Isolation of a Pseudomonas Stutzeri strain that degrades 1,2,4-trichlorobenzene and characterization of its degradative plasmid

The genetic information encoding metabolic pathways for xenobiotic compounds in bacteria often resides on catabolic plasmids. The aim of the present work was to know the location of the genes for degrading 1,2,4-trichlorobenzen. In this paper a 1,2,4-trichlorobenzene-degrading strain THSL-1 was isolated from the soil of Tianjin Chemical Plant using 1,2,4-trichlorobenzene as the sole carbon source. The strain was identified as Pseudomonas stutzeri through morphologic survey and 16S rDNA sequence determination. A plasmid was discovered from strain THSL-1 by using the alkali lysis method. When the plasmid was transformed into E. coli. JM109 by the CaCl₂ method, the transformant could grow using 1,2,4-trichlorobenzene as the sole carbon source and had the degradation function of 1,2,4-trichlorobenzene. Therefore, it could be deemed that the plasmid carried the degradative genes of 1,2,4-trichlorobenzene. The average size of the plasmid was finally determined to be 40.2 Kb using selectively three kinds of restricted inscribed enzymes (HindIII, BamHI, and XholI) for single cutting and double cutting the plasmid pTHSL-1, respectively.     

Isolation and characterization of an Arthrobacter sp. strain HB-5 that transforms atrazine

A bacterial strain (HB-5) capable of utilizing atrazine as sole carbon and nitrogen source for growth was isolated from an industrial wastewater sample by enrichment culture. The isolate was identified as Arthrobacter sp. according to its phenotypic features, physiologic and biochemical characteristics, and phylogenetic analysis. The strain exhibited faster atrazine degradation rates in atrazine-containing mineral media than the well-characterized atrazine-degrading bacteria Pseudomonas sp. ADP. The broad optimum pH and temperature ranges observed for strain HB-5 indicate that it has potential for remediation of atrazine-contaminated sites. Strain HB-5 first metabolizes atrazine to yield hydroxyatrazine. Then, the bacterium metabolizes hydroxyatrazine to cyanuric acid, but could not mineralize atrazine.     

Biodegradation of paclobutrazol by a microbial consortium isolated from industrially contaminated sediment

Biodegradability of the plant growth retardant paclobutrazol by a microbial consortium in which Pseudomonas was the predominant strain was investigated in batch culture. The consortium which had been isolated from an industrially contaminated sediment was proven to be useful for the treatment of effluents containing paclobutrazol. Paclobutrazol was degraded by the pure isolated strain of Pseudomonas sp. as well as the microbial consortium. Paclobutrazol was utilized as the sole source of carbon and energy. Sixty percent of the paclobutrazol was degraded by the microbial consortium from an initial concentration of 54 mg L^?1 within 48 h and more than 98% of an initial concentration of 3.4 mg L^?1 was degraded within 36 h. The optimum temperature and pH were determined to be 30°C and 7.0, respectively. A pure strain of a bacterium, isolated from the enrichment culture was identified as Pseudomonas sp. The microbial consortium was tolerant of high pH and could degrade paclobutrazol faster than the pure strain. The degradation rate of this plant growth regulator in an aerobic environment was greater than that under anaerobic conditions.     

间苯二酚的微生物降解研究

从土壤中筛选的一株降解活性较高的细菌Ｔ１，当以间苯二酚为唯一碳源时，降解率可达９８％，研究表明，菌体接种量的大小对间苯二酚降解速度有一定的影响；降解酶为胞内酶，经鉴定，Ｔ１菌为黄单胞菌，其某些特性与已知菌种有差异，需进一步研究。     

