化酸酶的结构导向工程:重新塑造基质接入道和结合口袋,以高效合成金纳米德
Dong Ma1, Zhongyi Cheng1, Laichuang Han1
1Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, Jiangsu, China.
International journal of biological macromolecules
|November 2, 2023
概括
工程化烯酸酸酶 (NHase) 变体对像肉酸这样的大型烯酸具有显著增强的活性. 这一突破改善了从具有挑战性的基板中工业生产有价值的胺.
科学领域:
- 生物催化和酶工程 生物催化和酶工程
- 工业生物技术 工业生物技术
- 蛋白质化学 蛋白质化学
背景情况:
- 肉胺衍生物是重要的天然产品构建块.
- 酸盐酸酶 (NHase) 对于工业胺的生产至关重要,但与巨大的酸盐如肉酸盐抗争.
研究的目的:
- 为了设计一个酸酶 (NHase),提高了对大型酸基质的催化效率.
- 调查增强NHase活动的结构和动态基础.
主要方法:
- Pseudonocardia热NHase (PtNHase) 的位点定向突变产生 βF37P/L48P/F51N 变异体.
- 酶活性测定,结构分析,分子动力学模拟和DCCM分析.
主要成果:
- 与野生型PtNHase.相比,βF37P/L48P/F51N变体显示了肉二的催化效率增加了14.88倍.
- 突变扩大了基质接入道和结合口袋,增强了酶动态和产品释放.
- 完成了 cinnamonitrile 到 cinnamamide 的完全转化,获得了高产量和生产率.
结论:
- 同时发展基质接入道和绑定口袋是NHase工程的有效策略.
- 工程NHases显示出从重的烯酸中大规模工业化生产胺的巨大潜力.
相关概念视频
Preparation of Nitriles
2.1K
One of the common methods to prepare nitriles is the dehydration of amides. This method requires strong dehydrating agents like phosphorous pentoxide or boiling acetic anhydride for converting amides to nitriles. Another reagent namely, thionyl chloride also accomplishes the dehydration of amides, where amide acts as a nucleophile. The first step of the mechanism involves the nucleophilic attack by the amide on the thionyl chloride to form an intermediate. In the next step, the electron pairs...
2.1K
Nitriles to Carboxylic Acids: Hydrolysis
3.9K
Nitriles undergo acid-catalyzed hydrolysis or base-catalyzed hydrolysis to form a carboxylic acid. These reactions proceed via an amide intermediate.
3.9K
Preparation of Carboxylic Acids: Hydrolysis of Nitriles
4.3K
Nitriles (R–CN) can be converted into carboxylic acids (R–COOH) upon treatment with aqueous acids, i.e., upon hydrolysis of nitriles. Under base-catalyzed conditions, carboxylate anions (R–COO−) are formed.
4.3K
Preparation of Amines: Reduction of Amides and Nitriles
2.4K
Nitriles can be reduced to primary amines using reducing agents like lithium aluminum hydride or catalytic hydrogenation. The reduction introduces an amino group with an extra carbon in the skeleton. Nitriles are formed from the reaction between alkyl halides and sodium cyanide through the SN2 mechanism. Primary alkyl halides are the preferred substrates to prepare nitriles.
Amides can be reduced to primary, secondary, and tertiary amines using catalytic hydrogenation, active metals like Fe,...
Amides can be reduced to primary, secondary, and tertiary amines using catalytic hydrogenation, active metals like Fe,...
2.4K
Preparation of Amines: Reduction of Oximes and Nitro Compounds
3.7K
Oximes can be reduced to primary amines using catalytic hydrogenation, hydride reduction, or sodium metal reduction. The reduction of aliphatic and aromatic nitro compounds to primary amines takes place by either catalytic hydrogenation or by using active metals like Fe, Zn, and Sn in the presence of an acid.
Though catalytic hydrogenation can reduce nitrobenzenes, the reduction is nonselective in the presence of other functional groups. For instance, if nitrobenzene contains an aldehyde group,...
Though catalytic hydrogenation can reduce nitrobenzenes, the reduction is nonselective in the presence of other functional groups. For instance, if nitrobenzene contains an aldehyde group,...
3.7K
Preparation of Amides
3.0K
Amides are synthesized by treating carboxylic acids with amines in the presence of dehydrating agents like dicyclohexylcarbodiimide (DCC).
The DCC-promoted synthesis of amides begins with the protonation of DCC by carboxylic acid. The protonation makes it a better acceptor. Next, the addition of carboxylate to the protonated carbodiimide gives a reactive acylating agent.
Subsequently, the amine acts as a nucleophile that attacks the acylating agent to form a tetrahedral intermediate. In the...
The DCC-promoted synthesis of amides begins with the protonation of DCC by carboxylic acid. The protonation makes it a better acceptor. Next, the addition of carboxylate to the protonated carbodiimide gives a reactive acylating agent.
Subsequently, the amine acts as a nucleophile that attacks the acylating agent to form a tetrahedral intermediate. In the...
3.0K


