アミド合成のためのリガゼの発見,特徴付け,工学
Michael Winn1, Michael Rowlinson1, Fanghua Wang1,2
1Department of Chemistry, Manchester Institute of Biotechnology, The University of Manchester, Manchester, UK.
Nature
|May 20, 2021
まとめ
コロナチンおよび関連する細菌の植物毒素は,ホルモンジャスマニル-L-イソルエシン (JA-Ile) の模倣剤である. 研究者らは,JA-Ileと様々なアミドを合成するコロナファシウム酸リガゼ (CfaL) を特定し,新しい除草剤と医薬品の可能性を提示した.
科学分野:
- 生物化学
- 分子生物学
- 植物病理学
背景:
- コロナチンおよび関連する細菌の植物毒素は,植物ホルモンジャスマニル-L-イソルエシン (JA-Ile) を模倣し,重要な植物信号伝達経路を妨害する.
- これらの植物毒素は,植物生理に干渉する能力があるため,新しい選択的除草剤を開発する可能性を秘めています.
研究 の 目的:
- コロナファシウム酸をアミノ酸と結合させる酵素を特定し,特徴づけること.
- コロナファシウム酸リガゼ (CfaL) の酵素活性と構造特性を調査する.
- 様々な産業のための様々なアミドを合成する際のCfaLの潜在的応用を調査する.
主な方法:
- コロナファシウム酸リガゼ (CfaLs) の酵素特性および構造解明
- JA-Ile合成を含む基質の特異性と産物形成を決定する生化学的測定.
- 構造誘導型変異は,改良されたCfaLの変異を設計する.
主要な成果:
- 決定的な結合ステップを触媒する酵素として,コロナファシウム酸リガゼ (CfaLs) を特定し,特徴づけました.
- CfaLは植物酵素Jar1と類似してJA-Ileを合成することが示され,この結合活動の独立した進化を示唆している.
- 選択的運動解像度を含む幅広いアミドを合成するCfaLの能力を示し,ホモキラル製品を得ました.
結論:
- CfaLは植物酵素から独立して進化し,同様の結合反応を触媒化し,植物毒素と重要な植物ホルモンを生成する細菌酵素である.
- エンジニアリングされたCfaL変種は活性性が向上し,バイオテクノロジーの応用の可能性を強調しています.
- CfaL酵素は,様々なアミドを合成するための汎用性のあるプラットフォームを提供し,農業化学および医薬品開発に重大な影響を及ぼします.
関連する概念動画
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...
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01:07Acid Halides to Amides: Aminolysis
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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...
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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...
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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...
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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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