硫氧化细菌的筛选及其脱硫性能研究

从含硫土壤中分离得到一株高效的硫氧化细菌A62.通过间歇性试验对菌株A62的特性进行了研究.结果表明,该菌单个细胞为杆状、革兰氏阴性,好氧菌,且当温度30℃,pH7.5,S2-≤300mg/L时,硫化物的去除率最高达92.6%,它对工业脱硫有一定的应用价值.     

包埋培养法筛选白酒窖泥中一株功能细菌

采用微包埋培养法结合流式细胞分选技术,对白酒窖泥中的细菌进行分离和筛选,分离到95株细菌。经菌株生长实验从中筛选出8株生长较缓慢的菌株LS4、LS27、LS32、LS37、LS64、LS66、LS69’、LS85,经气相色谱分析,LS66菌株产白酒香味物质较为丰富,确定其为出发菌株。对其进行形态学观察、生理生化和16S r DNA鉴定,该菌株可能为一株窖泥未培养功能菌株uncultured bacterium clone N8-7,对该菌株的培养条件进行初步研究,菌株生长的最适温度、最适p H值和最佳发酵接种量分别为35℃、p H 6.5和4%。     

絮凝-Fenton氧化处理E段漂白废水的研究

采用絮凝-Fenton氧化处理E段漂白废水。通过正交实验确定了最佳操作参数。Al2（SO4）5-Fen％on氧化配合处理E段漂白废水的最佳操作条件为：Al2（SO4）3用量0．6g／L,氧化反应的pH值为5,H2O2用量1．Og／L,FeSO4用量0．8g／L,在此条件下废水COD的去除率可达90．64％。CPAM-Fenton氧化配合处理E段漂白废水的最佳操作条件为：CPAM用量0．8mg／L,氧化反应pH值6,H202用量1．5g／L,FeSO4用量1．2g／L,在此条件下废水COD的去除率可达82．43％。     

Fenton氧化和生物法结合深度处理硫化黑印染废水

采用Fenton氧化和生物氧化结合的方法,研究硫化黑印染废水的COD去除率和处理成本。探讨了Fenton氧化的条件包括氧化时间、m(H2O2)∶m(COD)、n(H2O2)∶n(Fe2+)以及Acinetobacter sp.DS-9生物氧化法二级串联处理系统的脱硫和COD去除效果。结果表明,最佳条件为:pH=3,m(H2O2)∶m(COD)=1∶2,n(H2O2)∶n(Fe2+)=10∶1,反应90 min后,按5%的接种量接入高效硫氧化菌株Acinetobacter sp.DS-9。废水脱硫效率提高了34.5%,COD去除率提高了74%。     

烟区土壤高效氨化芽孢杆菌菌株的筛选鉴定及特性研究

为提高有机肥中有机氮的分解效率，满足烤烟生长前期氮素需求，利用氨化细菌培养基从贵州烟区土壤中分离、纯化获得了5株氨化细菌菌株，通过纳氏试剂比色法测定培养液中的氨氮浓度，5株菌株的氨氮产量在297~460 mg/L之间；筛选出氨化作用最强的2株菌株，通过形态、生理生化特征、16S rRNA基因测序及系统发育分析，鉴定AMM-2菌株为Bacillus toyonensis，AMM-5为Bacillus mobilis。氨化特性研究显示，在pH 7.0的氨化细菌培养基中，30℃条件下培养48 h，2株菌产生的NH₄⁺-N浓度分别可达514和505 mg/L，在pH 4.0~10.0和25~30℃温度范围内均表现较强的氨化能力，其中AMM-5菌株的温度适应性更广、耐盐性更高。研究获得了分解有机氮能力强的芽孢杆菌菌株，为提高有机肥的肥效提供了菌株资源。     

土壤中砷氧化细菌的分离及其培养条件研究

通过外加砷源驯化肇庆市鼎湖山自然保护区土壤中细菌,采用富集、稀释平板、硝酸盐漫过方法从土壤中分离出2株砷氧化细菌。氧化砷实验表明:As-I、As-Q能高效将As(Ⅲ)氧化为As(Ⅴ),其氧化率可达99%。培养条件研究表明,2菌株最适生长温度为30℃,As-I、As-Q最适生长pH分别为8和7。     

