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Bioproduction of l-Aspartic Acid and Cinnamic Acid by l-Aspartate Ammonia Lyase from Pseudomonas aeruginosa PAO1

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Abstract

Aspartase (l-aspartate ammonia lyase, EC 4.3.1.1) catalyses the reversible amination and deamination of l-aspartic acid to fumaric acid which can be used to produce important biochemical. In this study, we have explored the characteristics of aspartase from Pseudomonas aeruginosa PAO1 (PA-AspA). To overproduce PA-AspA, the 1425-bp gene was introduced in Escherichia coli BL21 and purified. A 51.0-kDa protein was observed as a homogenous purified protein on SDS-PAGE. The enzyme was optimally active at pH 8.0 and 35 °C. PA-AspA has retained 56% activity after 7 days of incubation at 35 °C, which displays the hyperthermostablility characteristics of the enzyme. PA-AspA is activated in the presence of metal ions and Mg2+ is found to be most effective. Among the substrates tested for specificity of PA-AspA, l-phenylalanine (38.35 ± 2.68) showed the highest specific activity followed by l-aspartic acid (31.21 ± 3.31) and fumarate (5.42 ± 2.94). K m values for l-phenylalanine, l-aspartic acid and fumarate were 1.71 mM, 0.346 μM and 2 M, respectively. The catalytic efficiency (k cat/K m) for l-aspartic acid (14.18 s−1 mM−1) was higher than that for l-phenylalanine (4.65 s−1 mM−1). For bioconversion, from an initial concentration of 1000 mM of fumarate and 30 mM of l-phenylalanine, PA-AspA was found to convert 395.31 μM l-aspartic acid and 3.47 mM cinnamic acid, respectively.

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Authors and Affiliations

  1. Department of Biochemistry, P. D. Patel Institute of Applied Sciences, CHARUSAT, Changa, 388421, Gujarat, India

    Arti T. Patel, Rekha C. Akhani, Manisha J. Patel, Samir R. Dedania & Darshan H. Patel

  2. Dr. K.C.Patel Research and Development Center, CHARUSAT, Changa, 388421, Gujarat, India

    Darshan H. Patel

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Patel, A.T., Akhani, R.C., Patel, M.J. et al. Bioproduction of l-Aspartic Acid and Cinnamic Acid by l-Aspartate Ammonia Lyase from Pseudomonas aeruginosa PAO1. Appl Biochem Biotechnol 182, 792–803 (2017). https://doi.org/10.1007/s12010-016-2362-7

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