简单芽孢杆菌产高效微生物絮凝剂

通过从绿化植物根际土壤和污水处理厂的活性污泥中分离筛选絮凝剂产生菌,得到一株稳定高效的微生物絮凝剂产生菌PS1,根据形态学特征、生理生化实验以及16S rDNA序列分析将其鉴定为简单芽孢杆菌(Bacillus simplex)。对菌株PS1产生絮凝剂的最佳培养时间、絮凝活性分布以及pH、CaCl2、絮凝剂投量对絮凝效果的影响进行了研究,并考察了其对实际废水的絮凝效果。结果表明,菌株PS1产絮凝剂的最佳培养时间为36 h,产生的絮凝活性物质全部存在于发酵液离心后的上清液中;当pH为7.0～8.5、CaC12投量为0.25～0.35 g/L、发酵液投加量的体积分数为1.5%～2.5%时,菌株PS1发酵液对4 g/L的高岭土悬浊液的絮凝效果最佳,絮凝率达到97%。菌株PS1所产絮凝剂对城市污水、啤酒废水、淀粉废水、医院废水的絮凝率可达90%以上。     

聚合物驱采出水中聚丙烯酰胺的微生物联合降解作用研究

通过对2株细菌的培养降解实验研究聚丙烯酰胺(hydrolyzed polyacrylamide,HPAM)降解菌对水环境下聚丙烯酰胺的降解作用,讨论协同降解机理。2株降解聚丙烯酰胺的菌株假单胞菌CJ419、枯草芽孢杆菌FA16在初始30℃废水样品上培养,定期测量细菌生物量和HPAM降解率。培养30 d后CJ419和FA16对聚合物的降解率最大值分别达到30.4%和25%,而以1∶1比例的混合菌降解率最大值达到80.3%。对2株菌胞外各组分研究表明:混合菌降解HPAM的机理主要由胞外降解酶系水解聚合物侧链基团导致HPAM降解为小分子物质,同时生长过程中降解菌还会释放非蛋白还原性物质引发氧化反应共同参与HPAM降解。     

Reduction of selenite to red elemental selenium by moderately halotolerant Bacillus megaterium strains isolated from Bhitarkanika mangrove soil and characterization of reduced product

Two Gram (+) bacterial strains, BSB6 and BSB12, showing resistance and potential for Se(IV) reduction among 26 moderately halotolerant isolates from the Bhitarkanika mangrove soil were characterized by biochemical and 16S rDNA sequence analyses. Both of them were strictly aerobic and able to grow in a wide range of pH (4-11), temperature (4-40 °C) and salt concentration (4-12%) having an optimum growth at 37 °C, pH ∼7.5 and 7% salt (NaCl). The biochemical characteristics and 16S rDNA sequence analysis of BSB6 and BSB12 showed the closest phylogenetic similarity with the species Bacillus megaterium. Both the strains effectively reduced Se(IV) and complete reduction of selenite (up to 0.25 mM) was achieved within 40 h. SEM with energy dispersive X-ray and TEM analyses revealed the formation of nano size spherical selenium particles in and around the bacterial cells which were also supported by the confocal micrograph study. The UV-Vis diffuse reflectance spectra and XRD of selenium precipitates revealed that the selenium particles are in the nanometric range and crystalline in nature. These bacterial strains may be exploited further for bioremediation process of Se(IV) at relatively high salt concentrations and green synthesis of selenium nanoparticles.     

The presence of Bacillus thuringiensis (Bt) protein in earthworms Eisenia fetida has no deleterious effects on their growth and reproduction

Earthworms Eisenia fetida, bred in substances with stover of two genetically-engineered Bacillus thuringiensis (Bt) corns (5422Bt1 (Event Bt11) and 5422CBCL (MON810)) expressing Cry1Ab and their near-isogenic non-Bt corn (5422), were used to investigate the non-target effects of Bt corn on soil-dwelling organisms. Cry1Ab concentrations in substances, casts and guts of E. fetida were also investigated by Enzyme-linked immunosorbent assay (ELISA). More than 90% individuals of E. fetida survived over a period of 30 d, irrespective of whether they received Bt corn or non-Bt corn. Compared to 5422 treatments, significantly higher relative growth rate and more number of new offspring and cocoons of E. fetida were found in 5422Bt1 and 5422CBCL treatments. These results were unlikely to be directly caused by Cry1Ab released from Bt corns but rather by differences in other factors of plants such as plant components (soluble sugar, total organic carbon, total protein and available phosphorus of Bt corns were more than 5422). ELISA results indicated immunoreactive Cry1Ab was detectable in substances, and the casts, guts of E. fetida from Bt corns treatments, of which the highest levels were detected in substances under the corresponding experimental conditions. With the increase of treated time, a strong decline was observed in Cry1Ab from substances and casts of E. fetida, whereas Cry1Ab in guts of E. fetida from 5422Bt1 treatments gradually increased and that from 5422CBCL treatments increased between 14 and 30 d. Therefore, the presence of Cry1Ab in E. fetida had no deleterious effects on their growth and reproduction.     