Fenton氧化法处理亚麻纤维加工废水的工艺研究

本实验采用了Fenton氧化法处理亚麻加工废水。考察了影响Fenton氧化处理的因素即H2O2投加量、FeSO4.7H2O投加量、pH值和反应时间。通过正交实验确定Fenton试剂处理该废水的最佳氧化条件为H2O2投加量10mL,FeSO4.7H2O投加量1.0g,pH=3,反应时间为35min。COD去除率可达70.12%,色度去除率可达81.42%,浊度去除率可高达82.96%。实验证明采用Fenton氧化实验处理该亚麻纤维加工废水具有一定的可行性。     

海南黑山羊瘤胃液中乳酸菌的分离和鉴定

以海南黑山羊瘤胃液为研究对象,对其进行乳酸菌的分离和鉴定,为黑山羊饲料的青贮提供优质的乳酸菌菌株。通过形态学观察及H2O2酶、糖发酵等生理生化试验对所分得菌株进行初步鉴定,并对分离得到的菌株进行16S r DNA基因扩增与测序。结果表明从黑山羊胃液中筛选出15株革兰氏阳性、H2O2酶阴性的乳酸菌疑似菌株,除菌株HBG20不能在p H 3.0条件下生长,其余菌株均能生长;三株代表菌株HBG11、HBG46和HBG86在低温5℃和高温55℃条件以及p H2.5时均能生长,同时在盐浓度为6.5%时也能生长。16S r DNA基因测序结果表明,菌株HBG11与耐久肠球菌(Enterococcus durans)同源性最高,菌株HBG46与植物乳杆菌(Lactobacillus plantarum)同源性最高,菌株HBG86与干酪乳杆菌(Lactobacillus casei)同源性最高。     

一株硫化物氧化细菌的分离、鉴定和脱硫效果初步验证

从无锡市某硫酸厂采集的土样中分离到一株具有硫化物氧化能力的细菌,对该细菌的16S rDNA序列进行测定,并根据16S rDNA构建了系统发育树.结果表明,TL-1与硫杆菌属有较高的同源性,与那不勒斯硫杆菌 (Thiobacillus neapolitanus) 同源性为95.3%,结合菌落、细菌形态鉴定菌株为硫杆菌属( Thiobacillus ),并命名为TL-1.同时,对TL-1的脱硫效率进行了初步研究.结果表明,在硫化物为惟一硫源且其质量浓度分别为60,120,180,240 mg/L时,4 h时硫化物去除率分别达到87.92%、71.25%、63.45%和13.31%.     

产细菌素乳酸菌的筛选与鉴定

利用溶钙圈法及牛津杯双层平板法,从欧洲萨拉米香肠、新疆奶酪、自发酵泡菜和韩式辣白菜中分离筛选产细菌素的乳酸菌,通过形态学、生理生化鉴定、16S rDNA将菌株鉴定到种,并对其细菌素基因进行克隆,确定细菌素种类。结果表明:从分离纯化到的206株乳酸菌中筛选出抑菌效果较好的菌株SA102、SA104、N105、N107、N108,在排除酸、过氧化氢等干扰因素后,菌株所产抑菌物质对蛋白酶敏感,初步判定起抑菌作用的是细菌素。16S r DNA结果表明这5株产细菌素的菌株2株为乳酸乳杆菌,3株为植物乳杆菌,通过设计引物对菌株细菌素基因进行PCR快速筛选,结果表明菌株N107、N108含有IIb类细菌素Plantaricin KJ基因编码序列,生理生化指标测试证实这些产细菌素菌株不产气、不产氨、不产硫化氢、不产粘、不产色素,具有较好的生物安全性。最后对发酵液中细菌素粗提物的酸碱稳定性和温度稳定性进行了初步探究,以期为将来细菌素的进一步实际应用提供一定的理论借鉴。     

