微生物学通报 SEP 20, 2009, 36(9): 324~328 Microbiology tongbao@im.ac.cn 研究报告 2009 by Institute of Microbiology, CAS 脱硫弧菌 SRB7 对重金属铬 Cr(VI) 的还原特性 马小珍 费保进 金楠 赵锐 兰贵红 陈涛 2 乔代蓉 * (. 60064) (2. 6004) 摘要 : 利用化工厂污泥废水中分离到的一株耐酸脱硫弧菌 (Desulfovibrio SRB7), 对不同 ph 温度 碳源 菌废比等条件下 SRB7 还原 Cr(VI) 能力进行研究 并从 H 2 S 还原途径 电子传递途径以及胞外聚合物 (EPS) 吸附途径研究 SRB7 菌体整体去除 Cr(VI) 的特性 结果表明 : 当 Cr(VI) 起始浓度为 50 mg/l 时, ph 7.5, 培养温度 36 C, 碳源为乳酸钠, 混合菌液和 Cr(VI) 溶液的菌废比为 :5(V/V) 能获得很好的还原效果 在 SRB7 去除 Cr(VI) 的特性中, 吸附途径对 Cr(VI) 的去除几乎不起作用 ; 电子传递途径在 Cr(VI) 的还原过程中不占优势, 24 h 去除率为 5.42%; 而 H 2 S 途径在 Cr(VI) 的还原过程中占主导地位, 24 h 去除率为 78.02% 关键词 : 脱硫弧菌 SRB7, 还原条件, H 2 S 途径, 电子传递途径, 吸附途径 Characteristics of Reduce Cr(VI) by Desulfovibrio SRB7 MA Xiao-Zhen FEI Bao-Jin JIN Nan ZHAO Rui LAN Gui-Hong CHEN Tao 2 QIAO Dai-Rong * (. College of Life Science, Sichuan University, Chengdu, Sichuan 60064, China) (2. Shanghai Tianmei Scientific Instrument Limited Company Chengdu Branch Office, Chengdu, Sichuan 6004, China) Abstract: Pollution of heavy metal chromium Cr(VI) in environment would cause cancer. Chromium Cr(VI) was defined as one of 29 important pollutants by American EPA. This study was carried out on the conditions of removal Cr(VI) including different originated ph, temperature, carbon source and the volume ratio of bacteria to wastewater using an acidproof Desulfovibrio (SRB7) which was departed from sewage sludge effluent water of a chemical plant. Meanwhile the characteristics of reducing Cr(VI) through sulfureted hydrogen approach, electron transfer approach and adsorption approach were studied. The results showed that the conditions of removal Cr(VI) were as follows: ph 7.5, temperature 36 C, using lactate sodium as the carbon sources and the volume ratio of bacteria to wastewater at :5(V/V), when the influent concentration was 50 mg/l. In this study for characteristics of reducing Cr(VI) by SRB7, adsorption approach couldn t remove Cr(VI); electron transfer approach which removed 5.42% of the Cr(VI) at 24 h, was not preponderant in the process of reduce Cr(VI); sulfureted hydrogen approach which removed 78.02% of the Cr(VI) at 24 h, was preponderant in the process of reduce Cr(VI). Keywords: Desulfovibrio SRB7, Reduce conditions, Sulfureted hydrogen approach, Electron transfer approach, Adsorption approach 基金项目 : (No. 2005DKA2208-7); (No. 07PT-03); (No. 2008GZ002); (No. SB2007FY400) * 通讯作者 :Tel: 86-28-8542842; : qiaodr@sohu.com 收稿日期 :2008-2-25; 接受日期 :2009-02-26
