微生物学通报 FEB 20, 2009, 36(2): 255~260 Microbiology tongbao@im.ac.cn 2009 by Institute of Microbiology, CAS 专论与综述 异养硝化细菌脱氮特性及研究进展 * 苟莎黄钧 ( 610041) 摘要 : 异养硝化细菌能够在利用有机碳源生长的同时将含氮化合物硝化生成羟胺 亚硝酸盐 硝酸盐等产物, 多数还能同时进行好氧反硝化作用, 直接将硝化产物转化为含氮气体 因此, 这类细菌已成为废水处理中生物脱氮新工艺的重要研究对象 本文综述了目前所分离出的一些异养硝化菌的脱氮特性, 分析了各种环境条件如温度 ph 溶解氧 碳源类型 C/N 以及抑制剂等对异养硝化菌的影响, 并介绍了异养硝化菌的应用现状及前景 关键词 : 异养硝化细菌, 异养硝化, 好氧反硝化 Advances in Denitrification Characteristics of Heterotrophic Nitrification Bacteria GOU Sha HUANG Jun * (Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu, Sichuan 610041, China) Abstract: Heterotrophic nitrification bacteria are able to utilize organic carbon sources to grow and produce hydroxylamine, nitrite and nitrate from nitrogen compounds, and most of them can also denitrify these products to gaseous nitrogen compounds simultaneously. Therefore, more and more attentions are paid to heterotrophic nitrification bacteria for wastewater treatment. This paper reviews the denitrification characteristics of some isolated heterotrophic nitrification bacteria, and analyzes the influence of various conditions to the heterotrophic nitrification bacteria, such as temperature, ph, DO, carbon sources, C/N, and inhibitors, etc. At last, the present situation and potential applications of wastewater treatment by heterotrophic nitrification bacteria are introduced. Keywords: Heterotrophic nitrification bacteria, Heterotrophic nitrification, Aerobic denitrification,,,,,,, [1],, [2],, COD [1],,,, 基金项目 : 863 (No. 2006AA06Z330) * 通讯作者 :Tel: 86-28-85244876; Fax: 85222753; : huangjun@cib.ac.cn 收稿日期 :2008-06-17; 接受日期 :2008-08-29
256 微生物学通报 2009, Vol.36, No.2, 1 异养硝化菌的发现及其脱氮特性 ( ),,,,, [3,4],,,, [5], [5,6],, COD,,,, [7,8],,, 1 Table 1 表 1 一些异养硝化菌的脱氮特性 The denitrification characteristics of some heterotrophic nitrification bacteria Strain type Original place Nitrification product NO 2 Denitrification product NO 3 N 2 NO N 2 O Acinetobacter sp. [9] ABS (1) / + + / / / Acinetobacter sp. YY4* [10] / + / / / Alcaligenes faecalis NH17* [11] / + + + Alcaligenes faecalis No. 4* [12,13] + + + + + Alcaligenes faecalis subsp.parafaecalis* [14] / / + + / + + Arthrobacter sp. (strain9006) [15] / + / / / Arthrobacter B D [6] / + + / / / Bacillus sp. LY * [16] MBR + + + + Bacillus subtilis [17] / + Corynebacterium sp. [9] ABS (1) / + + / / / Diaphorobacter sp.* [18] / + / / / Pseudomonas azalaica PB16* [19] / / + + Pseudomonas alcaligenes AS-1* [20] / + + Pseudomonas putida [21] + + + / + Pseudomonas putida sp. DN1. 2* [22] / / / / Rhodococcus sp. HN* [8] / + / / / Thiosphaera pantotropha* [23] / + + / + Thermus sp.* [24], / + / / / Xanthomonas sp. [9] ABS (1) / + + / / / +: ; : ; /: ; : ; *: ; ABS (1) : (styrene). Note: +: Product detected; : No product detected; /: No mention in the reference; : Anaerobic denitrification; *: Aerobic or facultative aerobic denitrification; ABS (1) correspond to the wastewater contains acrylonitrile, butadiene and styrene.