单核增生性李斯特菌hly缺失菌株的构建及生物特性的初步鉴定

为研究单核增生性李斯特菌(Listeria monocytogenes,Lm)hly基因对毒力的影响,利用同源重组原理,使用穿梭载体,构建单核细胞增生性李斯特菌野生菌株EGD-e的hly基因缺失菌株EGD-eΔhly。细菌活性实验证明缺失菌株生长状态与亲本无差异,但EGD-eΔhly丧失溶血活性,细胞侵袭能力降低约90%,在10~7 cells/m L腹腔注射条件下不表现动物毒性。在转录水平上EGD-eΔhly的毒力基因inl C、prf A的表达量分别下降了81%和76%,act A、plc B的表达量分别提高了2.7倍和1.8倍。hly缺失菌株的成功构建及生物特性的初步研究结果为研究Lm致病机理提供依据。     

一株高浓度氨氮耐受的除氨氮菌筛选、鉴定及发酵条件优化

利用摇瓶富集培养以及平板驯化筛选的方法,从受发酵工业污水污染严重的土壤中分离筛选得到高氨氮耐受的除氨氮菌株N-2。根据形态特征及基于16S rDNA序列的系统发育分析进行菌株鉴定;以高浓度氨氮模拟废水试验验证菌株最高氨氮耐受浓度;并以氨氮去除率为评价指标,通过单因素及正交试验优化确定菌株最佳氨氮去除条件,为其工业化应用奠定理论基础。结果表明:筛选得到一株综合性能优良的N-2菌株,经鉴定为球形赖氨酸芽孢杆菌（Lysinibacillus sphaericus）;通过氮素形态测定表明该菌经细菌同化作用可在10 h内对氨氮实现快速去除,无硝酸盐氮与亚硝酸盐氮积累,且在氨氮初始浓度为6000 mg/L的模拟氨氮废水中仍能进行正常生长繁殖;优化确定最佳氨氮去除条件为C/N为10、接种量5%、pH8.0、温度31 ℃、转速180 r/min和装液量62.5 mL/250 mL;且在最优条件下对50 mg/L氨氮10 h去除率可达97.7%,对100 mg/L氨氮10 h去除率为91.0%,对500 mg/L氨氮10 h去除率为71.7%。由此可见,菌株N-2在高氨氮浓度发酵工业废水的氨氮去除具有广阔发展前景。     

Functional genomics of commercial baker's yeasts that have different abilities for sugar utilization and high-sucrose tolerance under different sugar conditions

In the modern baking industry, high-sucrose-tolerant (HS) and maltose-utilizing (LS) yeast were developed using breeding techniques and are now used commercially. Sugar utilization and high-sucrose tolerance differ significantly between HS and LS yeasts. We analysed the gene expression profiles of HS and LS yeasts under different sucrose conditions in order to determine their basic physiology. Two-way hierarchical clustering was performed to obtain the overall patterns of gene expression. The clustering clearly showed that the gene expression patterns of LS yeast differed from those of HS yeast. Quality threshold clustering was used to identify the gene clusters containing upregulated genes (cluster 1) and downregulated genes (cluster 2) under high-sucrose conditions. Clusters 1 and 2 contained numerous genes involved in carbon and nitrogen metabolism, respectively. The expression level of the genes involved in the metabolism of glycerol and trehalose, which are known to be osmoprotectants, in LS yeast was higher than that in HS yeast under sucrose concentrations of 5-40%. No clear correlation was found between the expression level of the genes involved in the biosynthesis of the osmoprotectants and the intracellular contents of the osmoprotectants. The present gene expression data were compared with data previously reported in a comprehensive analysis of a gene deletion strain collection. Welch's t-test for this comparison showed that the relative growth rates of the deletion strains whose deletion occurred in genes belonging to cluster 1 were significantly higher than the average growth rates of all deletion strains.     

