Identification and Metabolic Mechanism of Non-fermentative Short-cut Denitrifying Phosphorus-removing Bacteria

doi:10.1016/S1004-9541(13)60465-6

Chin.J.Chem.Eng. ›› 2013, Vol. 21 ›› Issue (3): 332-340.DOI: 10.1016/S1004-9541(13)60465-6

• ENERGY, RESOURCES AND ENVIRONMENTAL TECHNOLOGY • Previous Articles

Identification and Metabolic Mechanism of Non-fermentative Short-cut Denitrifying Phosphorus-removing Bacteria

LIU Hui^1,2,3, SUN Yanfu², JIA Xiaoshan¹, LI Jun³, ZHOU Kangqun², QU Xiangdong^1,2, TAO Xueqin², CHEN Yu³

1 School of Environmental Science and Engineering, Sun Yat-sen University, Guangzhou 510275, China ;
2 Department of Environment Science and Engineering, Zhongkai University of Agriculture and Engineering, Guangzhou 510225, China ;
3 Key Laboratory of Beijing Water Quality Science and Water Environment Recovery Engineering, Beijing University of Technology, Beijing 100124, China

Received:2011-12-05 Revised:2012-04-05 Online:2013-04-01 Published:2013-03-28
Supported by:
Supported by the National Natural Science Foundation of China (51078008), the Natural Science Foundation of Guangdong Province (06022869, 07003251), and the National Key Scientific and Technological Project Water Pollution Control and Treatment (2008ZX07211-003, 2009ZX07314-009-003).

Identification and Metabolic Mechanism of Non-fermentative Short-cut Denitrifying Phosphorus-removing Bacteria

刘晖^1,2,3, 孙彦富², 贾晓珊¹, 李军³, 周康群², 屈向东^1,2, 陶雪琴², 陈瑜³

1 School of Environmental Science and Engineering, Sun Yat-sen University, Guangzhou 510275, China ;
2 Department of Environment Science and Engineering, Zhongkai University of Agriculture and Engineering, Guangzhou 510225, China ;
3 Key Laboratory of Beijing Water Quality Science and Water Environment Recovery Engineering, Beijing University of Technology, Beijing 100124, China

通讯作者: JIA Xiaoshan
基金资助:
Supported by the National Natural Science Foundation of China (51078008), the Natural Science Foundation of Guangdong Province (06022869, 07003251), and the National Key Scientific and Technological Project Water Pollution Control and Treatment (2008ZX07211-003, 2009ZX07314-009-003).

Abstract

Abstract: To investigate the characteristics and metabolic mechanism of short-cut denitrifying phosphorus-removing bacteria (SDPB) that are capable of enhanced biological phosphorus removal (EBPR) using nitrite as an electron acceptor, an aerobic/anoxic sequencing batch reactor was operated under three phases. An SDPB-strain YC was screened after the sludge enrichment and was identified by morphological, physiological, biochemical properties and 16S rDNA gene sequence analysis. Denitrifying phosphorus-removing experiments were conducted to study anaerobic and anoxic metabolic mechanisms by analyzing the changes of chemical oxygen demand (COD), phosphate, nitrite, poly-β-hydroxybutyrate (PHB), and glycogen. The results show that strain YC is a non-fermentative SDPB similar to Paracoccus denitrificans. As a kind of non-fermentative bacteria, the energy of strain YC was mainly generated from phosphorus release (96.2%) under anaerobic conditions with 0.32 mg P per mg synthesized PHB. Under anoxic conditions, strain YC accumulated 0.45 mg P per mg degraded PHB, which produced most of energy for phosphate accumulation (91.3%) and a little for glycogen synthesis (8.7%). This metabolic mechanism of strain YC is different from that of traditional phosphorus-accumulating organisms (PAOs). It is also found that PHB, a kind of intracellular polymer, plays a very important role in denitrifying and accumulating phosphorus by supplying sufficient energy for phosphorous accumulation and carbon sources for denitrification. Therefore, monitoring ΔP/ΔPHB and? ΔNO₂^--N/ΔPHB is more necessary than monitoring ΔP/ΔCOD,?ΔNO₂^--N/ΔCOD, or ΔNO₂^--N.

