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Characterization of Five Chromium-Removing Bacteria Isolated from Chromium-Contaminated Soil

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Characterization of Five Chromium-Removing Bacteria Isolated from Chromium-Contaminated Soil Water Air Soil Pollut (2014) 225:1904 DOI 10.1007/s11270-014-1904-2 Characterization of Five Chromium-Removing Bacteria Isolated from Chromium-Contaminated Soil Zhiguo He & Shuzhen Li & Lisha Wang & Hui Zhong Received: 21 January 2013 /Accepted: 10 February 2014 /Published online: 21 February 2014 # Springer International Publishing Switzerland 2014 Abstract The potential for bioremediation of chromi- Keywords Chromium-removing bacteria . um pollution using bacteria was investigated in this Pseudochrobactrum saccharolyticum . Aerobic process . study. Five chromium-removing bacteria strains were Biotransformations . Bioremediation . Waste treatment successfully isolated from Cr(VI)contaminated soils and identified by their 16S rRNA gene sequences. The optimum growth temperature (30–40 °C) and pH (8.5– 1 Introduction 11) for the five isolates were investigated. The effect of initial Cr(VI) concentrations (0–1,575 mg L−1)onbac- Among heavy-metal pollutants, chromium (Cr) is con- terial growth was also studied. Results showed that sidered to be toxic and one of the main pollutants Pseudochrobactrum saccharolyticum strain W1 had (Yewalkar et al. 2007). Chromium is widely used in high chromium-removing ability and could grow at − industrial operations such as leather tanning, pigment Cr(VI) concentrations from 0 to 1,225 mg L 1.Toour production, electroplating, paints, steel manufacture, knowledge, this is the first report of chromium removal and automobile production (Wang and Xiao 1995; by a member of the Pseudochrobactrum genus. Pattanapipitpaisal et al. 2001). Intensive industrial ap- Sporosarcina saromensis W5 had the highest − − plications of chromium and releases of associated waste chromium-removing rate of 0.79 mg h 1 mg 1 biomass. have caused substantial soil contamination. Chromium Exopolysaccharide (EPS) production and components exists in several oxidation states, from −2 to 6 (Jacobs of the five bacteria strains were also investigated, and a and Testa 2005). However, in the environment, the most positive relationship was found between the bacterial stable and common forms are the trivalent [Cr(III)] and chromium removal and EPS production. hexavalent [Cr(VI)] species (Fendorf 1995). The Cr(VI) : : form is more reactive and harmful than the trivalent Z. He S. Li L. Wang (Francisco et al. 2002) which is, in comparison, less School of Minerals Processing and Bioengineering, Central South University, toxic, less soluble, and less mobile than the hexavalent Changsha 410083, People’s Republic of China form (Stanin 2005). In recent years, more and more : : attention has been focused on the bioremediation of Z. He S. Li L. Wang Cr(VI) contamination with chromate-resistant bacteria Key Laboratory of Biohydrometallurgy of Ministry of Education, Central South University, (Zhu et al. 2008). Changsha, Hunan, China 410083 In this work, plating method was used to isolate Cr(VI)-removing bacteria from chromium- * H. Zhong ( ) contaminated soil samples adjacent to a chromium land- School of Life Science, Central South University, Changsha, Hunan, China 410012 fill in Changsha, Hunan province, China. The growth e-mail: [email protected] conditions, such as pH, temperature, and the initial 1904, Page 6 of 10 Water Air Soil Pollut (2014) 225:1904 A number of previous studies have shown that many Cr(VI) concentration of 1,050 mg L−1.Inverylimited microorganisms possess Cr(VI) tolerance/resistance. As previous studies, Sporosarcina sp. have been found outlined in Table 1, P. saccharolyticum strain W1, capable of growing in Cr(VI) concentration of 5 ppm Oceanobacillus sp. W4, and S. saromensis W5 had (5 mg L−1) (Fein et al. 2002) and tolerate 2,900 μM higher Cr(VI)-resistance ability than most other previ- (=150.8 mg L−1)ofCr(Bafana2011), while ously identified strains, which highlight the potential of S. saromensis W5 could tolerate Cr(VI) concentration these three isolates as bioremediators of Cr(VI) from of up to 1, 400 mg L−1. Most previous studies have chromium-polluted areas. Pseudochrobactrum was re- found that Cr(VI) inhibits bacterial growth at any Cr(VI) cently proposed by Kämpfer et al. (2006) and comprises concentration (He et al. 2009; Middleton et al. 2003; five species to date: P. saccharolyticum (Kampfer et al. Camargo et al. 2003). It has been reported that the 2006), Pseudochrobactrum asaccharolyticum (Kampfer growth of Arthrobacter sp. and Bacillus sp. was stimu- et al. 2006), Pseudochrobactrum kiredjianiae (Kampfer lated by Cr(VI) concentrations of 50 and 5 mg L−1, et al. 2007), Pseudochrobactrum lubricantis (Kampfer