氮离子注入选育高自溶干酪乳杆菌及其肽聚糖水解酶的RT-qPCR分析

通过采用低能N^+注入对干酪乳杆菌116-a进行诱变,以筛选得到高自溶的干酪乳酸菌突变体,并采用实时荧光定量PCR检测N-乙酰胞壁质酶和N-乙酰氨基葡糖胺糖苷酶在野生菌体和突变体中的表达差异。结果显示:经过离子能量30 keV N^+注入诱变,在注入剂量0.5×10^15 ions/cm^2时,筛选得到遗传稳定性良好的最大正突变菌株,其自溶度为56.3%,最大正突变菌株自溶度比野生型菌株的自溶度提高了25.7%。实时荧光定量PCR检测发现,N-乙酰氨基葡糖苷酶和N-乙酰胞壁质酶在突变体中的表达量比野生菌体中的表达量均有所上调,其中N-乙酰氨基葡糖苷酶表达量上调了约55倍,而N-乙酰胞壁质酶的表达量仅上调2.8倍。因此,N-乙酰氨基葡糖苷酶在干酪乳杆菌116-a自溶中起着重要作用,该酶表达量的上调可以有效提高菌体的自溶。     

一株SOD生产菌的分离、初步鉴定和产酶条件优化

为得到高产超氧化物歧化酶(SOD)的菌株,通过稀释平板法从土壤中进行SOD生产菌株分离和筛选,利用非变性聚丙烯酰胺凝胶电泳和氯化硝基四氮唑蓝(NBT)染色法鉴定SOD,采用抑制剂实验分析SOD类型,利用生理生化性质分析和16S rDNA亲缘关系分析进行种属鉴定,并进一步研究碳源、氮源、碳氮比、接种量和装液量对发酵产酶的影响。结果表明：SOD活性染色显示一条条带,说明只产一种SOD同工酶。该SOD对氯仿-乙醇敏感,对H2O2不敏感,为Mn-SOD。理化性质和16S rDNA序列分析初步鉴定该菌为肠杆菌科沙雷氏菌属。发酵条件优化结果表明产酶最佳条件为：最佳碳源为玉米粉,最佳氮源为蛋白胨,碳氮比4:1,接种量8%,250mL三角瓶装液量70mL。     

Evaluation of the nifH gene marker of Methanobrevibacter smithii for the detection of sewage pollution in environmental waters in Southeast Queensland, Australia

This study aimed at evaluating the host-specificity and -sensitivity of the nifH gene marker of Methanobrevibacter smithii by screening 272 fecal and wastewater samples from 11 animal species including humans in Southeast Queensland (SEQ), Australia. In addition, environmental water samples (n = 21) were collected during the dry and wet weather conditions and tested for the presence of the nifH marker along with other sewage-associated markers, namely, enterococci surface protein (esp) found in Enterococci faecium, Bacteroides HF183, adenoviruses (AVs), and polyomaviruses (PVs). The overall host-specificity of the nifH marker to differentiate between human and animal feces was 0.96 (maximum value of 1), while the overall sensitivity of this marker in human sourced feces and wastewater was 0.81 (maximum value of 1). Among the 21 environmental water samples tested, 2 (10%), 3 (14%), 12 (57%), 6 (29%), and 6 (29%) were positive for the nifH, esp, HF183, AVs and PVs markers, respectively. The prevalence of the nifH marker in environmental water samples, however, was low compared to other markers, suggesting that the use of this marker alone may not be sensitive enough to detect fecal pollution in environmental waters. The nifH marker, however, appears to be sewage-specific in SEQ, Australia, and therefore, it is recommended that this marker should be used as an additional marker in combination with the HF183 or viral markers such as AVs or PVs for accurate and sensitive detection of fecal pollution in SEQ waterways.     

酸性土壤中大豆优势根瘤菌的分离、鉴定及其生物学特性

从两个酸性土壤生态实验点（江西鹰潭、浙江金华）种植的不同大豆品种根瘤中,分离纯化出29个分离物,并对其进行了分子鉴定和生物学特性分析。从其中16个分离物中扩增出nifH基因,测序结果表明,分离纯化出的菌株主要为慢生根瘤属（Bradyrhizobium）菌株。10个分离自浙江金华的菌株,在大豆品种之间的种属差异不大。在6个分离自江西鹰潭的菌株中,耐铝和敏感的大豆品种根瘤菌株存在明显的基因型差异。通过基于nifH基因的系统发育分析选取代表性菌株,进行16S rDNA序列测定和菌株回接结瘤实验,进一步明确和验证了其种属特性。同时,对经过初筛能结瘤的4株根瘤菌进行了生长曲线和生理生化特性的测定。本文结果表明在酸性土壤中,优势根瘤菌存在地域差异性。     

