Bioprocess Development for the Synthesis of 4-Aminophenylacetic Acid Using Nitrilase Activity of Whole Cells of Alcaligenes faecalis MTCC 12629
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Nitrilases are important industrial enzymes to synthesize commercially useful acids from nitriles. In the present study, an aryl specific inducible nitrilase was produced from Alcaligenes faecalis MTCC 12629 which hydrolyzed 4-aminophenylacetonitrile to 4-aminophenylacetic acid. A bioprocess was developed to synthesize 4-aminophenylacetic acid from 4-aminophenylacetonitrile at 500 ml scale under optimized reaction conditions. The maximum activity of nitrilase for biotransformation of 4-aminophenylacetonitrile was recorded in 50 mM KH2PO4/K2HPO4 buffer of pH 7 at 45 °C. Operational stability of present enzyme was found at 45 °C for fed batch reaction. The bioprocess designed at 500 ml scale included 17 U/ml catalyst amount, 50 mM substrate addition after 30 min in total five feedings for 150 min at 45 °C. The approach of substrate feeding at lower amount resulted in accumulation of 175 mM product in 500 ml reaction after five feedings of substrate. The rate of product formation was found to be 2.5 g/gdcw/h with the volumetric productivity of 26.6 g/l.
KeywordsNitrilase 4-Aminophenylacetonitrile 4-Aminophenylacetic acid Alcaligenes faecalis MTCC 12629
The authors acknowledge University Grants Commission (UGC) New Delhi, India, for financial support in the form of UGC SAP SRF to Ms. Neerja Thakur and Faculty Fellowship to Prof. T. C. Bhalla under UGC-BSR scheme.
Compliance with Ethical Standards
Conflict of interest
The authors declare no conflict of interest.
- 8.Zhu X, Gong J, Li H, Lu Z, Zhou Z, Shi J, Xu Z (2014) Screening, identification and culture optimization of a newly isolated aromatic nitrilase-producing bacterium–Pseudomonas putida CGMCC3830. Sheng Wu Gong Cheng Xue Bao 30:412–424Google Scholar
- 17.Ekwuribe NN, Rodger Liddle (2011) Methods and compositions employing 4-aminophenylacetc acid compounds. United States Patent US 8,048,924 B2Google Scholar
- 18.Bedair AH, Ali FM, El-Agrody AM, Eid FA, El-Nassag MAA, El-Sherbeny G (2006) Preparation of 4-aminophenylacetic acid derivatives with promising antimicrobial activity. Acta Pharm 56:273–284Google Scholar
- 29.Sun HH, Gao WY, Fan HY, Wang HL, Wei DZ (2015) Cloning, purification and evaluation of the enzymatic properties of a novel arylacetonitrilase from Luminiphilus syltensis NOR5-1B: a potential biocatalyst for the synthesis of mandelic acid and its derivatives. Biotechnol Lett 37:1655–1661CrossRefGoogle Scholar
- 32.Nagasawa T, Nakamura T, Yamada H (1990) e-Caprolactam, a new powerful inducer for the formation of Rhodococcus rhodochrous J1 nitrilase. Arch Micobiol 155:13–17Google Scholar
- 45.Fan H, Chen L, Sun H, Wang H, Liu Q, Wei YRD (2017) Development of nitrilase-mediated process for phenylacetic acid production from phenylacetonitrile. Chem Pap 1985-1992Google Scholar