反应条件对苛求芽孢杆菌胞内尿酸酶与嘌呤衍生物作用的影响

用底物尿酸的紫外吸收变化跟踪反应,用单或双底物米氏方程和初速度估算苛求芽孢杆菌胞内尿酸酶最大反应速度,按Arrenius经验公式得到该尿酸酶在pH 9.2和pH 7.4的活化能,但这些活化不能解释其在两个pH下的催化活性差异.该酶同常见嘌呤衍生物氧嗪酸、黄嘌呤和尿酸有亲和力,但对尿酸亲和力最低,且亲和力与嘌呤衍生物解离成阴离子的能力相关.在生理条件下,该酶对尿酸的亲和力相对于最适条件有显著下降.据此推测,优化静电相互作用选择性增强该酶对尿酸的亲和力是提高其成药性的可能措施之一.     

高效絮凝菌的鉴定及结构研究

从活性污泥中筛选到一株高效絮凝剂产生菌RL-2。在1%CaCl2存在下,其发酵液对高岭土悬液具有较高的絮凝活性。根据形态及生理生化特征,利用16S rRNA部分序列分析确定该菌归属于Bacillus brevis。溶解性,茚三酮反应,Molish反应,电性试验及薄层色谱分析表明,RL-2所产高聚物为一阴离子型多糖,其结构单体为葡萄糖、半乳糖醛酸和甘露糖;由乌氏粘度法测得分子量大约为3.64×106Da。     

蜡状芽胞杆菌对芘的降解特性及降解酶研究

考察蜡状芽胞杆菌(Bacillus cereus)对水体中芘的降解特性,分析其代谢产物和降解酶的活性.结果表明,1 mg·L-1Mn2+、0.1 mg·L-1Fe3+和10 mg·L-1葡萄糖混合物对芘的降解有明显促进作用;1 g·L-1菌体在120 h内对2.5μmol·L-1芘的降解率达到61.4%.利用LC-MS/MS分析芘代谢产物,检测到1-萘酚、2-萘酚、9-羟基菲和1-羟基芘4种单羟基多环芳烃,表明芘在单加氧酶作用下开环降解,且B.cereus能有效分解利用4种代谢产物,其最高利用率分别为100%、90.3%、98.3%和52.7%.酶活力分析实验结果表明,B.cereus具有的水杨酸羟化酶,邻苯二酚1,2-双加氧酶和邻苯二酚2,3-双加氧酶在芘的降解中起关键作用,其酶活力经芘诱导后均有明显提高.结合产物分析及酶活测定,推断B.cereus对芘的降解途径以及降解过程是由单加氧酶和双加氧酶联合起作用.     

Application of Bacillus subtilis strain for microbial-enhanced oil recovery

In order to improve the oil recovery, injection of exogenous bacteria into the oil reservoir is one of the most widely used microbial flooding methods. In this study, a screened strain of Bacillus subtilis (B. subtilis) was introduced to perform the microbial flooding. The biosurfactants produced by B. subtilis was one kind of cyclic lipopeptides, which could reduce the surface tension of the culture solution from 68 mN/m to 25 mN/m and also decrease the interfacial tension of water/oil from 25.6 to 4.6 mN/m. Emulsification tests indicated that the strain and the biosurfactants could degrade and emulsify the crude oil. In the oil displacement experiments, oil recovery was increased by 32.4% by injecting fermentation broth into the simulated formation. By respectively performing the emulsification and oil displacement tests, it was demonstrated that the biosurfactants and degradation of the microbes in the heavy components of the crude oil are the main factors to enhance the oil recovery. Besides, the optimal cultural temperature for strain of B. subtilis was set as 40°C. Nevertheless, the strain was inappropriate for the oil displacement under acidic conditions. In addition, the hydrophilic sands and an optimal culture solution volume of 0.7 pore volume (PV) would be in favor of the oil recovery. It was further confirmed that the efficiency of microbial flooding was much higher than that of the chemical oil displacement.     

Droplet size spectra and deposits of two aerially sprayed Bacillus thuringiensis formulations on artificial samplers and live foliage

Abstract

Two commercial formulations of Bacillus thuringiensis var. kurstaki (BTK), Foray^® 48B and Thuricide^® 48LV, were applied aerially over nine spray blocks in a hardwood forest in West Virginia in 1991. Droplet spectra and spray mass deposits were determined using water‐sensitive paper strips (WSPS), glass micro‐fiber filters (GMFFs), glass plates and castor oil. Mass deposits of BTK were also assessed on natural foliage by two bioassay methods, i.e., feeding of homogenized foliage containing a starch‐sucrose solution and force‐feeding bioassay of foliar extracts containing re‐dissolved protein precipitate. Deposits on canopy foliage and ground samplers were also assessed by total protein assay and enzyme‐linked immunosorbent assay (ELISA). Droplet spectra on the WSPS were different from those on castor oil. Droplets on horizontal ground WSPS were larger than those on vertical ground WSPS. WSPS placed at canopy level collected more droplets than those at ground level. The total protein deposits (ng/cm²) were consistently higher on all blocks than the delta‐endotoxin protein deposits. Spray mass recovery on the ground samplers were low, and ranged from 2.9 to 8.0% of the applied rates.     