类胡萝卜素高产菌Y11的鉴定及培养条件的优化

Y1 1菌株分离自高原湖泊深水层 ,生长 pH范围广 ,利用的碳、氮源种类多 ,且耐受一定的NaCl、磷酸盐和硫化物 ,经鉴定为类球红细菌 (Rhodobactersphaeroides)。通过正交实验 ,确定了优化的培养基 1 2 # 成分 :NH4 Cl 2 g ;乙酸钠 2g ;KH2 PO4 0 1 g ;MgSO4 ·7H2 O 0 0 1 g;Na2 CO30 1 g ;酵母膏 0 5 g ;无机盐 0 0 5mL ,水 1 0 0 0mL ,pH 7 0。在 1 2 # 培养基中 ,Y1 1菌株的生长量较高 ,为类胡萝卜的提取打下基础     

农用废弃物固态发酵生产衣康酸工艺研究

以土曲霉AS32811为发酵菌种,农用废弃物麸皮7.5g,玉米芯6.0g为出发培养基,在自然pH值条件下采用单因素试验研究碳源、氮源、无机盐对衣康酸产酸效果的影响。结果显示,该菌可以利用实验所选择的碳源生产衣康酸,葡萄糖为碳源时得率最高;氮源实验结果表明,NH4NO3是最适的产酸氮源。在单因素试验的基础上进行了五因素四水平的正交试验以优化产酸工艺。衣康酸生产的优化条件为:250mL三角瓶中加入玉米芯7.5g、麸皮6.0g、葡萄糖4g、营养液25mL(NH4NO3,4g/L;KH2PO4,0.1g/L;MgSO4,6g/L;C6H12O6,4g;ZnSO4.7H2O,0.015g/L),接种量为1mL(1×107孢子/mL)、温度36℃、湿度为65%条件下自然发酵72h,衣康酸产率可达63.5%。以土曲霉AS32811为发酵菌种,农用废弃物麸皮7.5g,玉米芯6.0g为出发培养基,在自然pH值条件下采用单因素试验研究碳源、氮源、无机盐对衣康酸产酸效果的影响。结果显示,该菌可以利用实验所选择的碳源生产衣康酸,葡萄糖为碳源时得率最高;氮源实验结果表明,NH4NO3是最适的产酸氮源。在单因素试验的基础上进行了五因素四水平的正交试验以优化产酸工艺。衣康酸生产的优化条件为:250mL三角瓶中加入玉米芯7.5g、麸皮6.0g、葡萄糖4g、营养液25mL(NH4NO3,4g/L;KH2PO4,0.1g/L;MgSO4,6g/L;C6H12O6,4g;ZnSO4.7H2O,0.015g/L),接种量为1mL(1×107孢子/mL)、温度36℃、湿度为65%条件下自然发酵72h,衣康酸产率可达63.5%。     

Ammonia accumulation in culture broth by the novel nitrogen-fixing bacterium, Lysobacter sp. E4

This is the first report that Lysobacter fixes nitrogen under free-living conditions, as shown by its ability to grow on nitrogen-free medium and accumulate relatively high amounts of ammonia in the culture broth. Growth of the E4 Lysobacter strain, isolated in a screen for nitrogen-fixing and ammonia-producing bacteria, resulted in higher ammonia accumulation (0.53 mM ammonium ion concentration) in media containing glucose rather than other tested carbon sources. The optimum glucose concentration was 0.30% at an initial medium pH of 7.0 and incubation temperature of 30 °C. From time-course experiments, when the glucose in the culture was exhausted, ammonia began to be accumulated, and maximum ammonia accumulation (∼ 1.60 mM) was reached after 8 days of incubation. Ammonia accumulation by this strain required molybdenum, manganese, and iron.