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Evolution of New Traits in Microbes01:24

Evolution of New Traits in Microbes

Microorganisms evolve rapidly due to their large population sizes and short generation times, often exhibiting measurable changes within days under laboratory conditions. Natural selection acts on standing genetic variation, enabling the retention and amplification of beneficial traits that confer fitness advantages in changing environments.Adaptive Pigment Regulation in RhodobacterIn Rhodobacter, a genus of purple non-sulfur bacteria, light-harvesting pigments such as bacteriochlorophyll and...
Conservative Site-specific Recombination and Phase Variation02:53

Conservative Site-specific Recombination and Phase Variation

Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
The recognition sites for Cre recombinase called LoxP...
Bioreactor Controls-III01:22

Bioreactor Controls-III

Strain improvement is a foundational strategy in industrial microbiology aimed at maximizing microbial productivity, particularly because natural isolates typically yield commercially valuable products in very low concentrations. Although optimizing the culture medium and environmental conditions can improve yields, these adjustments are inherently limited by the organism’s genetic potential. As a result, the focus shifts toward genetic modifications to enhance biosynthetic capacity. The...
Allosteric Proteins-ATCase01:19

Allosteric Proteins-ATCase

Binding sites linkages can regulate a protein's function.  For example, enzyme activity is often regulated through a feedback mechanism where the end product of the biochemical process serves as an inhibitor.
Aspartate transcarbamoylase (ATCase) is a cytosolic enzyme that catalyzes the condensation of L-aspartate and carbamoyl phosphate to  N-carbamoyl-L-aspartate. This reaction is the first step in pyrimidine biosynthesis. UTP and CTP, the end products of the pyrimidine synthesis pathway,...
Catalytically Perfect Enzymes01:07

Catalytically Perfect Enzymes

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.

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Updated: May 28, 2026

Mutagenesis and Functional Selection Protocols for Directed Evolution of Proteins in E. coli
09:01

Mutagenesis and Functional Selection Protocols for Directed Evolution of Proteins in E. coli

Published on: March 16, 2011

変異した基質選択性プロファイルを持つソルターゼA変異体の方向進化.

Kirill Piotukh1, Bernhard Geltinger, Nadja Heinrich

  • 1Department of Protein Engineering, Leibniz-Institut für Molekulare Pharmakologie, Robert-Rössle-Strasse 10, 13125 Berlin, Germany.

Journal of the American Chemical Society
|October 8, 2011
PubMed
まとめ

研究者は,誘導進化を用いて,幅広い基質選択性を有する変異型ソルターゼA酵素を設計した. このブレークスルーにより,痕跡のないタンパク質半合成が可能になり,タンパク質工学とバイオテクノロジーの応用が進んでいます.

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A New Screening Method for the Directed Evolution of Thermostable Bacteriolytic Enzymes
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A New Screening Method for the Directed Evolution of Thermostable Bacteriolytic Enzymes

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関連する実験動画

Last Updated: May 28, 2026

Mutagenesis and Functional Selection Protocols for Directed Evolution of Proteins in E. coli
09:01

Mutagenesis and Functional Selection Protocols for Directed Evolution of Proteins in E. coli

Published on: March 16, 2011

In Vitro Directed Evolution of a Restriction Endonuclease with More Stringent Specificity
09:16

In Vitro Directed Evolution of a Restriction Endonuclease with More Stringent Specificity

Published on: March 25, 2020

A New Screening Method for the Directed Evolution of Thermostable Bacteriolytic Enzymes
13:30

A New Screening Method for the Directed Evolution of Thermostable Bacteriolytic Enzymes

Published on: November 7, 2012

科学分野:

  • バイオケミストリー バイオケミストリー
  • プロテイン工学は,タンパク質の
  • 分子生物学は分子生物学である.

背景:

  • ソルターゼA (SrtA) は,タンパク質工学におけるタンパク質結合に不可欠な細菌のトランスペプチダゼである.
  • 現在のSrtA方法はLPxTGモチーフを必要とし,しばしばタンパク質半合成中に望ましくない変異を導入します.
  • タンパク質の組立を改善するために,変異された基板特異性を持つソルターゼ変種が必要である.

研究 の 目的:

  • 変異したソルターゼA酵素を開発するために,幅広い基質選択性を持つ.
  • LPxTGモチーフの限界を克服することによって,痕跡のないタンパク質半合成を可能にします.
  • 異なる結合能力を持つエンジニアリングされたソルターゼのためのプラットフォームを確立する.

主な方法:

  • 誘導進化は,突然変異したソルターゼAライブラリを作成するために使用されました.
  • フェーグディスプレイスクリーニングは,変異した基質認識ループを持つソルターゼ変異体を分離するために使用されました.
  • エンジニアリングされたソルターゼは,ヒストンH3.3の無痕半合成に適用されました.

主要な成果:

  • 基質選択性が著しく拡大した変異性ソルターゼAを成功裏に分離した.
  • エンジニアリングされたソルターゼは,ヒストンH3の無痕半合成を促進し,LPxTGモチーフの必要性を排除しました.
  • これは,ソルターゼの活性と汎用性を高める機能獲得変異を表しています.

結論:

  • ソルターゼAの誘導進化により,基板特異性を合わせた酵素が得られます.
  • トレースレスタンパク質半合成は,エンジニアリングされたソルターゼで達成可能であり,タンパク質のエンジニアリング効率を改善します.
  • この研究は,バイオテクノロジーにおけるエンジニアリングされたソルターゼを使用した汎用性のあるタンパク質組立プラットフォームの道を開く.