Characterization and biodegradation kinetics of a new cold-adapted carbamazepine-degrading bacterium, Pseudomonas sp. CBZ-4

Carbamazepine is frequently detected in waters and hardly eliminated during conventional wastewater treatment processes due to its complicated chemical structure and resistance to biodegradation. A carbamazepine-degrading bacterium named CBZ-4 was isolated at a low temperature （10℃） from activated sludge in a municipal wastewater treatment plant. Strain CBZ-4, which can use carbamazepine as its sole source of carbon and energy, was identified as Pseudomonas sp. by the 16S rRNA gene sequence. The composition and percentage of fatty acids, which can reveal the cold-adaptation mechanism of strain CBZ-4, were determined. Strain CBZ-4 can effectively degrade carbamazepine at optimal conditions： pH 7.0, 10℃, 150 r/min rotation speed, and 13% inoculation volume. The average removal rate of carbamazepine was 46.6% after 144 hr of incubation. The biodegradation kinetics of carbamazepine by CBZ-4 was fitted via the Monod model. Vmax and Ks were found to be 0.0094 hr^-1 and 32.5 mg/L, respectively.     

Bioremediation of Soils by Plant–Microbe Systems

Abstract

Sustainable ecosystems can be designed to eliminate environmental toxins and reduce pathogen loads through the direct and indirect consequences of plant and microbial activities. We present an approach to the bioremediation of disturbed environments, focusing on petroleum hydrocarbon (PHC) contaminants. Treatment consists of incorporating a plant-based amendment to enhance ecosystem productivity and physiochemical degradation followed by the establishment of plants to serve as oxidizers and foundations for microbial communities. Promising amendments for widespread use are entire plants of the water fern Azolla and seed meal of Brassica napus (rapeseed). An inexpensive byproduct from the manufacture of biodiesel and lubricants, rapeseed meal is high in nitrogen (6% wt/wt), stimulates >100-fold increases in populations of resident Streptomyces species, and suppresses fungal infection of roots subsequently cultivated in the amended soil. Synergistic enzymatic and chemical activities of plant and microbial metabolism in root zones transform and degrade soil contaminants. Emphasis is given to mechanisms that enable PHC functionalization via reactive molecular species.     

溴氰菊酯降解菌的筛选及其降解特性研究

以过期变质的溴氰菊酯农药为供试材料,采用细菌培养基初筛后,依据气相色谱法测定溴氰菊酯选择性培养基中溴氰菊酯降解率的复筛结果,筛选出2株(X_(09)、X_(20))溴氰菊酯降解菌.结果表明,过期变质的农药同样是筛选农药降解菌的重要资源;筛选出的2株溴氰菊酯降解菌X_(09)、X_(20)分别隶属于肠杆菌属(Enterobacter sp.)和假单胞菌属(Pseudomonas sp.).其降解温度为20～35℃,降解pH值为6～10.2株降解菌在培养温度为30℃条件下,溴氰菊酯降解率分别为65.6%和48.4%;在pit值为7.0条件下,溴溴氰菊酯的降解率分别为68.1%和49.5%;加大接种量(20%-25%)可以提高X_(09)的溴氰菊酯降解率(69.1%),添加1%的牛肉膏、葡萄糖、蔗糖可显著提高X_(20)溴氰菊酯降解率(69.3%).     

Bacterial Poly(Hydroxyalkanoate) Polymer Production from the Biodiesel Co-product Stream

A co-product stream from soy-based biodiesel production (CSBP) containing glycerol, fatty acid soaps, and residual fatty acid methyl esters (FAME) was utilized as a fermentation feedstock for the bacterial synthesis of poly(3-hydroxybutyrate) (PHB) and medium-chain-length poly(hydroxyalkanoate) (mcl-PHA) polymers. Pseudomonas oleovorans NRRL B-14682 and P. corrugata 388 grew and synthesized PHB and mcl-PHA, respectively, when cultivated in up to 5% (w/v) CSBP. In shake flask culture, P. oleovorans grew to 1.3 ± 0.1 g/L (PHA cellular productivity = 13–27% of the bacterial cell dry weight; CDW) regardless of the initial CSBP concentration, whereas P. corrugata reached maximum cell yields of 2.1 g/L at 1% CSBP, which tapered off to 1.7 g/L as the CSBP media concentration was increased to 5% (maximum PHA cellular productivity = 42% of the CDW at 3% CSBP). While P. oleovorans synthesized PHB from CSBP, P. corrugata produced mcl-PHA consisting primarily of 3-hydroxyoctanoic acid (C_8:0; 39 ± 2 mol%), 3-hydroxydecanoic acid (C_10:0; 26 ± 2 mol%) and 3-hydroxytetradecadienoic acid (C_14:2; 15 ± 1 mol%). The molar mass (M_n) of the PHB polymer decreased by 53% as the initial CSBP culture concentration was increased from 1% to 5% (w/v). In contrast, the M_n of the mcl-PHA polymer produced by P. corrugata remained constant over the range of CSBP concentrations used.     

