Biodegradation of 2-Chlorobenzoic Acid by Enterobacter cloacae: Growth Kinetics and Effect of Growth Conditions

Abstract

ABSTRACTEnterobacter cloacae was originally isolated from soil irrigated with wastewater on the basis of its ability to grow with linear alkylbenzene sulfonate (LAS) as the sole source for carbon and energy. The isolated bacterium was grown in batch cultures using a 2-chlorobenzoic acid (2-CBA)-containing minimal salt medium (MSM). 2-CBA was found to be the sole source for carbon and energy. 2-CBA inhibited the growth rate with a maximum concentration of 10 mM, after which no growth occurred. The Haldane model was used to predict the specific growth rate concentration data. 2-CBA degradation by starved E. cloaca cells was faster than that of nonstarved cells. The maximum growth rates on 2-CBA (2 mM) for starved and nonstarved cells reached only 0.34 and 0.28 h−1, respectively. Glucose, lactose, sucrose, maltose, succinic acid, and mannitol as additional carbon sources at a fixed concentration (0.2%) caused the degradation rate of 2-CBA to proceed faster at ranges between 1.08- and 1.5-fold higher than that of the control. In contrast, using only fructose and sorbitol as the carbon sources showed catabolic repression of the degradation activity of 2-CBA by E. cloaca cells, although their cell mass was improved. All nitrogen sources supplied caused an increase in cell mass, whereas only lysine, alanine, glutamine, casein, and yeast extract caused a decrease in the degradation rate of 2-CBA, with a range between 12% and 28%. The activity of C120 could be detected in a crude extract of E. cloacae cells, indicating that the chloroaromatic ring fission occurs through the ortho pathways, not through the meta pathways. The data showed that different initial cell (inocula) densities did not affect the induction time for 2-CBA degradation. However, doubling the initial cell densities reduced the time required for reaching the complete degradation. 2-CBA degradation was optimally achieved at a 37°C incubation temperature and a pH of 7.5.