毒死蜱降解菌株Bacillus latersprorus DSP的降解特性及其功能定位

从土壤中分离到一株能有效降解毒死蜱的细菌DSP，该分离株鉴定为侧芽孢杆菌（Bacillus latersprorus）。在纯培养条件下测定了分离株DSP对毒死蜱的降解性能。在接种量为菌浓度OD415=0．2，pH7．0、25℃条件下，测得1、10mg L^-1毒死蜱的降解符合一级动力学特征，其降解半衰期分别为1．48d、5．00d，100mg L^-1毒死蜱对DSP菌有明显的抑制作用；分离株DSP在不同pH及温度下对毒死蜱的降解作用为pH7．0〉pH5．0〉pH9．0，35℃〉25℃〉15℃。该菌株含有一个20kb左右的质粒，通过吖啶橙与升温法对质粒消除实验证实，随着质粒的丢失，菌株利用毒死蜱的能力也丧失，用热击法和CaCl2法将菌株质粒转人大肠杆菌JM109和质粒消除处理的DSP^-菌中，随着质粒的获得，这些转化子获得了降解毒死蜱的能力。研究结果表明，Bacillus latersprorus DSP降解毒死蜱的功能和质粒有关。     

普施特降解菌Bacillus sp.zx2和zx7生长及降解特性

芽孢杆菌zx2和zx7是普施特高效降解菌,研究其生长和降解特性旨在为普施特污染土壤的生物修复提供科学依据。采用瓶培养法,对芽孢杆菌zx2和zx7的生长特性及单菌和复合菌对普施特的降解特性进行了研究。结果表明,zx2和zx7均可在普施特初始浓度≤200mg·L-1的无机盐培养液中生长良好,zx2在温度25～35℃和pH4.0～7.0时生长良好,而zx7适宜在温度30～35℃和pH5.0～8.0时生长,可见在适应性上二者互补。在最佳条件（温度32℃、pH6.0和普施特初始浓度为200mg·L-1）下,zx2和zx7在无机盐培养液中对普施特降解动态均符合阻滞动力学,半衰期分别为3.8d和2.8d,培养6d时普施特降解率分别为85.81%和90.27%。在培养过程中,zx2的pH是降低的,而zx7的pH基本不变,可初步表明二者降解机理不同;zx2和zx7复合菌（1∶1）对普施特降解率比单菌低,为82.70%,这可能是因为zx2或zx7降解普施特的过程中利用了对方产生的降解产物。     

沼灌土壤17β-雌二醇专性降解菌的分离筛选及降解特性

集约化畜禽养殖场产生的沼液通常就地回用,在循环利用有机物的同时也会带来类固醇雌激素（Steroid Estrogens,SEs）的累积及污染。为降低沼灌后SEs对水土环境的污染风险,该研究采用富集和纯化培养法,对西南地区某奶牛养殖场沼灌区土壤中雌激素降解菌进行分离及筛选,获得一株利用17β-雌二醇（17β-E2）为唯一碳源生长繁殖的降解菌。通过16S rDNA 基因序列进行同源性比对以确定种属,并研究其降解特性。分别研究了菌株在不同温度、pH值、底物浓度三种单因素条件下的降解特性,然后利用三因素三水平正交试验继续优化菌株最适降解条件。结果表明：分离出的优势菌为生丝微菌属（Hyphomicrobium sp.）,命名为Hyphomicrobium sp.SS-1,该菌株在10～40 ℃、pH值为5～9、底物浓度为1～10 mg/L的条件下,均能不同程度降解17β-E2。其中菌株在温度为30 ℃、pH值为7、底物浓度5 mg/L的条件下,培养7 d对17β-E2的降解率可达71%,并伴随毒性低于E2的降解产物E1和E3生成,总雌激素去除率为56.8%。正交试验结果显示,各因素对菌株降解能力的影响顺序从小到大为：底物浓度、温度、pH值,且都为显著影响（P<0.05）;菌株最适降解条件为温度35 ℃、pH值为7、底物浓度5 mg/L,该条件下培养7 d,菌株对17β-E2的降解率可达97.09%。研究结果可为复杂基质环境中微生物降解SEs提供优质菌种资源,并为沼液灌溉区土壤的雌激素污染修复提供有效途径。     

