泽拉伦降解酶进化以提高在酸性条件下的水解效率,通过合理的设计
Xingyue Xing1, Xiaowei Chen1, Xihuo You2
1National "111" Center for Cellular Regulation and Molecular Pharmaceutics, Cooperative Innovation Center of Industrial Fermentation (Ministry of Education & Hubei Province), Key Laboratory of Fermentation Engineering (Ministry of Education), Hubei University of Technology, Wuhan 430068, PR China.
Food chemistry
|June 15, 2024
概括
研究人员设计了一种zearalenone降解酶,以改善酸性水解. 进化的酶在降解泽拉伦方面显示出显著增强的活性和有效性,特别是在酸性环境中,如猪胃.
科学领域:
- 生物技术是生物技术.
- 酶工程是什么? 酶工程是什么?
- 菌毒素降解 菌毒素降解
背景情况:
- 泽拉伦是一种普遍存在的真菌毒素,在牲畜中造成了重大经济损失.
- 在酸性条件下,当前的泽拉伦排毒方法往往缺乏有效性.
- 酶工程为开发有针对性的真菌毒素降解解决方案提供了一个有希望的策略.
研究的目的:
- 理性地设计和发展一种在酸性pH下增强水解效率的zearalenone降解酶.
- 描述进化的酶的运动性质和降解能力.
- 为了研究改善酶活性背后的分子相互作用.
主要方法:
- 理性酶设计和定向进化引入突变.
- 在酸性pH (4.2) 和37°C下进行酶活性测定.
- 动力参数的确定 (Km).
- 使用猪胃菌进行降解实验.
- 分子对接模拟. 分子对接模拟.
主要成果:
- 进化的泽拉伦降解酶在pH4.2和37°C时,其活性增加了大约5倍 (从7.69U/mg增加到38.67U/mg).
- 迈克利斯常数 (Km) 从283.61μM下降到75.33μM,表明基质结合得到改善.
- 进化的酶在模拟的猪胃中有效降解了zearalenone.
- 分子对接揭示了酶和zearalenone之间增加的键和π-sigma相互作用.
结论:
- 理性的设计和进化成功地在酸性条件下提高了zearalenone降解酶的效率.
- 进化后的酶在减轻动物料中泽拉伦污染的实际应用方面具有显著的潜力.
- 这项研究提供了对酶基质相互作用的见解,以优化酸性环境中的酶功能.
相关概念视频
Esters to Carboxylic Acids: Acid-Catalyzed Hydrolysis
2.8K
Hydrolysis of esters under acidic conditions proceeds through a nucleophilic acyl substitution. In the presence of excess water, the reaction proceeds in a reversible manner, forming carboxylic acids and alcohols.
During hydrolysis, the ester is first activated towards nucleophilic attack through the protonation of the carboxyl oxygen atom by the acid catalyst. The protonation makes the ester carbonyl carbon more electrophilic. In the next step, water acts as a nucleophile and adds to the...
During hydrolysis, the ester is first activated towards nucleophilic attack through the protonation of the carboxyl oxygen atom by the acid catalyst. The protonation makes the ester carbonyl carbon more electrophilic. In the next step, water acts as a nucleophile and adds to the...
2.8K
Catalytically 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
Reduction of Alkenes: Asymmetric Catalytic Hydrogenation
3.3K
Catalytic hydrogenation of alkenes is a transition-metal catalyzed reduction of the double bond using molecular hydrogen to give alkanes. The mode of hydrogen addition follows syn stereochemistry.
The metal catalyst used can be either heterogeneous or homogeneous. When hydrogenation of an alkene generates a chiral center, a pair of enantiomeric products is expected to form. However, an enantiomeric excess of one of the products can be facilitated using an enantioselective reaction or an...
The metal catalyst used can be either heterogeneous or homogeneous. When hydrogenation of an alkene generates a chiral center, a pair of enantiomeric products is expected to form. However, an enantiomeric excess of one of the products can be facilitated using an enantioselective reaction or an...
3.3K
Acid-Catalyzed Dehydration of Alcohols to Alkenes
19.5K
In a dehydration reaction, a hydroxyl group in an alcohol is eliminated along with the hydrogen from an adjacent carbon. Here, the products are an alkene and a molecule of water. Dehydration of alcohols is generally achieved by heating in the presence of an acid catalyst. While the dehydration of primary alcohols requires high temperatures and acid concentrations, secondary and tertiary alcohols can lose a water molecule under relatively mild conditions.
19.5K
E1 Reaction: Stereochemistry and Regiochemistry
9.4K
One of the critical aspects of the E1 reaction mechanism, as also observed in E2, is the regiochemistry, with multiple regioisomers obtained as products. In the example discussed, the presence of water as a weak base favors elimination over substitution to generate two alkenes. Given that alkenes’ stability increases with the number of alkyl groups across the double bond, typically, E1 reactions lead to the Zaitsev product, for this is more substituted and stable than the Hofmann product.
9.4K
E2 Reaction: Stereochemistry and Regiochemistry
11.5K
Elimination reactions of alkyl halides can yield one or more alkenes depending on the specific regiochemical and stereochemical considerations. While the regiochemistry of the reaction governs the location of the double bond in the product, the stereochemical requirements often influence the geometry.
When a substrate with two different β hydrogens undergoes an E2 elimination, the presence of a strong base can yield two regioisomeric alkenes. The more-substituted alkene is the major...
When a substrate with two different β hydrogens undergoes an E2 elimination, the presence of a strong base can yield two regioisomeric alkenes. The more-substituted alkene is the major...
11.5K


