不同环境条件下Pseudomonas sp.脱氮特性及功能基因表达差异研究
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摘要
从活性污泥中分离获得一株PCL降解菌,经形态学和16S rDNA鉴定后命名为Pseudomonas sp.JQ-H3.经过脱氮实验验证,该菌能够以PCL为唯一碳源,分别以氨氮或硝酸盐氮为氮源进行异养硝化好氧反硝化.该菌能够在36h内去除93.11%的氨氮(初始氨氮浓度为102.41mg/L),氨氮最大降解速率为5.77mg/(L·h);并且能够在48h内去除93.93%的硝酸盐氮(初始氨氮浓度为99.01mg/L),硝酸盐氮最大降解速率为4.12mg/(L·h).对PCL膜的降解实验结果表明,菌株能够在60d内将初始重量为100mg的PCL薄膜降解94.03%,且胞外脂肪酶活性在30d时达到最大值9.18mU/mL.另外,Q-PCR实验结果表明,弱碱性环境促进了amoA和nirS基因的表达; napA、cnorB、nosZ基因的成功表达,进一步证明了菌株的异养硝化好氧反硝化能力.
A PCL-degrading strain was isolated and identified as Pseudomonas sp. JQ-H3 through morphology observation and phylogenetic analysis. The strain showed the heterotrophic nitrification-aerobic denitrification(HN-AD) ability using PCL as a sole carbon source. About 93.11% of initial ammonium(102.41 mg/L) was degraded within 36 h with the maximum ammonium removal rate of 5.77 mg/(L·h), while approximately 93.93% of initial nitrate(99.01 mg/L) was removed within 48 h with the maximum ammonium removal rate of 4.12 mg/(L·h). 94.03% of PCL film with initial weight of 100 mg was decomposed after 60 days, and the extracellular lipases activity reached the maximum value of 9.18 mU/mL at 30 th days. In addition, the expression levels of amo A and nirS genes were promoted in the weak alkaline environment. The successful expression of napA, cnor B and nosZ genes further proved the HN-AD ability of the strain JQ-H3.
A PCL-degrading strain was isolated and identified as Pseudomonas sp. JQ-H3 through morphology observation and phylogenetic analysis. The strain showed the heterotrophic nitrification-aerobic denitrification(HN-AD) ability using PCL as a sole carbon source. About 93.11% of initial ammonium(102.41 mg/L) was degraded within 36 h with the maximum ammonium removal rate of 5.77 mg/(L·h), while approximately 93.93% of initial nitrate(99.01 mg/L) was removed within 48 h with the maximum ammonium removal rate of 4.12 mg/(L·h). 94.03% of PCL film with initial weight of 100 mg was decomposed after 60 days, and the extracellular lipases activity reached the maximum value of 9.18 mU/mL at 30 th days. In addition, the expression levels of amo A and nirS genes were promoted in the weak alkaline environment. The successful expression of napA, cnor B and nosZ genes further proved the HN-AD ability of the strain JQ-H3.
引文
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[2]Lloyd D,Boddy L,Davies K J P.Persistence of bacterial denitrification capacity under aerobic conditions:The rule rather than the exception.[J].FEMS Microbiology Letters,1987,45(3):185-190.
[3]Yan L,Liu S,Liu Q,et al.Improved performance of simultaneous nitrification and denitrification via nitrite in an oxygen-limited SBRby alternating the DO[J].Bioresource technology,2019,275:153-162.
[4]Nitrogen removal characteristics of a heterotrophic nitrifier Acinetobacter junii YB and its potential application for the treatment of high-strength nitrogenous wastewater[J].Bioresource Technology,2015,193:227-233.
[5]Li C,Yang J,Wang X,et al.Removal of nitrogen by heterotrophic nitrification-aerobic denitrification of a phosphate accumulating bacterium Pseudomonas stutzeri YG-24[J].Bioresource Technology,2015,182:18-25.
[6]Wang T,Dang Q,Liu C,et al.Heterotrophic nitrogen removal by a newly-isolated alkalitolerant microorganism,Serratia marcescens W5[J].Bioresource Technology,2016,211:618-627.
[7]Zhang Q,Liu Y,Ai G M,et al.The characteristics of a novel heterotrophic nitrification-aerobic denitrification bacterium,Bacillus methylotrophicus strain L7[J].Bioresource Technology,2012,108:35-44.
