一株絮凝剂产生菌的筛选及其絮凝特性研究

目的:筛选并研究对有毒物质有一定耐受性的絮凝剂产生菌。方法:利用含苯酚、邻苯二甲酸二丁酯和Pb2(SO4)3的分离培养基从土壤和活性污泥中分离筛选絮凝剂产生菌,对所得的菌种进行摇瓶发酵试验,分别考察其产絮凝剂的周期、絮凝活性分布以及对有毒物质的耐受性等特征,通过提取絮凝剂,将其絮凝活性与其它絮凝剂进行比较。结果:得到一株对苯酚具有一定耐受性的絮凝剂产生菌B2(Serratiasp.),其产絮凝剂的最佳培养时间为48h,絮凝率高于80%。苯酚浓度达0.6g/L时,B2菌的絮凝活性仍高于70%。其90%的絮凝物质集中于菌体,且热稳定性好,对多种悬浊液的絮凝活性高于硫酸铝、PAC。结论:新型絮凝剂产生菌B2对苯酚耐受性强,且絮凝剂提取简便,具有重要的研究价值。     

产微生物絮凝剂的菌株筛选、鉴定及发酵条件优化

微生物絮凝剂是一类新型的水处理制剂,因其具有高效、无毒、易降解等特点,已成为水处理剂开发的研究热点。为了扩大微生物絮凝剂的种类,本研究从活性污泥和土壤样品中筛选絮凝剂产生菌,最终获得一株具有絮凝活性的菌株,通过16S rDNA鉴定,该菌株为蒙氏假单胞菌(Pseudomonas monteilii),因此命名为P. monteilii YR-1。利用单因素实验和正交试验对该菌株的发酵条件进行了优化,其最佳碳、氮源分别为25 g/L葡萄糖和2 g/L复合氮源(酵母浸粉:尿素:硫酸铵=5:5:2),培养基初始pH值为10,培养温度为28℃,发酵时间为72 h。在最佳培养条件下,其发酵液的絮凝率从37.68%提高到63.52%。这是关于蒙氏假单胞菌产絮凝剂的首次报道。     

高效微生物絮凝剂D6对屠宰废水的应用研究

筛选出一株针对屠宰废水有高效絮凝活性的微生物絮凝剂产生菌,并对其所产微生物絮凝剂D6进行单因子絮凝条件优化和正交试验优化得到最佳絮凝条件。絮凝条件包括:微生物絮凝剂D6投加量、pH、金属离子种类、1%CaCl₂投加量。试验中发现碱性环境是微生物絮凝剂D6发挥絮凝活性的前提,表明微生物絮凝剂D6分子链的充分伸展对絮凝作用起决定因素,因此微生物絮凝剂D6的絮凝机理为吸附架桥作用。其最佳絮凝条件为微生物絮凝剂D6的投加量为25mL·L^-1,1%CaCl₂投加量55mL·L^-1,pH为8。在最佳絮凝条件下,絮凝率为96.6%,浊度去除率为97.8%,SS去除率为92.6%。     

微生物絮凝剂产生菌的筛选及其絮凝特性初步研究

目的:从海口市白沙门污水处理厂活性淤泥中分离微生物絮凝剂产生菌,并对其絮凝特性进行初步研究。方法:稀释平板法进行菌株分离,高岭土悬浊法进行菌株的筛选以及菌株产絮凝剂特性、絮凝剂活性成分和热稳定性的研究;通过形态学和生理生化试验进行菌株的初步鉴定。结果:筛选到2株具有高絮凝活性的产生菌XN-6和XN-8,分别属于乳杆菌属(Lactobacillus sp.)和不动杆菌属(Acinetobater sp.)。菌株XN-6和XN-8发酵液絮凝率最高时,菌株XN-6的最佳pH值、最适温度和最佳转速分别为7、30℃、120r/min;XN-8的最佳pH值、最适温度和最佳转速分别为8、32℃、120r/min;菌株产絮凝剂活性成分热稳定性较差。菌株XN-6和XN-8单独处理污水的絮凝率分别为81.01%和76.78%,联合处理污水的絮凝率为79.08%,基本上不存在协同作用。结论:菌株XN-6和XN-8可作为原始菌株用于开发微生物絮凝剂,有望能应用于污水的处理。     