Key words: short-cut denitrifying phosphorus removing bacteria, Paracoccus denitrificans, non-fermentative bacteria, metabolic mechanism, poly-&, beta, -hydroxybutyrate

摘要： To investigate the characteristics and metabolic mechanism of short-cut denitrifying phosphorus-removing bacteria (SDPB) that are capable of enhanced biological phosphorus removal (EBPR) using nitrite as an electron acceptor, an aerobic/anoxic sequencing batch reactor was operated under three phases. An SDPB-strain YC was screened after the sludge enrichment and was identified by morphological, physiological, biochemical properties and 16S rDNA gene sequence analysis. Denitrifying phosphorus-removing experiments were conducted to study anaerobic and anoxic metabolic mechanisms by analyzing the changes of chemical oxygen demand (COD), phosphate, nitrite, poly-β-hydroxybutyrate (PHB), and glycogen. The results show that strain YC is a non-fermentative SDPB similar to Paracoccus denitrificans. As a kind of non-fermentative bacteria, the energy of strain YC was mainly generated from phosphorus release (96.2%) under anaerobic conditions with 0.32 mg P per mg synthesized PHB. Under anoxic conditions, strain YC accumulated 0.45 mg P per mg degraded PHB, which produced most of energy for phosphate accumulation (91.3%) and a little for glycogen synthesis (8.7%). This metabolic mechanism of strain YC is different from that of traditional phosphorus-accumulating organisms (PAOs). It is also found that PHB, a kind of intracellular polymer, plays a very important role in denitrifying and accumulating phosphorus by supplying sufficient energy for phosphorous accumulation and carbon sources for denitrification. Therefore, monitoring ΔP/ΔPHB and? ΔNO₂^--N/ΔPHB is more necessary than monitoring ΔP/ΔCOD,?ΔNO₂^--N/ΔCOD, or ΔNO₂^--N.

关键词: short-cut denitrifying phosphorus removing bacteria, Paracoccus denitrificans, non-fermentative bacteria, metabolic mechanism, poly-&, beta, -hydroxybutyrate

LIU Hui, SUN Yanfu, JIA Xiaoshan, LI Jun, ZHOU Kangqun, QU Xiangdong, TAO Xueqin, CHEN Yu. Identification and Metabolic Mechanism of Non-fermentative Short-cut Denitrifying Phosphorus-removing Bacteria[J]. Chin.J.Chem.Eng., 2013, 21(3): 332-340.

刘晖, 孙彦富, 贾晓珊, 李军, 周康群, 屈向东, 陶雪琴, 陈瑜. Identification and Metabolic Mechanism of Non-fermentative Short-cut Denitrifying Phosphorus-removing Bacteria[J]. Chinese Journal of Chemical Engineering, 2013, 21(3): 332-340.

References

1 The Nutrient Removal Task Force of the Water Environment Federation, Nutrient Removal WEF Manual of Practice No. 34, WEF Press, Alexandria Virginia, 2-3 (2010).

2 Kerrn-Jesperson, J.P., Henze, M., “Biological phosphorus uptake under anoxic and aerobic conditions”, Water Res., 27 (4), 617-624 (1993).

3 Oehmen, A., Lemos, P.C., Carvalho, G., Yuan, Z.G., “Advances in enhanced biological phosphorus removal: from micro to macro scale”, Water Res., 41 (11), 2271-2300 (2007).

4 Cui, Y.W., Wang, S.Y., Li, J., “On-line monitoring for phosphorus removal process and bacterial community in sequencing batch reactor”, Chin. J. Chem. Eng., 17 (3), 484-492 (2009).

5 Sun, H.W., Yang, Q., Peng, Y.Z., “Nitrite accumulation during the denitrification process in SBR for the treatment of pre-treated landfill leachate”, Chin. J. Chem. Eng., 17 (6), 1027-1031 (2009).

