Tag Archives: enzymes

Nanozymes as an antidote for pesticides

Should you have concerns about exposure to pesticides or chemical warfare agents (timely given events in Syria as per this April 4, 2017 news item on CBC [Canadian Broadcasting News Corporation] online) , scientists at the Lomonosov Moscow State University have developed a possible antidote according to a March 8,, 2017 news item on phys.org,

Members of the Faculty of Chemistry of the Lomonosov Moscow State University have developed novel nanosized agents that could be used as efficient protective and antidote modalities against the impact of neurotoxic organophosphorus compounds such as pesticides and chemical warfare agents. …

A March 7, 2017 Lomonosov Moscow State University press release on EurekAlert, which originated the news item, describes the work in detail,

A group of scientists from the Faculty of Chemistry under the leadership of Prof. Alexander Kabanov has focused their research supported by a “megagrant” on the nanoparticle-based delivery to an organism of enzymes, capable of destroying toxic organophosphorous compounds. Development of first nanosized drugs has started more than 30 years ago and already in the 90-s first nanomedicines for cancer treatment entered the market. First such medicines were based on liposomes – spherical vesicles made of lipid bilayers. The new technology, developed by Kabanov and his colleagues, uses an enzyme, synthesized at the Lomonosov Moscow State University, encapsulated into a biodegradable polymer coat, based on an amino acid (glutamic acid).

Alexander Kabanov, Doctor of Chemistry, Professor at the Eshelman School of Pharmacy of the University of North Carolina (USA) and the Faculty of Chemistry, M. V. Lomonosov Moscow State University, one of the authors of the article explains: “At the end of the 80-s my team (at that time in Moscow) and independently Japanese colleagues led by Prof. Kazunori Kataoka from Tokyo began using polymer micelles for small molecules delivery. Soon the nanomedicine field has “exploded”. Currently hundreds of laboratories across the globe work in this area, applying a wide variety of approaches to creation of such nanosized agents. A medicine on the basis of polymeric micelles, developed by a Korean company Samyang Biopharm, was approved for human use in 2006.”

Professor Kabanov’s team after moving to the USA in 1994 focused on development of polymer micelles, which could include biopolymers due to electrostatic interactions. Initially chemists were interested in usage of micelles for RNA and DNA delivery but later on scientists started actively utilizing this approach for delivery of proteins and, namely, enzymes, to the brain and other organs.

Alexander Kabanov says: “At the time I worked at the University of Nebraska Medical Center, in Omaha (USA) and by 2010 we had a lot of results in this area. That’s why when my colleague from the Chemical Enzymology Department of the Lomonosov Moscow State University, Prof. Natalia Klyachko offered me to apply for a megagrant the research theme of the new laboratory was quite obvious. Specifically, to use our delivery approach, which we’ve called a “nanozyme”, for “improvement” of enzymes, developed by colleagues at the Lomonosov Moscow State University for its further medical application.”

Scientists together with the group of enzymologists from the Lomonosov Moscow State University under the leadership of Elena Efremenko, Doctor of Biological Sciences, have chosen organophosphorus hydrolase as a one of the delivered enzymes. Organophosphorus hydrolase is capable of degrading toxic pesticides and chemical warfare agents with very high rate. However, it has disadvantages: because of its bacterial origin, an immune response is observed as a result of its delivery to an organism of mammals. Moreover, organophosphorus hydrolase is quickly removed from the body. Chemists have solved this problem with the help of a “self-assembly” approach: as a result of inclusion of organophosphorus hydrolase enzyme in a nanozyme particles the immune response becomes weaker and, on the contrary, both the storage stability of the enzyme and its lifetime after delivery to an organism considerably increase. Rat experiments have proved that such nanozyme efficiently protects organisms against lethal doses of highly toxic pesticides and even chemical warfare agents, such as VX nerve gas.

Alexander Kabanov summarizes: “The simplicity of our approach is very important. You could get an organophosphorus hydrolase nanozyme by simple mixing of aqueous solutions of anenzyme and safe biocompatible polymer. This nanozyme is self-assembled due to electrostatic interaction between a protein (enzyme) and polymer”.

