Screening and Purification of a Novel Transaminase Catalyzing the Transamination of Aryl ${\beta}-Amino$ Acid from Mesorhizobium sp. LUK

  • Kim, Ju-Han (Institute for Molecular Biology and Genetics, and School of Chemical and Biological Engineering, Seoul National University) ;
  • Kyung, Do-Hyun (Institute for Molecular Biology and Genetics, and School of Chemical and Biological Engineering, Seoul National University) ;
  • Yun, Hyung-Don (Institute for Molecular Biology and Genetics, and School of Chemical and Biological Engineering, Seoul National University) ;
  • Cho, Byung-Kwan (Institute for Molecular Biology and Genetics, and School of Chemical and Biological Engineering, Seoul National University) ;
  • Kim, Byung-Gee (Institute for Molecular Biology and Genetics, and School of Chemical and Biological Engineering, Seoul National University)
  • Published : 2006.11.30

Abstract

Mesorhizobium sp. LUK, which utilizes 3-amino-3-phenylpropionic acid as the sole source of nitrogen with high enantioselectivity (E(S)>100), was isolated using enrichment culture. The enzyme involved in the utilization of (S)-3-amino-3-phenylpropionic acid was confirmed to be a transaminase and was purified by 235-folds with a specific activity of 0.72 U/mg. The molecular weight of the purified protein was ca. 47 kDa and the active enzyme was determined as a dimer on gel filtration chromatography. The N-terminal sequence was obtained from the purified protein. Spontaneous decarboxylation of produced ${\beta}-keto$ acids was observed during the chiral resolution of 3-amino-3-phenylpropionic acid.

