Production of cocoa butter-like fat from interesterification of vegetable oils

Cocoa butter-like fat was prepared from completely hydrogenated cottonseed and olive oils by enzymatic interesterification. The optimum reaction time to produce the major-component of cocoa butter, 1(3)-palmitoyl-3(1)-stearoyl-2-monoolein (POS), was 4 hr. The cocoa butter-like fat was isolated from the reaction mixture by two filtration steps. The yield of cocoa butter-like fat was 19%, based on the weight of the original oils. Chromatographic analysis of the product by reversephase high-performance liquid chromatography (HPLC) has shown it contains triglyceride components similar to those of cocoa butter, but that it has slightly more diglycerides. The melting point of this product, as measured by a differential scanning calorimeter, is 39°C, which compares well to the 36°C melting point of natural cocoa butter. Presented in part at the AOCS meeting in Cincinnati, Ohio, in 1989.     

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.     

A predicted three-dimensional structure of human cytochrome P450: implications for substrate specificity

A three-dimensional structure for human cytochrome P450IA1 waspredicted based on the crystal coordinates of cytochrome P450camfrom Pseudomonas putida. As there was only 15% residue identitybetween the two enzymes, additional information was used toestablish an accurate sequence alignment that is a prerequisitefor model building. Twelve representative eukaryotic sequenceswere aligned and a net prediction of secondary structure wasmatched against the known -helices and ß-sheets ofP450cam. The cam secondary structure provided a fixed main-chainframework onto which loops of appropriate length from the humanP450IA1 structure were added. The model-built structure of thehuman cytochrome conformed to the requirements for the segregationof polar and nonpolar residues between the core and the surface.The first 44 residues of human cytochrome P450 could not bebuilt into the model and sequence analysis suggested that residues1–26 formed a single membrane-spanning segment. Examinationof the sequences of cytochrome P450s from distinct gene familiessuggested specific residues that could account for the differencesin substrate specificity. A major substrate for P450IA1, 3-methyl-cholanthrene,was fitted into the proposed active site and this planar aromaticmolecule could be accommodated into the available cavity. Residuesthat are likely to interact with the haem were identified. Thesequence similarity between 59 eukaryotic enzymes was representedas a dendrogram that in general clustered according to genefamily. Until a crystallographic structure is available, thismodel-building study identifies potential residues in cytochromeP450s important in the function of these enzymes and these residuesare candidates for site-directed mutagenesis.     

Periplaneta americana perception of phytochemical naphthoquinones as allelochemicals

Electrochemistry of malePeriplaneta americana perception of naphthoquinones as allelochemicals is presented. Importance of the oxidation-reduction potential and hydrogen-bonding capabilities of the naphtho-quinone messenger to its exchange of energy with the information-encoding, redox protein of the chemosensory neuron is considered. Evidence that the required information is initially encoded by an electroreactive protein in dendritic membrane is analyzed. A sulfhydryl-disulfide protein-based electrochemical mechanism for the demonstrated linear conversion of a molecular messenger energy state into whole insect behavior (avoidance or feeding inhibition) is elucidated. Even though the information-encoding mechanism is genetically conserved (kept electrochemically similar) within the species, behavioral biotypical differences regarding sensitivity to naphthoquinones are allowed through a group of proteins in the outer aqueous medium in the chemosensory sensillum. Such proteins electrochemically interface the dendritic membrane of the chemosensory neuron in the sensillum with the environment external to the sensillum. These interfacing proteins bind (complex) with chemical messengers (e.g., naphthoquinones) and/or variously degrade them, and thus determine the amount (moles) that is required in the environment to result in stimulation of the underlying sensory neurons and in a changed behavior.     

Rapid detection of antigen binding by antibody fragments expressed in the periplasm of Escherichia coli

Bacterial expression systems can greatly facilitate proteinengineering of antibodies. We have developed a system for high-levelexpression of antibodies, antibody fragments, or hybrid antibodieswith novel effector functions in the periplasm of Escherichiacoli. From 5 ml of cells, a simple extraction yields sufficientmaterial for SDS-gel electro-phoresis, detection and characterizationof hapten binding. To demonstrate our system, heavy-chain variableregions and 1 light chains of a mouse anti-NP antibody weresynthesized as hybrid proteins with a bacterial signal peptide(Omp F). Each chain is secreted into the periplasm where processing(cleavage of the signal peptide), folding and heterodimer associationtake place. Periplasmic proteins are released by cold osmoticshock, and hapten-binding activity is easily detected withoutfurther manipulation. The ease of genetic engineering in thissystem will facilitate the production of immunoglobulin derivativesdesigned for specific applications, and expression of thesemolecules in a native state will allow the rapid screening ofcombinatorial libraries and the results of mutagenesis.     