According to the scientist’s words the simplicity and technological effectiveness of the approach along with the obtained promising results of animal experiments bring hope that this modality could be successful and in clinical use.

Members of the Faculty of Chemistry of the Lomonosov Moscow State University, along with scientists from the 27th Central Research Institute of the Ministry of Defense of the Russian Federation, the Eshelman School of Pharmacy of the University of North Carolina at Chapel Hill (USA) and the University of Nebraska Medical Center (UNC) have taken part in the Project.

Here’s a link to and a citation for the paper,

A simple and highly effective catalytic nanozyme scavenger for organophosphorus neurotoxins by Elena N. Efremenko, Ilya V. Lyagin, Natalia L. Klyachko, Tatiana Bronich, Natalia V. Zavyalova, Yuhang Jiang, Alexander V. Kabanov. Journal of Controlled Release Volume 247, 10 February 2017, Pages 175–181  http://dx.doi.org/10.1016/j.jconrel.2016.12.037

This paper is behind a paywall.

Treating bandages with enzymes and polyethylene glycol or cellulase* could make antibacterial nanoparticles better adhere

It’s been a while since I’ve featured research from Iran. This work is focused on bandages, burns, and nanoparticles according to an Oct. 18, 2016 news item on Nanowerk (Note: A link has been removed),

Pre-treating the fabric surface of the bandages used to treat burns with enzymes and polyethylene glycol or cellulase may promote the adhesion of antibacterial nanoparticles and improve their bacteria-repelling ability. These are the findings of a group of scientists from the Islamic Azad University, Iran, published in The Journal of The Textile Institute (“NiO-/cotton- modified nanocomposite as a medication model for bacterial-related burn infection”).

An Oct. 18, 2016 Taylor & Francis (Publishing) Group press release (received via email), which originated the news item, expands on the theme,

Injuries caused by burns are a global health problem, with the World Health Organisation citing 195,000 deaths per year worldwide as a result of burns from fires alone. Burn injuries are particularly susceptible to infections, hospital-acquired or otherwise, with the bacteria Pseudomonas aeruginosa accounting for over half of all severe burn infections.

Noble metal (particularly silver) antimicrobials have long been identified as having potential for combating bacterial infection; however, there are concerns about dressings adhering to wounds and toxic effects on skin cells. Currently, scientists are researching nanoparticles which can be used to introduce these antimicrobial properties into the textiles used in dressings.

The authors of this paper have studied 150 cases to identify the most common infections in burns. In the paper, they also identified a method for giving cotton bandages antibacterial properties by coating the fabric surface with a Nickel oxide (NiO)/organic polymer/enzyme matrix in order to promote their bacteria-resistant qualities and suitability for use on burn victims.

Pseudomonas and Staphylococci infections emerged as the two most common pathogens in the Iran Burn Centre, where the study took place, and the authors evaluated their design of the bandage against these as well as fifteen other strains of bacteria. They conclude by proposing further studies into the combination of bactericidal polymers with bacteria-killing metal-oxide nanoparticles in cotton fabrics. Whilst their current design does not meet the criteria for a susceptibility test, they are hopeful that further studies will reveal the clinical relevance of their design.

Here’s a link to and a citation for the paper,

NiO-/cotton- modified nanocomposite as a medication model for bacterial-related burn infections by Azadeh Basiri, Nasrin Talebian & Monir Doudi. The Journal of The Textile Institute http://dx.doi.org/10.1080/00405000.2016.1222863
Pages 1-9 Published online: 12 Sep 2016

This paper is behind a paywall.

*’Cellulase’ changed to ‘Cellulose’ Nov. 15, 2016 at 1832 PT and changed back again on Nov. 16, 2016. Sorry for the confusion but by the time I published this piece I’d forgotten checking to confirm the existence of cellulase.