6 Zhang, S.J., Peng, Y.Z., WANG, S.Y., “Short-cut removal of nitrogen from mature municipal landfill leachate at ambient temperature”, J. Chem. Ind. Eng., 58 (4), 1042-1047 (2007).

7 Meinhold, J., Filipe, C.D.M., Daigger, G.T., Isaacs, S., “Characterization of the denitrifying fraction of phosphate accumulating organisms in biological phosphate removal”, Water Sci. Technol., 39 (1), 31-42 (1999).

8 Hu, J.Y., Ong, S.L., Ng, W.J., “A new method for characterizing denitrifying phosphorus removal bacterial by using three different types of electron acceptors”, Water Res., 37 (14), 3463-3471 (2003) .

9 Mino, T., van Loosdrecht, M.C.M., Heijnen, J.J., “Microbiology and biochemistry of the enhanced biological phosphate removal process”, Water Res., 32 (1), 3193-3207 (1998).

10 Kong, Y.H., Nielsen, J.L., Nielsen, P.H., “Microautoradiographic study of Rhodocyclus-related polyphosphate-accumulating bacteria in full-scale enhanced biological phosphorus removal plants”, Appl. Environ. Microbiol., 70 (9), 5383-5390 (2004).

11 Oehmen, A., Zeng, R.J, Yuan, Z.G., Keller, J., “Anaerobic metabolism of propionate by polyphosphate-accumulating organisms in enhanced biological phosphorus removal systems”, Biotechnol. Bioeng., 91 (1), 43-53 (2005).

12 Ahn, J., Daidou, T., Tsuneda, S., “Metabolic behavior of denitrifying phosphate-accumulating organisms under nitrate and nitrite electron conditions”, Biotechnol. Bioeng., 92 (5), 442-446 (2001).

13 Meihold, J., Filipe, C.D.M., Dagger, G.T., “Characterization of the denitrifying fraction of phosphate accumulating organisms in biological phosphate removal process”, Water Sci. Technol., 39 (1), 31-42 (1999).

14 Johwan, A., Tomotaka, D., Satoshi, T., “Metabolic behavior of denitrifying phosphate-accumulating organisms under nitrate and nitrite electron acceptor conditions”, Biotechnol. Bioeng., 92 (5), 442-446 (2001)．

15 Li, H.J., Chen, Y.G., Gu, G.W., “The effect of propionic to acetic acid ratio on anaerobic-aerobic (low dissolved oxygen) biological prosphorus and nitrogen removal”, Biores. Technol., 99, 4400-4407 (2008).

16 Puig, S., Coma, M., Monclus, H., van Loosdrecht, M.C.M., “Balaguer selection between alcohols and volatile fatty acids as external carbon sources for EBPR”, Water Res., 42, 557-566 (2008).

17 Fuhs, G.W., Chen, M., “Microbiological basis of phosphate removal in the activated sludge process for the treatment of wastewater”, Microb. Ecol., 2 (2), 119-138 (1975).

18 Nakamura, K., Hiraishi, A., Yoshimi, Y., Kawaharasaki, M., Masuda, K., Kamagata, Y., “Microlunatus phosphovorus gen-nov, sp-nov, a new gram-positive polyphosphate-accumulating bacterium isolated from activated-sludge”, Int. J. Syst. Evo. Microbiol., 45 (1), 17-22 (1995).

19 Seviour, R.J., Mino, T., Onuki, M., “The microbiology of biological phosphorus removal in activated sludge systems”, FEMS Microbiol. Rev., 27 (1), 99-127 (2003).

20 Johwan, A., Tomotaka, D., Satoshi, T., Akira, H., “Characterization of denitrifying phosphate-accumulating organisms cultivated under different electron acceptor conditions using polymerase chain reaction-denaturing gradient gelel ectrophoresis assay”, Water Res., 36, 403-412 (2002).

21 Kong, Y.H., Nielsen, J.L., Nielsen, P.H., “Identity and ecophysiology of uncultured actinobacterial polyphosphate-accumulating organisms in full-scale enhanced biological phosphorus removal plants”, Appl. Environ. Microbiol., 71 (7), 4076-4085 (2005).

