Bambang Sunarko, Nunik Sulistinah
May 18, 2017
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Abstract
ABSTRACT Characterization of Nitrile Hydratase and Amidase of Pseudomonas sp BP3 in Bioconversion of Adiponitrile to Adipic Acid. Adipic acid is a commercially important compound, primarily used as precursor for the production of nylon 6.6. It is also used for plasticizer, fibers, and food additive. Synthesis of adipic acid by chemical means requires large amount of energy and concentrated acid. It also produces N2O as by product, which is very toxic and suspected causes depletion of the ozone layer. The purpose of this research was to study the bioconversion of adiponitrile by Pseudomonas sp. BP3 and to characterize the involved enzymes in the whole cell. Pseudomonas sp. BP3 was able to utilize adiponitrile as the sole source of carbon and nitrogen. It’s doubling time (td) and growth rate constant (?) during the growth in adiponitrile were 2 hours and 0.346/h, respectively. The optimum pH and temperature of nitrilehydratase were pH 7.0 and 30°C, respectively, while those of amidase were pH 6 and 50°C. Vmax and Ks of nitrile hydratase were 8.3 nM/ml.min. and 55.56 mM, respectively, and of amidase were 5,9 nM/ml.min and 50 mM. The rate of adiponitrile consumption was 0.245 mM/ h and of adipic acid formation was 0.181 mM/h. The yield of bioconversion of adiponitrile and adipamide were about 50 % and 25%, respectively. Key words: Bioconversion, adiponitrile, adipic acid, Pseudomonas sp. BP3, nitrile hydratase, amidase