respectively (Megharaj et al. 2003). In this study, growth et al. 2009), and Pseudochrobactrum glaciei of the five isolates was promoted by Cr(VI) when under (Romanenko et al. 2008). Pseudochrobactrum sp. certain Cr(VI) concentrations. P. saccharolyticum strain (Kampfer et al. 2006; Kampfer et al. 2007; Kampfer W1 and S. saromensisW5 were stimulated by Cr(VI) et al. 2009; Romanenko et al. 2008) was able to grow at concentration from 0 to 875 mg L−1 and from 175 to 15–40 °C (optimum temperature was 25–30 °C) and the 1,400 mg L−1, respectively. This mechanism was not optimum pH value was about 7.1–7.5. In this study, the explicit and need to be further studied. optimum temperature for P. saccharolyticum strain W1 was 35 °C and it could grow at a pH range of 8.0–11.0, 3.3 Cr(VI) Removal by the Five Isolates with optimum pH value of 9.5. To our knowledge, this is the first report of chromium resistance by a strain from The ability of the five isolates to remove Cr(VI) when Pseudochrobactrum sp. incubated in their respective optimum initial Cr(VI) Oceanobacillus sp. was initially reported to grow at concentrations for growth was studied. Abiotic removal pH 9–10 and at 15–40 °C with the optimum temperature of Cr(VI) has also been evaluated and was found to be at 30–36 °C and was identified as a halotolerant obligate neglectable. The changes in Cr(VI) concentration in the alkaliphile isolated from the skin of a rainbow trout medium with time for the five isolates were shown in (Yumoto et al. 2005). Molokwane et al. (2008) also Fig. 3. A decrease of the chromium concentration in a reported that a mixed culture of bacteria, containing solution was observed with time, with the maximum Oceanobacillus sp., could remove Cr(VI) under anaer- change generally observed within the initial 15 h for all obic condition. In this study, Oceanobacillus sp. W4 isolates. Among the five isolates, S. saromensis W5 had was observed to have the ability to remove chromate the highest chromium-removal rate of 0.79 mg h−1 mg−1 under aerobic conditions and could grow at a wider biomass, as the Cr(VI) concentration decreased from range of temperature and pH value than those previously 1,050 to 165 mg L−1 within the first 15 h. The concen- reported. tration of Cr(VI) decreased from 350 to 111.76 mg L−1 As described by Ilhan et al. (2004), the optimum (a rate of 0.21 mg h−1 mg−1 biomass) in incubations with temperature for chromium removal by a strain of P. saccharolyticum strain W1 within the first 15 h. The S. saprophyticus was 27 °C and the optimum pH value removal rate of Cr(VI) by S. saprophyticus strain W2 was found to be at 2.0. Mistry et al. (2010)reportedthe and Lysinibacillus sp. strain W3 was both about optimum pH value was 7.0 for the chromate removal of 0.07 mg h−1 mg−1 biomass within 15 h at the initial astrainofS. saprophyticus. In this study, Cr(VI) concentration of 175 mg L−1. According to Fein S. saprophyticus W2 could grow at a pH range of 8.0– et al. (2002), a strain of Sporosarc inaureae was capable 10.0 even though the 16S rRNA gene sequences share of removing Cr from a medium containing 5 mg L−1 99 % identity, which may indicate that strain W2 and the Cr(VI); calculation based on data got in the first 20 h strain of S. saprophyticus described in Ilhan et al. (2004) showed a removal rate of 1.15×10−5 mg h−1 mg−1 may belong to different subspecies. S. saromensis W5 biomass. S. saromensis W5 had much greater ability had the highest resistance to chromium among the five to remove Cr(VI) when compared with that. As de- isolates, as it reached the maximum cell density at scribed by Ilhan et al. (2004), the Cr(VI) bio-removing 1904, Page 8 of 10 Water Air Soil Pollut (2014) 225:1904 removal and EPS production was investigated, and the References results are shown in Table 2. The EPS content of the five isolates ranged from 68.03 to 189.19 mg in 1 g of dried Badar, U., Ahmed, N., Beswick, A., Pattanapipitpaisal, P., & cell material. P. saccharolyticum strain W1 and Macaskie, L. (2000). Reduction of chromate by microorgan- S. saromensis W5, which had higher chromium- isms isolated from metal contaminated sites of Karachi, – removing ability as compared with the other isolates, Pakistan. Biotechnology Letters, 22(10), 829 836. Bafana, A. (2011). Mercury resistance in Sporosarcina sp. G3. produced much more EPS, with 178.60 and Biometals, 24(2), 301–309. − 189.19 mg g 1, respectively. S. saprophyticus strain Camargo, F., Okeke, F., & Frankenberger, B. (2003). Chromate W2 (82.15 mg g−1), Lysinibacillus sp. strain W3 reduction by chromium-resistant bacteria isolated from soils −1 contaminated with dichromate. Journal of Environmental (68.03 mg g ), and Oceanobacillus sp. W4 – −1 Quality, 32(4), 1228 1233. (140.40 mg g ) had lower EPS contents which Camargo, F. A. O., Okeke, B. C., Bento, F.
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