一株新的多菌灵高效降解茵的筛选与降解特性分析

从长期施用多菌灵农药的土壤中,通过富集筛选,获得1株新的多菌灵高效降解菌株。通过生理生化实验和16SrDNA序列同源性分析鉴定该菌株,应用高效液相色谱法对纯培养条件下菌株的降解特性和粗酶提取液的降解性能进行了分析。结果表明,筛选所获得的菌株与Raoultella菌属的亲缘关系最近,将其命名为Raoultellasp．MBC,该菌株能在以多菌灵为唯-碳源的无机盐培养基中生长;25℃、pH7．0、200r·min。的最适生长条件下避光振荡培养72h,多菌灵的降解率达到100％;在最适培养条件下外加氮源和碳源在培养后期均可以提高多菌灵的降解率,外加氮源对多菌灵的降解效果优于外加碳源;该菌体的粗酶提取液具有降解多菌灵活性,且多菌灵降解酶为诱导酶。研究结果为多菌灵污染土壤的生物修复和酶修复提供了材料和理论依据。     

哒螨灵农药在土壤中的降解吸附和移动特性

采用室内模拟试验方法,研究了哒螨灵在3种土壤中的降解、吸附和移动特性。结果表明,25℃下,哒螨灵在江西红壤、河南二合土和东北黑土中的降解半衰期分别为41．0、55．9和72．2d,属于易降解农药,其降解速率依次为江西红壤〉河南二合土〉东北黑土。酸性条件有利于哒螨灵的降解,土壤pH值对哒螨灵降解的影响比土壤有机质含量大。3种土壤对哒螨灵农药的吸附均较好地符合Freundich方程,吸附系数鼠值分别为3．35×10^3,6．17×10^3和8．48×10^3,具有极强的吸附性,且土壤有机质含量越高,对哒螨灵的吸附性越强。土壤对哒螨灵的吸附自由能变化均小于40kJ·mol^-1,属于物理吸附。哒螨灵在土壤中不易移动,3种土壤薄层移动试验的Rf值均仅为0．05,正常条件下不会造成对地下水的污染。     

假丝酵母菌降解养殖水体氨氮的特性研究

为筛选降解养殖水体氨氮的功能菌,从海水中取样,用212mg/L氨氮浓度的分离培养基进行培养,筛选获得了一株能高效降解养殖水体氨氮功能的菌株,经鉴定为假丝酵母菌。通过研究养殖水体氨氮浓度、pH值、温度等因素对菌株降解氨氮作用的影响,结果表明,菌株降解氨氮较适宜的条件为：养殖水体氨氮≤20 mg/L,pH值6～7,温度25～30℃,盐浓度0～1％,溶氧2 mg/L以上。假丝酵母菌在这种水体生态条件下接种5％,氨氮降解率接近80％。     

具有产表面活性剂功能石油降解茵的筛选及其发酵条件优化

从广州某炼油厂附近石油污染的土壤中筛选出一株可高效产表面活性剂的原油降解菌株MZ01,结合菌株形态观察、革兰氏染色和16SrDNA序列同源性进行分析鉴定其属于假单胞菌属（Pseudomonassp．MZ01）,该菌9d对原油的降解率达54．7％。通过正交实验优化其产表面活性剂的环境因子并进行发酵培养,结果表明,MZ01最佳发酵条件为：酵母膏（3g·L^-1）作为氮源,玉米油（2g·L^-1）作为碳源,温度为25℃,pH值为9．0和含盐量为5％。该条件下3d的发酵产物经提纯后得到表面活性剂产量为2．27g·L^-1,测得该产物CMC值为0．1g·L^-1,可将水的表面张力从初始的72mN·m^-1降至30mN·m^-1。     