微嗜酸寡养单胞菌EFS1的产电性能及在污水处理中的应用

【背景】微生物燃料电池(Microbial Fuel Cell,MFC)作为一种新型的燃料电池资源,在产电的同时可应用于污水处理领域,达到资源最大化的目的。【目的】从MFC中分离获得一株可培养微生物,研究其产电特性及在污水处理中的微生物絮凝、重金属耐受、苯酚降解性能,为扩展产电菌资源库提供理论基础。【方法】利用WO_3纳米探针从MFC阳极中筛选获得一株具备产电和絮凝性能的菌株,命名为EFS1。运用循环伏安分析结合扫描电镜观测阳极电极;改变外电阻测定极化曲线和功率密度曲线。测定菌株的絮凝、重金属耐受及苯酚降解性能。【结果】经16S rRNA基因序列分析,结合形态学和生理生化鉴定菌株EFS1为微嗜酸寡养单胞菌(Stenotrophomonas acidaminiphila)。菌株EFS1具有稳定的产电周期,周期电压最高可达300m V,功率密度可达56.25m W/m~2;扫描电镜发现菌株存在直接接触电极及分泌电子中介体传递电子的方式;MFC内阻为1 000Ω左右。有氧条件下菌株的絮凝率可达到70%,存在电子受体的无氧环境中可达到80%;该菌株还具有良好的Cd~(2+)、Cu~(2+)、Mn~(2+)耐受性及苯酚降解性能,在48 h、2–4 mg/L时苯酚降解率达到了100%。【结论】研究验证了产电菌EFS1具备絮凝能力、重金属耐受、苯酚降解的可能性,为产电菌的开发及污水处理方面提供理论依据。     

微生物絮凝剂产生菌的筛选及培养研究

从活性污泥中筛选出3株能产生高效絮凝活性的微生物,对其中1株TJ3进行其最佳培养条件及废水絮凝实验研究。结果表明TJ3的最佳培养条件为：以葡萄糖、蛋白胨作碳,氮源,初始pH值6～7,温度25～30℃,摇床转速120～160r/min,培养时间72h,可产生高絮凝活性的絮凝剂,对高岭土悬浊液絮凝率达92.4%,同时在最佳培养条件下,该菌所产絮凝剂对多种废水净化效果明显,尤其对净化分散兰染液,酵母菌菌液分离最为突出,絮凝率均在95%以上,具有较高的絮凝性能。     

絮凝剂产生菌培养条件的研究及在废水处理中的应用

通过菌种富集、分离、纯化,从含有大量微生物菌群的土壤和活性污泥中筛选到一株对高岭土悬浊液具有较高絮凝活性的絮凝剂产生菌,将其命名为B23.通过研究该菌株在不同培养时间的生长情况和发酵液的絮凝活性,从而得出发酵液絮凝活性与菌体生长量呈正相关.通过对该菌株培养条件的研究表明,在培养时间为24h、培养温度为30℃、培养基初始pH值为8.0,以葡萄糖为碳源、(NH4)2SO4为氮源时,发酵液絮凝活性最强.用B23菌株所产絮凝剂处理废水后,废水中CODCr的去除率为62.48%,SS的去除率为84.47%,表明该菌株有良好的实际应用前景.     

