草鱼铜绿假单胞菌灭活疫苗发酵培养基的优化及其在发酵罐的生长试验

为培养优质的铜绿假单胞菌Pseudomonas aeruginosa菌液,通过单因素试验对草鱼铜绿假单胞菌灭活疫苗发酵培养基的氮源、碳源和磷酸盐成分进行了筛选,采用正交试验法对培养基各主要成分的用量进行了优化组合,并经过验证试验绘制出了铜绿假单胞菌JP802在优化培养基条件下的5 L发酵罐生长曲线。结果表明:草鱼赤皮病铜绿假单胞菌JP802发酵培养基中最佳氮源为蛋白胨+牛肉膏+酵母膏,最佳碳源为葡萄糖,最佳磷酸盐为磷酸氢二钾;确立了培养基的优化配方为蛋白胨10 g/L、牛肉膏5.0 g/L、酵母膏2.5 g/L、葡萄糖5.0 g/L、磷酸氢二钾0.75 g/L、氯化钠5.0 g/L,JP802菌株在此培养基中发酵14 h菌体浓度达到最大(OD_(600 nm)值为6.44)。研究表明,通过对发酵培养基的优化,可以获得更高产量的铜绿假单胞菌JP802发酵菌液。     

用黄色隐球酵母发酵生产辅酶Q10最佳条件的选择

从黄色隐球酵母CryptococcusluteolusL3302中提取辅酶Q10,经过对氮源、碳源、初始pH、发酵温度等的研究分析,得到最佳的发酵条件。通过最优化试验,确定培养基的碳源为蔗糖和葡萄糖各1 25g/L;氮源为酵母膏和玉米浆,各0 3g/L;pH值6 5,温度28℃,接种量5%,500mL锥形瓶中的装液量为50mL;生长因子以蛋白水解液为优。按此发酵条件上罐发酵,发酵液中菌体生长量为13 4g/L,辅酶Q10的产量为18 2mg/L。     

纤维素酶基因重组乳酸菌的优化培养

试验采用逐步确定单一变量的设计,对影响重组乳酸菌生长的条件进行优化,确定其最佳生长培养基成分为:蔗糖10 g,蛋白胨10 g,牛肉膏10 g,酵母浸出物5 g,吐温80 1 m L,柠檬酸二铵2 g,CH_3COONa·3H_2O5 g,磷酸氢二钾1 g,盐液甲5.0 m L,蒸馏水1 000 m L;最佳生长环境为:pH为2.0～6.4,其中pH 3.0最佳,培养温度为37℃,培养时间为26 h,厌氧培养。     

一株沿海滩涂细菌的降油及生长特性研究

采用以原油为唯一碳源的基础培养基,从原油污染的沿海滩涂土壤中分离筛选具有降油性能的细菌;采用柴油培养基测定分离菌株对柴油的原始降解率;通过在柴油培养基中添加不同浓度的葡萄糖(0、1、2、4、8、16 g/L),及不同浓度的酵母膏、蛋白胨、尿素、硫酸铵(以氮计,浓度为0.5、1、2 g/L)和磷酸二氢钠(0、4、8、12 g/L)后,测定分离菌株的降油性能;鉴定分离菌株并研究其生长特性。结果表明:共分离到5株能以原油为唯一碳源生长的细菌,其中1株原始降油率最高(19.0%),编号为Y-3;在添加1 g/L以上葡萄糖时,Y-3菌株降油率升高,添加4 g/L葡萄糖时达最高(79.9%);酵母膏和蛋白胨可提高Y-3菌株的降油率,尿素、硫酸铵和磷酸盐对降油率影响不明显;根据形态学、生理生化鉴定以及16S rDNA序列分析,确定Y-3菌株为恶臭假单胞菌Pseudomonas putida,Y-3菌株的最适生长温度为30℃,最适生长pH为8,适宜生长NaCl浓度为0～30 g/L。     

Paracoccus sp. QD15-1发酵培养基优化

Paracoccus sp. QD15-1是一株高效酞酸酯降解菌,在酞酸酯污染修复方面具有较好的利用前,而廉价发酵基质是应用于生产实践的前提。本实验以Paracoccus sp. QD15-1为研究对象,用爬坡法来确碳源、氮源和微量元素的最佳范围,通过响应面中心合成设计法优化发酵培养基。研究结果表明,最优的碳源为玉米面粉,氮源为大豆面粉;经爬坡实验确碳源、氮源和痕量元素分别为10,20,1.5mL/L;响应面实验确最优培养基基质为玉米面粉9.5 g/L、大豆面粉18.6 g/L和痕量元素1.64 mL/L,预测Paracoccus sp. QD15-1的菌落数4.67×109 CFU/mL,实测菌落数为4.5×109 CFU/mL。相较于DMP无机盐培养基菌量提高22.22%,廉价发酵培养基大幅降低生产成本,为大规模发酵生产Paracoccus sp. QD15-1提供重要技术支持。     