Enzyme-based sustainable sensing devices

This story about a sustainable sensing device involves sweat. A July 28, 2016 news item on ScienceDaily describes the sweaty situation,

It may be clammy and inconvenient, but human sweat has at least one positive characteristic — it can give insight to what’s happening inside your body. A new study published in the ECS [Electrochemical Society] Journal of Solid State Science and Technology aims to take advantage of sweat’s trove of medical information through the development of a sustainable, wearable sensor to detect lactate levels in your perspiration.

Caption: Depiction of patch sensor via CFDRC. Credit: Sergio Omar Garcia/CFDRC

Caption: Depiction of patch sensor via CFDRC. Credit: Sergio Omar Garcia/CFDRC

The patch in that image doesn’t seem all that wearable but presumably there will be some changes made. A July 28, 2016 Electrochemical Society news release on EurekAlert, which originated the news item, provides more detail about the technology,

“When the human body undergoes strenuous exercise, there’s a point at which aerobic muscle function becomes anaerobic muscle function,” says Jenny Ulyanova, CFD Research Corporation (CFDRC) researcher and co-author of the paper. “At that point, lactate is produce at a faster rate than it is being consumed. When that happens, knowing what those levels are can be an indicator of potentially problematic conditions like muscle fatigue, stress, and dehydration.”

Utilizing green technology

Using sweat to track changes in the body is not a new concept. While there have been many developments in recent years to sense changes in the concentrations of the components of sweat, no purely biological green technology has been used for these devices. The team of CFDRC researchers, in collaboration with the University of New Mexico, developed an enzyme-based sensor powered by a biofuel cell – providing a safe, renewable power source.

Biofuel cells have become a promising technology in the field of energy storage, but still face many issues related to short active lifetimes, low power densities, and low efficiency levels. However, they have several attractive points, including their ability to use renewable fuels like glucose and implement affordable, renewable catalysts.

“The biofuel cell works in this particular case because the sensor is a low-power device,” Ulyanova says. “They’re very good at having high energy densities, but power densities are still a work in progress. But for low-power applications like this particular sensor, it works very well.”

In their research, entitled “Wearable Sensor System Powered by a Biofuel Cell for Detection of Lactate Levels in Sweat,” the team powered the biofuel cells with a fuel based on glucose. This same enzymatic technology, where the enzymes oxidize the fuel and generate energy, is used at the working electrode of the sensor which allows for the detection of lactate in your sweat.

Targeting lactate

While the use of the biofuel cell is a novel aspect of this work, what sets it apart from similar developments in the field is the use of electrochemical processes to very accurately detect a specific compound in a very complex medium like sweat.

“We’re doing it electrochemically, so we’re looking at applying a constant load to the sensor and generating a current response,” Ulyanova says, “which is directly proportional to the concentration of our target analyte.”

Practical applications

Originally, the sensor was developed to help detect and predict conditions related to lactate levels (i.e. fatigue and dehydration) for military personnel.

“The sensor was designed for a soldier in training at boot camp,” says Sergio Omar Garcia, CFDRC researcher and co-author of the paper, “but it could be applied to people that are active and anyone participating in strenuous activity.”

As for commercial applications, the researchers believe the device could be used as a training aid to monitor lactate changes in the same way that athletes use heart rate monitors to see how their heart rate changes during exercise.

On-body testing

The team is currently working to redesign the physical appearance of the patch to move from laboratory research to on-body tests. Once the scientists optimize how the sensor adheres to the skin, its sweat sample delivery/removal, and the systems electronic components, volunteers will test its capabilities while exercising.

“We had actually talked about this idea to some local high school football coaches,” Ulyanova says, “and they seem to really like it and are willing to put forth the use of their players to beta test the idea.”

After initial data is gathered, the team will be able to work with other groups to interpret the data and relate it to the physical condition of the person. With this, predictive models could be built to potentially help prevent conditions related to individual overexertion.

Future plans for the device include implementing wireless transmission of results and the development of a suite of sensors (a hybrid sensor) that can detect various other biomolecules, indicative of physical or physiological stressors.

