INVESTIGATION OF THE ACTIVE SITE INTEGRITY AND REACTIVITY OF PROTEINS USED IN IN

中所用蛋白质活性位点完整性和反应性的研究

• 批准号: 7722032
• 负责人: SANG-KYU LEE
• 金额: $ 0.02万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-03-01 至 2009-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7722032
• 关键词: Active Sites; Biological Warfare; Biotechnology; Calpain; Chelating Agents; Chemistry; Cobalt; Computer Retrieval of Information on Scientific Projects Database; Condition; Electronics; Environment; Enzymes; Funding; Glucose; Grant; Health; High temperature of physical object; Human; Institution; Investigation; Magnesium; Manganese; Methods; Numbers; Pesticides; Process; Proteins; Research; Research Personnel; Resources; Source; Spectrum Analysis; Structure; Sweetening Agents; System; Thermolysin; United States National Institutes of Health; VX nerve gas; Zinc; aryldialkylphosphatase; desire; glucose isomerase; interest; metalloenzyme; remediation

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. There is growing interest in the use of metalloenzymes in industrial applications, as they can often provide an efficient means of carrying out catalytic processes, while reducing the impact on human health and the environment. However, for these enzymes to be effective, the active sites must be able to withstand harsh conditions, which may include high temperature, extreme pH, and/or the presence of chelators. The integrity of the protein active sites must be maintained to preserve the desired catalytic activity. Hence the optimization of metalloenzymes for industrial use requires a means of characterizing the active site geometric and electronic structure under the required application conditions. XAS edge and EXAFS spectroscopy provides an ideal means for examining changes in the active site of biocatalysts, and hence we propose to apply these methods to a number of enzyme systems of industrial interest. Our initial studies will focus on the zinc- and calcium-dependent neutral protease (NPR) and thermolysin (THR), and their optimization for sweeteners and other industrial applications. Our XAS studies will later be extended to include the zinc-, manganese- or cobalt-dependent organophosphorus hydrolases (OPH) which have been developed for bio-remediation of pesticides and the breakdown of biological warfare agents such as VX nerve gas; and the magnesium-, cobalt- or manganese-dependent D-glucose isomerases (GI) (also used in sweetener applications). These studies should make an important contribution to industrial biotechnology and will further a sustainable chemistry approach to industrial processes.

该副本是利用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这不一定是调查员的机构。 在工业应用中使用金属酶的兴趣越来越大，因为它们通常可以提供有效的方法来进行催化过程，同时减少对人类健康和环境的影响。但是，要使这些酶有效，活动部位必须能够承受恶劣的条件，这可能包括高温，极端pH和/或螯合剂的存在。 必须保持蛋白质活性位点的完整性以保持所需的催化活性。因此，用于工业使用的金属酶优化需要一种在所需的应用条件下表征活性位点几何和电子结构的方法。 XAS Edge和EXAFS光谱学为检查生物催化剂的主动部位变化提供了理想的方法，因此我们建议将这些方法应用于许多工业兴趣的酶系统。 我们的最初研究将集中于锌和钙依赖性的中性蛋白酶（NPR）和热溶酶素（THR），以及它们对甜味剂和其他工业应用的优化。 我们的XAS研究后来将扩展到包括锌，锰或钴依赖性的有机磷水解酶（OPH），这些（OPH）是为农药的生物修复而开发的，以及对VX神经气体等生物战剂的分解；以及镁，钴或锰依赖性的D-葡萄糖异构酶（GI）（也用于甜味剂施用）。这些研究应该为工业生物技术做出重要贡献，并将进一步采取可持续的化学方法来实现工业过程。