VHB在假单胞菌中的表达及其在环境中的应用

简介了透明颤菌血红蛋白(VHB)的结构,特性和作用机理,并对近年来国外对VHB在假单胞菌属表达及在环境治理中的研究进行了评述,最终展望了VHB在生物修复中的应用前景。     

1,2,4-三氯苯降解菌的分离及其降解质粒的研究

 以1,2,4-三氯苯为唯一碳源,从天津化工厂氯苯生产车间的土壤中分离到1株1,2,4-三氯苯的降解菌THSL-1.通过形态观察和16SrDNA序列测定,该菌株被初步鉴定为施氏假单胞菌(Pseudomonasstutzeri).经质粒检测,在菌株THSL-1中发现1条质粒条带,将所获得质粒转化到E.coli.JM109中,转化子能以1,2,4-三氯苯为唯一碳源生长,且对1,2,4-三氯苯有降解作用.因此,可以认为该质粒携带降解1,2,4-三氯苯的基因.选用HindIII、BamHI、XholI3种限制性内切酶分别对质粒pTHSL-1进行单酶切、双酶切,最终确定质粒平均大小为40.2kb.     

假单胞菌摄磷和释磷条件的研究

考察了碳、硫等物质对废水生物脱磷优势菌假单胞菌磷代谢的影响．结果表明，缺磷环境导致假单胞菌产生过量摄磷现象；假单胞菌无论是经过无碳厌氧、无碳好氧还是无硫厌氧、无硫好氧培养后，转接到富磷培养基内好氧培养时，都产生过量摄磷现象．随着在富磷培养基内的好氧培养时间延长，细菌的摄磷量下降．     

不同底物和环境条件对两株铜绿假单胞菌接触烷烃方式及降解活性的影响

考察了不同烷烃底物和温度、酸度、盐度等环境条件对两株铜绿假单胞菌接触烷烃方式和降解活性的影响.结果表明,微生物接触烷烃的两种方式——直接接触方式和乳化接触方式同时存在于菌体降解烷烃过程中且受底物和环境条件的影响.底物烷烃碳数的增加对菌株两种接触方式降解烷烃都有利,而盐度增加对它们都不利;高温、偏碱性环境以及延长降解时间有利于菌体以乳化方式降解烷烃;低温、中性溶液环境有利于菌体以直接接触方式降解烷烃.     

耐冷嗜碱蒙氏假单胞菌H97的鉴定及其好氧反硝化特性

结合形态、磷脂脂肪酸及16S rRNA基因序列分析鉴定了分离自贵州冬水田的好氧反硝化菌株H97,采用模拟废水探讨了不同温度、接菌量、C/N、初始p H和碳源种类对菌株H97反硝化能力的影响.结果表明,菌株H97为蒙氏假单胞菌(Pseudomonas monteilii),目前国内外尚无蒙氏假单胞菌具有耐冷嗜碱好氧反硝化作用的研究报道,是对好氧反硝化菌的补充;该菌的最适脱氮条件为15℃、p H 9.0、C/N 15、接种量1.5×106CFU·(100 m L)-1,碳源丁二酸钠.此外,菌株H97在p H为7.0~11.0时,均表现出良好的脱氮能力,对硝酸盐氮和总氮的去除率分别可达91.21%和79.10%以上,初始p H为12.0时,对硝酸盐氮和总氮的去除率仍分别达64.75%和36.78%,表现出了较强耐碱能力.在初始硝酸盐氮浓度为50.0 mg·L-1和最适脱氮条件下,菌株H97在48 h内对硝酸盐氮和总氮的去除率可达97.69%和96.32%.同时,H97对温度适应范围广,在15~40℃均具有较强好氧反硝化能力,是1株耐冷嗜碱好氧反硝化细菌,在碱性氮污染水体处理中具有较好的应用潜力.     

Pseudomonas flava WD-3固定化技术及其强化SBR污水处理的应用研究

由南四湖底泥中分离纯化出一株低温菌Pseudomonas flava WD-3,为提高其处理污水的效率并实现重复利用,本试验选取在低温(6~8℃)条件下,采用天然材料海藻酸钠(SA)和人工合成材料聚乙烯醇(PVA)共同作用的复合材料为包埋载体,利用正交试验探索其最优的固定化条件;并将最优的Pseudomonas flava WD-3固定化小球投入到SBR污水处理工艺中,探究其投加量和水力停留时间对生活污水处理效果的影响.结果发现,当SA质量分数为4%,PVA质量分数为6%,Ca Cl_2质量分数为5%,交联时间为24 h时,Pseudomonas flava WD-3固定化小球的物理性能最好,且对污水的处理效果最佳,分别是游离菌对COD、TP和NH+4-N去除效率的1.11、1.17、1.14倍.在SBR工艺中,通过连续3个月的监测数据发现,Pseudomonas flava WD-3的固定化小球对污水NH+4-N、TP、COD和TN的去除率分别介于72.78%~77.48%、62.04%~69.26%、89.08%~92.72%和85.08%~89.08%,且平均去除效率分别为未投加该菌的1.13、1.91、1.25、1.56倍,最终采取的最佳水力停留时间为8 h.并且,Pseudomonas flava WD-3在不同投加量及不同水力停留时间条件下,对污水中各污染物的降解过程符合一级动力学模型.