微生物絮凝剂产生菌的最佳培养条件

从重庆城南污水厂的回流污泥中筛选得到一株具有稳定絮凝活性的菌株RL-2，初步鉴定为假单胞菌属。通过优化试验确定菌株RL-2产絮凝剂的最佳培养基及培养条件为：以葡萄糖为碳源、酵母膏 (NH4)2SO4为氮源，初始pH值为8，培养温度为30℃，通气量(以摇床转速表示)为120～150r／min，培养2d可获得具有高絮凝活性的生物絮凝剂。且在最佳操作条件下，该菌株对高岭土悬浊液的絮凝率达98．1％。对多种实际废水有明显的净化效果。     

产κ-卡拉胶酶菌株的筛选及其发酵条件的优化

对κ-卡拉胶酶产生菌进行富集培养,以κ-卡拉胶为唯一碳源的平板初筛,从多种海藻上分离纯化获得32株具有卡拉胶酶活性的菌株,经摇瓶复筛,从亮管藻上分离的HC4菌株酶活力最高,为50.75U/mL。测定了HC4菌株的16S rRNA基因序列1436 bp,在核苷酸序列数据库(NCBI)中进行同源性检索,发现它与Tamlana agarivorans的相似性为98%。通过单因子筛选和正交试验,确定该菌株产酶的最佳培养基组成为:碳源κ-卡拉胶5 g/L;氮源蛋白胨3 g/L和NaNO31 g/L;无机盐NaCl 20 g/L、K2HPO4.3H2O 1 g/L、MgSO4.7H2O 0.5 g/L和CaCl20.1 g/L。最佳培养条件为:摇床转速150 r/min,250 mL三角瓶中发酵培养基的装液量50 mL,接种量4%,发酵培养基起始pH7.5,温度28℃,培养时间28 h。经培养基组成及培养条件优化后,HC4菌株的κ-卡拉胶酶酶活力提高到602.30 U/mL,是优化前的11.87倍。     

利用糖蜜废水生产微生物絮凝剂及其絮凝条件优化

为了提高微生物絮凝剂的产量,增强其絮凝效果和稳定性,降低培养成本,以糖蜜废水替代葡萄糖作为培养基的碳源和能源,对荧光假单胞菌C-2进行发酵培养来制取微生物絮凝剂,并用其处理造纸中段废水,考察了最佳培养条件：结果表明,荧光假单胞菌C-2产生微生物絮凝剂的最佳培养条件为：糖蜜废水的COD为10000mg／L,培养基的初始pH值为8．0,接种量为2．5mL／50mL,培养温度为30℃,摇床转速为160r／min。在最佳培养条件下对废水中COD和浊度的去除率分别可达87．27％和94．95％,说明利用糖蜜废水取代葡萄糖作为培养基的碳源和能源来制备微生物絮凝剂是完全可行的。     

Thermobifida fusca海藻糖合成酶基因的克隆表达及发酵优化

将来源于嗜热单孢菌(Thermobifida fusca)的海藻糖合成酶基因进行克隆表达,构建重组菌E.coli BL 21(DE3){pET-24a(+)/Tres}。并对基因工程菌的发酵产酶条件进行优化,得到最优培养基成分为:甘油12 g/L,酵母粉24 g/L,蛋白胨12g/L,磷酸盐浓度60mmol/L,Zn2+2mmol/L。最佳培养条件为:装液量20 ml(250 ml摇瓶),发酵2 h后25℃诱导并添加终浓度为0.4 mmol/L的IPTG。优化后的最终发酵酶活达到28.6 U/ml,为未优化前的3.4倍,是目前国内报道大肠杆菌摇瓶发酵产海藻糖合成酶的最高表达水平,为该酶的工业化生产奠定了基础。     

重组大肠杆菌产Streptomyces sp.FA1来源木聚糖酶的摇瓶发酵优化

为了实现来源于Streptomyces sp.FA1的木聚糖酶的高效胞外分泌表达,对E.coli BL21(DE3)/pET20b(+)/coe/xynA基因工程菌的发酵产酶诱导条件进行优化,获得最优的诱导条件为25℃发酵6 h后添加终浓度为0.4 mmol/L的IPTG。在此基础上对发酵培养基进一步优化,得到最优培养基成分为:甘油11 g/L,酵母粉24 g/L,蛋白胨8 g/L,磷酸盐浓度89 mmol/L,镁离子4mmol/L。最终酶活达到780.2 U/ml,为未优化前的2.2倍,是目前大肠杆菌摇瓶发酵产木聚糖酶的最高表达水平,为实现该酶的工业化生产奠定基础。     

浅析高速公路工程招投标中的最低评标价法

结合目前高速公路施工项目招投标活动的实际情况,阐述了最低评标价法的概念,归纳了其优点及存在问题,对完善最低评标价法提出几点策略,以期促进招标投标市场更加规范有序。     

