一株冢村菌的筛选及其对邻苯二甲酸二丁酯的降解特性研究

为了获得邻苯二甲酸酯(PAEs)生物降解菌，从农田土壤筛选出一株能够以 PAEs为唯一碳源和能源生长的微生物，命名为 SD2。采用聚合酶链式反应(PCR)扩增得到其 16S rDNA并构建分子系统发育树，可鉴定出其为冢村菌(Tsukamurella)。研究该菌株在不同的温度、 pH值和转速条件下对邻苯二甲酸二丁酯(DBP)的降解情况，同时采用高效液相色谱(HPLC)测定该菌株对 DBP的降解能力。结果表明， SD2对 DBP的最佳降解条件为：温度为 30℃， pH值为 9，转速为 150 r/min。在该最佳条件下， SD2在 72 h内可以完全降解 500 mg/L的 DBP。底物广谱试验表明，该菌能够降解长短链的 PAEs化合物及常见的代谢产物。同时采用气相质谱联用仪(GC-MS)检测发现， DBP在降解的过程中产生了中间代谢产物邻苯二甲酸丁基甲酯和邻苯二甲酸(PA)，预示着该菌具备完全降解 DBP的能力。这是首次关于冢村菌降解 PAEs污染物的报道，能够为 PAEs污染环境的生物降解及修复提供新的微生物资源。

Abstract

In order to obtain phthalic acid esters (PAEs) biodegradation strain, a bacterial strain capable of growing with PAEs as the sole carbon source and energy was selected from contaminated soil and named as SD2. The 16S rDNA obtained by PCR ampli.cation and the construction of molecular phylogeny trees can be identi.ed as Tsukamurella sp. The degrada-tion of DBP under different temperature, pH value, and rotational speed conditions was investigated. The degradation abil-ity of the strain to DBP was determined by high performance liquid chromatography. The results showed that the optimum degradation condition of SD2 is temperature of 30℃, pH value of 9 and rotational speed of 150 r/min. Under this optimum condition, SD2 can completely degrade 500 mg/L DBP within 72 h. The intermediate metabolites, butylmethyl phthalate and phthalic acid, were detected by GC-MS during the degradation of DBP, and phthalic acid could be further completely degrad-ed into CO2 and H2O. To our knowledge, this is the.rst report about the biodegradation of PAEs by genus Tsukamurella.

参考文献

[1] 杨捷,姚炎华,尹大强,等.邻苯二甲酸酯高效降解菌的筛选及表征[J].同济大学学报 (自然科学版), 2018, 46(7): 944-950. YANG Jie, YAO Yanhua, YIN Daqiang, et al. Screening and char-acterization of phthalate degrading bacteria[J]. Journal of Tongji University (Natural Science Edition), 2018, 46(7): 944-950.

[2] BENJAMIN S, PRADEEP S, JOSH M S, et al. A monograph on the remediation of hazardous phthalates[J]. Journal of Hazardous Materials, 2015, 298: 58-72.

[3] 严佳丽,陈湖星,杨扬,等.一株高效 DEHP降解菌的分离、鉴定及其降解特性[J].微生物学通报, 2014, 41(8): 1532-1540. YAN Jiali, CHEN Huxing, YANG Yang, et al. Isolation, iden-tification and degradation characteristics of an efficient DEHP degrading bacterium[J]. Bulletin of Microbiology, 2014, 41(8): 1532-1540.

[4] LIAO C S, CHEN LC, CHEN B S, et al. Bioremediation of en-docrine disruptor di-n-butyl phthalate ester by Deinococcus ra-diodurans and Pseudomonas stutzeri[J]. Chemosphere, 2010,78(3): 342-346.

[5] 吴琼,张晓峰,任伟,等.邻苯二甲酸酯的污染现状及微生物降解研究进展[J].微生物学杂志, 2018, 38(2): 122-128. WU Qiong, ZHANG Xiaofeng, REN Wei, et al. Pollution status and microbial degradation of phthalates[J]. Journal of Microbiol-ogy, 2018, 38(2): 122-128.

[6] 于晓章,乐东明,任燕飞.邻苯二甲酸酯在环境中的降解机制[J].生态科学, 2015, 34(4): 180-187. YU Xiaozhang, LE Dongming, REN Yanfei. Degradation mecha-nism of phthalate esters in the environment[J]. Ecological Sci-ence, 2015, 34(4): 180-187.

[7] 金德才,梁任星,王洋洋,等.一株 DBP高效降解菌的筛选及其降解特性[J].中南大学学报 (自然科学版), 2010, 41(1): 8-14. JIN Decai, LIANG Renxing, WANG Yangyang, et al. Screening and degradation characteristics of a DBP degrading bacterium[J] Journal of Central South University (Natural Science Edition),.2010, 41(1): 8-14.

[8] LU Y, TANG F, WANG Y, et al. Biodegradation of dimethyl phthalate, diethyl phthalate and di-n-butyl phthalate by Rhodococcus sp. L4 isolated from activated sludge[J]. Journal of Hazard-ous Materials, 2009, 168(2/3): 938-943.

[9] 杨婧,郭楚玲,刘沙沙,等.邻苯二甲酸酯降解菌的筛选、降解特性及土壤修复研究[J].农业环境科学学报, 2018, 37(5): 933-940. YANG Jing, GUO Chuling, LIU Shasha, et al. Screening, degra-dation characteristics and soil remediation of phthalate degrading bacteria[J]. Journal of Agricultural Environmental Sciences, 2018, 37(5): 933-940.