[8]王秀杰,王维奇,李军,等.异养硝化菌Acinetobacter sp.的分离鉴定及其脱氮特性[J].中国环境科学,2017,37(11):4241-4250.
[9]Goodwin D G,Boyer I,Devahif T,et al.Biodegradation of Carbon Nanotube/Polymer Nanocomposites using a Monoculture[J].Environmental Science&Technology,52(1):40-51.
[10]Shah A A,Nawaz A,Kanwal L,et al.Degradation of poly(ε-caprolactone)by a thermophilic bacterium Ralstonia sp.strain MRL-TL isolated from hot spring[J].International Biodeterioration&Biodegradation,2015,98:35-42.
[11]Luo G,Xu G,Gao J,et al.Effect of dissolved oxygen on nitrate removal using polycaprolactone as an organic carbon source and biofilm carrier in fixed-film denitrifying reactors[J].Journal of Environmental Sciences,2016,43:147-152.
[12]Yao S,Ni J,Ma T,et al.Heterotrophic nitrification and aerobic denitrification at low temperature by a newly isolated bacterium,Acinetobacter sp HA2[J].Bioresource Technology,2013,139:80-86.
[13]Suzuki M,Tachibana Y,Oba K,et al.Microbial degradation of poly(ε-caprolactone)in a coastal environment[J].Polymer Degradation and Stability,2018,149:1-8.
[14]Khatiwala V K,Shekhar N,Aggarwal S,et al.Biodegradation of poly(ε-caprolactone)(PCL)film by Alcaligenes faecalis[J].Journal of Polymers and the Environment,2008,16(1):61-67.
[15]Lowe S E,Jain M K,Zeikus J G.Biology,ecology,and biotechnological applications of anaerobic bacteria adapted to environmental stresses in temperature,p H,salinity,or substrates[J].Microbiological Reviews,1993,57(2):451-509.
[16]Liu Y,Hu T,Song Y,et al.Heterotrophic nitrogen removal by Acinetobacter sp.Y1isolated from coke plant wastewater[J].Journal of Bioscience and Bioengineering,2015,120(5):549-554.
[17]Jung J H,Ree M,Kim H.Acid-and base-catalyzed hydrolyses of aliphatic polycarbonates and polyesters[J].Catalysis Today,2006,115(1-4):283-287.
[18]Loh X J.The effect of p H on the hydrolytic degradation of poly(ε-caprolactone)-block-poly(ethylene glycol)copolymers[J].Journal of Applied Polymer Science,2013,127(3):2046-2056.
[19]Chen Q,Ni J.Ammonium removal by Agrobacterium sp.LAD9capable of heterotrophic nitrification-aerobic denitrification[J].Journal of Bioscience and Bioengineering,2012,113(5):619-623.
[20]Ren Y,Yang L,Liang X.The characteristics of a novel heterotrophic nitrifying and aerobic denitrifying bacterium,Acinetobacter junii YB[J].Bioresource Technology,2014,171:1-9.
[21]Duan J,Fang H,Su B,et al.Characterization of a halophilic heterotrophic nitrification-aerobic denitrification bacterium and its application on treatment of saline wastewater[J].Bioresource Technology,2015,179:421-428.
[22]Yang J,Wang Y,Chen H,et al.Ammonium removal characteristics of an acid-resistant bacterium Acinetobacter sp.JR1from pharmaceutical wastewater capable of heterotrophic nitrification-aerobic denitrification[J].Bioresource Technology,2019,274:56-64.
[23]Mevel G,Prieur D.Heterotrophic nitrification by a thermophilic Bacillus species as influenced by different culture conditions[J].Canadian Journal of Microbiology,2000,46(5):465-473.
[24]Huang X,Li W,Zhang D,et al.Ammonium removal by a novel oligotrophic Acinetobacter sp.Y16 capable of heterotrophic nitrification-aerobic denitrification at low temperature[J].Bioresource Technology,2013,146:44-50.
[25]Luo G,Xu G,Tan H,et al.Effect of dissolved oxygen on denitrification using polycaprolactone as both the organic carbon source and the biofilm carrier[J].International Biodeterioration&Biodegradation,2016,110:155-162.
[26]Shoda M,Ishikawa Y.Heterotrophic nitrification and aerobic denitrification of high-strength ammonium in anaerobically digested sludge by Alcaligenes faecalis strain No.4[J].Journal of Bioscience and Bioengineering,2014,117(6):737-741.
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