Place-Making in Megaprojects in Melbourne

This article analyses state-led place-making practices in Melbourne. The two levels of government that influence planning in Melbourne make much of the city as ‘culturally vibrant’ and ‘creative’, and have incorporated creative city-inspired place-making principles into many layers of the planning system. An examination of the development of two mixed-use megaprojects in central Melbourne reveals however that ideals of culturally engaging public places, and indeed of creative landscapes for middle-class consumption, wither in the face of more basic imperatives for economic development. In these case studies the ‘creative city’, no matter how the idea is interpreted, has little traction either as a set of inclusive place-making principles or as a gentrification strategy. The article concludes that the creative city-inspired place-making objectives in the planning system at both state and local levels are ambiguous in their overall intents and completely unsupported by statutory controls. Such place-making objectives as are realised are the compromised results of the interplay of uncoordinated decisions, delivered at the pleasure of the developer.     

Pathogens in septage in Vietnam

Septage is widely acknowledged as a major source of infectious pathogens while disposal of septage, and the operation and maintenance of septic tanks, is not regulated in many developing countries. Twenty untreated septage and septage sludge samples were taken from Can Tho City, Vietnam to examine their pathogen content, and indicator micro-organisms. Escherichia coli and Enterococcus spp. were detected in all samples, regardless of sludge storage time. Phages were detected in 80% of samples. Salmonella spp. were detected in 70% of the untreated septage and 60% of septage sludge samples. Concentrations of phages and bacteria tested in septage sludge after many years of tank storage were much higher than the expected levels. Helminth ova were present in 95% of untreated septage samples with an average of 450 ova l^− 1, and were detected in all septage sludge samples with an average of 16,000 ova l^− 1. Twelve varieties of helminth ova were identified. More helminth ova varieties in higher concentrations were found in septage than those reported from stool samples. The varieties' frequency ranged from 10% to 50% and Ascaris lumbricoides predominated. Results show that pathogens and indicator micro-organisms, especially helminth ova, accumulate in sludge. Thus helminth ova should be considered when septage sludge is treated and used for agriculture. Proper health protection measures must be applied for people handling septage.     

在市场中实践在实践中学习在学习中进步--工程公司市场化进程的回顾与展望

近两年来，工程公司在提高市场竞争意识、推动市场化进程中做了大量工作，其中坚持走出去的方针是我们自我加压、推行加快市场化步伐的一个重要战略步骤。     

新时期广东城市化的新特点

新时期广东城市化出现以下特点:知识源导向、城市由单体向群体化发展、产业集群推动、政府主导自上而下的集约化发展、城镇化向城市化转变、由重“数量”发展向重“质量”发展的转变。     

新形势下高校强化基建财务管理的探讨

在市场条件下高校基建财务人员在加强日常财务管理的同时,必须树立起高度的工作责任心,抓好事前监督、事中监督、事后监督三个环节,全面理解、贯彻执行《高等学校财务制度》,管好用好各项基本建设资金,提高基建财务管理水平,更好地完成高校各项基建任务,以有限的投资,最快的建设速度,获得最大的投资效益。     

2005年北京办公物业市场七大趋势

趋势一:CBD区域写字楼集中放量从各区域规划方案上看,CBD区域多数项目在03～04年处于项目的报批手续及拆阶段,预计2005年正式推向市场,因此2005年CBD区域将成为北京市场供应集中的域,区域即将供应的项目有北京财富中心(二期)、金地国际花园、万达广场、SOHO尚都、万通中心、世纪财富中心、怡禾国际中心、光华国际等。除去未知项目,2005CBD写字楼办公面积的供应量为60万平方米左右。中关村区域经过一段时间的集中供后开始逐步放缓,2005年供应的体量相对较大的项目有中关村金融中心、新中关。金街区域可能开盘的项目只有2个:英蓝国际中…     

浅谈我国工程项目建设成本控制存在的问题

结合目前我国施工企业工程项目成本管理现状,对工程项目成本管理过程中存在的问题进行了较详细的分析,并提出了一些解决问题的办法,以做好项目成本控制工作,进而增强企业经济实力。     

Variation in property taxation in Tulsa

Conclusions It may be reasonably concluded from this study that the range of variation in these ratios of assessed to actual value are of a reasonable size for the great majority of cases. The frequency distribution of these sample ratios was not normal, however, as was assumed a process of this type would produce. On the other hand, the failure of the regression analysis to identify any significant variables which would explain these variations would tend to re-enforce the assumption that the fluctuations were indeed of a random nature.To sum it up in one paragraph, aside from a few extreme cases, the assessing process appears to give estimates which are more consistent than would be expected under the assumption that any errors would be of a random nature, but the regression analysis was unable to isolate any particular socioeconomic characteristics which would explain the variation which was present.Financial support for this research project was furnished by the University of Tulsa.