: SRB7 Cr(VI) 325 Cr, Cr(III) Cr(VI), Cr(VI), EPA 29, Cr(VI) [],, Cr(III) Cr(VI) [2,3],, Cr(VI),, (Sulfate-reducing bacterium, SRB), [4], 2,, : Cr(VI) Mn(IV) U(VI),, Cu 2+ Zn 2+, Cr 2 O 2 7 3, : H 2 S (EPS), Cr(VI),, Cr(VI) (Desulfovibrio) [5] SRB7, ph (5.0~8.5), SRB7, Cr(VI), H 2 S (SRB) Cr(VI),, SRB7 材料与方法. 材料.. 菌株 : (SRB7)..2 培养基 : L : NaCl 2.0 g, NH 4 Cl.0 g, MgSO 4 7H 2 O 2.0 g, Na 2 SO 4 0.5 g, K 2 HPO 4 0.5 g, 70% 5 ml,.0 g, ph 7.5.2 硫酸盐还原菌的培养 500 ml, N 2, 0 5 Pa 20 min, SRB7 0%, 36 C 3 d,,, SRB.3 铬 Cr(VI) 的测定 (GB/T 5555.5 995) Cr(VI).4 脱硫弧菌 SRB7 对 Cr(VI) 的还原 50 mg/l Cr(VI) 00 ml, 5 ml,,, 36 C 24 h, 2 h Cr(VI) Cr(VI).5 脱硫弧菌 SRB7 对 Cr(VI) 的还原条件研究.5. 不同温度对 SRB7 还原 Cr(VI) 的影响 : 25 C 36 C 45 C 55 C, Cr(VI).5.2 不同 ph 对 SRB7 还原 Cr(VI) 的影响 : ph 4.0 5.0 5.5 6.0 6.5 7.0 7.5 8.0 8.5 9.0 9.5, Cr(VI).5.3 不同碳源对 SRB7 还原 Cr(VI) 的影响 : 5 SRB7, Cr(VI).5.4 不同菌废比对 SRB7 还原 Cr(VI) 的影响 : :0.5:0 :5 :4, Cr(VI).6 脱硫弧菌 SRB7 还原铬 Cr(VI) 特性的初步研究.6. SRB7 还原 Cr(VI) 吸附途径的研究 : EPS [6], 5 ml EPS, Cr(VI).6.2 SRB7 还原 Cr(VI) 电子转移途径的研究 : [7], 5 ml, Cr(VI).6.3 SRB7 还原 Cr(VI) H 2 S 途径的研究 : 00 ml
326 微生物学通报 2009, Vol.36, No.9, 4000 r/min 30 min H 2 S 5 ml, Cr(VI) 2 结果与分析 2. 脱硫弧菌 SRB7 对 Cr(VI) 的还原 SRB7 Cr(VI), 2 h Cr(VI) 69.84%,, 24 h 96.86% 图 SRB 对铬 Cr(VI) 的还原 Fig. Reduction of Cr(VI) by SRB 2.2 脱硫弧菌 SRB7 还原铬 Cr(VI) 条件的优化 2.2. 不同温度条件下 SRB7 对 Cr(VI) 的还原 : SRB7 36 C, 45 C, OD 55 C, SRB7 [5] 36 C Cr(VI) 96.64% 45 C Cr(VI) 36 C, 93.92% 25 C Cr(VI) 36 C 45 C 55 C ( ), SRB7 Cr(VI) 36 C 2.2.2 不同 ph 条件下 SRB7 对 Cr(VI) 的还原 : SRB7 ph 2, ph 5.0~8.5, SRB7 Cr(VI), ph 7.5 96.86%, ph 5.0 85.96% Cr(VI) 表 不同温度下 Cr(VI) 的去除率 Table Removal rate of Cr(VI) under different temperature ( C) Temperature ( C) 25 89.32 36 96.64 45 93.92 55 7.64 表 2 不同 ph 条件下 Cr(VI) 的去除率 Table 2 Removal rate of Cr(VI) under different ph ph 4.0 7.22 5.0 85.96 6.0 92.68 6.5 95.60 7.0 96.24 7.5 96.86 8.0 94.76 9.0 0.6 2.2.3 不同碳源条件下 SRB7 对 Cr(VI) 的还原 : SRB7, : > > > > [5], SRB7 Cr(VI) 5, H 2 S, Cr(VI) 24 h, 96.44%,, Cr(VI) ( 3), SRB7 Cr(VI) 表 3 Table 3 不同碳源条件下 Cr(VI) 的去除率 Removal rate of Cr(VI) under different carbon sources Carbon source Sodium formate 9.00 Sodium acetate 89.32 Ethanol 93.08 Sodium propionate 94.56 Sodium lactate 96.44 2.2.4 不同菌废比条件下 SRB7 对 Cr(VI) 的还原 : 4 SRB7 Cr(VI) ( 2), :0, Cr(VI) 77.8%.5:0 :5 :4, Cr(VI) 00% 4 Cr(VI), :5, Cr(VI) 0.00952 mg/(ml h) :4,, 99.38%, Cr(VI) :5 Cr(VI) 50 mg/l, :5 2.3 脱硫弧菌 SRB7 还原铬 Cr(VI) 特性的初步研究 2.3. 胞外聚合物 (EPS) 吸附途径的研究 : (EPS) 24 h, Cr(VI)
