一株产耐高温碱性蛋白酶菌株的筛选

通过形态观察和 16SrDNA序列同源性比较 ,将一株产耐高温碱性蛋白酶菌株GXD990 1初步鉴定为枯草芽胞杆菌 (Bacillussubtilis)。对GXD990 1菌株产蛋白酶的培养条件进行分析表明 ,最佳产酶培养基为 :麦芽糖 3%、NH4 H2 PO4 0 3%、MnCl2 0 0 5%、KCl0 0 5%、Fe2 (SO4 ) 30 0 0 1%、K2 HPO4 0 1% ,pH11 2。在 37℃培养 6 0h后 ,蛋白酶活力达到546 5U/mL发酵液     

高温大曲中细菌总DNA提取方法比较

研究了5种基于不同裂解机理的提取方法对高温大曲中总基因组提取效果的影响.结果表明,基于SDS-酶法裂解原理的提取方法获得的结果最优,此方法获得的总基因组片段大约为21 kb;纯度为:A260/A80=1.762,A260/A230=1.587;并且能不经纯化直接用于16S rDNA的扩增.为利用16S rDNA分析等后续一系列的分子生物学技术操作打好基础,使得利用先进的分子生物学技术对大曲微生物的研究成为可能.     

一株蛋白酶产生菌的分离鉴定

从郑州奶牛场土壤样品中分离到一株芽孢杆菌,命名为A—19菌株。该菌株有较高的蛋白酶活性,37℃、pH7．0培养48h后,其蛋白酶活力可达63．5U/mL。菌体生长专性需氧,有运动性,粗糙型菌落,氧化酶试验阳性,DNA（G＋C）的摩尔分数为47％。该菌株的16SrDNA序列与芽孢杆菌属有很高的同源性,Bacillus vallismortis（99．61％）,B．subtilis（98．81％）,B．amyloliquefaciens（98．95％）,B．licheniformis（97．93％）。通过其生理生化特征和16S rDNA序列相似性的系统发育分析,确定A—19菌株属于芽孢杆菌属。该菌的16S rDNA序列已被GenBank收录,序列登陆号为EU561347。     

0癌症进展Oncology Progress5R-5E0546E

ieved in the study of gynecological cancers,early diagnosis,early treatment and valid recognition of precancerous stage are still the priority of therapy.The standardization of treatment on gynecological cancers remains to be generalized and the surgeries are prone to be humanize and individuate.The technique of clinical treatment should be based on evidence-based medicine and cooperation of multi-ceneri和L.hilgardii无法加以区分,但是仍有助于将未知杆菌归入这一群。     

碱性果胶酶产生菌Bacillus claussi XJU-8的分离鉴定及酶学特性分析

从新疆石河子市一处工业污水(pH11.0)中分离到一株碱性果胶酶产生菌,编号XJU-8。应用形态学检测、生理生化实验、16S rDNA序列分析、系统进化分析对菌株进行多相鉴定。XJU-8可在pH5.0～13.0的LB培养基上生长,最适生长温度37℃。16S rDNA序列构建的系统进化树表明XJU-8与Bacillus claussi类聚在一起,而且序列同源性高达99%以上。XJU-8与B.claussi GMBAE 42在氧化酶,柠檬酸盐利用和水解Tween-40、Tween-60上有差异,且XJU-8有宽范围pH值耐受性,被认为是B.claussi的一个新品系。该菌产生的碱性果胶酶最适pH10.0,最适温度50℃,在pH9.0～12.0有较高活性和稳定性。酶活性可被Ca2+盐激活。XJU-8所产碱性果胶酶特性良好,工业应用潜能巨大。     

Lactic acid bacteria isolated from ethnic preserved meat products of the Western Himalayas

We used culture- and molecular-biology-based methods to investigate the diversity of lactic acid bacteria (LAB) in the ethnic chevon (goat) meat products chartayshya, jamma and arjia of the Western Himalayas. In six chartayshya, six jamma and four arjia samples, LAB were the predominant microbial component involved in the fermentation of these samples, and the total LAB population in arjia (7.8 ± 0.1 log cfu g⁻¹; mean ± SD) was significantly higher (P < 0.05) than in chartayshya (6.9 ± 0.1 log cfu g⁻¹) and jamma (7.5 ± 0.1 log cfu g⁻¹). We identified 53 LAB samples by 16S rRNA and phenylalanyl-tRNA synthase (pheS) genes sequencing. The LAB isolates were identified as Enterococcus durans, Enterococcus faecalis, Enterococcus faecium, Enterococcus hirae, Leuconostoc citreum, Leuconostoc mesenteroides, Pediococcus pentosaceus, and Weissella cibaria. These results revealed that there is a high level of diversity of LAB in the Himalayan ethnic preserved meat products.     

