Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
권호 표지

Engineering Recombinant Streptomyces coelicolor Malate Synthase with Improved Thermal Properties by Directed Mutagenesis

  • Koh, Ro-Sita (Department of Microbiology, Faculty of Medicine, National University of Singapore) ;
  • Goh, Liuh-Ling (Department of Microbiology, Faculty of Medicine, National University of Singapore) ;
  • Sim, Tiow-Suan (Department of Microbiology, Faculty of Medicine, National University of Singapore)
  • Published : 2004.06.01
Download PDF
⟨ Previous Next ⟩

Abstract

Streptomyces thermovulgans malate synthase (stMS) is known to be more thermostable and thermoactive than S. coelicolor malate synthase (scMS). Therefore, based on the amino acid sequence of stMS, 3 scMS mutants, namely P186R, T8PL9P, and T8PL9PP186R, were created by site-directed mutagenesis in an attempt to engineer a more thermoactive and thermostable enzyme. An enzymatic analysis of the wild-type and mutant MS revealed that P186R and T8PL9PP186R were more thermoactive than the wild-type scMS and T8PL9P. Furthermore, all 3 mutants exhibited a greater thermo stability than scMS, thereby suggesting that both R186 and P8P9 can cause increased thermo stability in scMS.

Keywords

References

  1. Prot. Eng. v.9 no.8 Enzyme thermostability and thermoactivity Danson,M.J.;D.W.Hough;R.J.M.Russel;G.L.Taylor;L.Pearl https://doi.org/10.1093/protein/9.8.629
  2. Funct. Integr. Genomics v.1 The stability of thermophilic proteins:A study based on comprehensive genome comparison Das,R.;M.Gerstein https://doi.org/10.1007/s101420000003
  3. J. Ind. Microbiol. Biotech. v.30 Thermostable malate synthase of S.thermovugaris Goh,L.L.;R.Koh;P.Loke;T.S.Sim https://doi.org/10.1007/s10295-003-0082-9
  4. Biochem. J. v.99 The role and control of the glyoxylate cycle in Escherichia coli Kornberg,H.L. https://doi.org/10.1042/bj0990001
  5. Prot. Eng. v.13 no.3 Factors enhancing protein thermostability Kumar,S.;C.J.Tsai;R.Nussinov https://doi.org/10.1093/protein/13.3.179
  6. Prot. Eng. v.15 no.1 The effect of proline insertions on the thermostability of a barley alpha-glucosidase Muslin,E.H.;S.E.Clarke;C.A.Henson https://doi.org/10.1093/protein/15.1.29
  7. J. Bacteriol. v.98 Regulation of glyoxylate metabolism in Escherichia coli Ornston,L.N.;M.K.Ornston
  8. Nature Struc. Biol. v.7 no.5 Two exposed amino acid redidues confer thermostability on a cold shock protein Perl,D.;U.Mueller;U.Heinemann;F.X.Schmid https://doi.org/10.1038/75151
  9. Eur J. Biochem. v.145 no.3 The amino-acid sequence of copper/zinc superoxide dismutase from swordfish liver. Comparison of copper/zinc superoxide dismutase sequences Rocha,H.A.;W.H.Bannister;J.V.Bannister https://doi.org/10.1111/j.1432-1033.1984.tb08580.x
  10. Nucleic Acids Res. v.22 CLUSTAL W:Improving the sensitivity of progressive multiple sequence alignment through sequence weighting,position-specific gap penalties and weight matrix choice Thompson,J.D.;D.G.Higgins;T.J.Gibson https://doi.org/10.1093/nar/22.22.4673
  11. J. Mol. Catalysis B. v.4 Protein thermostabilization by proline substitution Watanabe,K.;Y.Suzuki https://doi.org/10.1016/S1381-1177(97)00031-3
  12. Prot. Eng. v.12 no.8 Increasing the thermostability of D-xylose isomerase by introduction of a proline into the turn of a random coil Zhu,G.;P.C.Xu;M.K.Teng;L.M.Tao;X.Y.Zhu;C.J.Wu;J.Hang;L.W.Niu;Y.Z.Wang https://doi.org/10.1093/protein/12.8.635