1株杀鲑气单胞菌杀鲑亚种对甲酸乙酯的降解研究

doi:10.13560/j.cnki.biotech.bull.1985.2019-0178

摘要/Abstract

摘要： 甲酸乙酯广泛存在于化工行业的废水中,对生物体及环境都具有很大的危害。微生物降解是处理甲酸乙酯的重要方法之一。采用富集培养法和平板划线法从活性污泥中筛选出一株能降解甲酸乙酯的菌株ETH-3。经形态观察、生理生化试验和16S rDNA鉴定,确定该菌株为杀鲑气单胞菌杀鲑亚种（Aeromonas salmonicida subsp.salmonicida）。通过摇瓶实验测定菌株ETH-3的最佳生长环境条件以及其对甲酸乙酯的降解能力,结果表明,该菌株最佳生长温度为25-30℃、最佳生长pH为8左右。该菌株具有较强降解甲酸乙酯能力,在30℃时,36 h内对0-10 500 mg/L的甲酸乙酯可达完全降解。经GC-MS对代谢产物的分析发现,甲酸乙酯会先被微生物代谢为甲酸和乙醇,后甲酸和乙醇会被微生物进一步降解为CO₂和H₂O。

关键词: 甲酸乙酯, 生物降解, 杀鲑气单胞菌杀鲑亚种

Abstract: Ethyl formate is widely present in wastewater from the chemical industry and is very harmful to both organisms and the environment. Microbial degradation is one of the important approaches for the treatment of ethyl formate. In this study,a strain of ETH-3 capable of degrading ethyl formate was screened from activated sludge by enrichment culture method and plate scribing method. Then,the strain was identified as Aeromonas salmonicida subsp. salmonicida by morphological observation,physiological and biochemical tests and 16S rDNA identification. The optimal growth conditions of strain ETH-3 and its ability to degrade ethyl formate were determined by shake flask test. The results showed that the optimal growth temperature of the strain was 25-30℃,and the optimal growth pH was about 8. The strain had strong ability to degrade ethyl formate. At 30℃,the degradation of 0-10 500 mg/L ethyl formate was completely degraded within 36 h. Analysis of metabolites by GC-MS revealed that ethyl formate was first metabolized by microorganisms to formic acid and ethanol,and then formic acid and ethanol were further degraded by microorganisms into CO₂ and H₂O.

Key words: ethyl formate, biodegradation, Aeromonas salmonicidal subsp salmonicida

张珊珊, 姚小龙, 王珂, 尤雅. 1株杀鲑气单胞菌杀鲑亚种对甲酸乙酯的降解研究[J]. 生物技术通报, 2019, 35(5): 109-117.

ZHANG Shan-shan, YAO Xiao-long, WANG Ke, YOU Ya. Degradation of Ethyl Formate by Aeromonas salmonicida subsp.salmonicida[J]. Biotechnology Bulletin, 2019, 35(5): 109-117.