应用好氧生物菌剂深度处理消化污泥

为评估应用好氧生物菌剂-地衣芽孢杆菌深度处理消化污泥的可行性,比较了不同接种比（分别为2.7×10^-3,2.7×10^-2和2.7×10^-1,以总固体之比计）条件下,菌解处理过程中消化污泥液相的溶解性有机碳DOC,溶解性总氮DN,氨氮,荧光性有机物浓度和消化污泥絮体中的蛋白质,氨氮浓度,以及以模化CST值表征的脱水性能的变化,同时分析了机械破碎法和菌解法对消化污泥再次厌氧消化产气性能的影响.结果表明,需要达到较高的接种比,投加地衣芽孢杆菌才能显著促进消化污泥胞内物质溶出,大幅提高消化污泥的生物可降解性.接种比为2.7×10^-1时,菌解过程中消化污泥的液相DOC,DN,氨氮和消化污泥絮体中的氨氮含量达到最大值,分别是对照组的6.23,2.83,5.93和4.94倍;并且,消化污泥絮体的蛋白质平均降解速率最高;同时,消化污泥的产甲烷潜力优于其它接种比,是机械破碎处理后消化污泥产甲烷潜力的5.96倍.但是,在菌解处理结束后,消化污泥的模化CST值是对照组的2.69倍,说明经菌解后消化污泥的脱水性能有所劣化.     

精喹禾灵降解菌株Bacillus subtilis H的分离鉴定及降解特性

以精喹禾灵为唯一碳源,从长期受精喹禾灵污染土壤中分离筛选出一株精喹禾灵降解菌株H.通过形态观察、生理生化试验和16S rRNA基因序列分析,将菌株H鉴定为枯草芽孢杆菌（Bacillus subtilis ）.菌株H在温度为30~42℃和pH值7~9范围内,72h内对100mg/L精喹禾灵的降解率均可达95%以上.利用超高效液相色谱-质谱联用（UPLC-MS）鉴定菌株H降解精喹禾灵的产物为精喹禾灵酸.这是首次报道枯草芽孢杆菌对精喹禾灵的降解.     

苏云金芽孢杆菌抗东亚飞蝗(Locusta migratoria manilensis)的活性

采用毒力生物测定的方法,测定了600株从我国各地土壤和死虫等样品中分离的苏云金芽孢杆菌对东亚飞蝗的毒力.这些菌株几乎分布于已知的70个H血清型.在600株苏云金芽孢杆菌中,有560株(占93.4%)对东亚飞蝗无毒(死亡率小于20%),只有3株(占0.5%)对东亚飞蝗有较高毒性(死亡率大于70%).同时对筛选出的特异性菌株的杀虫活性进行了进一步研究.图2表2参8     

苏云金芽胞杆菌菌株WB9编码活性因子的基因分析

采用PCR—RFLP及PCR技术对苏云金芽胞杆菌新分离菌株WB9的ICP、VIP、几丁质酶和肠毒素4种活性因子的编码基因进行了分析．结果表明，该菌株含有编码ICP的cry1Aa、cry1Ab、cry1Cb、cry1Fa和cry1GaS个cry1基因型，编码Vip3A蛋白的vip3A基因以及编码3种肠毒素的hblA、bceT和entS基因；不含编码几丁质酶的基因．将vip3A、hblA、bceT和entS基因PCR产物回收纯化后直接测序，经在线的BLAST软件进行同源性分析，前两个基因片段与Bt已公布基因序列的相应片段同源性分别为99％和96％～98％，bceT基因片段与蜡质芽孢杆菌bceT相应片段同源性为97％．     

抗真菌多肽——捷安肽素高产菌的选育

从新疆棉株上分离得到一株细菌ZK ,经培养物性状和生理生化鉴定 ,确定该菌为芽孢杆菌属 (Bacillussp.)菌 ,其代谢产物为一种抗病原真菌的肽类物质———捷安肽素 .以此株菌作为出发菌株 ,进行紫外线、微波和亚硝基胍诱变 ,诱变处理后获得高产突变株Mv2 8,在摇瓶试验中 ,该变异株产捷安肽素活性比出发菌株提高 31.6 % .图 5表 5参 12