![Quantitative SERS Detection of Uric Acid via Formation of Precise Plasmonic Nanojunctions within Aggregates of Gold Nanoparticles and Cucurbit[n]uril](/_next/image?url=https%3A%2F%2Fcloudfront.jove.com%2FCDNSource%2Fteasers%2F61682.jpg&w=3840&q=50)
针对尿酸氧化酶的高效特征,以提高其溶解度
Mohammad Reza Rahbar1, Navid Nezafat1,2, Mohammad Hossein Morowvat1,2
1Pharmaceutical Sciences Research Center, Shiraz University of Medical Sciences, Shiraz, Iran.
Applied biochemistry and biotechnology
|February 3, 2024
概括
蛋白质工程可以改善大规模生产的尿酸氧化酶 (UOX) 溶解度. 针对阳性表面补丁,特别是氨酸残留物,在实验室测试中显著提高了UOX溶解度.
科学领域:
- 生物化学 生物化学
- 蛋白质工程是指蛋白质的工程.
- 生物技术是生物技术.
背景情况:
- 蛋白质药物大规模生产面临着重要的溶解性挑战.
- 尿酸氧化酶 (UOX) 是一种同位素的无辅因子酶,具有不够的溶解性.
- 确定影响UOX溶解性的关键因素对于蛋白质工程至关重要.
研究的目的:
- 确定影响尿酸氧化酶 (UOX) 溶解度的最有效因素.
- 确定蛋白质工程的最佳目标,以提高UOX溶解度.
- 为了验证向突变对UOX溶解性的有效性.
主要方法:
- 收集并减少了来自各种生物体的同源尿酸氧化酶 (UOX) 序列的库.
- 分析了序列和结构衍生元素,以确定最多和最少可溶性UOX变体.
- 采用比较常数方法确定表面特征,如氨酸分布,作为工程目标.
- 在最不溶解的UOX中引入Arg到Ala突变,以评估可溶性增强.
主要成果:
- 发现Metabacillus fastidiosus UOX是最容易溶解的,而Agrobacterium globiformis UOX是最难溶解的.
- 发现阳性表面斑块,特别是那些富含氨酸残留物的表面斑块,对于溶解性至关重要.
- 两个Arg到Ala突变显著增加了工程化尿酸氧化酶的可溶性.
- 溶解度预测算法之间的差异凸显了多算法方法和区域表面分析的必要性.
结论:
- 阳性表面斑块,特别是富含氨酸的区域,是工程尿酸氧化酶 (UOX) 溶解度的有效目标.
- 向突变,例如Arg到Ala的替代,可以显著提高蛋白质的溶解性.
- 数据减少和比较常数方法对于设计其他生物技术产品以提高溶解度是有价值的.
更多相关视频
08:24Optimization of the Ugi Reaction Using Parallel Synthesis and Automated Liquid Handling
Published on: November 11, 2008
16.4K
12:05U2O5 Film Preparation via UO2 Deposition by Direct Current Sputtering and Successive Oxidation and Reduction with Atomic Oxygen and Atomic Hydrogen
Published on: February 21, 2019
8.1K
相关概念视频
02:44Oxidation of Alkenes: Syn Dihydroxylation with Osmium Tetraoxide
10.2K
Alkenes are converted to 1,2-diols or glycols through a process called dihydroxylation. It involves the addition of two hydroxyl groups across the double bond with two different stereochemical approaches, namely anti and syn. Dihydroxylation using osmium tetroxide progresses with syn stereochemistry.
10.2K
01:14Drug Metabolism: Phase II Reactions
3.8K
Phase II reactions are essential for the detoxification and elimination of drugs from the body. These reactions involve the conjugation of parent drugs or their phase I metabolites with endogenous molecules, resulting in more hydrophilic drug conjugates. The primary conjugation reactions in this phase are sulfation and glucuronidation. Both sulfation and glucuronidation typically produce biologically inactive metabolites. However, in some cases involving prodrugs, active metabolites may be...
3.8K
01:24Phase II Reactions: Glucuronidation
428
Glucuronidation, a pivotal phase II biotransformation process, involves the coupling of glucuronic acid to a drug or xenobiotic. Given its widespread occurrence and critical role in drug metabolism, it's considered the most crucial phase II reaction. It enhances the water solubility of substances, aiding their expulsion from the body. The driving force behind these reactions is a group of enzymes known as UDP-glucuronosyltransferases (UGTs). UGTs facilitate the transfer of a glucuronic acid...
428
01:09Colloidal precipitates
582
The high insolubility of some precipitates can result in an unfavorable relative supersaturation. This can lead to colloidal particles with a large surface-to-mass ratio, where adsorption is promoted. For instance, in the precipitation of silver chloride, silver ions are adsorbed on the surface of the colloidal particles, forming a primary layer. This layer attracts ions of opposite charge (such as nitrate ions), forming a diffuse secondary layer of adsorbed ions. This electric double layer...
582
02:50Catalysis
26.9K
The presence of a catalyst affects the rate of a chemical reaction. A catalyst is a substance that can increase the reaction rate without being consumed during the process. A basic comprehension of a catalysts’ role during chemical reactions can be understood from the concept of reaction mechanisms and energy diagrams.
26.9K
01:07Catalytically Perfect Enzymes
4.0K
The theory of catalytically perfect enzymes was first proposed by W.J. Albery and J. R. Knowles in 1976. These enzymes catalyze biochemical reactions at high-speed. Their catalytic efficiency values range from 108-109 M-1s-1. These enzymes are also called 'diffusion-controlled' as the only rate-limiting step in the catalysis is that of the substrate diffusion into the active site. Examples include triose phosphate isomerase, fumarase, and superoxide dismutase.
Most enzymes...
Most enzymes...
4.0K