Here’s a link to and a citation for the paper,

Wearable Sensor System Powered by a Biofuel Cell for Detection of Lactate Levels in Sweat by S. O. Garcia, Y. V. Ulyanova, R. Figueroa-Teran, K. H. Bhatt, S. Singhal and P. Atanassov. ECS J. Solid State Sci. Technol. 2016 volume 5, issue 8, M3075-M3081 doi: 10.1149/2.0131608jss

This paper is behind a paywall.

An enzyme’s atoms are in a subtle dance that can affect protein function

This research comes from Québec’s Institut national de la recherche scientifique (INRS) according to a Dec. 10, 2015 news item on ScienceDaily,

Infinitesimal fluctuations occurring on the milli- and even nano-second time scales within the three-dimensional structure of enzymes may be one of the keys to explaining protein function. Professor Nicolas Doucet’s team at INRS has demonstrated that even when certain amino acids are far from the active site of an enzyme, a change in their flexibility and atomic fluctuations can significantly impact enzyme activity. This phenomenon, which has been underestimated up to now, could explain certain protein engineering failures and help improve the way synthetic functional enzymes are designed.

A Dec. 10, 2015 INRS news release on EurekAlert, which originated the news item, provides an explanation of an enzyme’s functions and what the researchers found out,

Enzymes are nanomachines that are exceptionally efficient at catalyzing a chemical reaction. They play a role in all cellular mechanisms. Like all proteins, they are made up of amino acid chains that are folded and assembled in a very precise 3D structure. Some enzymes, like ribonuclease A, are so efficient that they catalyze the transformation of chemical molecules thousands of times per second.

In this study, Donald Gagné, a researcher in Professor Doucet’s lab holding a PhD in biology from INRS, analyzed the impact of removing a methyl group located near a loop distant from the reaction site of ribonuclease A–a very slight change that presumably would have no effect. The mutation does not perturb the 3D structure of the enzyme. However, it did result in a four-fold reduction in the affinity of ribonuclease A for nucleotides (molecules to which it must bind to carry out its function). How is this possible?

Using crystallography techniques and nuclear magnetic resonance to examine the enzyme at atomic resolution, Donald Gagné compared normal ribonuclease A with the mutated enzyme. He observed that when ribonuclease A is modified, the nucleotides do not position themselves correctly and have a harder time binding to the active site. It appears that this repositioning is due to an increase in enzyme fluctuations caused by the elimination of this distant methyl group, which we can picture as creating vibrations that spread through the enzyme structure all the way to the site of catalysis.

This demonstration of the importance of enzyme dynamics could change our understanding of protein and enzyme mechanisms. While it remains a challenge to measure fluctuations at this atomic scale, researchers have studied the three-dimensional structure of proteins to understand how they function. Despite the staggering complexity of this phenomenon, we now know that proteins are increasingly regulated by the subtle dance of their atoms.

Here’s a link to and a citation for the paper,

Perturbation of the Conformational Dynamics of an Active-Site Loop Alters Enzyme Activity by Donald Gagné, Rachel L. French, Chitra Narayanan, Miljan Simonović, Pratul K. Agarwal, Nicolas Doucet. Structure Volume 23, Issue 12, p2256–2266, 1 December 2015 DOI: http://dx.doi.org/10.1016/j.str.2015.10.011

This paper is behind a paywall.

Northwestern University’s (US) International Institute for Nanotechnology (IIN) rakes in some cash

Within less than a month Northwestern University’s International Institute for Nanotechnology (IIN) has been granted awarded two grants by the US Department of Defense.

4D printing

The first grant, for 4D printing, was announced in a June 11, 2015 Northwestern news release by Megan Fellman (Note: A link has been removed),

Northwestern University’s International Institute for Nanotechnology (IIN) has received a five-year, $8.5 million grant from the U.S. Department of Defense’s competitive Multidisciplinary University Research Initiative (MURI) program to develop a “4-dimensional printer” — the next generation of printing technology for the scientific world.

Once developed, the 4-D printer, operating on the nanoscale, will be used to construct new devices for research in chemistry, materials sciences and U.S. defense-related areas that could lead to new chemical and biological sensors, catalysts, microchip designs and materials designed to respond to specific materials or signals.