高效微生物絮凝剂产生菌TJ-3的絮凝特性分析

从活性污泥中分离筛选出一株高效絮凝剂产生菌TJ-3, 经生理生化试验检测其属于革兰氏阴性菌, 短杆状, 16S rDNA测序鉴定为铜绿假单胞菌。生长曲线表明, TJ-3的生长稳定期较长, 所产微生物絮凝剂(MBF)的稳定性良好。TJ-3产MBF对高岭土悬液具有良好的絮凝效果, 最佳条件下的絮凝率为98.2%。絮凝活性分布实验结果表明, TJ-3所产MBF的活性物质大部分存在于离心后的沉淀物中。处理100 mL高岭土悬液, pH值8.5、1%(质量分数)CaCl2溶液投加量3.5 mL、菌液投加量1.5 mL时, 絮凝效果最佳。     

生物絮凝剂高产菌株的选育

以野生菌谷氨酸棒杆菌Corynebacterium glutamicumCCTCCM 2 0 10 0 5为出发菌株 ,进行紫外和甲基磺酸乙酯逐级诱变 ,获得一株絮凝剂高产菌XMU YH1111,通过条件优化实验 ,获得突变株XMU YH1111合成生物絮凝剂的最佳发酵条件 :葡萄糖为碳源 ,尿素和酵母膏为氮源 ,培养基初始 pH 4～ 8,种子最佳种龄 16h ,接种量 5 % ,发酵罐通风量 1L/ (L·min) ,搅拌转速 10 0r/min。在此发酵条件下 ,絮凝活性最高可达到 892U/mL ,比原出发菌株CCTCCM 2 0 10 0 5的絮凝活性提高 2倍以上。     

微生物絮凝剂的研制——菌种选育、絮凝效果及提取工艺

从土壤、河泥、活性污泥中分离出 75 2株细菌 ,以发酵液对高岭土悬浮液絮凝效果中国石油化工股份有限公司科学技术研究开发资金资助项目 (No . 990 0 1 8)收稿日期 :2 0 0 0 0 4 0 3 ,修改日期 :2 0 0 0 1 2 3 0为指标 ,筛选出 1株絮凝剂产生高效菌。该菌在实验室培养条件下 ,以 0 2mL/3 0mL接种量 ,5h种龄的种子液接种 ,2 5℃、pH7摇床培养 3d可达最高絮凝活性。最佳培养基配方为 :葡萄糖 2 0g ,尿素 0 3g ,酵母膏 0 6g ,Na     

3-D models of the antigenomic ribozyme of the hepatitis delta agent with eight new contacts suggested by sequence analysis of 188 cDNA clones.

We mapped 359 mutations at 25 positions in synthetic variants of the antigenomic ribozyme of the hepatitis delta agent by analyzing the sequences of 188 cDNA clones. These data were used to identify three features of the ribozyme: highly conserved nucleotides, positions with restricted nucleotide substitutions and three-dimensional relationships between nucleotides. The distribution of mutations at the 25 positions was as follows: G-11 (the eleventh nucleotide from the cleavage site) was mutated in 56 clones; G-12 in 36; U-15 in 33; C-13 in 26; G-28 in 23; C-27 in 21; C-29 in 19; U-26 in 17; C-18 in 14; A-14 in 13; C-16 in 13; C-19 in 12; U-17 in 11; A-20 in 10; G-42 in 9; G-40 in 7; G-41 in 7; C-24 in 6; U-32 in 6; U-23 in 5; C-25 in 4; C-21 in 3; G-30 in 3; G-31 in 3; C-22 in 1. All clones containing a mutation at C-25 had an A at this position, suggesting that the extra cyclic amino group present in adenine and cytosine may function during the cleavage event. Mutations at certain positions were common in simple clones (containing only one or two mutations), while mutations at other positions were over-represented in more complex clones. Both compensatory base changes and co-mutational frequencies were used to identify eight pairs of nucleotides which may interact with each other: G-11 and C-18, G-12 and C-27, C-13 and G-28, C-21 and U-23/C-24, C-21 and G-30, U-23 and G-31/U-32, C24 and G-30, C-27 and G-42. These pairs, which involve some of the most conserved positions in the molecule, suggest interactions among nucleotides previously depicted in open-loop structures. The newly proposed points of contact between pairs of nucleotides are compatible with both the axehead and pseudoknot secondary structural models and were combined with previously proposed Watson-Crick base paired helices to produce two three dimensional models. In both of these, C-25 and C-76 are placed near the cleavage site.     

