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Purification and Characterization of the Recombinant Arabidopsis thaliana Acetolactate Synthase

  • Published : 1997.06.20

Abstract

Acetolactate synthase was purified from Escherichia coli MF2000/pTATX containing Arabidopsis thaliana acetolactate synthase gene. Purification steps included DEAE cellulose ion exchange column chromatography, phenyl sepharose hydrophobic column chromatography, hydroxylapatite affinity column chromatography, and Mono-Q HPLC. Molecular weight was estimated to be ∼65 KDa and purification fold was 109 times. The enzyme showed a pH optimum of 7 and the $K_M$ value was 5.9 mM. The purified enzyme was not inhibited by any of the end products, valine, leucine, and isoleucine.

Keywords

References

  1. Plant physiol. v.85 Mazur, B. J.;Chui, C. F.;Smith, J. K.
  2. J. Bacteriol. v.157 Eoyang, L.;Silverman, P. M.
  3. Nucleic Acids Res. v.7 Lawther, R. P.;Nichols, B.;Zurawski, G.;Hatfield, G. W.
  4. Nucleic Acids Res. v.11 Squires, C. H.;DeFelice, M.;Devereux, J.;Calvo, J. M.
  5. Nucleic Acids Res. v.13 Wek, R. C.;Hauser, C. A.;Harifield, G. W.
  6. Genetics v.109 Falco, S. C.;Dumas, K. D.
  7. Nucleic Acids Res. v.13 Falco, S. C.;Dumas, K. D.;Livak, K. J.
  8. Curr. Genet. v.24 Bekkaoui, F.;Nadin-Davis, S. A.;Crosby, W. L.
  9. Science v.224 Chaleff, R. S.;Mauvais, C. J.
  10. Plant Physiol. v.76 Shaner, D. L.;Anderson, P. C.;Stidham, M. A.
  11. 1989 BCPC Monograph no.42 Hawkes, T. R.;Copping, L. G.(ed.);Dalziel, J.(ed.);Dodge, A. D.(ed.)
  12. Naturforsch. v.40c Huppartz, J. L.;Casida, J. E. Z.
  13. Abstr. Pap. Am. Chem. Soc., 198 Meet., AGRO No. 39 Crews, A. D.;Alvarado, S. I.;Brady, T. E.;Doehner, R. F.;Gange, D. M.;Little, D. L.
  14. Herbicides and Plant Metabolism Hawkes, T. R.;Howard, J. I.;Pontin, S. E.;AD Dodge(ed.)
  15. Plant Physiol. v.100 Christopher, J. T.;Powles, S. B.;Holtum, J. A. M.
  16. Plant Physiol. v.102 Richter, J.;Powles, S. B.
  17. Physiologia Plantalum v.91 Tardif, F. J.;Powles, S. B.
  18. Plant Physiol. v.94 Landstein, D.;Chipman, D. M.;Arad, S(M).;Barak, Z.
  19. Planta v.191 Landstein, D.;Arad, S(M).;Barak, Z.;Chipman, D. M.
  20. Planta v.197 Landstein, D.;Epelbaum, S.;Arad, S(M).;Barak, Z.;Chipman, D. M.
  21. Plant Physiol. v.103 Schmitzer, P. R.;Eilers, R. J.;Cseke, C.
  22. J. Biol. Chem. v.270 Bernasconi, P.;Woodworth, A. R.;Rosen, R. A.;Subramanian, M. V.;Siehl, D. L.
  23. J. Biol. Chem. v.271 Bernasconi, P.;Woodworth, A. R.;Rosen, B. A.;Subramanian, M. V.;Siehl, D. L.
  24. Science v.229 Meyerowitz, E. M.;Pruitt, R. E.
  25. Methods Enzymol. v.166 Eoyang, L.;Silverman, P. M.
  26. Methods Enzymol. v.166 Schloss, J. V.;Van Dyk, D. E.
  27. Methods Enzymol. v.166 Barak, Z. E.;Calvo. J. M.;Schloss,J. V.
  28. Eur. J. Biochem. v.185 Poulsen, C.;Stougaard, P.
  29. Naturforsch v.43c Durner, J.;Boger, P. Z.
  30. Physiol. v.97 Singh, B. J.;Schmitt, G. K.;Lillis, M.;Hand, J. M.;Misra, R.
  31. Proc. Natl. Acad. Sci. USA v.86 Smith, J. K.;Schloss, J. V.;Mazur, B. J.
  32. Molecular cloning; A laboratory manual Maniatis, T.;Fritsch, E. F.;Sambrook, J.
  33. J. Biol. Chem. v.161 Westerfeld, W. W.
  34. Anal. Biochem. v.72 Bradford, M. M.
  35. Gel Electrophoresis of Protein (2nd edition) Hames, B. D.
  36. Methods Enzymol. v.182 Gorbunoff, M. J.
  37. Anal. Biochem. v.171 Singh, B. J.;Stidham, M. A.;Shaner, D. L.
  38. Biochemistry v.28 Gollop, N.;Damri, B.;Barak, Z.;Chipman, D. M.
  39. Plant Physiol. v.99 Singh, B. J.;Szamosi, I.;Hand, J. M.;Misra, R.
  40. Eur. J. Biochem. v.3 Magee, P.;de Robichon-Szulmajster, H.