一株从金钱树上分离的产纤维素降解酶的菌株的分离及特性鉴定

本研究从金钱树（Zamioculcas zamiifolia）白绢病（Southern blight）病株上分离到一株产纤维素酶菌株SM,经形态观察和rDNA—ITS序列分析鉴定为齐整小核菌（Sclerotium rolfsii Sacc）,经以羧甲基纤维素钠为唯一碳源的培养基培养和刚果红染色测定,证明SM菌株可产生高活性纤维素降解酶。本文对SM菌株产纤维素降解酶的液态发酵条件进行了研究,结果表明：在起始pH6．0,无机氮：有机氮：纤维素碳源之比为0．07g：1．4g：1．6g,温度为30℃,摇床转速为160r／min,发酵培养时间为5d的条件下,该菌株发酵液的CMC酶活性达到11．7U／mL,滤纸酶活性达到2．156U／mL,β-葡萄糖苷酶活性达到3．911U／mL。     

甲磺隆降解菌FLDA的分离鉴定及其降解特性研究

从生产甲磺隆的农药厂内采取污泥，经驯化富集后筛选到一株能高效降解甲磺隆的细菌FLDA，根据表型特征、生理生化特性及16S rDNA序列同源性分析，将FLDA初步鉴定为假单胞菌（Pseudomonas sp）。该菌能在含甲磺隆（30mgL^-1）的基础盐液体培养基中降解甲磺隆，5d降解率达72．6％，该菌降解甲磺隆的最适pH为7．0，最适温度为30℃，该菌降解甲磺隆的速率和起始接种量呈正相关。酶的定域实验表明，该菌中甲磺隆水解酶为胞内酶。FLDA投加土壤，可提高土壤中甲磺隆的降解速率。     

恶唑菌酮土壤降解影响因子研究

探讨了土壤环境中的主要因素:土壤微生物、温度、含水量、pH值以及施用有机肥对恶唑菌酮降解的影响。结果表明:土壤微生物对恶唑菌酮在土壤中的降解起着重要作用,相同条件下灭菌土壤的降解半衰期是非灭菌土壤的27.6倍。环境温度、土壤含水量等对恶唑菌酮降解也有影响,在15℃～40℃的试验条件下,随着温度升高,恶唑菌酮的降解速率加快,特别是15℃～25℃温度范围内降解速率上升较快;过高和过低的土壤含水量都不利于土壤中恶唑菌酮的降解,土壤含水量为50?～100?时适宜恶唑菌酮的降解;此外施用有机肥会加速恶唑菌酮的降解;而土壤pH值对降解的影响不显著。     

Biodegradation of chloroacetamide herbicides by Paracoccus sp. FLY-8 in vitro

A butachlor-degrading strain, designated FLY-8, was isolated from rice field soil and was identified as Paracoccus sp. Strain FLY-8 could degrade and utilize six chloroacetamide herbicides as carbon sources for growth, and the degradation rates followed the order alachlor > acetochlor > propisochlor > butachlor > pretilachlor > metolachlor. The influence of molecular structure of the chloroacetamide herbicides on the microbial degradation rate was first analyzed; the results indicated that the substitutions of alkoxymethyl side chain with alkoxyethyl side chain greatly reduced the degradation efficiencies; the length of amide nitrogen's alkoxymethyl significantly affected the biodegradability of these herbicides: the longer the alkyl was, the slower the degradation efficiencies occurred. The phenyl alkyl substituents have no obvious influence on the degradation efficiency. The pathway of butachlor complete mineralization was elucidated on the basis of the results of metabolite identification and enzyme assays. Butachlor was degraded to alachlor by partial C-dealkylation and then converted to 2-chloro-N-(2,6-dimethylphenyl)acetamide by N-dealkylation, which subsequently transformed to 2,6-diethylaniline, which was further degraded via the metabolites aniline and catechol, and catechol was oxidized through an ortho-cleavage pathway. This study highlights an important potential use of strain FLY-8 for the in situ bioremediation of chloroacetamide herbicides and their metabolite-contaminated environment.     

