通过对同类蛋白质块的结构引导的体内重组来设计非特异性过氧酶
Alejandro Beltran-Nogal1, Ivan Mateljak2, David Gonzalez-Perez1
1Institute of Catalysis, CSIC, Marie Curie 2, Madrid, Spain.
Methods in enzymology
|April 27, 2025
概括
这项研究引入了一种创新的方法,通过计算设计和DNA混合来创建多种真菌非特异性过氧酶 (UPO). 这种方法扩大了酶工程和探索UPO功能工具包.
科学领域:
- 生物催化和酶工程 生物催化和酶工程
- 蛋白质化学和有针对性的进化.
- 菌酶研究 菌酶研究
背景情况:
- 的非特异性过氧酶 (UPO) 是C-H氧功能化的有价值的生物催化剂.
- 以前的努力集中在定向进化上,以增强UPO属性.
- 使用UPO的酶基因生成仍未得到充分探索.
研究的目的:
- 描述一种用于构建功能多样化的UPO模拟器的方法.
- 探索将计算设计与DNA混合用于UPO工程的潜力.
- 扩大工程UPO用于生物催化物的目录.
主要方法:
- 应用SCHEMA-RASPP计算算法用于奇米拉设计.
- 在体内进行DNA混合以产生遗传多样性.
- 构建不同类型的异构无特异性过氧酶的构造.
主要成果:
- 展示一个可行的策略,以创建功能多样化的UPO幻象.
- 计算预测和实验性DNA混合的成功结合.
- 建立一个平台来生成新的UPO变体.
结论:
- 描述的方法可以构建多种UPO仿真模型.
- 这种方法显著扩大了UPO工程的范围.
- 这些发现促进了针对特定生物催化剂应用量身定制的UPO的开发.
相关概念视频
Conservative Site-specific Recombination and Phase Variation
5.9K
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...
The recognition sites for Cre recombinase called LoxP...
5.9K
Homologous Recombination
49.7K
The basic reaction of homologous recombination (HR) involves two chromatids that contain DNA sequences sharing a significant stretch of identity. One of these sequences uses a strand from another as a template to synthesize DNA in an enzyme-catalyzed reaction. The final product is a novel amalgamation of the two substrates. To ensure an accurate recombination of sequences, HR is restricted to the S and G2 phases of the cell cycle. At these stages, the DNA has been replicated already and the...
49.7K
Protein Import into the Peroxisomes
3.3K
Cells contain membrane-bound organelles called peroxisomes that oxidize organic molecules by transferring hydrogen atoms to oxygen, producing hydrogen peroxide. Peroxisomes enzymatically convert the released hydrogen peroxide into water and oxygen.
Peroxisomal Protein Import:
Peroxisomes lack the genetic machinery required to code for their own proteins. Hence, most peroxisomal membrane, lumenal and transmembrane proteins are synthesized in the cytoplasm or ER and transported to the peroxisome...
Peroxisomal Protein Import:
Peroxisomes lack the genetic machinery required to code for their own proteins. Hence, most peroxisomal membrane, lumenal and transmembrane proteins are synthesized in the cytoplasm or ER and transported to the peroxisome...
3.3K
Exon Recombination
3.5K
The evolution of new genes is critical for speciation. Exon recombination, also known as exon shuffling or domain shuffling, is an important means of new gene formation. It is observed across vertebrates, invertebrates, and in some plants such as potatoes and sunflowers. During exon recombination, exons from the same or different genes recombine and produce new exon-intron combinations, which might evolve into new genes.
Exon shuffling follows “splice frame rules.” Each exon...
Exon shuffling follows “splice frame rules.” Each exon...
3.5K
Restriction Enzymes
29.4K
Restriction enzymes are bacterial enzymes used to cut DNA in a sequence-specific manner. To cleave DNA, they bind to specific palindromic sequences called restriction sites. Such palindromic DNA sequences or inverted repeats are commonly found in regions of functional significance, such as the origin of replication, gene operator sites, and regions containing transcription termination signals.
The host bacteria protect their own genomic DNA from these enzymes by methylating these sites. Some...
The host bacteria protect their own genomic DNA from these enzymes by methylating these sites. Some...
29.4K
Mismatch Repair
4.6K
Organisms are capable of detecting and fixing nucleotide mismatches that occur during DNA replication. This sophisticated process requires identifying the new strand and replacing the erroneous bases with correct nucleotides. Mismatch repair is coordinated by many proteins in both prokaryotes and eukaryotes.
The Mutator Protein Family Plays a Key Role in DNA Mismatch Repair
The human genome has more than 3 billion base pairs of DNA per cell. Prior to cell division, that vast amount of genetic...
The Mutator Protein Family Plays a Key Role in DNA Mismatch Repair
The human genome has more than 3 billion base pairs of DNA per cell. Prior to cell division, that vast amount of genetic...
4.6K


