Biochemical characterization of a novel cycloisomaltooligosaccharide glucanotransferase from Paenibacillus sp. 598K

Cycloisomaltooligosaccharide glucanotransferase (CITase; EC 2.4.1.248), a member of the glycoside hydrolase family 66 (GH66), catalyzes the intramolecular transglucosylation of dextran to produce cycloisomaltooligosaccharides (CIs; cyclodextrans) of varying lengths. Eight CI-producing bacteria have been found; however, CITase from Bacillus circulans T-3040 (CITase-T3040) is the only CI-producing enzyme that has been characterized to date. In this study, we report the gene cloning, enzyme characterization, and analysis of essential Asp and Glu residues of a novel CITase from Paenibacillus sp. 598K (CITase-598K). The cit genes from T-3040 and 598K strains were expressed recombinantly, and the properties of Escherichia coli recombinant enzymes were compared. The two CITases exhibited high primary amino acid sequence identity (67%). The major product of CITase-598K was cycloisomaltoheptaose (CI-7), whereas that of CITase-T3040 was cycloisomaltooctaose (CI-8). Some of the properties of CITase-598K are more favorable for practical use compared with CITase-T3040, i.e., the thermal stability for CITase-598K (≤ 50 °C) was 10 °C higher than that for CITase-T3040 (≤ 40 °C); the k_cat/K_M value of CITase-598K was approximately two times higher (32.2 s^− 1 mM^− 1) than that of CITase-T3040 (17.8 s^− 1 mM^− 1). Isomaltotetraose was the smallest substrate for both CITases. When isomaltoheptaose or smaller substrates were used, a lag time was observed before the intramolecular transglucosylation reaction began. As substrate length increased, the lag time shortened. Catalytically important residues of CITase-598K were predicted to be Asp144, Asp269, and Glu341. These findings will serve as a basis for understanding the reaction mechanism and substrate recognition of GH66 enzymes.     

北京大学与中南大学神经生物学硕士研究生课程设置分析

通过对北京大学与中南大学神经生物学研究生培养方案及课程设置进行分析,探寻两所高校之间的差别。结果显示两所高校神经生物学硕士研究生课程设置基本符合培养目标,北京大学课程设置较中南大学更加全面合理,尤其在学术道德、实验技能、学术交流等方面的课程设置优于中南大学,建议中南大学新设或加强部分课程。     

1996—2008年广东省城市化进程与生态环境的协调性分析

在构建城市化和城市生态环境评价指标体系、并对广东省城市化发展水平与城市生态环境质量分别进行综合评价的基础上,将模糊数学中的隶属度引入到城市化与市生态环境协调度的计算中,并对两者协调发展程度的变化趋势进行了分析。结果发现,1996-2008年间,广东省城市综合发展与城市环境的协调度具有明显的阶段性和波动性特征,总体均呈现下降趋势;静态协调度可大体上划分为两个阶段,其中前9年均处于总体协调阶段,以1998年的静态协调度为最高,但自2005年后,两者的静态协调度逐年降低,处于不协调阶段;与静态协调发展度相比较,广东省的城市化发展与城市生态环境的动态协调度变化略为平滑。说明广东省生态环境对城市化的响应较为明显,但生态环境质量发展指数的上升滞后于城市化发展的进程。随着城市化进程的不断发展,城市化与生态环境的矛盾逐渐加剧,生态环境对城市化的约束作用逐渐增加,系统的协调性也有所下降。     

Structural behavior of RNA-binding proteins in the free state and in complex with RNA: <Emphasis Type="Italic">Escherichia coli</Emphasis> ribosomal protein L25 and 5S rRNA

A novel approach was proposed to evaluate the steadiness of polar clusters containing the RNA-binding sites on the protein surface. The degree of clustering of RNA-binding polar residues was used as a measure of the steadiness of the corresponding polar clusters. Escherichia coli ribosomal protein L25 utilizes two binding sites, S1 and S2, to complexate with a 5S rRNA fragment. The cluster distribution of RNA-contacting polar residues on the protein surface was studied using the structural data on the complex (in crystal and in solution) and the free state (in solution). The degree of polar residue clustering in S1 and S2 in crystal was estimated at 71.4 and 100%, respectively. For the free state in solution, the degree of clustering of the two sites was 22.8 and 68.6%, respectively. Thus, the steadiness was quantitatively estimated for the RNA-binding sites of two different types, one preexisting in the protein and the other induced by the RNA structure upon complexation. The difference between the protein structures in crystal and in solution was found to be functionally significant. The results can be extrapolated to numerous complexes of proteins with double-stranded RNA and DNA.     

具有一般形式饱和接触率SEIS模型的周期解

利用重合度的延拓定理,导出了具有一般形式饱和接触率SEIS模型周期解的存在性准则.     

