Researchers engineer E. coli to produce new forms of popular antibiotic

In Science Advances, University at Buffalo researchers will report that they have managed to turn E. coli into tiny factories for producing new forms of the popular antibiotic erythromycin -- including three that were shown in the lab to kill drug-resistant bacteria.

    ter disks holding The white filantibiotics sit on petri dishes housing erythromycin-resistant Bacillus subtilis. The filter disks circled in red hold new forms of erythromycin created by University at Buffalo researchers, and the dark halo around them indicates that the drug has seeped out of the disk to kill the surrounding bacteria.

Like a dairy farmer tending to a herd of cows to produce milk, researchers are tending to colonies of the bacteriaEscherichia coli (E. coli) to produce new forms of antibiotics -- including three that show promise in fighting drug-resistant bacteria.

The research, which will be published May 29 in the journalScience Advances, was led by Blaine A. Pfeifer, an associate professor of chemical and biological engineering in the University at Buffalo School of Engineering and Applied Sciences. His team included first author Guojian Zhang, Yi Li and Lei Fang, all in the Department of Chemical and Biological Engineering.

For more than a decade, Pfeifer has been studying how to engineer E. coli to generate new varieties of erythromycin, a popular antibiotic. In the new study, he and colleagues report that they have done this successfully, harnessing E. coli to synthesize dozens of new forms of the drug that have a slightly different structure from existing versions.

Three of these new varieties of erythromycin successfully killed bacteria of the species Bacillus subtilis that were resistant to the original form of erythromycin used clinically.

'We're focused on trying to come up with new antibiotics that can overcome antibiotic resistance, and we see this as an important step forward,' said Pfeifer, Ph.D.

'We have not only created new analogs of erythromycin, but also developed a platform for using E. coli to produce the drug,' he said. 'This opens the door for additional engineering possibilities in the future; it could lead to even more new forms of the drug.'

The study is especially important with antibiotic resistance on the rise. Erythromycin is used to treat a variety of illnesses, from pneumonia and whooping cough to skin and urinary tract infections.

E. coli as a factory

Getting E. coli to produce new antibiotics has been something of a holy grail for researchers in the field.

That's because E. coli grows rapidly, which speeds experimental steps and aids efforts to develop and scale up production of drugs. The species also accepts new genes relatively easily, making it a prime candidate for engineering.

While news reports often focus on the dangers of E. coli, most types of this bacteria are actually harmless, including those used by Pfeifer's team in the lab.

Over the past 11 years, Pfeifer's research has focused on manipulating E. coli so that the organism produces all of the materials necessary for creating erythromycin. You can think of this like stocking a factory with all the necessary parts and equipment for building a car or a plane.

With that phase of the research complete, Pfeifer has turned to the next goal: Tweaking the way his engineered E. coli produce erythromycin so that the drug they make is slightly different than versions used in hospitals today.

That's the topic of the new Science Advances paper.

The process of creating erythromycin begins with three basic building blocks called metabolic precursors -- chemical compounds that are combined and manipulated through an assembly line-like process to form the final product, erythromycin.

To build new varieties of erythromycin with a slightly different shape, scientists can theoretically target any part of this assembly line, using various techniques to affix parts with structures that deviate slightly from the originals. (On an assembly line for cars, this would be akin to screwing on a door handle with a slightly different shape.)

In the new study, Pfeifer's team focused on a step in the building process that had previously received little attention from researchers, a step near the end.

The researchers focused on using enzymes to attach 16 different shapes of sugar molecules to a molecule called 6-deoxyerythronolide B. Every one of these sugar molecules was successfully adhered, leading, at the end of the assembly line, to more than 40 new analogs of erythromycin -- three of which showed an ability to fight erythromycin-resistant bacteria in lab experiments.

'The system we've created is surprisingly flexible, and that's one of the great things about it,' Pfeifer said. 'We have established a platform for using E. coli to produce erythromycin, and now that we've got it, we can start altering it in new ways.'

What are cranial neuralgias, facial pain, and other headaches?