“This research promises to bring transformative advancement to the development of biosensors, adaptive optics, artificially engineered tissues and more by utilizing nanotechnology,” said IIN director and chemist Chad A. Mirkin, who is leading the multi-institution project. Mirkin is the George B. Rathmann Professor of Chemistry in the Weinberg College of Arts and Sciences.

The award, issued by the Air Force Office of Scientific Research, supports a team of experts from Northwestern, the University of Miami, the University of California, San Diego, and the University of Maryland.

In science, “printing” encodes information at specific locations on a material’s surface, similar to how we print words on paper with ink. The 4-dimensional printer will consist of millions of tiny elastomeric “pens” that can be used individually and independently to create nanometer-size features composed of hard or soft materials.

The information encoded can be in the form of materials with a defined set of chemical and physical properties. The printing speed and resolution determine the amount and complexity of the information that can be encoded.

Progress in fields ranging from biology to chemical sensing to computing currently are limited by the lack of low-cost equipment that can perform high-resolution printing and 3-dimensional patterning on hard materials (e.g., metals and semiconductors) and soft materials (e.g., organic and biological materials) at nanometer resolution (approximately 1,000 times smaller than the width of a human hair).

“Ultimately, the 4-D printer will provide a foundation for a new generation of tools to develop novel architectures, wherein the hard materials that form the functional components of electronics can be merged with biological or soft materials,” said Milan Mrksich, a co-principal investigator on the grant.

Mrksich is the Henry Wade Rogers Professor of Biomedical Engineering, Chemistry and Cell and Molecular Biology, with appointments in the McCormick School of Engineering and Applied Science, Weinberg and Northwestern University Feinberg School of Medicine.

A July 10, 2015 article about the ‘4D printer’ grant  by Madeline Fox for the Daily Northwestern features a description of 4D printing from Milan Mrksich, a co-principal investigator on the grant,

Milan Mrksich, one of the project’s five senior participants, said that while most people are familiar with the three dimensions of length, width and depth, there are often misconceptions about the fourth property of a four-dimensional object. Mrksich used Legos as an analogy to describe 4D printing technology.

“If you take Lego blocks, you can basically build any structure you want by controlling which Lego is connected to which Lego and controlling all their dimensions in space,” Mrksich said. “Within an object made up of nanoparticles, we’re controlling the placement — as we use a printer to control the placement of every particle, our fourth dimension lets us choose which nanoparticle with which property would be at each position.”

Thank you Dr. Mrksich and Ms. Fox for that helpful analogy.

Designing advanced bioprogrammable nanomaterials

The second grant, announced in a July 6, 2015 Northwestern news release by Megan Fellman, is apparently the only one of its kind in the US (Note: A link has been removed),

Northwestern University’s International Institute for Nanotechnology (IIN) has been awarded a U.S. Air Force Center of Excellence grant to design advanced bioprogrammable nanomaterials for solutions to challenging problems in the areas of energy, the environment, security and defense, as well as for developing ways to monitor and mitigate human stress.

The five-year, $9.8 million grant establishes the Center of Excellence for Advanced Bioprogrammable Nanomaterials (C-ABN), the only one of its kind in the country. After the initial five years, the grant potentially could be renewed for an additional five years.

“Northwestern University was chosen to lead this Center of Excellence because of its investment in infrastructure development, including new facilities and instrumentation; its recruitment of high-caliber faculty members and students; and its track record in bio-nanotechnology and cognitive sciences,” said Timothy Bunning, chief scientist at the U.S. Air Force Research Laboratory (AFRL) Materials and Manufacturing Directorate.

Led by IIN director Chad A. Mirkin, C-ABN will support collaborative, discovery-based research projects aimed at developing bioprogrammable nanomaterials that will meet both military and civilian needs and facilitate the efficient transition of these new technologies from the laboratory to marketplace.

