Influence of amino acids, organic solvents and surfactants for phenylalanine ammonia lyase activity in recombinant Escherichia coli |
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Authors: | J.D. Cui S.R. Jia A.Y. Sun |
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Affiliation: | Tianjin Key Laboratory of Industry Microbiology, Tianjin University of Science and Technology, Tianjin, China; State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, China |
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Abstract: | Aim: To improve phenylalanine ammonia lyase (E.C.4·3·1·5-PAL) activity in recombinant Escherichia coli . Some methods for enrichment of PAL activity in recombinant E. coli JM109 were described. In an effort to create a rich enzyme source these methods would lead to improvements in the production of l -phenylalanine. Methods and Results: The possibilities of enriching PAL activity in recombinant E . coli was investigated by using individual and combinations of amino acids, organic solvents and surfactants. PAL activity was induced by adding combination of l -phenylalanine and l -tyrosine, activities as high as 64·3 U g−1of cells were obtained and enzyme activity was enriched by over 3·5-fold in comparison with the control. Permeabilization with cetyl trimethyl ammonium bromide or the acetone significantly enriched cellular PAL activity, which improved over 8·2- and 9·0-fold compared with the control, as high as 148·5 and 164·5 U g−1of cells respectively. Conclusion: These efforts may provide some effective methods for enhancing l -phenylalanine ammonia lyase activity. Significance and Impact of the Study: These approaches for manipulating recombinant E . coli in an effort to create a rich enzyme source would serve as a biotechnologically important protocol for production of l -phenylalanine. |
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Keywords: | amino acid organic solvents phenylalanine ammonia lyase recombinant Escherichia coli surfactants. |
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