Enhancement of the enzymatic activity of Escherichia coli acetyl esterase by a double mutation obtained by random mutagenesis.

@article{Kobayashi2012EnhancementOT,
  title={Enhancement of the enzymatic activity of Escherichia coli acetyl esterase by a double mutation obtained by random mutagenesis.},
  author={Ryuichi Kobayashi and Nobutaka Hirano and Shigenori Kanaya and Mitsuru Haruki},
  journal={Bioscience, biotechnology, and biochemistry},
  year={2012},
  volume={76 11},
  pages={2082-8}
}
A double mutant of Escherichia coli acetyl esterase (EcAE) with enhanced enzymatic activity was obtained by random mutagenesis using error-prone PCR and screening for enzymatic activity by observing halo formation on a tributyrin plate. The mutant contained Leu97Phe (L97F) and Leu209Phe (L209F) mutations. Single mutants L97F and L209F were also constructed and analyzed for kinetic parameters, as well as double mutant L97F/L209F. Kinetic analysis using p-nitrophenyl butyrate as substrate… CONTINUE READING

From This Paper

Figures, tables, and topics from this paper.

References

Publications referenced by this paper.
Showing 1-2 of 2 references

Experiments in Molecular Genetics,’

  • JH Miller
  • ed. Miller JH, Cold Spring Harbor Laboratory…
  • 1972

Similar Papers

Loading similar papers…