High-sensitivity analyses of metabolites in biological samples by capillary electrophoresis using dynamic pH junction-sweeping

Emerging fields of biochemical research, such as metabolomics, present challenges to current separation technologies because of the large number of metabolites present in a cell and their often low (submicromolar) concentration. Although capillary electrophoresis (CE) holds great promise as the method of choice for high-resolution separations of biological samples, it suffers from poor concentration sensitivity, especially with the use of UV detection. In CE, sweeping and dynamic pH junction represent two complementary on-line focusing techniques that have been used for sensitivity enhancement of hydrophobic and weakly acidic analytes, respectively. However, the application of either the sweeping or dynamic pH junction technique alone might, in some cases, be less effective for the analysis of certain sample mixtures. Recent work in the development of a hyphenated dynamic pH junction-sweeping technique is presented as an effective on-line method of preconcentration suitable for both hydrophilic (anionic) and hydrophobic (neutral) analytes. Sensitive analyses of flavin metabolites by CE with laser-induced fluorescence (LIF) detection is demonstrated in various biological matrixes, including cell extracts of Bacillus subtilis, pooled human plasma, as well as heat-deproteinized flavoenzymes. Enhanced analyte band narrowing and improved sensitivity is achieved for flavins using dynamic pH junction-sweeping compared to either sweeping or dynamic pH junction alone. This results in over a 1200-fold improvement in sensitivity relative to conventional injection methods, giving a limit of detection (LOD, defined as S/N = 3) of about 4.0 x 10(-12) M. Strategies for sensitive and more comprehensive analyses of other cell metabolites, including nucleotides, coenzymes, and steroids, are also discussed when using on-line focusing techniques in conjunction with multiplexed CE and UV detection.     

Production and purification of CGTase of alkalophylicBacillus isolated from Brazilian soil

Alkalophylic bacilli that produce cyclodextringlycosyltransferase (CGTase) were isolated from Brazilian soil, with a scheme of two plating steps. In the first step, the bacterial isolate forms a halo in the cultivation medium that contains γ-cyclodextrin (CD) complexing dyes. The CGTase of an isolate was purified 157-fold by biospecific affinity chromatography, with β-CD showing a mol wt of 77,580 Daltons. It produces a γ- to β-CD ratio of 0.156 and a small amount of α-CD, using maltodextrin 10% as substrate, at 50°C, pH 8.0 and 22 h reaction time, reaching 21.4% conversion of the substrate to cyclodextrins. In the second screening step, the isolates chosen give larger halos with β-CD complexing dyes, and smaller halos with β-CD complexing dyes, leading to a 30% improvement in γ-CD selectivity, although at lower total yield for cyclodextrins (11.5%).     

CW2M3分泌新型淀粉酶的最适条件及酶的应用条件

从原筛选的产酶菌株CW₂出发， 经多次紫外诱变、 初筛和复筛后， 获得了变异菌株CW₂M₃， 其产酶水平为原菌株的332.7％， 且具有良好的遗传稳定性. 薄层层析证明，CW₂M₃分泌的新型淀粉酶能有效地催化淀粉降解产生异麦芽低聚糖， 产物主要包括异麦芽糖、 潘糖、 异麦芽三糖等. 用正交法结合薄层层析法确定了各因素对CW₂M₃产酶水平的影响， 结果表明， 培养温度是显著因素，CW₂M₃的最适产酶条件为： 用牛肉膏培养基（牛肉膏0.7%、 蛋白胨0.7%、 NaCl 0.5%、 pH 7.0）培养， 温度40 ℃， 时间12　h. 用该酶催化淀粉转化为异麦芽低聚糖时， 酶促反应时间和温度均是显著影响因素， 酶催化淀粉降解的最适条件为: 温度65 ℃， 酶促反应1.0　h， 体系pH为7.0.     

两株芽孢杆菌对玉门油田原油的降解作用

以石油为唯一碳源，从被石油污染的玉门油田土壤中，分离、筛选出2株高效降解石油的茵株A和B，研究了它们对原油的降解能力．结果表明，经12d后，A、B对原油的降解率分别可迭61．71％和53．12％．GC-MS分析表明，饱和烃中的正构烷烃、烷基环己烷、烷基苯和二环烷均能被明显降解，芳香烃中的萘和烷基萘经降解后几乎消失，菲和烷基菲经菌A作用后发生明显降解．茵株经鉴定，二者均为芽孢杆茵．     

细菌性痢疾的菌型分布及药敏调查

目的 :了解细菌性痢疾的菌型分布及细菌对抗生素的敏感率 ,为经验性使用抗生素提供依据。方法 :对2000年1月—2003年8月我院细菌性痢疾患者做大便培养 ,对阳性菌株做药敏分析。结果 :本地菌痢以福氏占优势 ,占88.9 % ,宋内氏占11.1 %。药敏结果显示细菌对抗生素的敏感率依次是菌必治91.7 % ,头孢塞肟90.2 % ,先锋必89.9%,左氧氟沙星89.5% ,……氟派酸42 % ,氨苄青霉素35%。结论 :第三代头孢菌素对痢疾杆菌普遍敏感 ,而对既往常用的氨苄青霉素、氟派酸耐药性较高。     

酶化苏云金芽孢杆菌固态发酵培养基研究

在生物农药生产过程中，粉碎能耗占据了生产成本的10％以上。采用纤维素酶对苏云金芽孢杆菌固体发酵的培养基进行前处理，得到一种易粉碎的培养基，用该培养基发酵所得产品后处理能耗明显降低，且和原培养基发酵产品的效价相当，证明纤维素酶的加入没有影响菌种的生长。     

用中药治疗绿脓杆菌感染的急性肾盂炎的探讨

本文采用西医检查，中医治疗绿脓杆茵感染的急性肾孟炎，疗程短，疗效高，治愈了一例临床典型病例．     

Bt防治栗实象甲的试验

Bt(Bacillus thuryingieusis)已成为大规模生产和运用的生物杀虫制剂，通过试验，发现湖北康欣实业有限公司生产的生绿Bt悬浮剂在给定施用范围内稀释800倍杀灭栗实象甲效果最好。