    Street, A. G. and S. L. Mayo (1999) Computational protein design. Structure 7: 105–109.CrossRefGoogle Scholar
  32. 32.
    Yun, H., B. Y. Hwang, J. H. Lee, and B. G. Kim (2005) Use of enrichment culture for directed evolution of the vibrio fluvialis JS17 ω-transaminase, which is resistant to product inhibition by aliphatic ketones. Appl. Environ. Microbiol. 71: 4220–4224.CrossRefGoogle Scholar
  33. 33.
    Truppo, M. D., J. D. Rozzell, J. C. Moore, and N. J. Turner (2009) Rapid screening and scale-up of transaminase catalysed reactions. Org. Biomol. Chem. 7: 395–398.CrossRefGoogle Scholar
  34. 34.
    Truppo, M. D. and N. J. Turner (2010) Micro-scale process development of transaminase catalysed reactions. Org. Biomol. Chem. 8: 1280–1283.CrossRefGoogle Scholar
  35. 35.
    Hopwood, J., M. D. Truppo, N. J. Turner, and R. C. Lloyd (2011) A fast and sensitive assay for measuring the activity and enantioselectivity of transaminases. Chem. Commun. 47: 773–775.CrossRefGoogle Scholar
  36. 36.
    Schatzle, S. M. Hohne, E. Redestad, K. Robins, and U. T. Bornscheuer (2009) Rapid and sensitive kinetic assay for characterization of ω-transaminases. Anal. Chem. 81: 8244–8248.CrossRefGoogle Scholar
  37. 37.
    Cassimjee K. E., M. S. Humble, V. Miceli, C. G. Colomina, and P. Berglund (2011) Active site quantification of an ω-transaminase by performing a half transamination reaction. ACS Catal. 1: 1051–1055.CrossRefGoogle Scholar
  38. 38.
    Schatzle S., M. Hohne, K. Robins, and U. T. Bornscheuer (2010) Conductometric method for the rapid characterization of the substrate specificity of amine-transaminases. Anal. Chem. 82: 2082–2086.CrossRefGoogle Scholar
  39. 39.
    Sehl, T., R. C. Simon, H. C. Hailes, J. M. Ward, U. Schell, M. Pohl, and D. Rother (2012) TTC-based screening assay for omega-transaminases: A rapid method to detect reduction of 2-hydroxy ketones. J. Biotechnol. 159: 188–194.CrossRefGoogle Scholar
  40. 40.
    Martin, A. R., R. DiSanto, I. Plotnikov, S. Kamat, D. Shonnar, and S. Pannuri (2007) Improved activity and thermostability of (S)-aminotransferase by error-prone polymerase chain reaction for the production of a chiral amine Biochem. Eng. J. 37: 246–255.Google Scholar
  41. 41.
    Hwang, B. Y., S. H. Ko, H. Y. Park, J. H. Seo, B. S. Lee, and B. G. Kim (2008) Identification of ω-aminotransferase from caulobacter crescentus and site-directed mutagenesis to broaden substrate specificity. J. Microbiol. Biotechnol. 18: 48–54.Google Scholar
  42. 42.
    Kaulmann, U., K. Smithies, M. E. B. Smith, H. C. Hailes, and J. M. Ward (2007) Substrate spectrum of ω-transaminase from Chromobacterium violaceum DSM30191 and its potential for biocatalysis. Enz. Microb. Technol. 41: 628–637.CrossRefGoogle Scholar
  43. 43.
    Park, E., M. Kim, and J. S. Shin (2010) One-pot conversion of LThreonine into L-homoalanine: Biocatalytic production of an unnatural amino acid from a natural one. Adv. Synth. Catal. 352: 3391–3398.CrossRefGoogle Scholar
  44. 44.
    Park, E. S., M. Kim, and J. S. Shin (2012) Molecular determinants for substrate selectivity of ω-transaminases. Appl. Microbiol. Biotechnol. 93: 2425–2435.CrossRefGoogle Scholar
  45. 45.
    Hohne, M., S. Schatzle, H. Jochens, K. Robins, and U. T. Bornscheuer (2010) Rational assignment of key motifs for function guides in silico enzyme identification. Nat. Chem. Biol. 6: 807–813.CrossRefGoogle Scholar
  46. 46.
    Schatzle, S., F. Steffan-Munsberg, A. Thontowi, M. Hohne, K. Robins, and U. T. Bornscheuer (2011) Enzymatic asymmetric synthesis of enantiomerically pure aliphatic, aromatic and arylaliphatic amines with (R)-Selective amine transaminases. Adv. Synth. Catal. 353: 2439–2445.CrossRefGoogle Scholar

Copyright information

© The Korean Society for Biotechnology and Bioengineering and Springer-Verlag Berlin Heidelberg 2013

Authors and Affiliations

  • Sam Mathew
    • 1
  • Giyoung Shin
    • 1
  • Minsu Shon
    • 1
  • Hyungdon Yun
    • 1
  1. 1.School of BiotechnologyYeungnam UniversityGyeongsanKorea

Personalised recommendations