: 257 2 异养硝化菌脱氮影响因素,, [25], ( ) C/N ph 2.1 有机碳源类型, [26],,, Alcaligenes faecalis No. 4 [27] Arthrobacter BD α- [6] Arthrobacter sp., [15] Otani, Alcaligenes faecalis, [28] Hu Acinetobacter sp. Xanthomonas sp. 13 [9],, [17,26], [3,26,29] 2.2 底物浓度 ( 碳源 氮源 ), Mevel [26] C/N 3, ( ) ( ) 5, Bacillus MS30 5 mmol/l~30 mmol/l,, 0.65 μmol~0.87 μmol NO 2, ; 60 mmol/l, (1.80 μmol NO 2 /mg dry wt); 120 mmol/l,, 1.27 μmol NO 2 /mg dry wt [16] Bacillus sp. LY, 40 mg/l 80 mg/l 120 mg/l, (COD 400 mg/l); (COD 800 mg/l); (COD 1600 mg/l), 2.3 C/N C/NKim [17], C/N 8 C/N 4 Alcaligenes faecalis OKK17, C/NC/N 6,, [3] Alcaligenes faecalis No. 4, COD/TN 5,, ; COD/TN 20,, COD ; COD/TN 10,, COD [27], C/N, COD, C/N 2~10 [2,30] 2.4 ph, ph Arthrobacter B D ph 3~10 ph, ph<3 C, N, ph 3~7, ph 8.0~9.5 [6] Alcaligenes faecalis No. 4 ph 6 7 8 [12],, ph ph Arthrobacter B D, ph -, ph [6] ph ph Bacillus MS30 ph 6.0~6.5,
258 微生物学通报 2009, Vol.36, No.2 ph 7.5~8.0 ph 7.5, 75% [26], ph ph ph, ph 6~8 2.5 温度, 15 C 40 C, Alcaligenes faecalis subsp. parafaecalis N 2 O NO 28 C~35 C [31] Alcaligenes faecalis No. 4 30 C 37 C, 20 C,, [27], Acinetobacter sp. YY4 20 C~40 C,, 50 C, Acinetobacter sp. YY4, [10] Alcaligenes faecalis OKK17 35 C, 30 C [3] Bacillus MS30 55 C~60 C, 65 C, 75% [26], 28 C~37 C [10,27,31] 2.6 溶解氧浓度 (DO), Thiosphaera pantotropha (Nitrosomonas europaea), C/N Thiosphaera pantotropha Nitrosomonas europaea [4] Pseudonocardia ammonioxydans H9 DO 0.5 mg/l~1.5 mg/l (COD 1000 mg/l~3000 mg/l, 120 mg/l), ( >80%) [32], DO Robertson [23] Patureau DO (Microvirgula aerodenitrificans) DO ( DO 0.35 mg/l, Microvirgula aerodenitrificans DO 4.5 mg/l),, DO, [33] [4,23] Thiosphaera pantotropha DO, N 2 N 2 O DO, N 2 O, DO 95%, N 2 O [4],, 2.7 抑制剂, [21,23,29] (10 Pa) [21], Daum [21] 5% γ- (γ-aminobutyrate) Pseudomonas putida, NO, Paracoccus denitrificans, Thiosphaera pantotropha [23] Robertson [29] Thiosphaera pantotropha Pseudomonas denitrificans,,,, 3 异养硝化菌应用现状 3.1 异养硝化菌处理合成污水, [34] (105 mg/l) (95 mg/l) (69 mg/l)
: 259, (COD 1100 mg/l), 4 h 85% 60% 70% [35] MBR (COD 250 mg/l~750 mg/l, 45 mg/l~50 mg/l) COD/TKN 15, MBR 90.6% Gupta [30] (rotating biological contactor) Thiosphaera pantotropha COD 1000 mg/l, 22 mg/l, 82.5% Ahmad [32] Pseudonocardia ammonioxydans H9 (Air-Lift Bioreactor) (COD 0 mg/l~3000 mg/l, 120 mg/l), C/N, (70%~80%) C/N 4, COD (<60%) 3.2 异养硝化菌处理实际污水 [10] Acinetobacter sp. YY4 (COD 531.95 mg/l, 61.11 mg/l) (COD 875.72 mg/l, 221.01 mg/l), 12.4 g/l 89.54% (9 h) 95.79% (36 h) Joo [13] Alcaligenes faecalis No. 4 COD 12000 mg/l, 2000 mg/l, 65%, COD 100%,, [36] 4 结语,, C C/N ph DO,, 3 (1), ; (2),, ; (3),,,,,, COD, 参考文献 [1],,,.., 2006, 6: 41 42. [2],.., 2003, 25(5): 283 285. [3] Takayuki N, Taro Y, Hirosugu M, et al. Conditions for nitrification and denitrification by an immobilized heterotrophic nitrifying bacterium Alcaligenes faecalis OKK17. Journal of Fermentation