Enzymatic binding of the pollutant 2,6-xylenol to a humus constituent

Dimeric and polymeric hybrid oliogmers were obtained by the reaction of 2,6-xylenol and syringic acid in the presence of an enzyme from the fungus Trametes versicolor. Three hybrid dimers were formed in an assay at pH 4.5, while at pH 6.5, the formation of additional higher polymers with a phenyl-ether bond (tail to head linkage) was detected. The major product was identified as 3,5-dimethyl-3′,5′-dimethoxydiphenoquinone (4,4′), and it amounted to approximately 45% of the applied 2,6-xylenol after 3.5 hr incubation at pH 4.5. About 8% of the 2,6-xylenol was transformed to 4,4′-dihydroxy-3,5-dimethyl-3′,5′-dimethoxydiphenol and 22% to 3-methoxy-5-(2′,6′-dimethylphenoxy)-benzoquinone (1,2).     

乙草胺降解菌Y-4的分离鉴定及降解特性研究

从长期经乙草胺污染的污泥中分离到一株能以乙草胺为唯一碳源和能源生长的菌株Y-4,通过生理生化实验和16S rDNA同源性序列分析,鉴定为申氏杆菌属（Shinella sp.）。采用室内培养方法,研究了Y-4对乙草胺的降解特性。结果表明,Y-4能有效地降解浓度为5～200 mg.L-1的乙草胺,在48 h内对50 mg.L-1乙草胺的降解率达到83.3%。菌株Y-4降解乙草胺的最适pH值为8.0,最适温度为30℃,其对丙草胺和丁草胺等农药也有良好的降解效果。     

联苯菊酯降解菌的筛选、鉴定及降解特性研究

联苯菊酯是一种广谱高效杀虫剂,大规模的应用使其广泛残留在环境中,因此筛选联苯菊酯的高效降解菌具有重要意义。从扬州农药厂附近的地表土壤取样,利用富集驯化培养分离得到一株编号为S8的降解细菌,经表形特征、生理生化特性和16S rDNA序列分析其为醋酸钙不动杆菌（Acinetobacter calcoaceticus）,该菌株在pH7.0和30 ℃的条件下,对100 mg·L-1联苯菊酯的3 d降解率达56.4%,半衰期为60.7 h。其最适生长条件为：pH6.0~8.0,温度30~35 ℃,接种量5%。研究结果可为今后治理联苯菊酯残留污染提供理论参考。     

Determination of alachlor and its metabolite 2,6-diethylaniline in microbial culture medium using online microdialysis enriched-sampling coupled to high-performance liquid chromatography

In this study, a simple and novel microdialysis sampling technique incorporating hollow fiber liquid phase microextraction (HF-LPME) coupled online to high-performance liquid chromatography (HPLC) for the one-step sample pretreatment and direct determination of alachlor (2-chloro-2',6'-diethyl-N -(methoxymethyl)acetanilide) and its metabolite 2,6-diethylaniline (2,6-DEA) in microbial culture medium has been developed. A reversed-phase C-18 column was utilized to separate alachlor and 2,6-DEA from other species using an acetonitrile/water mixture (1:1) containing 0.1 M phosphate buffer solution at pH 7.0 as the mobile phase. Detection was carried out with a UV detector operated at 210 nm. Parameters that influenced the enrichment efficiency of online HF-LPME sampling, including the length of the hollow fiber, the perfusion solvent and its flow rate, the pH, and the salt added in sample solution, as well as chromatographic conditions were thoroughly optimized. Under optimal conditions, excellent enrichment efficiency was achieved by the microdialysis of a sample solution (pH 7.0) using hexane as perfusate at the flow rate of 4 μL/min. Detection limits were 72 and 14 ng/mL for alachlor and 2,6-DEA, respectively. The enrichment factors were 403 and 386 (RSD < 5%) for alachlor and 2,6-DEA, respectively, when extraction was performed by using a 40 cm regenerated cellulose hollow fiber and hexane as perfusion solvent at the flow rate of 0.1 μL/min. The proposed method provides a sensitive, flexible, fast, and eco-friendly procedure to enrich and determine alachlor and its metabolite (2,6-DEA) in microbial culture medium.     

