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Enhanced Activity of Phenylalanine Ammonia Lyase in Permeabilised Recombinant E. coli by Response Surface Method  

Cui, Jian-dong (Hebei Fermentation Engineering Research Center, College of Bioscience and Bioengineering, Hebei University of Science and Technology)
Li, Yan (Hebei Fermentation Engineering Research Center, College of Bioscience and Bioengineering, Hebei University of Science and Technology)
Jia, Shi-Ru (Key Laboratory of Industry Microbiology, Ministry of Education, Tianjin University of Science and Technology)
Publication Information
Food Science and Biotechnology / v.18, no.2, 2009 , pp. 494-499 More about this Journal
Abstract
To improve phenylalanine ammonia lyase (E.C.4.3.1.5-PAL) activity in recombinant Escherichia coli, Some approaches for improving phenylalanine ammonia lyase (PAL) activity in recombinant E. coli were developed following preliminary studies by means of response surface method. The results shown that permeabilization with combination of Triton X-100, cetyl trimethyl ammonium bromide (CTAB), and acetone enriched cellular recombinant PAL activity significantly, which improved over 10-fold as compared with the control (untreat cell), as high as 181.37 U/g. The optimum values for the tested variables were Triton X-100 0.108 g/L, CTAB 0.15 g/L, and acetone 45.2%(v/v). Furthermore, a second-order model equation was suggested and then validated experimentally. It was indicated that addition of surfactants and organic solvents made the cells more permeable and therefore allowed easier access of the substrate to the enzyme and excretion of the product, which increased the rate of transport of L-phenylalanine and trans-cinnamic acids. These improved methods of PAL activity enrichment could serve as a rich enzyme source, especially in the biosynthesis of L-phenylalanine.
Keywords
enzyme activity; recombinant Escherichia coli; optimization; response surface method; permeabilization;
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