Nonconventional hydrolytic dehalogenation of 1-chlorobutane by dehydrated bacteria in a continuous solid-gas biofilter

Benjamin Erable (1) , Isabelle Goubet (1) , Sylvain Lamare (1) , Amira Seltana (1) , Marie Dominique Legoy (1) , Thierry Maugard (1)

1 LBCBO - Laboratoire de Biotechnologies et de Chimie Bio-organique

Rhodococcus erythropolis NCIMB 13064 and Xanthobacter autotrophicus GJ10 are able to catalyze the conversion of halogenated hydrocarbons to their corresponding alcohols. These strains are attractive biocatalysts for gas phase remediation of polluted gaseous effluents because of their complementary specificity for short or medium and for mono-, di-, or trisubstituted halogenated hydrocarbons(C2-C8 for Rhodococcus erythropolis and C1-C4for Xanthobacter autotrophicus). After dehydration, these bacteria can catalyze the hydrolytic dehalogenation of 1-chlorobutane in a nonconventional gas phase system under a controlled water thermodynamic activity (aw). This process makes it possible to avoid the problems of solubility and bacterial development due to the presence of water in the traditional biofilters. In the aqueous phase, the dehalogenase activity of Rhodococcus erythropolis is less sensitive to thermal denaturation and the apparent Michaelis-Menten constants at 30jC were 0.4 mM and 2.40 Amol min�1 g�1 for Km and Vmax, respectively. For Xanthobacter autotrophicus they were 2.8 mM and 0.35 Amol min�1 g�1. In the gas phase, the behavior of dehydrated Xanthobacter autotrophicus cells is different from that observed with Rhododcoccus erythropolis cells. The stability of the dehalogenase activity is markedly lower. It is shown that the HCl produced during the reaction is responsible for this low stability. Contrary to Rhodococcus erythropolis cells, disruption of cell walls does not increase the stability of the dehalogenase activity. The activity and stability of lyophilized Xanthobacter autotrophicus GJ10 cells are dependant on various parameters. Optimal dehalogenase activity was determined for water thermodynamic activity (aw) of 0.85. A temperature of 30jC offers the best compromise between activity and stability. The pH control before dehydration plays a role in the ionization state of the dehalogenase in the cells. The apparent Michaelis-Menten constants Km and Vmax for the dehydrated Xanthobacter autotrophicus cells were 0.07 (1-chlorobutane thermodynamic activity) and 0.08 Amolmin�1 g�1 of cells, respectively. A maximal transformation capacity of 1.4 g of 1-chlorobutane per day was finally obtained using 1g of lyophilized Xanthobacter autotrophicus GJ10 cells.

Mots clés: Bioremediation; Haloalkane dehalogenase; Rhodococcus erythropolis NCIMB 13064; Solid-gas biofilter; Waste air treatment; Xanthobacter autotrophicus GJ10

Domaines: Génie chimique; Chimie organique; Biotechnologies; Biotechnologie

Benjamin Erable, Isabelle Goubet, Sylvain Lamare, Amira Seltana, Marie Dominique Legoy, et al.. Nonconventional hydrolytic dehalogenation of 1-chlorobutane by dehydrated bacteria in a continuous solid-gas biofilter. Biotechnology and Bioengineering, 2005, vol. 91, pp. 304-313. ⟨10.1002/bit.20437⟩. ⟨hal-00786202⟩