胺合成的连接酶的发现,表征和工程
Michael Winn1, Michael Rowlinson1, Fanghua Wang1,2
1Department of Chemistry, Manchester Institute of Biotechnology, The University of Manchester, Manchester, UK.
Nature
|May 20, 2021
概括
冠状腺素和相关的细菌植物毒素是雅士-L-异氨酸 (JA-Ile) 激素的模仿物. 研究人员发现了合成JA-Ile和各种胺的冠状腺酸酶 (CfaL),为新型除草剂和药物提供了潜力.
科学领域:
- 生物化学
- 分子生物学
- 植物病理学
背景情况:
- 冠状素和相关的细菌植物毒素模仿植物激素jasmonyl-L-isoleucine (JA-Ile),破坏关键的植物信号通路.
- 这些植物毒素具有开发新型选择性除草剂的潜力,因为它们能够干扰植物生理.
研究的目的:
- 识别和描述在冠状腺生物合成中负责将冠状腺酸与氨基酸结合的酶.
- 探索冠面酸链酶 (CfaLs) 的酶活性和结构特征.
- 研究CfaL在各种工业中合成多种胺的潜在应用.
主要方法:
- 冠面酸酶 (CfaLs) 的酶表征和结构分解.
- 生物化学试验以确定基质特异性和产品形成,包括JA-Ile合成.
- 通过结构引导的突变生成来设计增强的CfaL变体.
主要成果:
- 鉴定和表征冠面酸链酶 (CfaLs) 作为催化关键合步骤的酶.
- 证明CfaL可以合成JA-Ile,类似于植物酶Jar1,表明这种结合活动的独立演化.
- 展示了CfaL合成广泛的胺基的能力,包括选择性动态分辨率的血基质,以产生同质基质产品.
结论:
- CfaL 是细菌酶,它们与植物酶独立进化,催化类似的结合反应,产生植物毒素和必需的植物激素.
- 改造后的CfaL变种表现出更好的活性,突出显示了它们在生物技术应用中的潜力.
- CfaL酶为合成多种胺提供了多功能平台,对农业化学和制药发展具有重要意义.
相关概念视频
01:29Preparation of Amides
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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...
3.5K
01:07Acid Halides to Amides: Aminolysis
3.5K
Aminolysis is a nucleophilic acyl substitution reaction, where ammonia or amines act as nucleophiles to give the substitution product. Acid halides react with ammonia, primary amines, and secondary amines to yield primary, secondary, and tertiary amides, respectively.
In the first step of the aminolysis mechanism, the amine attacks the carbonyl carbon of the acyl chloride to form a tetrahedral intermediate. In the second step, the carbonyl group is re-formed with the elimination of a chloride...
In the first step of the aminolysis mechanism, the amine attacks the carbonyl carbon of the acyl chloride to form a tetrahedral intermediate. In the second step, the carbonyl group is re-formed with the elimination of a chloride...
3.5K
01:19Amines to Amides: Acylation of Amines
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Various carboxylic acid derivatives (such as acid chlorides, esters, and anhydrides) can be used for the acylation of amines to yield amides. The reaction requires two equivalents of amines. The first amine molecule functions as a nucleophile and attacks the carbonyl carbon to produce a tetrahedral intermediate. This is followed by the loss of the leaving group and restoration of the C=O bond.
Next, the second equivalent of amine serves as a Brønsted base and deprotonates the quaternary...
Next, the second equivalent of amine serves as a Brønsted base and deprotonates the quaternary...
2.9K
01:22Preparation of 1° Amines: Azide Synthesis
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Direct alkylation of ammonia produces polyalkylated amines, along with a quaternary ammonium salt. To exclusively prepare primary amines, the azide synthesis method can be used.
Azide ions act as good nucleophiles and react with unhindered alkyl halides to form alkyl azides. Alkyl azides do not participate in further nucleophilic substitution reactions, thereby eliminating the chances of polyalkylated products. Alkyl azides are reduced by hydride-based reducing agents, like lithium aluminum...
Azide ions act as good nucleophiles and react with unhindered alkyl halides to form alkyl azides. Alkyl azides do not participate in further nucleophilic substitution reactions, thereby eliminating the chances of polyalkylated products. Alkyl azides are reduced by hydride-based reducing agents, like lithium aluminum...
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01:28Amides to Carboxylic Acids: Hydrolysis
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Amides can undergo either acid-catalyzed hydrolysis or base-promoted hydrolysis through a typical nucleophilic acyl substitution. Each hydrolysis requires severe conditions.
Acid-catalyzed hydrolysis:
Hydrolysis of amides under acidic conditions yields carboxylic acids. Since the reaction occurs slowly, hydrolysis requires the conditions of heat.
The mechanism begins with the protonation of the carbonyl oxygen by the acid catalyst. The protonation makes the amide carbonyl carbon more...
Acid-catalyzed hydrolysis:
Hydrolysis of amides under acidic conditions yields carboxylic acids. Since the reaction occurs slowly, hydrolysis requires the conditions of heat.
The mechanism begins with the protonation of the carbonyl oxygen by the acid catalyst. The protonation makes the amide carbonyl carbon more...
3.7K
01:20Amides to Amines: LiAlH4 Reduction
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Amide reduction with strong reducing agents like lithium aluminum hydride proceeds through a nucleophilic acyl substitution to form amines. Primary, secondary, and tertiary amides yield primary, secondary, and tertiary amines, respectively.
Amide reduction requires two equivalents of the reducing agent, acting as a source of hydride ions. As shown in the figure, the reaction is initiated with a nucleophilic attack by the hydride ion at the carbonyl carbon to form a tetrahedral intermediate.
Amide reduction requires two equivalents of the reducing agent, acting as a source of hydride ions. As shown in the figure, the reaction is initiated with a nucleophilic attack by the hydride ion at the carbonyl carbon to form a tetrahedral intermediate.
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