参考文献

[1] Dojchinov G.Ethyl formate for farm stored grains[J]. Pesticide Outlook, 2004, 14(6):241-242.
[2] Lindinger C, Pollien P, Vos R CHD, et al.Identification of ethyl Formate as a quality marker of the fermented off-note in coffee by a nontargeted chemometric approach[J]. Journal of Agricultural and Food Chemistry, 2009, 57(21):9972-9978.
[3] Charles-Bernard M, Kraehenbuehl K, Rytz A, et al.Interactions between volatile and nonvolatile coffee components. 1. Screening of nonvolatile components[J]. Journal of Agricultural and Food Chemistry, 2005, 53(11):4417-4425.
[4] Opdyke DLJ.Ethyl formate[J]. Food & Cosmetics Toxicology, 1978, 16(Suppl 1):737-739.
[5] Gibson HW.Chemistry of formic acid and its simple derivatives[J]. Chemical Reviews, 1969, 69(5):673-692.
[6] Amoore JE, Hautala E.Odor as an aid to chemical safety:odor thresholds compared with threshold limit values and volatilities for 214 industrial chemicals in air and water dilution[J]. Journal of Applied Toxicology, 2010, 3(6):272-290.
[7] Netherlands HCot.Health-based reassessment of administrative occupational exposure limits. Committee on updating of occupational exposure limits. Ethyl formate[EB]. The Hague:Health Council of the Netherlands, 2002.
[8] Qian Q, Gong C, Zhang Z, et al.Removal of VOCs by activated carbon microspheres derived from polymer:a comparative study[J]. Adsorption, 2015, 21(4):333-341.
[9] Iranpour R, Cox HHJ, Deshusses MA, et al.Literature review of air pollution control biofilters and biotrickling filters for odor and volatile organic compound removal[J]. Environmental Progress & Sustainable Energy, 2010, 24(3):254-267.
[10] 王红丽, 景盛翱, 王倩, 等. 溶剂使用源有组织排放VOCs监测方法及组成特征[J]. 环境科学研究, 2016, 29(10):1433-1439.
[11] Ramalingam SG, Pré P, Giraudet S, et al. Different families of volatile organic compounds pollution control by microporous carbons in temperature swing adsorption processes[J]. Journal of Hazardous Materials, 2012, 221-222(4):242-247.
[12] 杨峰, 甲酸乙酯废水工艺研究[J]. 江苏科技信息, 2014, 12:25-27.
[13] 宋红改, 张亚雄. 微生物处理技术在有机污染物降解中的应用[J]. 环境工程, 2008(s1):205-208.
[14] Park IH, Ka JO.Isolation and characterization of 4-(2, 4-Dichlorophenoxy)butyric acid-degrading bacteria from agricultural soils[J]. Journal of Microbiology & Biotechnology, 2003, 13(2):243-250.
[15] 徐畅, 李琳. 气相色谱法测定环境空气中甲酸甲酯和甲酸乙酯[J]. 污染防治技术, 2016(3):85-87.
[16] 东秀珠, 蔡妙英. 常见细菌系统鉴定手册[M]. 北京:科学出版社, 2001.
[17] Meena SS, Sharma RS, Gupta P, et al.Isolation and identification of Bacillus megaterium YB3 from an effluent contaminated site efficiently degrades pyrene[J]. Journal of Basic Microbiology, 2016, 56(4):369-378.
[18] 宇盛好, 曹慧, 徐斐, 等. 烤鸭中金黄色葡萄球菌生长模型的建立[J]. 食品科学, 2015, 36(13):154-159.
[19] 李想, 张雪英, 周俊, 等. 1株铜绿假单胞菌对芘的降解特性及代谢途径[J]. 环境科学, 2018, 39(4):1794-1803.
[20] Chen YP, Chiang TK, Chung HY.Optimization of a headspace solid-phase microextraction method to quantify volatile compounds in plain sufu, and application of the method in sample discrimination[J]. Food Chemistry, 2019, 275:32-40.
[21] Barbusinski K, Kalemba K, Kasperczyk D, et al.Biological methods for odor treatment - A review[J]. Journal of Cleaner Production, 2017, 152:223-241.
[22] 丁庆伟, 霍丽娟, 杨改强, 等. 废水治理中微生物的生长规律研究[J]. 太原科技大学学报, 2011, 32(2):163-165.
[23] 何旭辉. 生物化学[M]. 第2版. 北京:人民卫生出版社, 2010:45-49.
[24] 郎咸勇, 张锦华, 刘宝生. 培养基高压蒸气灭菌后pH值的变化及其对细菌生长的影响[J]. 黑龙江畜牧兽医, 2016(4):30-33.
[25] 叶建生, 刘道成, 熊良伟, 等. pH值对光合细菌生长的影响[J]. 渔业致富指南, 2008(23):61-62.
[26] Zhou X, Jin W, Sun C, et al.Microbial degradation of N, N-dimethylformamide by Paracoccus, sp. strain DMF-3 from activated sludge[J]. Chemical Engineering Journal, 2018, 343:324-330.
[27] 诸葛健. 工业微生物实验与研究技术[M]. 北京:科学出版社, 2007:109-111.
[28] 曾恋之, 谭锋, 侯茂书. 温度对细菌在生菜基质中生长的影响[J]. 安徽农业科学, 2012(13):7916-7918.
[29] 田会芹, 李贤, 傅松哲, 等. 温度及pH对杀鲑气单胞菌生长的影响研究[J]. 海洋科学, 2015, 39(7):7-12.
[30] 董小娜, 杨洁, 陈泽慧, 等, 太湖激浪鱼内脏中藻毒素降解菌的筛选及其特性研究[J]. 湖北农业科学, 2017(16):3042-3047.
[31] 石成春. 有机污染物微生物共代谢降解及其动力学研究[J]. 化学工程与装备, 2010(7):164-167.
[32] 王家玲, 李顺鹏, 黄正. 环境微生物学[M]. 第2版. 北京:高等教育出版社, 2004:89-91.
[33] Tarakanova EG, Yukhnevich GV.Structure of proton disolvates formed in the acid hydrolysis of ethyl formate and methyl acetate[J]. Journal of Structural Chemistry, 2013, 54(3):492-498.
[34] 伦世仪. 生化工程[M]. 北京:中国轻工业出版社, 1993:101-104.
[35] Barbusinski K, Kalemba K, Kasperczyk D, et al.Biological methods for odor treatment—A review[J]. Journal of Cleaner Production, 2017, 152:223-241.
[36] 薛开先. 乙醇代谢酶遗传多态与癌易感性关系的研究[J]. 国际遗传学杂志, 2000(1):24-27.