Isolation and identification of lipase producing thermophilic Geobacillus sp. SBS-4S: cloning and characterization of the lipase

A thermophilic microorganism, SBS-4S, was isolated from a hot spring located in Gilgit, Northern Areas of Pakistan. It was found to be an aerobic, gram-positive, rod-shaped, thermophilic bacterium that grew on various sugars, carboxylic acids and hydrocarbons at temperatures between 45°C and 75°C. Complete 16S rRNA gene sequence of the microorganism exhibited homology to various species of genus Geobacillus. A highest homology of 99.8% was found with Geobacillus kaustophilus. A partial (0.7 kbp) chaperonin gene sequence also showed a highest homology of 99.4% to that of G. kaustophilus whereas biochemical characteristics of the microorganism were similar to Geobacillus uzenensis. Based on biochemical characterization, 16S rRNA and chaperonin gene sequences, we identified SBS-4S as a strain of genus Geobacillus. Strain SBS-4S produced several extracellular enzymes including amylase, protease and lipase. The lipase encoding gene was cloned, expressed in Escherichia coli and the gene product was characterized. The recombinant lipase was optimally active at 60°C with stability at wide pH range (6-12). The enzyme activity was enhanced remarkably in the presence of Ca(+2). The K(m) and the V(max) for the hydrolysis of p-nitrophenyl acetate were 3.8mM and 2273 μmol min(-1)mg(-1), respectively. The ability of the recombinant enzyme to be stable at a wide pH range makes it a potential candidate for use in industry.     

Microbiota of cocoa powder with particular reference to aerobic thermoresistant spore-formers

The microbiological criteria of commercial cocoa powder are defined in guidelines instituted by the cocoa industry. Twenty-five commercial samples were collected with the aim of assessing the compliance with the microbiological quality guidelines and investigating the occurrence and properties of aerobic Thermoresistant Spores (ThrS). Seventeen samples complied with the guidelines, but one was positive for Salmonella, five for Enterobacteriaceae and two had mould levels just exceeding the maximum admissible level. The treatment of the cocoa powder suspensions from 100 °C to 170 °C for 10 min, revealed the presence of ThrS in 36% of the samples. In total 61 ThrS strains were isolated, of which the majority belonged to the Bacillus subtilis complex (65.6%).Strains resporulation and spore crops inactivation at 110 °C for 5 min showed a wide diversity of heat-resistance capacities. Amplified fragment length polymorphism analysis revealed not only a large intraspecies diversity, but also different clusters of heat-resistant spore-forming strains. The heat-resistance of spores of six B. subtilis complex strains was further examined by determination of their D and z-values.We concluded that B. subtilis complex spores, in particular those from strain M112, were the most heat-resistant and these may survive subsequent preservation treatments, being potentially problematic in food products, such as chocolate milk.     

仿刺参肠道可培养微生物多样性研究

为了解大连地区仿刺参肠道可培养微生物种群的多样性信息,用2216、TCBS和葡萄糖琼脂培养基从大连地区冬、春两季的仿刺参肠道中共分离得到141株细菌,通过16SrDNA-RFLP分析后,得到17个不同的类群。从各个类群中随机选取代表菌株进行16SrDNA序列测定和系统发育分析,结果表明,大连地区仿刺参肠道细菌主要包括假单胞菌属（Pseudomonas）,弧菌属（Vibrio）,芽孢杆菌属（Bacillus）,希瓦氏菌属（Shewanella）,交替假单胞菌属（Pseudoalteromonas）,不动杆菌属（Acinetobacter）,气球菌属（Aerococcus）,发光菌属（Photobacterium）,葡萄球菌菌属（Staphylococcus）和Kushneria菌属共10个菌属。说明大连地区仿刺参肠道可培养微生物具有一定的多样性。