非正規のアミノ酸依存生物を生成するタンパク質におけるヒスティジンの機能的置換
Fei Gan1, Renhe Liu1, Feng Wang1
1California Institute for Biomedical Research (Calibr) , La Jolla , California 92037 , United States.
Journal of the American Chemical Society
|March 7, 2018
まとめ
研究者は,特定の非正規のアミノ酸を必要とする微生物を作り出す方法を開発し,ワクチンと生物学的封じ込めの安全性を高めました. これは,必須ヒスティジンアミノ酸を,機能的な類似体に置き換えることを含みます.
科学分野:
- 合成生物学
- バイオテクノロジー
- 微生物遺伝学
背景:
- 非正規のアミノ酸を必要とする生物は,生ワクチンと生物学的封じ込めに不可欠です.
- エッセンシャルアミノ酸残基の遺伝子操作は,オクストロフィーを達成するための経路を提供します.
研究 の 目的:
- 非正規のアミノ酸に 厳格な成長依存を伴う生物を作るための シンプルな戦略を開発する.
- メタロ酵素における重要なヒスティジン残基の機能的代替を非正規類型で実証する.
主な方法:
- ヒスティジン (His) の残留を亜鉛を含むマノース-6-リン酸同化酵素 (ManA) で3メチル-l-ヒスティジン (MeH) で遺伝的に置き換える.
- N262SとH264MeHの置換を備えた進化したManA変種 (Opt5) を含んだEscherichia coli株を設計する.
- エンジニアリングされた大腸菌株の成長依存性と逆転率の評価
主要な成果:
- 3-メチル-l-ヒスティジン (MeH) は,ManAにおける主要な金属結合リガンド (H264) を機能的に置換した.
- エンジニアリングされたManA変種 (Opt5) は,野生型酵素に匹敵する活性を示した.
- エンジニアリングされたEscherichia coli株は,非常に低いリバーション率でMeHに厳格な成長依存を示しました.
結論:
- この戦略は,オクストロフ生物を生成するための堅固な方法を提供します.
- このアプローチは,本質的なヒスティジン残基を持つ他の金属または非金属タンパク質に適用できる可能性がある.
- この方法は,再結合生物と生体ワクチンの生物学的封じ込めと安全性を高めます.
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