一株高效降解玉米赤霉烯酮的耐酸耐高温枯草芽孢杆菌的研究

为寻求更高效、实用的生物脱毒制剂,提高其应用范围,通过采集酸性、高温的温泉环境样品,筛选获得1株高效降解玉米赤霉烯酮(ZEN)的细菌菌株Y-33,并对该菌株生长和降解特性进行分析。结果表明,菌株Y-33在50℃、pH值5.0条件下孵育36 h,菌液对2 μg·mL^-1 ZEN的降解率高达93.79%,对20 μg·mL^-1 ZEN的降解率也达82.40%。从形态、生理生化特性及16S rDNA序列对菌株Y-33进行分析,初步鉴定为枯草芽孢杆菌(Bacillus subtilis)。该菌株发酵上清液在28℃条件下对ZEN降解率为62.02%。金属离子Mn²⁺能明显增强菌株Y-33发酵上清液对ZEN的降解能力,降解率达81.17%,而Zn²⁺严重抑制了Y-33发酵上清液对ZEN的降解能力,降解率仅为5.42%。本研究结果为ZEN解毒菌剂的开发与应用奠定了理论基础。     

采用Biolog, DGGE 和PLFA三种方法研究利用方式对土壤微生物群落结构的影响

Biolog, 16S rRNA gene denaturing gradient gel electrophoresis （DGGE）, and phospholipid fatty acid （PLFA） analyses were used to assess soil microbial community characteristics in a chronosequence of tea garden systems （8-, 50-, and 90- year-old tea gardens）, an adjacent wasteland, and a 90-year-old forest. Biolog analysis showed that the average well color development （AWCD） of all carbon sources and the functional diversity based on the Shannon index decreased （P 〈 0.05） in the following order： wasteland 〉 forest 〉 tea garden. For the DCCE analysis, the genetic diversity based on the Shannon index was significantly lower in the tea garden soils than in the wasteland. However, compared to the 90-year-old forest, the tea garden soils showed significantly higher genetic diversity. PLFA analysis showed that the ratio of Gram positive bacteria to Cram negative bacteria was significantly higher in the tea garden soils than in the wasteland, and the highest value was found in the 90-year-old forest. Both the fungal PLFA and the ratio of fungi to bacteria were significantly higher in the three tea garden soils than in the wasteland and forest, indicating that fungal PLFA was significantly affected by land-use change. Based on cluster analysis of the soil microbial community structure, all three analytical methods showed that land-use change had a greater effect on soil microbial community structure than tea garden age.     

降解菌HD接种和非接种根围土壤中丁草胺的降解动力学研究

测定了小麦、棉花、水稻和玉米根围土壤和非根围土壤中丁草胺的降解特征和降解菌变化动态。结果表明,种植作物丰富了土壤微生物,根围土壤丰富的微生物对丁草胺的降解具有显著的促进作用。根围土壤中丁草胺的降解是非根围土壤的1.63～2.34倍,相应的半衰期缩短为非根围土壤的 42.2％～72.8％。根围土壤接种处理后这种促进作用得到进一步加强,其降解速率是非根围土壤的1.68～2.83倍,半衰期为非根围土壤的34.4％～59.4％。试验结果表明,作物根围是丁草胺残留快速降解的微环境,作物根围接种处理可以强化丁草胺残留的微生物降解。     

Cry1Ac蛋白降解菌株FJSB3的分离鉴定及降解特性

从转Cry1Ac基因水稻种植田土壤中,分离纯化得到1株能高效降解Cry1Ac蛋白的细菌FJSB3。通过表型特征、16S rDNA扩增和电镜观察,初步鉴定FJSB3为寡养单胞菌（Stenotrophomonas sp.）。FJSB3发酵液能降解粗Bt蛋白。通过单因素试验确定FJSB3降解水稻秸秆中Cry1Ac蛋白的最适条...