孔雀石绿高效脱色菌株的筛选、鉴定与脱色特性研究

孔雀石绿应用广泛,但难以降解且对许多生物都具有致癌致畸性.从浙江温州皮革厂污泥中分离筛选到1株孔雀石绿高效脱色菌株DH-9,16S rRNA基因序列分析表明,该菌株属于Enterabacter sp.属.单因素实验结果表明:当pH值在3.0~9.0时,培养24 h以后,该菌株对孔雀石绿的脱色率均在90%以上;脱色的最适温度范围为30~40℃;多数所测试碳源对脱色没有显著影响,而多数所测试氮源则对脱色有显著的促进作用;所测金属离子中,仅Cu2+和Fe3+对脱色有显著的抑制效应;此外,当接种量达到3%(V∶V,菌体干重约0.23 g·L-1)以后,12 h的脱色率即可达到90%以上.响应面设计实验结果显示,菌株DH-9对孔雀石绿脱色的最优操作条件为:pH 6.0、1.0 g·L-1的半乳糖、1.0 g·L-1的酵母粉、3.0 mmol·L-1的氯化钙以及培养温度为34.5℃.验证实验结果表明:在最优条件下,该菌株在8 h内对孔雀石绿的脱色率可达99.4%.总体而言,菌株DH-9在孔雀石绿脱色中的实际应用潜能较大.     

高效聚磷菌Alcaligenes sp. ED-12菌株的分离鉴定及其除磷特性

利用经典的微生物筛选方法,从福州市闽侯县上街镇高岐村某排污口淤泥中分离出1株高效聚磷菌,并结合16S rRNA基因序列分析进行了菌株鉴定.结果表明,该菌株为产碱杆菌,将其命名为Alcaligenes sp.ZGED-12.理化因素实验显示,在以乙酸钠为碳源、NH4Cl为氮源,当C/N为3∶1,pH为8.0,温度和摇床转速分别为35℃和100 r·min-1时,该菌株的生长状态最好,对磷的去除能力也最强,最高除磷率可达80%.此外,该菌株能够耐受较高浓度的磷,当磷浓度超过45 mg·L-1时会产生抑制效应.同时,以聚乙烯醇(PVA)和海藻酸盐(SA)制备了聚磷微生物固定化小球,并考察了菌球对氮磷废水的净化效果.结果表明,氮磷的去除包括固定化材料的吸附作用及微生物的生长利用和/或贮存,显示出了良好的应用前景.     

Effect of commercial pesticides on plant growth-promoting activities of Burkholderia sp. strain L2 isolated from rhizosphere of Lycopersicon esculentum cultivated in agricultural soil

The in vitro study was carried out to examine the toxicity of four commercially used pesticides (phorate, mancozeb, chlorpyrifos, and endosulfan) at the recommended dose (1X) and higher field doses (1.5X, 2X, 2.5X, and 3X) on plant growth-promoting activities of Burkholderia sp. strain L2, isolated from pesticide-treated rhizosphere of Lycopersicon esculentum. At the recommended dose, level of toxicity of all the pesticides to phosphate solubilization activity and indole-3-acetic acid (IAA) production was less detrimental as compared to exceeding doses. At maximum dose rate of each pesticide, the percentage decrease in phosphate solubilization and IAA production over control was found in order of chlorpyrifos < mancozeb < endosulfan < phorate. Present study proves that Burkholderia sp. strain L2 could be used as a potential microbial inoculant in the agricultural soil having persistent pesticide residues.     

