Enhanced stability of tyrosine phenol-lyase from Symbiobacterium toebii by DNA shuffling

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Title
Enhanced stability of tyrosine phenol-lyase from Symbiobacterium toebii by DNA shuffling
Author(s)
Jin Ho Kim; Jae Jun Song; B G Kim; M H Sung; Sang Chul Lee
Bibliographic Citation
Journal of Microbiology and Biotechnology, vol. 14, no. 1, pp. 153-157
Publication Year
2004
Abstract
Tyrosine phenol-lyase (TPL) is a useful enzyme for the synthesis of pharmaceutical aromatic amino acids. In the current study, sequential DNA shuffling and screening were used to enhance the stability of TPL. Twenty-thousand mutants were screened, and several improved variants were isolated. One variant named A13V, in which the 13th amino acid alanine was substituted by valine, exhibited a higher temperature and denaturant stability than the wild-type TPL. The purified mutant TPL, A13V, retained about 60% of its activity at 76°C, whereas the activity of the wild-type TPL decreased to less than 20% at the same temperature. Plus, A13V exhibited about 50% activity with 3 M urea, while the wild-type TPL lost almost all its catalytic activity, indicating an increased denaturant tolerance in the mutant A13V. It is speculated that the substitution of Val for the Ala in the β-strand of the N-terminal arm was responsible for the heightened stabilization, and that the current results will contribute to further research on the structural stability of TPL.
Keyword
denaturant toleranceDNA shufflingthermostabilitytyrosine phenol-lyase
ISSN
1017-7825
Publisher
Korea Soc-Assoc-Inst
Type
Article
Appears in Collections:
Division of Bio Technology Innovation > SME Support Center > 1. Journal Articles
Division of Biomedical Research > Metabolic Regulation Research Center > 1. Journal Articles
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