Specific detection of Plesiomonas shigelloides isolated from aquatic environments, animals and human diarrhoeal cases by PCR based on 23S rRNA gene

Twenty-five strains of Plesiomonas shigelloides isolated from aquatic environment, 10 strains from human cases of diarrhoea and five strains from animals were identified by the polymerase chain reaction technique based on 23S rRNA gene. For this purpose, two primers targeted against part of the 5' half of the 23S rRNA gene of P. shigelloides (Escherichia coli number C-912, G-1195; Plesiomonas number C-906, G-1189) were designed. Results from our study indicated that this method might serve as a tool for a rapid and sensitive identification of P. shigelloides from different environmental and clinical sources.     

天然碱碱泥分离用微生物絮凝剂产生菌的筛选

为解决内蒙天然碱泥分离的问题,从土壤、污水和活性污泥中,经初筛得到的57株产絮凝剂菌株,均有絮凝活性。并通过复筛从中筛选出絮凝活性较高、性状相对更稳定的2株菌株W23和L42。细菌的全培养液对强碱性的天然碱碱泥具有较强絮凝作用,其平均絮凝率分别达到79.80%和87.96%。     

Influence of electron transport proteins on the reactions catalyzed by Fusarium fujikuroi gibberellin monooxygenases

The multifunctional cytochrome P450 monooxygenases P450-1 and P450-2 from Fusarium fujikuroi catalyze the formation of GA14 and GA4, respectively, in the gibberellin (GA)-biosynthetic pathway. However, the activity of these enzymes is qualitatively and quantitatively different in mutants lacking the NADPH:cytochrome P450 oxidoreductase (CPR) compared to CPR-containing strains. 3beta-Hydroxylation, a major P450-1 activity in wild-type strains, was strongly decreased in the mutants relative to oxidation at C-6 and C-7, while synthesis of C19-GAs as a result of oxidative cleavage of C-20 by P450-2 was almost absent whereas the C-20 alcohol, aldehyde and carboxylic acid derivatives accumulated. Interaction of the monooxygenases with alternative electron transport proteins could account for these different product distributions. In the absence of CPR, P450-1 activities were NADH-dependent, and stimulated by cytochrome b5 or by added FAD. These properties as well as the decreased efficiency of P450-1 and P450-2 in the mutants are consistent with the participation of cytochrome b5:NADH cytochrome b5 reductase as redox partner of the gibberellin monooxygenases in the absence of CPR. We provide evidence, from either incubations of GA12 (C-20 methyl) with cultures of the mutant suspended in [18O]H2O or maintained under an atmosphere of [18O]O2:N2 (20:80), that GA15 (C-20 alcohol) and GA24 (C-20 aldehyde) are formed directly from dioxygen and not from hydrolysis of covalently enzyme-bound intermediates. Thus these partially oxidized GAs correspond to intermediates of the sequential oxidation of C-20 catalyzed by P450-2.     

Study on the flocculability of the Arthrobacter sp., an actinomycete resuscitated from the VBNC state

A bioflocculant with high flocculating activity, LC13-SF, produced by strain LC13^T which was in a viable but nonculturable (VBNC) state, and which was woken up by Rpf (resuscitation promoting factor), was systematically investigated with regard to its fermentation conditions and flocculating activity. The key parameters influencing the bioflocculant LC13-SF were investigated through measuring the optical density at 660 (OD₆₆₀) of the fermentation liquid and the optical density at 550 (OD₅₅₀) of the centrifugal supernatant. The flocculating efficiency and the Zeta potentials were chosen as the response variables for the study of the flocculating activity. The results showed that the optimal conditions for bioflocculant LC13-SF production were a fermentation time of 72 h, an initial pH of 7.0, a fermentation temperature of 30°C and a shaking speed of 150 r/min. The optimized flocculating process was as follows: a final volume percentage of bioflocculant LC13-SF and 0.5% (w/w) CaCl₂ were 1.5 and 5%, respectively in a 4 g/L Kaolin suspension, and the system pH was adjusted to 8.0. Under these conditions, the flocculating efficiency and the absolute value of the Zeta potential reached 94.83% and 4.37, respectively.     