Neuralgia means nerve pain (neur= nerve + algia=pain). Cranial neuralgia describes inflammation of one of the 12 nerves that supply the motor and sensation function of the head and neck. Perhaps the most commonly recognized example is trigeminal neuralgia, which affects cranial nerve V (the trigeminal nerve) and can cause intense facial pain.

What causes tension headaches?

While tension headaches are the most frequently occurring type of headache, their cause is not known. The most likely cause is contraction of the muscles that cover the skull. When the muscles covering the skull are stressed, they may become inflamed, go into spasm, and cause pain. Common sites include the base of the skull where the trapezius muscles of the neck insert, the temples where muscles that move the jaw are located, and the forehead.

There is little research to confirm the exact cause of tension headaches. Tension headaches occur because of physical or emotional stress placed on the body. For example, these stressors can cause the muscles surrounding the skull to clench the teeth and go into spasm. Physical stressors include difficult and prolonged manual labor, or sitting at a desk or computer for long periods of time concentrating. Emotional stress also may cause tension headaches by causing the muscles surrounding the skull to contract.

What are the symptoms of tension headaches?

Common presentation of tension headaches includes the following:

• Pain that begins in the back of the head and upper neck and is described as a band-like tightness or pressure. It may spread to encircle the head.
• The most intense pressure may be felt at the temples or over the eyebrows.
• The pain can vary in intensity but usually is not disabling, meaning that the sufferer may continue with daily activities. The pain usually is bilateral (affecting both sides of the head).
• The pain is not associated with an aura (see below), nausea, vomiting, or sensitivity to light and sound.
• The pain occurs sporadically (infrequently and without a pattern) but can occur frequently and even daily in some people.
• The pain allows most people to function normally, despite the headache

• How are tension headaches diagnosed?

  The key to making the diagnosis of any headache is the history given by the patient. The health care professional will ask questions about the headache to try to help make the diagnosis. Those questions may include learning about the quality, quantity, and duration of the pain, and asking about any associated symptoms. The person with a tension headache will usually complain of pain that is mild-to-moderate, located on both sides of the head, described as a tightness that is not throbbing, and not made worse with activity. There will be no associated symptoms like nausea, vomiting, or light sensitivity.

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Biomedical sensors for disease detection made simple

Researchers have made a new protein detection platform using low-cost plastic and paper substrates. Their work could help reduce the cost and improve the accuracy of infectious disease diagnosis, they say.

Researchers in Korea have succeeded in making a new protein detection platform using low-cost plastic and paper substrates. Their work could help reduce the cost and improve the accuracy of infectious disease diagnosis.

Healthcare researchers are increasingly focused on the early detection and prevention of illnesses. Early and accurate diagnosis is vital, especially for people in developing countries where infectious diseases are the leading cause of death. One way to achieve early detection is by developing simple biomedical sensors.

One challenge, however, is to ensure that biomedical sensors are environmentally stable, disposable and cost-effective. Current biosensors are made of glass or very expensive. substrates that make it difficult to broaden their use and mobility in diagnostic and research areas.

Tackling the issue of expensive substrates, a research team at the Korea Advanced Institute of Science and Technology (KAIST) has developed a new protein detection platform that can be used with paper substrates. Not only is a paper substrate incredibly flexible and simple to handle, it is also 1,000 times less costly than the traditional glass substrate and 100 times less expensive than plastic substrates.

Called "initiated chemical vapour deposition" (iCVD), this process can cover surfaces without damaging substrates and can successfully immobilise proteins (such as fluorescent proteins or antibody fragments) that may indicate the presence, for example, of pathogens. Unlike previous methods, the iCVD process does not require solvents and therefore is much better at binding to different surfaces without degrading them.

So far, the KAIST team has successfully used the iCVD process not only on traditional substrates like glass, but also on flexible and low-cost alternatives such as paper and polyethylene films. The researchers hope this new technology will provide a cost-efficient platform to reduce the manufacturing costs of protein biochips. Further research will continue in 2015, they say, to improve the performance and operation of the paper-based biochip.