“大范围优先”对象形成的神经关联：前颞叶

知觉对象是什么？这事看起来很简单,却是认知科学许多领域中最核心的问题,日前为止还没有公认的解答。“大范围优先”的拓扑学方法把知觉对象的定义严格地联系到拓扑变换下的不变量。知觉对象的直观概念—形状改变时其整体一致性得以保持一可以精确地被拓扑不变量,如连接组分和洞,所刻画。知觉对象的拓扑学定义已经被大量的行为学实验所验证。这些实验一致表明,虽然对象的一致性能够在非拓扑变换下得以保持,却会被拓扑变换所破坏,从而一个新的对象被感知到。特别是进一步的功能磁共振成像实验揭示,前颞页区参与拓扑知觉和知觉对象的形成,而这一脑区本来是形式视觉通路的终点。行为学上“大范围优先”的结果与视觉通路神经解剖学结果的悖逆,提示我们应该注意对象表征形成的问题和更广泛的意义上,知觉到底在何处发生的基本问题。     

Potency and property of DNA conformational damage determine ATM activation: two new DNA topological probes

We have shown previously the intercalation geometry of a series of acenaphtho [1,2-b] pyrrole derivatives with DNA double helix in vitro. In this report we chose a couple of intercalating analogues and a Chinese traditional medicine Tanshinone IIA as probes to investigate the response of DNA damage sensor ataxia-telangiectasia mutated (ATM) protein toward the DNA topological change in vivo. The two analogues (1)a (3-(4-Methyl-piperazin)-8-oxo-8H-acenaphtho [1,2-b]pyrrole -9-carbonitrile) and (3)a (3-(3-Dimethylamino-propylamino)-8-oxo-8H-acenaphtho[1,2-b]pyrrole-9- carbonitrile) could unwind double helix to different extents, whereas Tanshinone IIA could wind the double helix. Using a combination of circular dichroism (CD) studies and immunoflurescence assays, we found for the first time that the ATM protein kinase can respond to the unwinding chromatin conformational damage caused by (1)a and (3)a, while it could not be activated by the winding effects caused by Tanshinone IIA. Moreover, the amount of ATM protein phosphorylation is consistent with the degree of unwinding conformational damage. The average number of ATM foci in an MCF-7 cell is 32 +/- 1.5 at 6 microM (1)a, which is significantly higher than the 8 microM (3)a exposure (15 +/- 0.5, p < 0.5). A new couple of DNA topological probes, (1)a and (3)a have been found for the future semi-quantitative investigation of factors involved in the DNA damage pathway.     

Electronic Nose with an Air Sensor Matrix for Detecting Beef Freshness

China is one of the largest meat producing countries in the wodd. With the growing concern for food safety more attention has been paid to meat quality. The application of conventional test methods for meat quality is limited by many factors, and subjectiveness, such as longer time to prepare samples and to test. A sensor matrix was constructed with several separate air sensors, and tests were conducted to detect the freshness of the beef. The results show that the air sensors TGS2610, TGS2600, TGS2611, TGS2620 and TGS2602 made by Tianjin Figaro Electronic Co, Ltd could be used to determine the degree of freshness but TGS2442 is not suitable. This study provides a foundation for designing and making an economical and practical detector for beef freshness.     

An enzyme family reunion — similarities,differences and eccentricities in actions on <Emphasis Type="Italic">α</Emphasis>-glucans

α-Glucans in general, including starch, glycogen and their derived oligosaccharides are processed by a host of more or less closely related enzymes that represent wide diversity in structure, mechanism, specificity and biological role. Sophisticated three-dimensional structures continue to emerge hand-in-hand with the gaining of novel insight in modes of action. We are witnessing the “test of time” blending with remaining questions and new relationships for these enzymes. Information from both within and outside of ALAMY_3 Symposium will provide examples on what the family contains and outline some future directions. In 2007 a quantum leap crowned the structural biology by the glucansucrase crystal structure. This initiates the disclosure of the mystery on the organisation of the multidomain structure and the “robotics mechanism” of this group of enzymes. The central issue on architecture and domain interplay in multidomain enzymes is also relevant in connection with the recent focus on carbohydrate-binding domains as well as on surface binding sites and their long underrated potential. Other questions include, how different or similar are glycoside hydrolase families 13 and 31 and is the lid finally lifted off the disguise of the starch lyase, also belonging to family 31? Is family 57 holding back secret specificities? Will the different families be sporting new “eccentric” functions, are there new families out there, and why are crystal structures of “simple” enzymes still missing? Indeed new understanding and discovery of biological roles continuously emphasize value of the collections of enzyme models, sequences, and evolutionary trees which will also be enabling advancement in design for useful and novel applications.     

Importance of the degree of antigen polymerization by detoxification in modulating the immunogenicity of acellular pertussis vaccine

For the acellular pertussis vaccine with a high immunogenicity, the concentration, composition and characteristics of acellular pertussis antigens are the crucial points to be considered. Nevertheless, it has not been proved yet whether or not the polymerization degree, one of the characteristics of formalin-detoxified acellular pertussis antigens, has an influence on vaccine potency. Thus, in the present study, the correlations among detoxification conditions of acellular pertussis bulks, their polymerization degrees and their immunogenicities were examined. In addition, the relative importance of pertussis toxoid in vaccine immunogenicity was also investigated. Results show that a lower lysine concentration during detoxification induces highly-polymerized antigens, the immunogenicity has a great dependency on the polymerization degree of antigens, and also pertussis toxoid has a relatively stronger influence on the immunogenicity than other antigens. Accordingly, in the aspect of the potency of detoxified acellular pertussis vaccine, it can be demonstrated that the polymerization of antigens and its degree are the major factors affecting the immunogenicity along with a relatively high content of pertussis toxoid.