MMP-2和TIMP-2基因启动子区多态性与卵巢上皮性癌关系的研究

为探讨MMP-2和TIMP-2基因启动子区单核苷酸多态性(SNPs)与卵巢上皮性癌发病风险的关系, 采用聚合酶链反应-限制性片段长度多态性(PCR-RFLP)方法检测了246例卵巢上皮性癌患者和324例对照妇女的MMP-2 C-1306T、C-735T和TIMP-2 G-418C 3个SNPs的基因型。结果显示, MMP-2 C-1306T SNP的等位基因及基因型频率分布在卵巢癌与对照组间无显著差异(P=0.55和P=0.42); 但卵巢癌组MMP-2 C-735T SNP的C等位基因和C/C基因型频率(80.7%和66.7%)明显高于对照组(75.5%和55.9%), 与T/T+C/T基因型比较, 携带C/C基因型可以显著增加卵巢癌的发病风险(OR=1.58, 95% CI=1.12~2.23), 进一步分层分析显示, C/C基因型主要与宫内膜样癌和年龄≥50岁妇女的发病风险显著相关, OR值分别为1.69(95%CI=1.03~2.79)和1.71(95% CI=1.14~2.57); 对MMP-2 C-1306T、C-735T 2个SNPs的单体型分析显示, 4种单体型频率(T_-1306-T_-735、T_-1306-C_-735、C_-1306-T_-735和C_-1306-C_-735)在两组间分布无显著差异(P=0.24); 虽然TIMP-2 G-418C SNP的等位基因及基因型频率在卵巢癌组与对照组间分布无显著性差异(P=0.33和P=0.47), 但以病理类型分层分析显示, 携带TIMP-2 G-418G/G基因型有增加宫内膜样癌发病风险的趋势(OR=1.62, 95%CI=0.94~2.78)。以上结果提示, MMP-2基因启动子区C-735T SNP的C/C基因型可能是卵巢上皮性癌发病的潜在危险因素, 而C-1306T SNP可能与卵巢上皮性癌的发病风险无关; TIMP-2 G-418C SNP可能与不同病理类型的卵巢上皮性癌发病风险有关。     

不同来源的尿卟啉原Ⅲ转甲基酶在脱氮假单胞菌中的表达及其对生产维生素B_(12)的影响

S-腺苷-L-甲硫氨酸依赖型尿卟啉原Ⅲ转甲基酶(S-adenosy-L-methionine uroprophyrinogen Ⅲ methyltransferase,SUMT)催化尿卟啉原Ⅲ(uroprophyrinogen Ⅲ,urogen Ⅲ)中心碳原子C-2和C-7甲基化生成前咕啉-2,是维生素B_(12)生物合成途径中的一步关键酶。本文分别克隆了荚膜红细菌来源的RCcob A1,RCcob A2和脱氮假单胞菌来源的PDcob A,并在VB_(12)生产菌株脱氮假单胞菌中过表达和发酵。通过对三株重组菌维生素B_(12)发酵结果分析可知,SUMT(PDcob A),SUMT1(RCcob A1)和SUMT2(RCcob A2)的表达有利于维生素B_(12)产量的提高,与对照菌株相比分别提高了16.48%,10.2%和31.86%。根据摇瓶发酵的结果在5 L发酵罐上进行了放大实验,p BBR122-PblaRCcob A2的产量为144.5 mg/L,相比对照菌p BBR122-Pbla(111.3 mg/L)产量提高了29.83%左右。结论:SUMT的表达可以在一定程度上解除维生素B_(12)合成途径中的瓶颈,提高维生素B_(12)产量。