6-Phospho-{beta}-galactosidases of Gram-positive and 6-phospho-{beta}-glucosidase B of Gram-negative bacteria: comparison of structure and function by kinetic and immunological methods and mutagenesis of the lacG gene of staphylococcus aureus

The 6-phospho-ß-galactosidase of Staphylococcus aureus,Lactococcus lactis and Lactobacillus casei and 6-phospho-ßglucosidaseB of Escherichia coli build a subfamily inside a greater enzymefamily, named the glycosal hydrolase family 1, which, hi addition,contains nine ß-glycosidases of different origins.Kinetic and immunological evidence is provided in this reportwhich strengthens the relationship of the four 6-phospho-ß-glycosidases.It is shown that the 6-phospho-ß-galactosidases and6-phospho-ß-glucosidase B are able to split aromaticß-galactoside phosphates and ß-glucosidephosphates. The turnover numbers of hydrolysis of substrateswith different epimerization at C-4 of the glycon vary up to15-fold only. Two polydonal antisera, one derived against thenative 6-phospho-ß-galactosidase from S.aureus andthe other derived against the 6-phospho-ß-glucosidaseB, cross-reacted with both enzymes. Peptides of the proteinswere separated by reverse phase HPLC. The cross-reacting peptideswere sequenced and shown to be localized at almost the sameposition in the aligned primary structures of both enzymes.An insertion of nine amino adds near these antigenic domainsis unique for the 6-phospho-ß-glycosidases and missingwithin the sequences of the ß-glycoside-specific membersof the family. The lacG gene of a 6-phospho-ß-galactosidasenegative S.aureus mutant was doned into E.coli and sequenced.In the totally inactive mutant protein only the glycine at position332 was changed to an arginine. This amino acid is part of thesequence insertion near the antigenic domain reacting with bothantisera. These data support the assumption that the regionis of great importance for the function of the enzymes and thatit is possible it determines the specificity of the phosphorylatedform of the substrates. In addition, the 6-phospho-ß-galactosidaseof S.aureus was modified by sitedirected mutagenesis of thecorresponding lacG gene hi order to replace residues Glul60and Glu375, which were suspected of being involved hi the generalacid catalysis of substrate hydrolysis, with glutamine residues.The mutant protein 160EQ retained some catalytic activity whilethe protein 375EQ was totally inactive. Glu375 is the activesite nudeophile of the 6-phospho-ß-galactosidase ofS.aureus. It is located in the sequence motif ENG where Glu358was identified as the catalytkally active nudeophile hi theß-glucosidase of Agrobacterium.     

Film morphology and reaction rate for the CVD of tungsten by the WF6—SiH4 reaction

Journal of Electronic Materials - Low pressure chemical vapor deposition (LPCVD) of tungsten (W) by SiH4 reduction of WF6 on Si(100) surfaces was studied in a single-wafer, cold-wall reactor over a...     

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6 kV变频电缆终端头的制作与安装

本文结合某铝带材热连轧机组现场安装实例，详细介绍6kV变频电缆终端头的制作与安装过程。     

Advances in titanium metal injection molding

The approaches to production of titanium powder injection molded parts are reviewed. Historically, oxygen levels have been too high for structural use (particularly with the Ti-6Al-4V alloy). However, recent advances in starting powders, binders and sintering facilities now allow oxygen levels in the Ti-6Al-4V alloy to be controlled to about 0.2 wt.% oxygen. This should result in significant expansion of the titanium PIM market place into aerospace, automobiles, surgical instruments, dentistry, communication devices (such as computers and cell phones), knives and guns. __________ Translated from Poroshkovaya Metallurgiya, Vol. 46, No. 5–6 (455), pp. 118–125, 2007.     

基于IPv6校园网的智能选课系统的J2EE架构的设计与实现

通过分析IPv6校园网自主学习模式下的自主选课问题,提出了利用J2EE技术构建基于IPv6校园网的智能选课系统的总体结构及具体实现.     

首钢6H3C单机架可逆式薄板冷轧机组技术特点

首钢6H3C单机架可逆式冷轧机组是目前世界上第一套通过中间辊交叉进行板型控制的轧机．也是世界上第一套投入工业生产的6H3C轧机。本文介绍了该机组的工艺、设备参数，技术特点，电气设备及自动化控制及生产状况，并提出了该机组可能存在的问题。     

浅谈下一代网络（NGN）

计算机网络的发展日新月异。下一代计算机网络（NGN）的探讨、研究已经成为当前的潮流。下一代网络是目前电信、互联网运营商和设备厂商都在讨论的热点技术，国内外许多网络运营商都在探讨，并正在或考虑建设试验网进行更深入的研究。本文介绍了NGN的定义、主要特点、分层结构和关键技术。