and Bioengineering, 1998, 86(4): 351 356. [4] Jetten MSM, Logemann S, Muyzer G, et al. Novel principles in the microbial conversion of nitrogen compounds, Antonie van Leeuwenhoek, 1997, 71: 75 93, 1997. [5] Pedersen H, Dunkin KA, Firestone MK. The relative importance of autotrophic and heterotrophic nitrification in a conifer forest soil as measured by N-15 tracer and pool dilution techniques. Biogeochemistry, 1999, 44(2): 135 150. [6] Brierley EDR, Wood M. Heterotrophic nitrification in an acid forest soil: isolation and characterisation of a nitrifying bacterium. Soil Biology & Biochemistry, 2001, 33(10): 1403 1409. [7] Schmidt I, Sliekers O, Schmid M, et al. New concepts of microbial treatment processes for the nitrogen removal in wastewater. FEMS Microbiology Reviews, 2003, 27(4): 481 492. [8],, 旸,.., 2003, 43(2):156 161. [9] Hu TL, Kung KT. Study of heterotrophic nitrifying bacteria from wastewater treatment systems treating acrylonitrile, butadiene and styrene resin wastewater. Water Science and Technology, 2000, 42(3-4): 315 321. [10],,,.. ( ), 2007, 37(3):
260 微生物学通报 2009, Vol.36, No.2 486 490. [11] Emiko M, Nobuhilko N, Toshiaki NK, et al. A simple screening procedure for heterotrophic nitrifying bacteria with oxygen-tolerant denitrification activity. Journal of Bioscience and Bioengineering, 2003, 95(4): 409 411. [12] Joo HS, Hirai M, Shoda M. Nitrification and denitrification in high-strength ammonium by Alcaligenes faecalis. Biotechnology Letters, 2005, 27(11): 773 778. [13] Joo HS, Hiraia M, Shoda M Piggery wastewater treatment using Alcaligenes faecalis strain No. 4 with heterotrophic nitrification and aerobic denitrification. Water Research, 2006, 40(16): 3029 3036. [14] Blagodatsky SA, Kesik M, Papen H, et al. Production of NO and N 2 O by the heterotrophic nitrifier Alcaligenes faecalis subsp. parafaecalis under varying conditions of oxygen saturation. Geomicrobiology Journal, 2006, 23(3-4): 165 176. [15] Witzel KP, Overbeck HJ. Heterotrophic nitrification by Arthrobacter sp. (strain 9006) as influenced by different cultural conditions, growth state and acetate metabolism. Archives of Microbiology, 1979, 122(2): 137 143. [16],,,. Bacillus sp. LY., 2007, 28(6): 1404 1408. [17] Kim JK, Park KJ, Cho KS, et al. Aerobic nitrification-denitrification by heterotrophic Bacillus strains. Bioresource Technology, 2005, 96(17): 1897 1906. [18] Khardenavis AA, Kapley A, Purohit HJ. Simultaneous nitrification and denitrification by diverse Diaphorobacter sp. Applied Microbiology and Biotechnology, 2007, 77: 403 409. [19] Jetten MSM, debruijn P, Kuenen JG. Hydroxylamine metabolism in Pseudomonas PB16: Involvement of a novel hydroxylamine oxidoreductase. Antonie Van Leeuwenhoek International Journal of General and Molecular Microbiology, 1997, 71(1-2): 69 74. [20] Su JJ, Yeh KS, Tsheng PW. A strain of Pseudomonas sp. isolated from piggery wastewater treatment systems with heterotrophic nitrification capability in Taiwan. Current Microbiology, 2006, 53(1): 77 81. [21] Daum M, Zimmer W, Papen H, et al. Physiological and molecular biological characterization of ammonia oxidation of the heterotrophic nitrifier Pseudomonas putida. Current Microbiology, 1998, 37(4): 281 288. [22] Huang J( ), Yang H( ), Li YJ( ).. : 200710049432.1. 