Isolation, Selection and Biodegradation Profile of Phenol Degrading Bacteria from Oil Contaminated Soil Bhavna Vishwas Mohite • Shraddha Pandurang Pawar • Ankush Morankar Received: 23 October 2010 / Accepted: 25 May 2011 / Published online: 4 June 2011 Ó Springer Science+Business Media, LLC 2011 Abstract In the present study, an aerobic bacterial strains OCS-A and OCS- B were isolated from an oil contaminated soil. The strains were identified to be Citrobacter freundi and Proteus mirabilis according to morphological, physiological and biochemical characteristics. The strains were able to degrade about 90% of 100 mg/L phenol within 80 h as sole carbon and energy source. The lag phase increased with increase in phenol concentration. Determination of metabolic intermediate 2-HMS, was done which indicate meta-cleavage pathway of phenol metabolism. Hence these isolates can be effectively used for bioremediation of phenol contaminated sites. Keywords Phenol Á Meta-cleavage pathway Á Bioremediation Á Microbial metabolism Á 2-HMS Introduction Now a day’s phenol is used by many industries in the manufacturing of products like insulation materials, adhe- sives, lacquers, paint, rubber, ink, dyes, illuminating gases, perfumes, soaps and toys. Phenol is one of the most widely used organic compounds in existence and is a basic structural unit for a variety of synthetic organic compounds including agricultural chemicals and pesticides, vegetables and coal. The German chemist, Runge isolated phenol from coal tar in 1834 and named karbolsaure (coal–oil acid or carbolic acid), though its composition was not known until 1841 (Nair et al. 2008). Biodegradation of phenol has increased strong attention throughout the world due to its poisonous properties and unrelenting nature (Leung et al. 1997). Microbial uptake and mineralization of phenol has been studied extensively (Dua et al. 2002; Lovely 2003; Watanabe 2001). The microorganisms that are normally used in phenol degra- dation include Pseudomonas sp, Candida tropicalis, Azo- tobacter sp, Alcaligenes sp, Acinetobacter sp etc. (Basha et al. 2010). The biological degradation is accomplished through benzene ring cleavage mediated by intracellular enzymatic reaction (Kumar et al. 2004). This study was aimed to evaluate the potential of phenol degraders from environment. Phenol degrading bacteria were isolated and their performance in treating the com- pound was investigated. Materials and Methods Two pure cultures used in this study were isolated from oil contaminated soil. The isolates were periodically subcultured on LB agar at 30°C and further maintained at 4°C. MP and MSM media were used in present study. MP medium com- posed of K 2 HPO 4 –2.75 gm, KH 2 PO 4 –2.25 gm (NH 4 )NO 3 – 1.0 gm, MgCl 2 Á6H 2 O–0.2 gm, NaCl–0.1 gm, FeCl 3 Á6H 2 O– 0.02 gm, CaCL 2 –0.01 gm (Watanabe et al. 1998). The minimal salt media (MSM) was used in this study. The medium was composed as: Na 2 HPO 4 (6 g), KH 2 PO 4 (3 g), NaCl (0.5 g), NH 4 Cl (1 g), CaCl 2 Á2H 2 O (1 M) and MgSO 4 7.H 2 O (1 M) in 1,000 mL double distilled water (Nagamani et al. 2009). The medium was autoclaved at 121°C for 15 min for sterilization. Phenol (10 mg/L) was separately sterilized and aseptically added into the sterile medium. Sample was collected from contaminated soil from oil industry and added into MP and MSM media in which B. V. Mohite (&) Á S. P. Pawar Á A. Morankar Department of Biotechnology, Moolaji Jaitha College, Jalgaon 425001, MS, India e-mail: mohite_bhavna@rediffmail.com 123 Bull Environ Contam Toxicol (2011) 87:143–146 DOI 10.1007/s00128-011-0322-z