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
- Enzymes Market to Exceed Valuation of US$ 16.3 Bn by 2031, TMR Studyon May 20, 2022 at 9:39 am
Widespread utilization of enzymes in food & beverage processing and manufacture of personal care products and pharmaceuticals propelling revenue prospects of enzymes market; commercialization of ...
- Demand For Lactase Enzymes Recorded A 3.7% YoY Growth Through 2031 – Fact.MR Studyon May 17, 2022 at 5:00 pm
Various regulations imposed by governments on the consumption of lactase enzyme in detail. Influence of modern technologies such as big data & analytics, artificial intelligence and social media ...
- Keep Your Gut Happy With the Best Digestive Enzymeson May 17, 2022 at 5:00 pm
The GNC Digestive Enzymes do not contain any artificial colors or flavors. Additionally, the serving size of this product is two tablets, making it hassle-free to get your daily dose. This pick ...
- Artificial proteins as specific and versatile neutralizing binders targeting the spike of SARS-CoV-2on May 13, 2022 at 12:16 pm
Researchers demonstrated that the biosynthetic proteins called αReps addressing the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike (S) protein could be novel SARS-CoV-2 antivirals ...
- Microbial jugglingon May 12, 2022 at 6:06 am
The enzyme the team studied is part of a family ... Erb’s research team had been working to develop bioreactors for artificial photosynthesis to convert carbon dioxide (CO 2) from the atmosphere ...
- AI-Designed Enzyme Devours Plastic Trash in Dayson May 7, 2022 at 4:59 pm
Now researchers have harnessed the power of artificial intelligence to design an enzyme that can degrade a commonly used plastic in just a day or two.
- How a soil microbe could rev up artificial photosynthesison April 29, 2022 at 9:59 am
But the carbon fixing champs are not plants, but soil bacteria. Some bacterial enzymes carry out a key step in carbon fixation 20 times faster than plant enzymes do, and figuring out how they do ...
- Fast and Efficient Plastic-Degrading Enzyme Developed Using AIon April 28, 2022 at 1:00 am
Enzymes that break down PET ... from synthetic biology to chemical engineering to artificial intelligence,” said Andrew Ellington, PhD, professor in the Center for Systems and Synthetic Biology ...
- This AI-Designed Enzyme Can Devour Plastic Trash In Hours: Videoon April 27, 2022 at 5:00 pm
Researchers at the University of Texas at Austin announced Thursday that they had used artificial intelligence to successfully engineer a type of enzyme, called a hydrolase, that can break down ...
via Bing News