2007.7.2. [23] Robertson LA, Kuenen JG. Combined heterotrophic nitrification and aerobic denitrification in Thiosphaera pantotropha and other bacteria. Antonie van Leeuwenhoek, 1990, [24] Mevel G, Prieur D. Thermophilic heterotrophic nitrifiers isolated from Mid-Atlantic Ridge deep-sea hydrothermal vents. Canadian Journal of Microbiology, 1998, 44(8): 723 733. [25] Vanniel EWJ, Braber KJ, Robertson LA, et al. Heterotrophic nitrification and aerobic denitrification in Alcaligenes faecalis strain TUD. Antonie van Leeuwenhoek, 1992, 62(3): 231 237. [26] Mevel G, Prieur D. Heterotrophic nitrification by a thermophilic Bacillus species as influenced by different culture conditions. Canadian Journal of Microbiology, 2000, 46(5): 465 473. [27] Joo HS, HiraiM, Shoda M. Characteristics of ammonium removal by heterotrophic nitrification-aerobic denitrification by Alcaligenes faecalis No. 4. Journal of Bioscience and Bioengineering, 2005, 100(2): 184 191. [28] Otani Y, Hasegawa K, Hanaki K. Comparison of aerobic denitrifying activity among three cultural species with various carbon sources. Water Science and Technology, 2004, 50(8): 15 22. [29] Robertson LA, Cornelisse R, Zeng R, et al. The effect of thiosulphate and other inhibitors of autotrophic nitrification on heterotrophic nitrifiers. Antonie Van Leeuwenhoek, 1989, 56(4): 301 309. [30] Gupta AB, Gupta SK. Simultaneous carbon and nitrogen removal from high strength domestic wastewater in an aerobic RBC biofilm. Water Research, 2001, 35(7): 1714 1722. [31] Kesik M, Blagodatsky S, Papen H, et al. Effect of ph, temperature and substrate on N 2 O, NO and CO 2 production by Alcaligenes faecalis p. Journal of Applied Microbiology, 2006, 101(3): 655 667. [32] Ahmad NUD, Xu HY, Chen LP, et al. Enhanced biological nutrient removal by the alliance of a heterotrophic nitrifying strain with a nitrogen removing ecosystem. Journal of Environmental Sciences-China, 2008, 20(2): 216 223. [33] Patureau D, Bernet N, Delgenes JP, et al. Effect of dissolved oxygen and carbon-nitrogen loads on denitrification by an aerobic consortium. Applied Microbiology and Biotechnology, 2000, 54(4): 535 542. [34],,,.., 2006, 22(21): 67 70. [35],,,. MBR., 2006, 29(1): 7 9. [36] Ho KL, Chung YC, Tseng CP. Continuous deodorization and bacterial community analysis of a biofilter treating nitrogen-containing gases from swine waste storage pits. Bioresour Technol, 2008, 99(8): 2757 2765. 57(3): 139 152.