Keywords

References

  1. Angelaud, R., Y.-L. Zhong, P. Maligres, J. Lee, and D. Askin. 2005. Synthesis of a ${\beta}$-amino acid pharmacophore via a ${\beta}$-lactam intermediate. J. Org. Chem. 70: 1949-1952 https://doi.org/10.1021/jo048249c
  2. Banerjee, A., M. Chase, R. A. Clayton, and B. Landis. 2005. Methods for the stereoselective synthesis and enantiomeric enrichment of ${\beta}$-amino acids. PCT Int. Appl. WO2005005633
  3. Choi, N. S., K. H. Yoo, J. H. Hahm, K. S. Yoon, K. T. Chang, B. H. Hyun, P. J. Maeng, and S. H. Kim. 2005. Purification and characterization of a new peptidase, bacillopeptidase DJ-2, having fibrinolytic activity: Produced by Bacillus sp. DJ-2 from Doen-Jang. J. Microbiol. Biotechnol. 15: 72-79
  4. Griffith, O. W. 1986. ${\beta}$-Amino acids: Mammalian metabolism and utility as alpha-amino acid analogues. Annu. Rev. Biochem. 55: 855-878 https://doi.org/10.1146/annurev.bi.55.070186.004231
  5. Guthrie, J. P. 2002. Uncatalyzed and amine catalyzed decarboxylation of acetoacetic acid: An examination in terms of No Barrier Theory. Bioorg. Chem. 30: 32-52 https://doi.org/10.1006/bioo.2001.1231
  6. Ito, S., A. Ota, K. Yamamoto, and Y. Kawashima. 1992. Resolution of the enantiomers of thiol compounds by reversed-phase liquid chromatography using chiral derivatization with 2,3,4,6-tetra-O-acetyl-${\beta}-_D$-glucopyranosyl isothiocyanate. J. Chromatogr. 626: 187-196 https://doi.org/10.1016/0021-9673(92)85409-M
  7. Juaristi, E. (ed), 1997. Enantioselective Synthesis of ${\beta}$-Amino Acids. Wiley-VCH, New York
  8. Lee, J.-S., H.-S. Baik, and S.-S. Park. 2005. Purification and characterization of two novel fibrinolytic proteases from mushroom, Fomitella fraxinea. J. Microbiol. Biotechnol. 16: 264-271
  9. Lelais, G. and D. Seebach. 2004. ${\beta}^2$-Amino acids-syntheses, occurrence in natural products, and components of ${\beta}-peptides^{1,2}$. Biopolymers 76: 206-243 https://doi.org/10.1002/bip.20088
  10. Liljeblad, A. and L. T. Kanerva. 2006. Biocatalysis as a profound tool in the preparation of highly enantiopure ${\beta}$-amino acids. Tetrahedron (in press)
  11. Murahashi, S.-I., Y. Imada, T. Kawakami, K. Harada, Y. Yonemushi, and N. Tomita. 2002. Enantioselective addition of ketene silyl acetals to nitrones catalyzed by chiral titanium complexes. Synthesis of optically active ${\beta}$-amino acids. J. Am. Chem. Soc. 124: 2888-2889 https://doi.org/10.1021/ja0176649
  12. Ojima, I., S. Lin, and T. Wang. 1999. Recent advances in the medicinal chemistry of taxoids with novel beta-amino acid side chains. Curr. Med. Chem. 6: 927-954
  13. Palko, M., L. Kiss, and F. Fulop. 2005. Synthesis of hydroxylated cyclic beta-amino acid derivatives. Curr. Med. Chem. 12: 3063-3083 https://doi.org/10.2174/092986705774933443
  14. Palomo, C., J. M. Aizpurua, I. Ganboa, and M. Oiarbide. 1999. From beta-lactams to alpha- and beta-amino acid derived peptides. Amino Acids 16: 321-343 https://doi.org/10.1007/BF01388175
  15. Seo, M.-J., J.-N. Kim, E.-A. Cho, H. Park, H.-J. Choi, and Y.-R. Pyun. 2005. Purification and characterization of a novel extracellular alkaline phytase from Aeromonas sp. J. Microbiol. Biotechnol. 15: 745-748
  16. Shin, J. S. and B.-G. Kim. 1997. Kinetic resolution of ${\alpha}$-methylbenzylamine with ${\omega}$-transaminase screened from soil microorganisms: Application of biphasic system to overcome product inhibition. Biotechnol. Bioeng. 55: 348-358 https://doi.org/10.1002/(SICI)1097-0290(19970720)55:2<348::AID-BIT12>3.0.CO;2-D
  17. Stewart, J. D. 2001. Dehydrogenase and transaminases in asymmetric synthesis. Curr. Opin. Chem. Biol. 5: 120-129 https://doi.org/10.1016/S1367-5931(00)00180-0
  18. Taylor, P. P., D. P. Pantaleone, R. F. Senkpeil, and I. G. Fotheringham. 1998. Novel biosynthetic approaches to the production of unnatural amino acids using transaminases. Trends Biotechnol. 16: 412-418 https://doi.org/10.1016/S0167-7799(98)01240-2
  19. Weidner, S., A. Puhler, and H. Kuster. 2003. Genomics insights into symbiotic nitrogen fixation. Curr. Opin. Biotechnol. 14: 200-205 https://doi.org/10.1016/S0958-1669(03)00022-3
  20. Yonaha, K., S. Toyama, and K. Soda. 1987. Omega-amino acid-pyruvate aminotransferase. Methods Enzymol. 143: 500-504 https://doi.org/10.1016/0076-6879(87)43090-5
  21. 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 ${\omega}$-transaminase which is resistant to product inhibition of aliphatic ketones. Appl. Environ. Microbiol. 71: 4220-4224 https://doi.org/10.1128/AEM.71.8.4220-4224.2005
  22. Yun, H., S. Lim, B.-K. Cho, and B.-G. Kim. 2004. ${\omega}$-Amino acid:pyruvate transaminase from Alcaligenes denitrificans Y2k-2: A new catalyst for kinetic resolution of ${\beta}$-amino acids and amines. Appl. Environ. Microbiol. 70: 2520-2534 https://doi.org/10.1128/AEM.70.4.2520-2524.2004
  23. Zahran, H. H. 2001. Rhizobia from wild legumes: Diversity, taxonomy, ecology, nitrogen fixation and biotechnology. J Biotechnol. 91: 143-153 https://doi.org/10.1016/S0168-1656(01)00342-X