Role of Cys221 and Asn116 in the zinc-binding sites of the Aeromonas hydrophila metallo-β-lactamase

The CphA metallo--lactamase produced by Aeromonas hydrophila exhibits two zinc-binding sites. Maximum activity is obtained upon binding of one zinc ion, whereas binding of the second zinc ion results in a drastic decrease in the hydrolytic activity. In this study, we analyzed the role of Asn116 and Cys221, two residues of the active site. These residues were replaced by site-directed mutagenesis and the different mutants were characterized. The C221S and C221A mutants were seriously impaired in their ability to bind the first, catalytic zinc ion and were nearly completely inactive, indicating a major role for Cys221 in the binding of the catalytic metal ion. By contrast, the binding of the second zinc ion was only slightly affected, at least for the C221S mutant. Mutation of Asn116 did not lead to a drastic decrease in the hydrolytic activity, indicating that this residue does not play a key role in the catalytic mechanism. However, the substitution of Asn116 by a Cys or His residue resulted in an approximately fivefold increase in the affinity for the second, inhibitory zinc ion. Together, these data suggested that the first zinc ion is located in the binding site involving the Cys221 and that the second zinc ion binds in the binding site involving Asn116 and, presumably, His118 and His196.Received 3 March 2003; received after revision 4 August 2003; accepted 25 August 2003     

NMR study on self-assembled cage complex of hexamethylenetetramine and cucurbit[n]urils

Self-assembled complexes between cage com-pounds cucurbit[n = 5—8]urils and hexamethylenetetramine were studied by using NMR techniques. Experimental results reveal that hexamethylenetetramine can lid cucurbit[5]uril to forming self-assembled capsules in which nothing is encap-sulated yet; the cavity of the cucurbit[7]uril can accommo-date a hexamethylenetetramine molecule to form a self- assembled host-guest inclusion. Moreover, both the cavity interaction of the cucurbit[7]uril with hexamethylenetetra-mine稨Cl and the portal interaction of the dipole carbonyl of the cucurbit[7]uril with hexamethylenetetramine稨Cl lead to form self-assembled capsules in which the hexamethylene-tetramine稨Cl are encapsulated in the hexamethylenetetra-mine稨Cl lidded cucurbit[7]uril. Although the structures of the portal and cavity to cucurbit[5]uril are similar, there is no obvious interaction between decamethylcucurbit[5]uril and hexamethylenetetramine, and also between cucurbit [6]uril or cucurbit[8]uril and hexamethylenetetramine.     

[CrMo_6O_(24)H_6]~(3-)掺杂聚苯胺材料的制备及荧光性质

在盐酸介质中,以(NH4)2S2O8为氧化剂,杂多阴离子[CrMo6O24H6]3-为掺杂剂合成出了聚苯胺掺杂材料,用FT-IR,XRD进行了表征,测定了该材料的电导率、常见溶剂中的溶解度及其荧光性质.该材料的电导率为1.4×10-2S·cm-1,最高溶解度可达到0.60mg·mL-1,在以260nm和332nm为激发波长,可分别在425nm和567nm处得到荧光发射峰。     

基于真实IPv6地址环境下的安全DNS系统设计

网络的安全保证变得日趋重要,在下一代互联网上这种要求更为迫切.以真实IPv6地址接入技术为平台,结合PK I的证书分发机制对密钥的分发实现安全便捷的管理,利用密钥机制对DNS全程数据通信进行签名加密保护进而可以实现下一代互联网络体系结构中DNS系统的安全,包括安全的域名更新、安全的域名查询以及抵御常见的DNS攻击.     

MIPv6切换延迟的优化方案

移动节点从一个接入路由器移到另一个接入路由器时会发生切换.由于链路交换延迟和IP层的相关操作使得移动节点在这个过程中不能够发送和接收数据.为减少移动IPv6切换延迟和满足时实需求,分析了快速移动IPv6和阶层移动IPv6切换协议特点,采用预先注册的方法给出了一种快速和阶层移动IPv6切换的融合方案,并详细的描述了该方案在宏观移动和微观移动情景下的原理架构结构和数据包的流程图.切换延迟分析结果,证明了方案的优越性.     

6-3-3并联机构工作空间分析与仿真

提出了6-3-3并联机构在给定姿态下工作空间分析的计算机仿真方法,该方法要点是：对给定的三维空间,不需拆解支链杆,只需使用三维搜索法遍历该空间,基于该机构逆位移解,结合滑块运动范围、球铰链半锥角最大值、奇异性条件和支链干涉动画检验,即可排除该三维空间中位姿不可能点,同时显示其中位姿可能点集——工作空间边界三维图形,作为推论还给出了使用工作空间截交面检验其内部是否存在空腔的方法。为6-3-3并联机构的设计和轨迹规划提供了基础。     

固定化优势菌降解2，6-二叔丁基酚的研究

用前期研究中获得的2,6-二叔丁基酚(简2,6-DTBP)降解菌(Aeromonas sp.),进行了对2,6-二叔丁基酚的降解性能等研究。结果表明菌株经固定化包埋后,降解底物2,6-DTBP的能力大大提高。在100.0mg/L的初始浓度下其降解率在十二天可达到81%。通过对固定化菌株的降解反应过程的动力学分析,其对底物的降解反应符合一级动力学特征,当2,6-DTBP初始浓度为100mg/L时,固定化菌种其动力学常数为0.1232,半衰期为5.63day。扫描电镜观察到菌种在海藻酸钙包埋载体中能良好地生长和繁殖。     

组合式生物脱氮除碳法处理尼龙6废水

用包含水解酸化、硝化和反硝化三个反应器的组合式生物脱氮除碳系统处理尼龙6废水,着重分析了该系统的回流比,溶解氧。结果显示系统的最佳工艺参数为:回流比为3,反应器的溶解氧分别为0.5～1.0mg·L-1、3.0～3.3mg.L-1、1.8～2.3mg·L-1、,同时CODCr和TN的去除率分别为95.85%、61.04%。     

基于IXP2400的双栈路由器的设计和实现

IPv4 over IPv6(简称4over6)机制是由清华大学提出来的一种用于解决IPv4网络通过IPv6 主干网而互通的机制.通过详细分析其在Solaris系统的实现,结合网络处理器的高速可编程特性,在IXP2400中实现了4over6机制,并且为了用户可以直接访问IPv6网络,添加了NAT-PT机制,实现了一整套的基于网络处理器的IPv4向IPv6的过渡机制.     

Max3100与AVR系列单片机的接口设计

介绍新型通用异步收发器Max3100的功能特点、引脚含义以及工作原理,给出用Max3100实现AVR系列单片机异步通讯接口设计实例.介绍通过Max3100分别将AVR的SPI接口和普通I/O接口转换为UART接口的硬件设计原理图和软件设计主要程序,对关键技术进行了说明.