Local polarity analysis: a sensitive method that discriminates between native proteins and incorrectly folded models

The evaluation of calculated protein structures is an importantstep in the protein design cycle. Known criteria for this assessmentof proteins are the polar and apolar, accessible and buriedsurface area, electrostatic interactions and other interactionsbetween the protein atoms (e.g. HO, S-S),atomic packing, analysisof amino acid environment and surface charge distribution. Weshow that a powerful test of accuracy of protein structure canbe derived by analysing the water contact of atoms and additionallytaking into account their polarity. On the basis of estimatedreference values of the polar fraction of typical globular proteinswith known structure (mean, SD and distribution), the evaluationof misfolded structures can be improved significantly. The referencevalues are derived by moving windows of different length (3–99amino acid residues) over the amino acid sequence. Model proteins,which are included in the Brookhaven protein structure databank,deliberately misfolded proteins, hypothetical proteins and predictedprotein structures are diagnosed as at least partially incorrectlyfolded. The local fault, mostly observed, is that polar groupsare buried too frequently in the interior of the protein. Thedatabase-derived quantities are useful in screening the designedproteins prior to experimentation and may also be useful inthe assessment of errors in the experimentally determined proteinstructures.     

芝麻渣中蛋白质的提取与纯化

采用超声波—微波协同提取法提取芝麻渣中蛋白质,并用超滤法进行纯化.实验考察了影响粗蛋白提取率的固液比、溶液pH值、微波功率、提取时间等因素,确定了最佳提取条件;同时考察了超滤膜的截留分子质量以及压力对超滤效果的影响.结果表明：超声波—微波协同提取最佳条件为超声波功率40 W,固液比1∶20,溶液pH11,微波功率250 W、提取时间90 s,提取率约55.32%;采用10万截留分子质量的超滤膜在0.18 MPa压力下对芝麻渣蛋白质纯化后,蛋白质纯度提高了15.67%.     

麻辣休闲食品的研制

对双螺杆蒸煮挤压机挤压面类麻辣休闲食品配方进行了研究。实验以小麦粉和大豆分离蛋白为蒸煮挤压产品的主要原料,利用正交设计方法将休闲麻辣食品的配方优化为：辣椒的添加量0.8%、孜然的添加量0.7%、麻椒的添加量为0.4%。     

Citrate-stabilized CdSe/CdS quantum dots as fluorescence probe for protein determination

A rapid, ultrasensitive and convenient fluorescence measurement technology based on the enhancement of the fluorescence intensity resulting from the interaction of functionalized CdSe/CdS quantum dots (QDs) with bovine serum albumin (BSA) was proposed. The citrate-stabilized CdSe/CdS (QDs) were synthesized by using Se powder and Na₂S as precursors instead of any pyrophoric organometallic precursors. The modified CdSe/CdS QDs are brighter and more stable against photobleaching in comparison with organic fluorophores. At pH 7.0, the fluorescence signal of CdSe/CdS is enhanced by increasing the concentration of BSA in the range of 0.1–10 μg/mL, and the low detection limit is 0.06 μg/mL. A linear relationship between the enhanced fluorescence peak intensity (ΔF) and BSA concentration (c) is established using equation ΔF=50.7c+16.4 (R=0.996 36). Results of determination for BSA in three synthetic samples are identical with the true values, and the recovery (98.9%–102.4%) and relative standard deviation (RSD, 1.8%–2.5%) are satisfactory.     

Application of ACO algorithm in protein structure prediction

The hydrophobic-polar (HP) lattice model is an important simplified model for studying protein folding. In this paper, we present an improved ACO algorithm for the protein structure prediction. In the algorithm, the "lone"ethod is applied to deal with the infeasible structures, and the "oint mutation and reconstruction"ethod is applied in local search phase. The empirical results show that the presented method is feasible and effective to solve the problem of protein structure prediction, and notable improvements in CPU time are obtained.     

Expression of green fluoscrescent protein gene in Sclerotinia sclerotiorum

Protoplasts of the pathogenic plant fungus,Sclerotinia sclerotiorum,were transformed using the pPGF plasmid,which contains green fluorescent protein gene,under the control of Aspergillus nidulans regulatory sequences. The pPGF plasmid was introduced by PEG/CaCl2 treatment. Positive transformants were harvested with hygromycin B (HYG) resistance as selective marker,and then were observed with green fluorescence phenomena in response to blue light,which suggested that GFP gene was cloned into genome DNA of S. sclerotiorum. The transformants were verified mitotically stable by Southern blotting analysis and passage culturing. This study is developed as an initial step for further research into infection mechanisms of S. sclerotiorum to plants and interactions with bio-control fungus.     

肽类消化吸收方式与机制研究进展

最新研究表明,寡肽可以被小肠直接吸收,但其吸收方式是与氨基酸不同的、相互独立的转运机制。寡肽的特殊载体,通过氢离子和钙离子浓度电导的消耗能量的主动转运系统、具pH依赖性的非耗能性钠离子和氢离子交换系统和谷胱甘肽（GSH）等三种系统进行转运。食物蛋白质的品质、肽链长度与肽的氨基酸组成、生理状况等因素都会影响寡肽的消化吸收。     

一株微生物采油菌的16S rDNA鉴定和绿色荧光蛋白分子标记

筛选到一株高效生产多糖的YX菌,油田先导试验证明该菌可以用于微生物调剖提高原油采收率（Microbial Enhanced Oil Recovery,MEOR）.经16SrDNA分析鉴定,YX菌属于肠内杆菌属（Enterobacter.）,为跟踪监测其动态信息,构建表达绿色荧光蛋白（green fluorescent protein,GFP）的重组质粒pET-30a（＋）-EGFP,利用CaCl2法将其转化至YX菌,利用荧光显微镜可以观察到重组YX（GFP）工程菌的绿色荧光.结果表明YX菌可以被绿色荧光蛋白基因标记,用于微生物采油研究.     

低温豆粕中磷脂对蛋白提取率的影响

通过3次酸沉的方法从大豆蛋白提取液中分离出7S、11S及磷脂膜蛋白组分,对比了两种低温豆粕所制得的蛋白产品各组分中脂质的含量,分析了磷脂在低温豆粕中的残留情况以及磷脂对低温豆粕中蛋白溶解性的影响,认为残留的磷脂对低温豆粕中蛋白提取是不利的.     

碱性蛋白酶水解豌豆蛋白的条件优化

采用碱性蛋白酶对豌豆蛋白进行水解,通过单因素试验,分析了温度、pH、酶与底物浓度比、底物浓度、水解时间对水解度影响,并在单因素试验的基础上,利用响应面分析法,优化确定的豌豆蛋白酶水解最佳条件为:温度54.5℃,pH7.9,酶与底物浓度比1.5%,底物浓度2.9%,水解时间3.0 h,最佳酶水解条件下的水解度为13.42%.