22 Martin, H.G., Ivanova, N., Kunin, V., Falk, W., Kerrie, W.B., “Metagenomic analysis of two enhanced biological phosphorus removal (EBPR) sludge communities”, Nature Biotech., 24 (10), 1263-1269 (2006).

23 Smolders, G.J.F., vandermeij, J., van Loosdrecht, M.C.M., “Stoichiometric model of the aerobic metabolism of the biological phosphorus removal process”, Biotech. Bioeng., 44 (7), 837-848 (1994).

24 Jorgense, K.S., Pauli, A.S., “Polyphospate accumulating among denitrifying bacteria in activated sludge”, Anaerobe, 1 (3), 161-168 (1995).

25 Don, J.B., Noel, R.K., James, T.S., BERGEY'S Manual of Systematic Bacteriology, 2nd edition, Springer, New York, 425-437(2004).

26 Dong, X.Z., Cai, M.Y., A Manual of Ordinary Bacteriology Identification, Science Press, Beijing, 353-391 (2001). (in Chinese)

27 Joseph, S., David, W.R., Molecular Cloning Volume 3, Cold Spring Harbor Laboratory Press, New York, 597-618 (2001).

28 Oehmen, A.B., Keller-Lehmann, R., Zeng, R.J., Yuan, Z.G., Keller, J., “Optimization of poly-beta-hydroxyalkanoate analysis using gas chromatography for enhanced biological phosphorus removal systems ”, Journal of Chromatography A, 1070,131-136 (2005).

29 Bond, P.L., Erhart, R., Wagner, M., Keller, J., Blackall, L.L., “Identification of some of the major groups of bacteria in efficient and nonefficient biological phosphorus removal activated sludge systems”, Appl. Environ. Microbiol., 65 (9), 4077-4084 (1999).

30 Donovan, P.K., Jean, P.E., Celia, F.G., “Redefining Paracoccus denitrificans and Paracoccus pantotrophus and the case for a reassessment of the strains held by international culture collections”, Int. J. Syst. Evo. Microbiol., 56, 2495-2500 (2006).

31 Nokhal, T.H., Schlegel, H.G., “Taxonomic Study of Paracoccus denitrificans”, Int. J. Syst. Evol. Microbiol., 33 (1), 26-37 (1983).

32 Ghosh, W., Mandal, S., Roy, P., “Paracoccus bengalensis sp. nov., a novel sulfur-oxidizing chemolithoautotroph from the rhizospheric soil of an Indian tropical leguminous plant”, Syst. Appl. Microbiol., 29, 396-403 (2006).

33 Yoram, B.A., Jaap, V.R., “Atypical polyphosphate accumulation by the denitrifying bacterium Paracoccus denitrificans”, Appl. Envrion. Microbiol., 66 (3), 1209-1212 (2000).

34 Smolders, G.J.F., vandermeij, J., van Loosdrecht, M.C.M., “Model of the anaerobic metabolism of the biological phosphorus removal process: stoichiometry and pH influence”, Biotech. Bioeng., 43 (6) , 461-470 (1994).

35 Smolders, G.J.F., vandermeij, J., van Loosdrecht, M.C.M., “Stoichiometric model of the aerobic metabolism of the biological phosphorus removal process”, Biotech. Bioeng., 44 (7), 837-848 (1994).

36 Smolders, G.J.F., vandermeij, J., van Loosdrecht, M.C.M., “A structured metabolic model for the anaerobic and aerobic stoichiometry and kinetics of the biological phosphorus removal process”, Biotech. Bioeng., 47 (3), 277-287 (1995).

37 Smolders, G.J.F., vandermeij, J., van Loosdrecht, M.C.M., “A metabolic model of the biological phosphorus removal process: effect of the sludge retention time”, Biotech. Bioeng., 48 (3), 222-233 (1995).