Bioprogrammable nanomaterials are structures that typically contain a biomolecular component, such as nucleic acids or proteins, which give the materials a variety of novel capabilities. [emphasis mine] Nanomaterials can be designed to assemble into large 3-D structures, to interface with biological structures inside cells or tissues, or to interface with existing macroscale devices, for example. These new bioprogrammable nanomaterials and the fundamental knowledge gained through their development will ultimately lead to the creation of wearable, portable and/or human-interactive devices with extraordinary capabilities that will significantly impact both civilian and Air Force needs.

In one research area, scientists will work to understand the molecular underpinnings of vulnerability and resilience to stress. They will use bioprogrammable nanomaterials to develop ultrasensitive sensors capable of detecting and quantifying biomarkers for human stress in biological fluids (e.g., saliva, perspiration or blood), providing means to easily monitor the soldier during times of extreme stress. Ultimately, these bioprogrammable materials may lead to methods to increase human cellular resilience to the effects of stress and/or to correct genetic mutations that decrease cellular resilience of susceptible individuals.

Other research projects, encompassing a wide variety of nanotechnology-enabled goals, include:

Developing hybrid wearable energy-storage devices;
Developing devices to identify chemical and biological targets in a field environment;
Developing flexible bio-electronic circuits;
Designing a new class of flat optics; and
Advancing understanding of design rules between 2-D and 3-D architectures.

The analysis of these nanostructures also will extend fundamental knowledge in the fields of materials science and engineering, human performance, chemistry, biology and physics.

The center will be housed under the IIN, providing researchers with access to IIN’s strong entrepreneurial community and its close ties with Northwestern’s renowned Kellogg School of Management.

This second news release provides an interesting contrast to a recent news release from Sweden’s Karolinska Intitute where the writer was careful to note that the enzymes and organic electronic ion pumps were not living as noted in my June 26, 2015 posting. It seems nucleic acids (as in RNA and DNA) can be mentioned without a proviso in the US. as there seems to be little worry about anti-GMO (genetically modified organisms) and similar backlashes affecting biotechnology research.

Bacteria and biobatteries

It’s more a possibility at the moment than anything else but researchers at Concordia University in Montréal, Canada have found a way to make an enzyme behave more like a battery. From the April 19, 2012 news item on Nanowerk,

Concordia Associate Professor László Kálmán — along with his colleagues in the Department of Physics, graduate students Sasmit Deshmukh and Kai Tang — has been working with an enzyme found in bacteria that is crucial for capturing solar energy. Light induces a charge separation in the enzyme, causing one end to become negatively charged and the other positively charged, much like in a battery.

In nature, the energy created is used immediately, but Kálmán says that to store that electrical potential, he and his colleagues had to find a way to keep the enzyme in a charge-separated state for a longer period of time.

“We had to create a situation where the charges don’t want to or are not allowed to go back, and that’s what we did in this study,” he says.

Kálmán and his colleagues showed that by adding different molecules, they were able to alter the shape of the enzyme and, thus, extend the lifespan of its electrical potential.

In the April 17, 2012 news item written by Luciana Gravotta for Concordia University, Kálmán provides an explanation of why the researchers were changing the enzyme’s shape,

In its natural configuration, the enzyme is perfectly embedded in the cell’s outer layer, known as the lipid membrane. The enzyme’s structure allows it to quickly recombine the charges and recover from a charge-separated state.

However, when different lipid molecules make up the membrane, as in Kálmán’s experiments, there is a mismatch between the shape of the membrane and the enzyme embedded within it. Both the enzyme and the membrane end up changing their shapes to find a good fit. The changes make it more difficult for the enzyme to recombine the charges, thereby allowing the electrical potential to last much longer.

“What we’re doing is similar to placing a race car on snow-covered streets,” says Kálmán. The surrounding conditions prevent the race car from performing as it would on a racetrack, just like the different lipids prevent the enzyme from recombining the charges as efficiently as it does under normal circumstances.

Apparently the researchers are hoping to eventually create biocompatible batteries with enzymes and other biological molecules replacing traditional batteries that contain toxic metals.