玉米浸泡液制备苏云金杆菌生物杀虫剂的影响因素研究

以玉米淀粉生产过程中的浸泡液为培养基,摇瓶发酵培养苏云金杆菌生物杀虫剂,通过一系列单因子试验,考察了不同培养条件(种子液的种龄、接种量、浸泡液的含固率、初始pH值、摇床转速、发酵温度及发酵时间)对苏云金杆菌在玉米浸泡液中的生长(菌数增长与芽孢形成)以及发酵液的生物毒效的影响.研究表明,在最佳摇瓶培养条件(种子液种龄10 h,接种量2%,浸泡液含固量3%,初始pH值7.0～7.5,摇床转速200 r/min,发酵温度30℃)下发酵48 h,活菌数和活芽孢数分别可达到7.9×108 CFU/mL和5.5×108 CFU/mL,毒力效价为698.0 IU/μL.本试验可为生物农药的工业化生产提供实用参数.     

四环素工业废水生化处理工艺的研究

本文报道了四环素生产废水絮凝处理的最佳组合是１５ｍｇ／ｋｇ聚丙烯酰胺＋３０００ｍｇ／ｋｇ硫酸亚铁；经驯化的接种水测出ＣＯＤ为２７６４ｍｇ／Ｌ时，废水的ＢＯＤ５为１８７３ｍｇ／Ｌ，ＢＯＤ５／ＣＯＤ≈６８％，属易生化处理范畴。生物膜处理可在２ｄ内将ＣＯＤ从４４８８ｍｇ／Ｌ降至５４９ｍｇ／Ｌ，处理中的优势菌群为Ｇ＾＋芽孢杆菌。经γ射线辐射后，废水对细菌不再有抑制作用。     

我国土壤中苏云金芽孢杆菌的分离与基因型的鉴定

从我国20个省市采集176份土壤中分离出苏云金芽孢杆菌51株.以cry1,cry2,cry3,cry4,cry-n,cry7,cry8,cry11,cry13和cyt的引物作PCR分析.结果表明:4株含有cry4,12株含有cry7,4株含有cry8,4株含有cry1,2株同时含有cry1和cry2,27株无PCR产物.未检测到其它基因型.经电镜扫描伴胞晶体形态多种多样,主要有菱形、球形、方形、多边形和不规则形.图3表3参15     

冰川环境耐冷菌的冷适蛋白酶分离纯化及酶学性质

采用离交和凝胶层析对“中国冰川1号”耐冷菌BacilluscereusSYPA23所产的蛋白酶进行分离纯化,并进行酶学性质研究.与中温酶相比,该蛋白酶具有较高的低温催化活力,其最适催化温度为42℃,适宜pH为7.0～8.5,SDSPAGE测定的分子量(Mr)为34.2×103.金属离子Mn2 、Ca2 对该酶有激活作用,Cu2 、Hg2 、Pb2 、Co2 对其活性有一定抑制作用.该酶属金属蛋白酶,其活性受EDTA强烈抑制,不受PMSF抑制.该蛋白酶具有低温酶典型的热不稳定性,0℃下半衰期24h,但Ca2 和一些低分子醇类物质能提高其稳定性.动力学数据分析表明,该蛋白酶在低温下对底物亲和能力高,在较宽温度范围内(25～45℃)均保持着较高的催化效能.图7表3参17     

产耐温蛋白酶苏云金芽孢杆菌FS140液体发酵条件优化

应用快速有效的数学统计方法对苏云金芽孢菌FS140耐温蛋白酶的发酵条件进行优化.首先采用二水平Plackett-Burman设计对影响产酶的8因素进行筛选,获得培养基成分中3个重要影响的因子:黄豆饼粉、酵母粉、葡萄糖;再利用响应面分析法对该3个因素进行3水平的优化,获得它们的最佳组合:黄豆饼粉1.8%、酵母粉0.36%、葡萄糖0.14%.优化后产酶水平达到837.71U mL-1,与响应面数学模型的预测值只有1.34%的误差.同时进行了发酵温度、初始pH、摇瓶装量与接种量等发酵条件的优化,FS140最终发酵产酶水平达918.91U mL-1.图2表5参14     

产木聚糖酶嗜碱细菌的筛选及产酶条件研究

利用透明圈法筛选到一株产木聚糖酶的嗜碱芽孢杆菌NT 9,该菌株产木聚糖酶为组成型。正交设计实验结果表明 ,合适的产酶条件为 :木糖 15 .0g/L ,硫酸铵 2 .5 g/L ,Tween 80 2 0 g/L ,K2 HPO4 1.0g/L ,MgSO4 ·7H2 O 0 .2g/L ,pH值 10 .0 ,3 7℃ ,2 0 0r/min ,振荡培养 72h。其木聚糖酶活力可达到 6.3 6IU/mL。