Pseudomonas sp.fz胞外分离物降解四溴双酚A

四溴双酚A(TBBPA)是全球消耗量最大的溴系阻燃剂。通过活性物质的定位、分离和TBBPA产物的分析等步骤,对Pseudomonas sp.fz胞外分离物降解TBBPA进行了研究。结果表明,存在于胞外的活性物质通过异丙基断裂和脱溴两条途径降解TBBPA。通过超滤,Sephadex G-10分离纯化得到了具有降解活性的小分子物质,分子量约为376~456 Da,初步鉴定为短肽类物质。这种小分子物质具有很好的热稳定性(30~80℃),在弱酸性条件下(2.0~5.0)活性较高,其活性还受金属离子、氧气和抑制剂(Na N3)的影响。此外,部分纯化的小分子物质在一定条件下能够产生羟基自由基(·OH)。     

产油脂微藻的分离鉴定及培养条件优化

为促进自养高产油微藻的工业化应用,从青海多种生境中分离到34株产油微藻,以筛选获得产油脂最高的青3-2-2株为出发株,18S rRNA分析表明其与栅列藻属(Scenedesmus sp.)同源性达到99%以上,确定该株属于栅列藻属.研究氮源、培养时间、培养温度、初始pH值等培养条件对微藻生物量及油脂含量的影响,优化培养条件为:硝酸钠为氮源,以10%(V/V)接种量接种于普通SE培养基,培养温度为25℃,初始pH 7.0,培养周期为20 d,可以获得较高的油脂产率,比优化前的产率增加了近70%.     

Pseudomonas sp. ZXY-1, a newly isolated and highly efficient atrazine-degrading bacterium, and optimization of biodegradation using response surface methodology

Atrazine, a widely used herbicide, is increasing the agricultural production effectively, while also causing great environmental concern. Efficient atrazine-degrading bacterium is necessary to removal atrazine rapidly to keep a safe environment. In the present study, a new atrazine-degrading strain ZXY-1, identified as Pseudomonas, was isolated. This new isolated strain has a strong ability to biodegrade atrazine with a high efficiency of 9.09 mg/L/hr.Temperature, p H, inoculum size and initial atrazine concentration were examined to further optimize the degradation of atrazine, and the synthetic effect of these factors were investigated by the response surface methodology. With a high quadratic polynomial mathematical model(R~2= 0.9821) being obtained, the highest biodegradation efficiency of 19.03 mg/L/hr was reached compared to previous reports under the optimal conditions(30.71°C, pH 7.14, 4.23%(V/V) inoculum size and 157.1 mg/L initial atrazine concentration).Overall, this study provided an efficient bacterium and approach that could be potentially useful for the bioremediation of wastewater containing atrazine.     

Microbial reduction of uranium (VI) by Bacillus sp. dwc-2: A macroscopic and spectroscopic study

The microbial reduction of U(VI) by Bacillus sp. dwc-2, isolated from soil in Southwest China, was explored using transmission electron microscopy (TEM), X-ray photoelectron spectroscopy (XPS) and X-ray absorption near edge spectroscopy (XANES). Our studies indicated that approximately 16.0% of U(VI) at an initial concentration of 100 mg/L uranium nitrate could be reduced by Bacillus sp. dwc-2 at pH 8.2 under anaerobic conditions at room temperature. Additionally, natural organic matter (NOM) played an important role in enhancing the bioreduction of U(VI) by Bacillus sp. dwc-2. XPS results demonstrated that the uranium presented mixed valence states (U(VI) and U(IV)) after bioreduction, which was subsequently confirmed by XANES. Furthermore, the TEM and high resolution transmission electron microscopy (HRTEM) analysis suggested that the reduced uranium was bioaccumulated mainly within the cell and as a crystalline structure on the cell wall. These observations implied that the reduction of uranium may have a significant effect on its fate in the soil environment in which these bacterial strains occur.     