: SRB7 Cr(VI) 327 2.3.3 H 2 S 途径的研究 : SRB7 H 2 S 24 h Cr(VI) SRB7 24 h Cr(VI) ( 4) 3 4, H 2 S, 80% 图 2 不同菌废比条件下 Cr(VI) 的去除率 Fig. 2 Removal rate of Cr(VI) under different volume ratio of bacteria to wastewater, 4 (EPS) Cr(VI) 2.3.2 电子传递途径的研究 : SRB7 24 h Cr(VI) 27.95 mg/l, 5.42%, SRB7 4 h Cr(VI) (2.4 mg/l) ( 3) 表 4 胞外聚合物 (EPS) 对 Cr(VI) 的吸附情况 Table 4 Adsorption condition of Cr(VI) by EPS (h) Time (h) Cr(VI) (mg/l) Remaining concentration of Cr(VI) (mg/l) 0 50 4 49.53 8 49.32 2 49.42 6 49.63 20 49.74 24 49.63 图 3 电子传递途径与 SRB7 原始菌液还原 Cr(VI) 的比较 Fig. 3 Comparison of reduction Cr(VI) by electron transfer approach and SRB7 initial culture 图 4 H 2 S 途径与 SRB7 原始菌液还原 Cr(VI) 的比较 Fig. 4 Comparison of reduction Cr(VI) by H 2 S approach and SRB7 initial culture 3 讨论 3. 影响脱硫弧菌 SRB7 还原重金属铬 Cr(VI) 的环境因素 Cr(VI),, Cr(VI) SRB7 36 C Cr(VI),, Cr(VI) Cr(VI) SRB7, Cr(VI), ph Cr(VI), ph [8],, SRB7 ph 7.5 Cr(VI), ph 5.0,, 85.96% Cr(VI) SRB7 Cr(VI),
328 微生物学通报 2009, Vol.36, No.9 [9],, Cr(VI) Cr(VI) :5,, Cr(VI), [0], SRB7 24 h Cr(VI) 96.86%,, Humphries [0] 3.2 脱硫弧菌 SRB7 对于重金属 Cr(VI) 可能的还原特性 3 : ) SRB S 2 ; 2) ; 3) (EPS) [7], (EPS) [ 3], Cu 2+ Zn 2+[6,4], (EPS), SRB7 (EPS) Cr 2 O 7 2 Cr(VI), [5] [9], Cr(VI), SRB7 Cr(VI), Cr(VI) H 2 S, H 2 S Cr(VI) SRB7 Cr(VI), 4 h, Cr(VI),, H 2 S Cr(VI), SRB7 Cr(VI), 4 h, H 2 S 4 h, H 2 S Cr(VI), SRB7 Cr(VI) H 2 S, 36 h Cr(VI) 98.54%,, SRB7 Cr(III), Cr(VI) 参考文献 [],,.., 997, 37(3): 3 5. [2].., 993, 4(4): 4 6. [3],,,. (VI)., 998, 7(): 85 87. [4],,,.. :, 993, pp.28 222. [5],,,.. ( ), 2006, 43(2): 45 457. [6],,. Cu 2+., 2005, 3(9): 25 28. [7] Payne RB, Gentry DM, Rapp-Giles BJ, et al. Uranium reduction by Desulfovibrio desulfricans strain G20 and a cytochrome C 3 mutant. Appl Environ Microb, 2002, 68(6): 329 332. [8] Reis MA, Almeida JS, Lemos PC, et al. Effect of hydrogen sulfide on growth of sulfate reducing bacteria. Biotechnol Bioeng, 992, 40(5): 593 600. [9],,,. Cr(VI)., 2005, : 05 0. [0] Humphries AC, Nott KP, Hall LD, et al. Reduction of Cr(VI) by immobilized cells of Desulfovibrio vulgaris NCIMB 8303 and Microbacterium sp. NCIMB 3776. Biotechnol Bioeng, 2005, 90(5): 589 596. [] Fukushi K, Kato S, Auntsuki T, et al. Isolation of copper binding proteins from activated sludge Znlture. Water Sci Technol, 200, 44: 453 459. [2] Beech IB, Cheung CWS. Interactions of exopolymers produced by sulphate reducing bacteria with metal ions. Int Biodeter Biodegr, 995, 35: 59 72. [3],,. (EPS) Cd., 2004, 7(5): 52 56. [4],,,. Zn 2+., 2005, 8(6): 53 55. [5] Smith WL, Gadd GM. Reduction and precipitation of chromate by mixed culture sulphate-reducting bacterial biofilms. J Appl Microbiol, 2000, 88: 983 99.