38 Smolders, G.J.F., van Loosdrecht, M.C.M., Heijnen, J.J, “Steady state analysis to evaluate the phosphate removal capacity and acetate requirement of biological phosphate removing mainstream and side stream process configuration”, Water Res., 30 (11), 2748-2760 (1996).

39 Smolders, G.J.F., van Loosdrecht, M.C.M., Heijnen, J.J., “A metabolic model for the biological phosphorus removal process”, Water Sci. Technol., 31 (2), 79-93 (1995).

40 Louie, T.M., Mah, T.J., Oldham, W., Ramey, W.D., “Use of metabolic inhibitors and gas chromatography/mass spectrometry to study poly-b-hydroxyalkanoates metabolism involving cryptic nutrients in enhanced biological phosphorus removal systems”, Water Res., 34, 1507-1514 (2000).

41 Lemos, P.C., Serafim, L.S., Santos, M.M., Reis, M.A., Santos, H., “Metabolic pathway for propionate utilization by phosphorus-accumulating organisms in activated sludge: ¹³C labeling and in vivo nuclear magnetic resonance”, Appl. Environ. Microbiol., 69, 241-251 (2003).

42 Pereria, H., Lemos, P.C., Reis, M.A., Crespo, J.P.S.G., “Model for carbon metabolism in biological phosphorus removal processes based on in vivo ¹³C-NMR labeling experiments”, Water Res., 30 (9), 2128-2138 (1996).

43 Elbehti, A., Brasseur, G., Lemesle-Meunier, D., “First evidence for existence of an uphill electron transfer through the bc(1) and NADH-Q oxidoreductase complexes of the acidophilic obligate chemolithotrophic ferrous ion-oxidizing bacterium Thiobacillus ferrooxidans”, J. Bacteriol., 182, 3602-3606 (2000).

[1]	Yefei Liu, Yang Zou, Hong Jiang, Huanxin Gao, Rizhi Chen. Deactivation mechanism of beta-zeolite catalyst for synthesis of cumene by benzene alkylation with isopropanol [J]. , 2017, 25(9): 1195-1201.
[2]	Bo Lü, Xiaogang Yang, Xudong Feng, Chun Li. Enhanced production of glycyrrhetic acid 3-O-mono-β-D-glucuronide by fed-batch fermentation using pH and dissolved oxygen as feedback parameters [J]. Chin.J.Chem.Eng., 2016, 24(4): 506-512.
[3]	Wei Ma, Mei Meng, Shumin Yan, Shufen Zhang. Salt-free reactive dyeing of betaine-modified cationic cotton fabrics with enhanced dye fixation [J]. Chin.J.Chem.Eng., 2016, 24(1): 175-179.
[4]	Daohong Xia, Shengjuan Jiang, Lantao Li, Yuzhi Xiang, Lijun Zhu. The biomimetic catalytic synthesis of acetal compounds using β-cyclodextrin as catalyst [J]. Chin.J.Chem.Eng., 2016, 24(1): 146-150.
[5]	REN Xiaoling, REN Jizhong, LI Hui, DENG Maicun. Permeation Characteristics of Light Hydrocarbons Through Poly(amide-6-β-ethylene oxide) Multilayer Composite Membranes [J]. Chin.J.Chem.Eng., 2013, 21(3): 232-237.
[6]	TANG Kewen, MIAO Jiabing, ZHOU Tao, LIU Yongbing. Equilibrium Studies on Liquid-Liquid Reactive Extraction of Phenylsuccinic Acid Enantiomers Using Hydrophilic β-CD Derivatives Extractants [J]. Chin.J.Chem.Eng., 2011, 19(3): 397-403.
[7]	YOU Xiaoyan, QIN Wei, DAI Youyuan. Phase Separation Behavior of Cocamidopropyl Betaine/Water/Polyethylene Glycol System [J]. , 2009, 17(5): 746-749.

Identification and Metabolic Mechanism of Non-fermentative Short-cut Denitrifying Phosphorus-removing Bacteria

Identification and Metabolic Mechanism of Non-fermentative Short-cut Denitrifying Phosphorus-removing Bacteria

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 7

Recommended Articles

Metrics

Comments