应用于矿山修复的高效菌株鉴定与溶岩机制:基于增强回归树分析

岩石矿区废弃地水土流失问题极度严峻,微生物对修复该问题造成的退化生境具有重要意义,为探究微生物对岩石矿区生境修复的有效性与作用机制,采用两次定向筛选,获得1株高效产铁载体及吲哚乙酸(IAA)的硅酸盐矿物分解细菌,利用16S rRNA基因序列分析鉴定该细菌为Pseudomonas protegens.通过该细菌对硅酸盐岩的岩石分解实验,测定培养液pH值,K、Al、Si这3种元素的释放量,检测岩石溶蚀的粒径变化,并分析培养液中不同有机酸,氨基酸及多糖的浓度变化,研究Pseudomonas protegens NLX-4对硅酸盐的溶蚀效果,同时引入增强回归树(boosted regression tree analysis,BRT)分析,探究其促溶机制.分析表明,该细菌通过代谢积累一定浓度的酒石酸(777 mg·L~(-1))以及多糖(8.21 g·L~(-1)),对硅酸盐岩产生释钾、释铝、脱硅的作用,从而有效促进岩石的分解.菌株Pseudomonas protegens NLX-4可作为修复废弃矿地被破坏生境的良好菌种资源.     

噻吩磺隆降解菌Staphylococcus sp.的分离及其降解特性与途径

采用富集培养技术从磺酰脲类除草剂污染土壤中筛选得到1株降解噻吩磺隆的细菌,命名为ZWS13.经形态特征和16S rRNA基因序列分析,初步鉴定为葡萄球菌属(Staphylococcus sp.).采用HPLC研究了初始底物浓度、温度、接种量、pH等因素对菌株ZWS13降解噻吩磺隆的影响.结果表明,菌株ZWS13对噻吩磺隆的降解具有较广的底物浓度范围,在7 d内对初始浓度5.0~100.0 mg·L~(-1)噻吩磺隆的降解率达到60%以上;菌株ZWS13降解噻吩磺隆的较适pH为8.0,较适温度为40℃,较适接种量为1%;其中,菌株培养温度为40℃时,菌株在10 d内对50.0mg·L~(-1)噻吩磺隆的降解率达到99%以上.降解谱测定结果表明,菌株对烟嘧磺隆、吡嘧磺隆和甲磺隆亦具有良好的降解效果.采用HPLC-MS分析确定了菌株对噻吩磺隆的5个降解产物,推测菌株对噻吩磺隆的降解途径主要为磺酰脲桥C—N键的断裂、脱甲基作用或脱酯作用的甲基丢失及三嗪环的开裂.研究表明,菌株ZWS13能够有效地降解噻吩磺隆,具有生物修复噻吩磺隆污染的潜力.     

溶藻活性物质对棕囊藻溶藻及其脂肪酸影响的模拟

为了探讨芽孢杆菌B1分泌的胞外溶藻活性物质对球形棕囊藻的溶藻特性和藻毒素物质脂肪酸的影响,比较了模拟自然水体中叶绿素a、p H、溶解氧DO、高锰酸盐指数和营养元素N、P浓度在溶藻前后的变化,并利用GC-MS检测了球形棕囊藻脂肪酸的成分和含量.用体积比1∶100的芽孢杆菌B1胞外活性物质处理模拟水体14 d,发现水体中叶绿素a、p H值和高锰酸盐指数随处理时间的增加而降低,DO和N、P浓度随处理时间的增加而增加.在第14 d时,处理组水体中p H值由8.50降低到7.51,叶绿素a降低82.3%(P0.05),DO增加29.5%(P0.05),高锰酸盐指数降低55.2%(P0.01).NH+4-N、NO-2-N、NO-3-N和PO3-4-P浓度分别增加了0.46、1.50、6.24和1.30倍.投加活性物质处理14 d后,球形棕囊藻藻毒素中的主要3种脂肪酸C18:2、C16:0和C18:1分别降低了100%、97.7%和85.4%(P0.01),总脂肪酸含量降低83.4%(P0.01).结果表明,芽孢杆菌B1胞外溶藻活性物质在模拟自然水体中能有效抑制球形棕囊藻的生长,并降低藻毒素脂肪酸的含量.研究结果为芽孢杆菌B1胞外活性物质的生态安全性应用提供理论基础.