Microcystin-LR biodegradation by Sphingopyxis sp. USTB-05

A promising bacterial strain for biodegrading microcystin-LR (MC-LR) as the sole carbon and nitrogen source was successfully isolated from Lake Dianchi, China. The strain was identified as Sphingopyxis sp. USTB-05, which was the first isolated MCs-biodegrading Sphingopyxis sp. in China. The average biodegradation rate of MC-LR by Sphingopyxis sp. USTB-05 was 28.8 mg??L^?1 per day, which was apparently higher than those of other bacteria reported so far. The optimal temperature and pH for both strain USTB-05 growth and MC-LR biodegradation were 30??C and 7.0, respectively. The release of MC-LR from the cyanobacterial cells collected from Lake Guishui and the biodegradation of MC-LR by both strain and cell-free extract (CE) were investigated. The results indicated that MC-LR with the initial concentration of 4.0 mg??L^?1 in water was biodegraded by Sphingopyxis sp. USTB-05 within 4 d, while MC-LR with the initial concentration of 28.8 mg??L^?1 could be completely removed in 3 h by CE of Sphingopyxis sp. USTB-05 containing 350 mg??L^?1 protein. During enzymatic biodegradation of MC-LR, two intermediate metabolites and a dead-end product were observed on an HPLC chromatogram. Moreover, the similar scanning profiles of MC-LR and its metabolic products indicate that the Adda side-chain of MC-LR was kept intact in all products.     

丁草胺高效降解细菌的分离

通过瓶培养法富集培养,从肥东县一块单晚稻田土中分离出一株丁草胺高效降解细菌ＷＹ３０６,经鉴定,该菌为节杆菌属菌Ａｒｔｈｒｏｂａｃｔｅｒ ｓｐ．ＷＹ３０６．Ａｒｔｈｒｏｂａｃｔｅｒ ｓｐ．ＷＹ３０６降解丁草胺的影响因素研究表明：丁草胺降解半衰期与初始菌量近似成反比;当丁草胺添加浓度为５、９ｍｇ／Ｌ时,其降解半衰期分别为０．９７ｈ和１．８６ｈ,随浓度的增大而增大,而当丁草胺添加浓度为０．８ｍｇ／Ｌ或其     

Isolation,identification, and the degradation characteristics of a thiobencarb-degrading bacterium Bacillus sp. T2北大核心CSCD

Thiobencarb, a thiocarbamate herbicide, is widely used to control weeds in rice paddies. Screening for highly efficient thiobencarb-degrading bacteria is important for the bioremediation of thiobencarb-contaminated environments. The aim of this study was to isolate and identify highly efficient thiobencarb-degrading bacteria and to identify the degradation pathway and the degrading properties. The thiobencarb-degrading strain was isolated using methods of microbiological acclimation and enrichment and was then identified using a 16S rRNA phylogenetic analysis. The degrading properties of the isolated bacterium were determined by single-factor experiments, and the degradation products were identified using gas chromatography-mass spectrometry (GC-MS). A thiobencarb-degrading strain T2, which can utilize thiobencarb as the sole source of carbon for energy and growth, was isolated from paddy soil. Strain T2 degraded more than 98.3% of 0.4 mmol/L of thiobencarb within 36 h. It was preliminarily identified as Bacillus sp. T2 according to the 16S rRNA gene analysis and from its morphological, physiological, and biochemical characteristics. The metabolic products of the thiobencarb degradation for strain T2 were identified as 4-chlorobenzyl mercaptan, 4-chlorobenzaldehyde, and 4-chlorobenzoic acid by the GC-MS. Based on metabolite identification, it was speculated that thiobencarb degradation in strain T2 was initiated by the hydrolysis of the thioester bond to produce 4-chlorobenzyl mercaptan, which was further oxidized to 4-chlorobenzaldehyde and 4-chlorobenzoic acid. The thiobencarb degradation that was initiated by the hydrolysis of the thioester bond by strain T2 is a new metabolic pathway, which provides valuable research material and reliable experimental data for revealing the metabolic process and mechanism of thiobencarb microbial degradation in soil. The strain Bacillus sp. T2 has a very high degradation efficiency, suggesting it is a good prospect for microbial remediation in thiobencarb-polluted environments. © 2018 Science Press. All rights reserved.     

Influence of environmental factors on biodegradation of quinalphos by <Emphasis Type="Italic">Bacillus thuringiensis</Emphasis>

Background

The extensive and intensive uses of organophosphorus insecticide—quinalphos in agriculture, pose a health hazard to animals, humans, and environment because of its persistence in the soil and crops. However, there is no much information available on the biodegradation of quinalphos by the soil micro-organisms, which play a significant role in detoxifying pesticides in the environment; so research is initiated in biodegradation of quinalphos.