[10] 温志丹,高大文,李喆,等.邻苯二甲酸酯降解菌的分离鉴定及降解特性[J].哈尔滨工业大学学报, 2013, 45(12): 38-42. WEN Zhidan, GAO Dawen, LI Zhe, et al. Isolation, identi.cation and degradation characteristics of phthalate degrading bacteria[J]. Journal of Harbin Institute of Technology, 2013, 45(12): 38-42.

[11] JIN D C, BAI Z H, ChANG D D, et al. Biodegradation of di-n-butyl phthalate by an isolated Gordonia sp. strain QH-11: genetic identi.cation and degradation kinetics[J]. Journal of Hazardous Materials, 2012, 221-222: 80-85.

[12] KONG X, JIN D C, TAI X, et al. Bioremediation of dibutyl phthalate in a simulated agricultural ecosystem by Gordonia sp. strain QH-11 and the microbial ecological e.ects in soil[J]. Sci-ence of the Total Environment, 2019, 667: 691-700.

[13] 李建龙,邵向丽,刘书亮,等.邻苯二甲酸二丁酯降解菌株的筛选鉴定及其降解特性研究[J].现代食品科技, 2014,10(30): 108-114. LI Jianlong, SHAO Xiangli, LIU Shuliang, et al. Screening, identi.cation and degradation characteristics of dibutyl phthalate degrading strain[J]. Modern Food Science and Technology, 2014,10(30): 108-114.

[14] 张付海,岳永德,花日茂,等.一株邻苯二甲酸酯降解菌降解特性研究[J].农业环境科学学报, 2007, 26(增刊): 79-83. ZHANG Fuhai, YUE Yongde, HUA Rimao, et al. Degradation characteristics of a phthalate degrading bacterium[J]. Journal of Agricultural Environmental Science, 2007, 26(S): 79-83.

[15] 吴学玲,金德才,赵维良,等. 4株邻苯二甲酸二丁酯降解菌的分离鉴定及其相关降解基因的克隆[J].环境科学, 2009,30(9): 2722-2727. WU Xueling, JIN Decai, ZHAO Weiliang, et al. Isolation and identi.cation of four DBP-degrading strains and molecular cloning of the degradation genes[J]. Environmental Science, 2009, 30(9): 2722-2727.

[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. DONG Xiuzhu, CAI Miaoying. Determinative manual for routine bacteriology[M]. Beijing: Science Press, 2001.

[17] YAMAMOTO S, HARAYAMA S. PCR amplification and direct sequencing of gyrb genes with universal primers and their applica-tion to the detection and taxonomic analysis of Pseudomonas pu-tida strains[J]. Applied and Environmental Microbiology, 1995,61(3): 1104-1109.

[18] YOON S H, HAS M, KWON S, et al. Introducing ezbiocloud: a taxonomically united database of 16S rRNA and whole genome assemblies[J]. International Journal of Systematic and Evolution-ary Microbiology, 2017, 67(5): 1613-1617.

[19] CHEN S, DONG Y H, CHANG C, et al. Characterization of a novel cyfluthrin-degrading bacterial strain Brevibacterium au-reum and its biochemical degradation pathway[J]. Bioresource Technology, 2013, 132: 16-23.

[20] 骆祝华,黄翔玲,叶德赞.环境内分泌干扰物 -邻苯二甲酸酯的生物降解研究进展[J].应用与环境生物学报, 2008, 14(6): 890-897. LUO Zhuhua, HUANG Xiangling, YE Dezan. Research progress on biodegradation of environmental endocrine disruptor phthalates[J]. Journal of Applied and Environmental Biology, 2008, 14(6): 890-897.

[21] SAVINI V, FAZII P, FAVARO M, et al. Tuberculosis-like pneumo-nias by the aerobic actinomycetes Rhodococcus, Tsukamurella and Gordonia[J]. Microbes and Infection, 2012, 14(5): 401-410.

[22] TEBYANIAN H, HASSANSHAHIAN M, KARIMINIK A. Hexa-decane-degradation by Teskumurella and Stenotrophomonas strains isolated from hydrocarbon contaminated soils[J]. Jundis-hapur Journal of Microbiology, 2013, 6(7): e9182.

[23] CHICIUDEAN I, NIE Y, T.NASE A M, et al. Complete genome sequence of Tsukamurella sp. MH1: a wide-chain length alkane-degrading actinomycete[J]. Journal of Biotechnology, 2018, 268: 1-5.

王雨婷, 靳拓, 胡芳, 黄仕林, 刘慧君, 金德才. 一株冢村菌的筛选及其对邻苯二甲酸二丁酯的降解特性研究[J]. 激光生物学报, 2020, 29(6): 570. WANG Yuting, JIN Tuo, HU Fang, HUANG Shilin, LIU Huijun, JIN Decai. Screening and Study on DBP Degradation Characteristics of a Tsukamurella sp. Strain[J]. Acta Laser Biology Sinica, 2020, 29(6): 570.

一株冢村菌的筛选及其对邻苯二甲酸二丁酯的降解特性研究

关于本站 Cookie 的使用提示

全站搜索

一株冢村菌的筛选及其对邻苯二甲酸二丁酯的降解特性研究

相关论文

相关资讯

关于本站 Cookie 的使用提示

全站搜索