title: Bioproduction of L-Aspartic Acid and Cinnamic Acid by L-Aspartate Ammonia Lyase from Pseudomonas aeruginosa PAO1.
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). Kmvalues for L-phenylalanine, L-aspartic acid and fumarate were 1.71 mM, 0.346 μM and 2 M, respectively. The catalytic efficiency (kcat/Km) for L-aspartic acid (14.18 s-1mM-1) was higher than that for L-phenylalanine (4.65 s-1mM-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.
authors.affiliation: Department of Biochemistry, P. D. Patel Institute of Applied Sciences, CHARUSAT, Changa, 388421, Gujarat, India.
authors.forename: Arti T
authors.initials: AT
authors.lastname: Patel
authors.affiliation: Department of Biochemistry, P. D. Patel Institute of Applied Sciences, CHARUSAT, Changa, 388421, Gujarat, India.
authors.forename: Rekha C
authors.initials: RC
authors.lastname: Akhani
authors.affiliation: Department of Biochemistry, P. D. Patel Institute of Applied Sciences, CHARUSAT, Changa, 388421, Gujarat, India.
authors.forename: Manisha J
authors.initials: MJ
authors.lastname: Patel
authors.affiliation: Department of Biochemistry, P. D. Patel Institute of Applied Sciences, CHARUSAT, Changa, 388421, Gujarat, India.
authors.forename: Samir R
authors.initials: SR
authors.lastname: Dedania
authors.affiliation: Department of Biochemistry, P. D. Patel Institute of Applied Sciences, CHARUSAT, Changa, 388421, Gujarat, India. darshanpatel.bt@charusat.ac.in.|Dr. K.C.Patel Research and Development Center, CHARUSAT, Changa, 388421, Gujarat, India. darshanpatel.bt@charusat.ac.in.
authors.forename: Darshan H
authors.initials: DH
authors.lastname: Patel
mesh_terms: D001220:Aspartate Ammonia-Lyase / Q000737:chemistry*; D001224:Aspartic Acid / Q000737:chemistry*; D001426:Bacterial Proteins / Q000737:chemistry*; D002934:Cinnamates / Q000737:chemistry*; D006358:Hot Temperature; D006863:Hydrogen-Ion Concentration; D011550:Pseudomonas aeruginosa / Q000201:enzymology*
chemical_list: D001426:Bacterial Proteins; D002934:Cinnamates; C029010:cinnamic acid; D001224:Aspartic Acid; D001220:Aspartate Ammonia-Lyase
keywords: Cinnamic acid; L-Aspartate ammonia lyase; L-Aspartic acid; Pseudomonas aeruginosa