Results

A soil bacterium strain, capable of utilizing quinalphos as its sole source of carbon and energy, was isolated from soil via the enrichment method on minimal salts medium (MSM). On the basis of morphological, biochemical and 16S rRNA gene sequence analysis, the bacterium was identified as to be Bacillus thuringiensis. Bacillus thuringiensis grew on quinalphos with a generation time of 28.38 min or 0.473 h in logarithmic phase. Maximum degradation of quinalphos was observed with an inoculum of 1.0 OD, an optimum pH (6.5–7.5), and an optimum temperature of 35–37 °C. Among the additional carbon and nitrogen sources, the carbon source—sodium acetate and nitrogen source—a yeast extract marginally improved the rate of degradation of quinalphos.

Conclusions

Display of degradation of quinalphos by B. thuringiensis in liquid culture in the present study indicates the potential of the culture for decontamination of quinalphos in polluted environment sites.

     

乐果降解菌LGX1的筛选及其降解特性研究

通过富集培养,从连续施用农药乐果的土壤中分离得到一株具有较强降解有机磷农药乐果能力的细菌菌株LGX1,通过菌落形态观察及细菌的16SrDNA测序,对其进行了鉴定,同时初步研究了其降解性能。结果表明：该菌株为蜡状芽孢杆菌（Bacillus cereus）,在接种量为20%,乐果初始质量浓度为100mg·L-1,外加碳源葡萄糖质量分数为2%,温度为35℃,初始pH值为4时该菌株对乐果的降解能力最强;并且能在以辛硫磷、毒死蜱和三唑磷为唯一碳源的基础盐培养基中生长,且均比在以乐果为唯一碳源的基础盐培养基中生长的OD600值要高,初步推断菌株LGX1对有机磷农药的降解有一定的广谱性。     

氯代酰胺类除草剂降解菌的分离及降解性能

从生产乙草胺的农药厂废水生物处理池活性污泥中分离到一株氯代酰胺类除草剂降解细菌,命名为Y3B-1.根据表型特征、生理生化特性和16S rDNA序列系统发育分析,将其鉴定为副球菌属(Paracoccus sp.).研究了菌株Y3B-1在不同条件下对多种氯代酰胺类除草剂的降解性能.结果表明:菌株Y3B-1能以乙草胺为碳源生长,并能降解乙草胺、丁草胺和丙草胺,3 d对这3种氯代酰胺类除草剂的降解率分别达到86.7%、65.5%和69.1%,不能降解异丙甲草胺.该菌降解乙草胺的最适温度为30℃,最适pH为7.0,对乙草胺的降解效果与接种量成正相关,对较低浓度的乙草胺有很好的降解效果,过高的起始浓度抑制其对乙草胺的降解,外加营养如酵母膏和土壤悬液则显著促进其对乙草胺的降解.图7参23     

luxAB基因标记甲基对硫磷降解菌DLL-1在土壤和植株根部的生态行为研究

发光酶标记是 1种有效的跟踪微生物在生态环境中动态行为的技术手段。采用luxAB基因标记技术对甲基对硫磷降解菌DLL - 1在土壤中的分布和植株内的定殖情况进行了研究。结果表明 ,DLL - 1可以较长时间的在植株根际定殖 ,3 0d后仍可用X -感光片检测到菌体的存在 ,而根际外未检测到DLL - 1。植株根剖开后在培养基上培养 2 4h后进行X -光片曝光 ,发现DLL - 1能够进入植株根内并定殖。菌株回收后采用测定其农药降解活力和检查质粒图谱 2种方法证实 ,所观测的回收菌株就是接种的DLL - 1菌株     

Isolation and characterization of a bacterial strain that efficiently degrades sex steroid hormones

A bacterial strain, ZY3, growing on sex steroid hormones as the sole source of carbon and energy was isolated from the sewage treatment plant of a prophylactic steroids factory. ZY3 degrades the 3-methoxy-17β-hydroxy-1,3,5(10),8(9)-δ-4-estren (MHE). This strain was preliminarily identified as Raoultella sp. ZY3 according to its morphology and its 16S rRNA gene sequence. During the experimental period (72 h), the optimum temperature, pH and 3-MHE concentration for the degradation of hydride by the strain ZY3 were 35°C, 10 and 10 mg/L, respectively. The degradation rate of the sex steroid hormones increased to 87% and 85% after the addition of maltose and peptone, respectively.     

