Scientists at the University of Basel, ETH Zurich, and NCCR Molecular Systems Engineering have developed an artificial metalloenzyme that catalyses a reaction inside of cells without equivalent in nature. This could be a prime example for creating new non-natural metabolic pathways inside living cells, as reported today in Nature.
The artificial metalloenzyme, termed biot-Ru–SAV, was created using the biotin–streptavidin technology. This method relies on the high affinity of the protein streptavidin for the vitamin biotin, where compounds bound to biotin can be introduced into the protein to generate artificial enzymes. In this study the authors introduced an organometallic compound, with the metal ruthenium at its base. Organometallic compounds are molecules containing at least one bond between a metal and a carbon atom, and are often used as catalysts in industrial chemical reactions. However, organometallic catalysts perform poorly, if at all, in aqueous solutions or cellular-like environments, and need to be incorporated into protein scaffolds like streptavidin to overcome these limitations.
“The goal was to create an artificial metalloenzyme that can catalyse olefin metathesis, a reaction mechanism that is not present among natural enzymes,” says Thomas R Ward, Professor at the Department of Chemistry, University of Basel, and senior author of the study. The olefin metathesis reaction is a method for the formation and redistribution of carbon-carbon double bonds widely used in laboratory research and large-scale industrial productions of various chemical products. Biot-Ru–SAV catalyses a ring-closing metathesis to produce a fluorescent molecule for easy detection and quantification.
Periplasm as reaction compartment
However, the environment inside a living cell is far from ideal for the proper functioning of organometallic-based enzymes. “The main breakthrough was the idea to use the periplasm of Escherichia coli as a reaction compartment, whose environment is much better suited for an olefin metathesis catalyst,” says Markus Jeschek, a researcher from the team of co-supervising author Sven Panke at the Department of Biosystems Science and Engineering, ETH Zurich in Basel. The periplasm, the space between the inner cytoplasmic membrane and the bacterial outer membrane in gram-negative bacteria, contains low concentrations of metalloenzymes inhibitors, such as glutathione.
Having found ideal in vivo conditions, the authors went a step forward and decided to optimize biot-Ru–SAV by applying principles of directed evolution, a method that mimics the process of natural selection to evolve proteins with enhanced properties or activities. “We could then develop a simple and robust screening method that allowed us to test thousands of biot-Ru–SAV mutants and identify the most active variant,” Ward explains.
Not only could the authors markedly improve the catalytic properties of biot-Ru–SAV, but they could also show that organometallic-based enzymes can be engineered and optimized for different substrates, thus producing a variety of different chemical products. “The exciting thing about this is that artificial metalloenzymes like biot-Ru–SAV can be used to produce novel high added-value chemicals,” Ward says. “It has a lot of potential to combine both chemical and biological tools to ultimately utilize cells as molecular factories.”
Learn more: Bringing artificial enzymes closer to nature
The Latest on: Artificial enzymes
via Google News
The Latest on: Artificial enzymes
- 2019 Study on Novel Enzyme Technologies for Pharmaceutical Applications - Waves of Technological Developments in Biocatalysis - ResearchAndMarkets.comon January 24, 2020 at 2:42 am
The current study discusses the novel enzyme technologies for enzyme discovery and improvements such as directed mutagenesis, metagenomics, artificial intelligence, and computational biology ...
- Odor Eliminator Market Precise Study on Factors, Market Drivers and Key Players Strategies Analyzed Till 2022|CAGR of 4.9%on January 17, 2020 at 12:30 am
The increasing awareness among people about the clean and hygienic environment has led to the increase in the usage of artificial environment contributors such as odor eliminators. Air odor ...
- Novel Enzyme Technologies for Pharmaceutical Applications, 2019 Research Studyon January 15, 2020 at 1:58 am
Dublin, Jan. 15, 2020 (GLOBE NEWSWIRE) -- The "Novel Enzyme Technologies for Pharmaceutical Applications" report has been added to ResearchAndMarkets.com's offering. This study discusses the novel ...
- Novel Enzyme Technologies for Pharmaceutical Applicationson January 14, 2020 at 8:15 am
/PRNewswire/ -- The current study discusses the novel enzyme technologies for enzyme discovery and improvement such as directed mutagenesis, ...
- Synthetic enzyme combats disorders caused by dysfunctional mitochondriaon January 14, 2020 at 7:38 am
The team added LOXCAT to a medium of cultured human cells with defective mitochondria and found that the artificial enzyme converted lactate to pyruvate. This enters cells and picks up electrons, ...
- Molecular factories: The combination between nature and chemistry is functionalon January 9, 2020 at 9:35 am
In this animal model, they produced the desired compound, which was catalyzed by the enzyme in the artificial organelle. The viability of the animal was not compromised by the injection.
- Catalytic protocells get zingyon January 8, 2020 at 8:58 am
used two different types of catalysts to develop a new type of artificial cell capable of decomposing hydrogen peroxide and generating oxygen. The team used a ruthenium-based inorganic catalyst in the ...
- Artificial cells act more like the real thingon December 5, 2019 at 11:19 pm
cell's motility, especially due to surface enzyme activity," said Darrell Velegol, distinguished professor of chemical engineering. "The research team developed a simple way to make an artificial cell ...
- Artificial cells act more like the real thingon December 5, 2019 at 1:24 pm
cell's motility, especially due to surface enzyme activity," said Darrell Velegol, distinguished professor of chemical engineering. "The research team developed a simple way to make an artificial ...
- Indian scientist's artificial enzymes may help treat heart diseaseson November 26, 2019 at 6:44 am
Professor G Mugesh and his team at the Indian Institute of Science are working on synthetic molecules such as artificial enzymes, which may help develop novel ways to tackle cardiovascular ...
via Bing News