基于图嵌入与拓扑结构信息的蛋白质复合物识别算法

蛋白质复合物是细胞结构和生化机制的研究基础,如何准确识别蛋白质复合物成为近年来的研究热点。针对传统算法根据结构信息对蛋白质复合物进行搜索存在敏感度和F-measure低的问题,以及现有监督学习算法根据人为构造特征进行蛋白质复合物识别存在特征构造不能较好地反映图的真实信息等不足,提出了graph2vec-SVM识别算法。将蛋白质复合物看作稠密子图并考虑子图模块度大小,利用graph2vec将图信息转换为向量,并进一步采用SVM分类器对蛋白质复合物进行识别,提高了蛋白质复合物识别的敏感度和F-measure。该算法分别与目前流行的4种非监督学习算法(ClusterOne、CMC、HC-PIN和COACH)和3种监督学习算法(SCI-BN、SCI-SVM和RM)进行比较,在精准度、敏感度和F-measure 3项指标上都显示出了良好的性能。     

Protein interaction networks: centrality,modularity, dynamics,and applications

In the post-genomic era, proteomics has achieved significant theoretical and practical advances with the development of high-throughput technologies. Especially the rapid accumulation of protein-protein interactions (PPIs) provides a foundation for constructing protein interaction networks (PINs), which can furnish a new perspective for understanding cellular organizations, processes, and functions at network level. In this paper, we present a comprehensive survey on three main characteristics of PINs: centrality, modularity, and dynamics. 1) Different centrality measures, which are used to calculate the importance of proteins, are summarized based on the structural characteristics of PINs or on the basis of its integrated biological information; 2) Different modularity definitions and various clustering algorithms for predicting protein complexes or identifying functional modules are introduced; 3) The dynamics of proteins, PPIs and sub-networks are discussed, respectively. Finally, the main applications of PINs in the complex diseases are reviewed, and the challenges and future research directions are also discussed.     

基于加权网络和局部适应度的蛋白质复合物识别算法

蛋白质复合物识别对分析蛋白质网络的结构特征和模块功能具有重要意义。通常在蛋白质网络中挖掘稠密子图或模块来识别其中的蛋白质复合物,限制了其应用范围和识别的准确性。针对该问题,提出了一种基于加权网络和局部适应度的蛋白质复合物识别算法,该算法综合稠密子图的密度指标和模块性定义了新的局部适应度函数,并基于边聚集系数构建加权的蛋白质网络,根据权值选择边,在加权蛋白质网络中将种子边不断聚类扩展,从而获取具有最大综合适应度的子图作为蛋白质复合物。在酵母蛋白质等多个实际网络中试验表明,该算法能够有效提升蛋白质复合物识别的准确性。     

Continuum electrostatic approach for evaluating positions and interactions of proteins in a bilayer membrane

Orientations of proteins in the membranes are crucial to their function and stability. Unfortunately the exact positions of these proteins in the lipid bilayer are mostly undetermined. Here, the spatial orientation of membrane proteins within the lipid membrane was evaluated using a Poisson–Boltzmann solvent continuum approach to calculate the electrostatic free energy of the protein solvation at various orientations in an implicit bilayer. The solvation energy was obtained by computing the difference in electrostatic energies of the protein in water and in lipid/water environments, treating each as an implicit solvent model. The optimal position of transmembrane proteins (TMP) in a lipid bilayer is identified by the minimum in the “downhill” pathway of the solvation energy landscape. The energy landscape pattern was considerably conserved in various TMP classes. Evaluation of the position of 1060 membrane proteins from the orientations of proteins in membranes (OPM) database revealed that most of the polytopic and β-barrel proteins were in good agreement with those of the OPM database. The study provides a useful scheme for estimating the membrane solvation energy made by lipid-exposed amino acids in membrane proteins. In addition, our results tested with the bacterial potassium channel model demonstrated the potential usefulness of the approach in assessing the quality of membrane protein models. The present approach should be applicable for constructing transmembrane proteins–lipid configuration suitable for membrane protein simulations and will have utility for the structural modeling of membrane proteins.