Methylobacterium sp. YAL-2对乙酰甲胺磷的降解特性

从有机磷生产厂家的下水道污泥中分离出一株对高浓度和低浓度乙酰甲胺磷都具有高效降解能力的寡营养菌YAL-2,根据形态、生理生化和16S rRNA基因系统发育分析,将菌株YAL-2鉴定为Methylobacterium sp.降解特性实验表明,菌株YAL-2能利用乙酰甲胺磷为唯一碳源生长和降解;在添加了甲醇的无机盐培养基中,84 h可完全降解300mg L-1乙酰甲胺磷,24 h将50 mg L-1和10 mg L-1乙酰甲胺磷降至非检测水平;4 d能完全去除100 mg L-1甲胺磷,5 d分别降解58.4%和40.6%的100 mg L-1乐果、敌敌畏.小青菜农药残留去除实验显示,菌株YAL-2可在7 d内将乙酰甲胺磷和甲胺磷将至限量水平.结果表明,将菌株YAL-2应用于保证果蔬等食品的食用安全是可行的.     

假单胞菌AEBL3对土壤中呋喃丹的生物降解

从农药污染的农田土壤中分离到一株假单胞菌AEBL3，该菌能够以呋喃丹为唯一的碳源和氮源生长。使用AEBL3作为生物强化剂对模拟呋喃丹污染的土壤环境进行了生物修复试验。结果表明，接种AEBL3能够明显增加土壤中呋喃丹的降解率。降解菌AEBL3在土壤中具有一定的移动性，当从土壤表层加菌时，对0-7cm深土层中的呋喃丹都有很好的降解效果。在各种投加方式的比较中，以投加降解菌原液修复效果最好，缓冲液悬浮的菌细胞次之，砂粒混合物的效果最差。16S rRNA的变性梯度电泳(DGGE)结果揭示了生物修复过程中土壤微生物菌群结构中的动态变化。     

一株焦化废水降酚菌的质粒初步研究

从处理焦化废水的活性污泥池中,分离得到了一株具有降酚能力的细菌, Ｊ Ａ． Ｊ Ａ 可以在含酚的 Ｈ Ｐ 复杂培养基中生长,也可以在以酚为唯一碳源的 Ａ Ｂ Ｐ 培养基中生长,且具有产黑色素的特点．用碱裂解提取质粒的方法发现它含有一个质粒,定名为ｐ Ｘ Ｈ１ ,并测定了这个质粒的大小约为２ ．６ ｋｂ ．用限制性内切酶 Ｂｇｌ Ｉ和 Ｒｓａ Ｉ进行双酶切,作出了初步的物理图谱．ｐ Ｘ Ｈ１ 经过２０ 次没有酚作为选择压传代培养后仍然能１００ ％ 保留,同时, Ｊ Ａ菌仍具有降酚能力,表明其具有很高的遗传稳定性     

一株降解除草剂膦化麦黄桐(PPT)菌的筛选与鉴定

从喷洒有除草剂的土壤中分离到一株能分解膦化麦黄桐(PPT)的细菌.该菌在以PPT为唯一碳源的培养基上生长,能利用PPT的最高浓度为2. 7g/L.采用常规生理生化鉴定方法,并结合16SrDNA序列分析法对该菌进行鉴定.结果表明,该菌与生癌肠杆菌(Enterobactercancerogenus)序列相似性为99. 3%,在细菌分类学上属于肠杆菌.将它命名为Enterobactersp. PPT. 图4表2参7     

烷烃降解菌的筛选、鉴定及优势菌株的降解特性

以正庚烷为唯一碳源,从长期受到石油污染的土壤中筛选获得可利用正庚烷的微生物14株.通过形态观察和16S rDNA序列比对,鉴定G2、G9、G14为红球菌属,G3、G27为人苍白杆菌属,G4、G7为芽孢杆菌属,G5、G10、G15、G25为节杆菌属,G16为缺陷短波单胞菌,G17、G22为嗜麦芽寡养单胞菌属.通过考察其降解烷烃的能力,确定Rhodococcus sp.G2为烷烃降解优势菌株.该菌株可代谢庚烷获得最大菌体浓度D600 nm=7.51.同时该菌对不同碳链长度的烷烃,如十二烷、十六烷、煤油和二甲苯均具有较强的降解能力,以十二烷为碳源的最大比生长速率为0.37 h-1,最高菌体浓度为D600 nm=12.00,在正十六烷中生长,最大比生长速率为0.23 h-1,在煤油中生长,最大比生长速率为0.14 h-1,在以二甲苯为唯一碳源时,D600 nm也可达到1.00左右.研究表明该菌株对于石油污染土壤的生物修复有很大的应用前景.图6表2参9     

多环芳烃降解菌ZL5分离鉴定及其降解质粒

通过选择性富集培养，从辽河油田石油污染土壤中分离到一株多环芳烃(PAHs)降解菌ZL5．它能以菲和芘为唯一碳源生长，但是不能利用萘．16S rDNA核苷酸序列分析结果表明，ZL5属于变形细菌α亚类中的鞘氨醇单胞菌属．该菌株含有一个大小约为60kb的质粒．丝裂霉素C消除实验表明，随着质粒的丢失，菌株利用菲和芘的能力也丧失．用电转化和氯化铷转化法分别将菌株ZL5的质粒导人大肠杆菌JM109和DH5α中，随着质粒的获得，这些转化子获得了降解菲和芘的能力．本研究结果表明，鞘氨醇单胞菌ZL5降解PAHs的功能和质粒有关。     

Effects of phosphorus sources on volatile organic compound emissions from <Emphasis Type="Italic">Microcystis flos</Emphasis>-<Emphasis Type="Italic">aquae</Emphasis> and their toxic effects on <Emphasis Type="Italic">Chlamydomonas reinhardtii</Emphasis>

There is diverse phosphorus (P) in eutrophicated waters, but it is considered as a crucial nutrient for cyanobacteria growth due to its easy precipitation as insoluble salts. To uncover the effects of complex P nutrients on the emission of volatile organic compounds (VOCs) from cyanobacteria and their toxic effects on other algae, the VOCs from Microcystis flos-aquae supplied with different types and amount of P nutrients were analyzed, and the effects of VOCs and their two main compounds on Chlamydomonas reinhardtii growth were investigated. When M. flos-aquae cells were supplied with K₂HPO₄, sodium pyrophosphate and sodium hexametaphosphate as the sole P source, 27, 23 and 29 compounds were found, respectively, including furans, sulfocompounds, terpenoids, benzenes, aldehydes, hydrocarbons and esters. With K₂HPO₄ as the sole P source, the VOC emission increased with reducing P amount, and the maximum emission was found under Non-P condition. In the treatments of M. flos-aquae VOCs under Non-P condition and two main terpenoids (eucalyptol and limonene) in the VOCs, remarkable decreases were found in C. reinhardtii cell growth, photosynthetic pigment content and photosynthetic abilities. Therefore, we deduce that multiple P nutrients in eutrophicated waters induce different VOC emissions from cyanobacteria, and P amount reduction caused by natural precipitation and algal massive growth results in more VOC emissions. These VOCs play toxic roles in cyanobacteria becoming dominant species, and eucalyptol and limonene are two toxic agents.     

降解氯氰菊酯光合细菌的分离鉴定及降解特性研究

从农药厂污水处理池的活性污泥中分离到一株高效氯氰菊酯农药降解菌,命名为PSB07-13。根据该菌体培养特征、菌落形态特征、活细胞光谱吸收特征、生理生化特性、光合作用内膜系统结构类型,并结合16S rRNA（Genebank Accession NO.EU366142）序列相似性分析,将其鉴定为沼泽红假单胞菌（Rhodopseudomonas palustris）。利用气相色谱对PSB07-13的降解能力进行了测定,结果表明：该菌培养6d后,对50mg·L-1的氯氰菊酯的降解率达到80.94%。降解特性研究结果表明：该菌在含氯氰菊酯培养基中的最适生长温度为30℃、pH为7.0及光照强度为7500lx;该菌不能以氯氰菊酯为唯一碳源和能源生长;该降解菌还能较好地降解甲氰菊酯、联苯菊酯、溴氰菊酯等菊酯类农药。该农药残留降解菌可以用于农药厂有机废水处理及农田农药残留降解,具有一定的应用前景。