Neutrons answer shampoo formulation puzzle

A research team at the Institut Laue-Langevin, the flagship centre for neutron science, has demonstrated quantitatively the science behind an anomaly in the surface tension of polyelectrolyte/surfactant mixtures. Their findings show that the dramatic increase in surface tension that affects the production of various pharmaceutical and cosmetic formulations is caused by the comprehensive aggregation of active ingredients. They have outlined a way to reload interfaces with functional components simply by tuning the way the materials are handled.

Surface tension is a property of liquids resulting from the cohesion of their molecules that helps them resist an external force. It is responsible for the shape of liquid droplets and the reason why insects can run on the surface of ponds.

Surfactants are substances that lower the surface tension of a liquid and can capture other substances, such as oil or grease in cleaning products. They are often combined with polyelectrolytes, made of long charged molecules, to improve the efficiency of detergents, wetting agents, emulsifiers, foaming agents, and dispersants in paints, shampoos and conditioners, and are used throughout the food industry. Also, the strong attractive interactions of surfactants with natural polyelectrolytes, such as proteins or DNA, play an important role in many biological processes, as well as in medical applications, such as drug and gene delivery.

The commercial production and performance of polyelectrolyte/surfactant mixtures, however, is affected by a peculiar phenomenon, first investigated in depth a decade ago. Whilst adding a surfactant to a polyelectrolyte solution initially causes the surface tension to decrease, as further surfactant is added the surface tension dramatically increases again. This feature, known as a ‘cliff edge peak’, is accompanied by a change in the appearance of the mixture, with the eventual loss of the cloudiness that is present as soon as the materials first interact.

From an industrial production point of view, this rise in surface tension reduces the performance of the additive, often requiring the introduction of further surfactant at extra cost. As a result, there is a lot of interest in understanding the interactions between these mixtures at the atomic level both in solutions and at surfaces. Of particular interest are the primary causes of the cliff edge peak and ways to prevent, lessen or delay its effects that could lead to more efficient formulations and reduce the effects of many pollutants in our environment.

To investigate this problem, Dr Richard Campbell (Institut Laue-Langevin), Dr Imre Varga (Eötvös-Loránd University, Hungary) and their co-workers looked at a system studied widely in the literature – an oppositely charged poly (diallyldimethylammonium chloride)/sodium dodecyl sulfate (Pdadmac/SDS) system.

The international research team, which also includes members from the UK and Sweden, used neutron reflectometry, a reflection technique used for measuring the composition and structure of thin films, to monitor the surface properties with respect to the slow generation of the cliff edge peak. The instrument used was the brand new FIGARO reflectometer (Fluid Interfaces Grazing Angles ReflectOmeter) at the Institut Laue-Langevin, which was constructed during the Institute’s innovative Millennium Programme. The researchers showed quantitatively for the first time that this striking feature in the surface tension results from the slow precipitation of particles into sediment from the aqueous solution. The precipitation depletes the solution and consequently the surface of its active ingredients, and also accounts for the loss of cloudiness observed.

As well as uncovering the reasons behind the rise in surface tension, the team were also keen to investigate methods to prevent its impact, which could directly benefit commercial applications. In the literature, researchers have suggested that the way these mixtures are handled could affect the nature of the material in the solution - a phenomenon called “non-equilibrium effects”.

To test whether the re-dispersion of surface-active material could actually switch off the cliff edge peak effect, the team carefully agitated a series of mixtures after the settling process had finished. A small mechanical stress provided just enough energy to re-disperse some of the sedimented particles and re-supplied the air/liquid interface with enough material to lower the surface tension once again.

“By approaching the problem in a different way, we have shown that the way you handle polyelectrolyte/surfactant systems can produce a variety of tuneable surface properties,” says Dr. Richard Campbell. “We hope that our findings will allow future industrial chemists across the pharmaceutical, detergency and cosmetic industries to generate better product output from their raw materials by learning to handle them in a smarter way, and create optimum surface properties on demand, rather than simply buying in more material to improve performance.”

There is hope also that this work can lead on to novel drug or gene delivery applications where one could apply an external stimulus to a stable biomacromolecule system in order to trigger the delivery of proteins or DNA to a given target.

More information: Re.:J. Phys. Chem. B, Article ASAP, DOI: 10.1021/jp2088803

Provided by Institut Laue-Langevin

Student brings home new expertise to answer question in antibiotic resistance

Working out the structure of a complex formed when a protein binds to DNA has proved to be key in understanding how an antibiotic-producing organism controls resistance to its own antibiotic, and may be an example of how other antibiotic producers regulate export to prevent self-toxicity.

The natural production of antibiotics by certain microorganisms is a complex and highly regulated process, not least because the organism making these compounds must protect itself from their toxic effects. Researchers at the John Innes Centre, which is strategically funded by the Biotechnology and Biological Sciences Research Council (BBSRC), have been studying the production of simocyclinone, produced by Streptomyces antibioticus, and in particular how the production of this potent antibiotic triggers an efficient pumping mechanism that exports the antibiotic from the cell.

Much of the work elucidating this protection mechanism has been carried out by Tung Le, a Vietnamese PhD student enrolled in the JIC's four-year rotation PhD programme. Tung, working under the supervision of Mark Buttner and David Lawson, showed that SimR, the protein Streptomyces antibioticus uses to regulate antibiotic export, can bind either to DNA or to the antibiotic itself, but crucially cannot bind to both. This means that when the antibiotic is around, SimR releases the DNA, which allows the expression of a gene that encodes a pump responsible for removing simocyclinone from the cell.

"This provides a mechanism that couples the potentially lethal biosynthesis of the antibiotic to its export, which has wider implications for resistance to clinically important antibiotics," commented Prof. Buttner. "However, we needed to know more detail about the interaction between SimR and DNA."

In this latest research, published in the journal Nucleic Acids Research, they show that the SimR protein has a novel 'arm' and that cutting off this arm unexpectedly weakened SimR binding to DNA. To determine the function of this arm, the researchers needed to work out the crystal structure of the protein bound to DNA, something which hadn't been achieved in Norwich before.

To overcome this skills gap, Tung won both a Korner Travelling Fellowship and an EMBO Short-Term Travel Fellowship to visit the University of Texas M.D. Anderson Cancer Center, home to one of the leading laboratories specialising in this technique. Tung spent three months working in the labs of Richard Brennan and Maria Schumacher, learning how to solve the structures of protein-DNA complexes. He was then able to apply this to his own project.

"I learned a lot and it was a great experience," said Tung. "The knowledge I brought back was not only useful for my project but will also be beneficial for others, and I feel very proud about that."

Usually, the SimR arm is unstructured, but in the presence of DNA they saw that it becomes ordered and binds into the minor groove of the DNA molecule. The crystal structure also shows how other parts of the SimR protein form sequence-specific interactions with a binding site in front of the export pump gene.

SimR is a member of a large family of regulatory proteins found in bacteria, and is the fifth one to have its structure solved when bound to DNA. The way these proteins recognise their target DNA sequences differs. This new example has wider implications, as a bioinformatic search of this family of regulators showed that many of them also have arms similar to the one characterised in this study.

Tung submitted his PhD thesis in early August, and his research has already produced four first-author papers. He is due to take up a post-doctoral position at the Massachusetts Institute of Technology in January.

"It can be very difficult for non-EU students to find the funding to study for a PhD in the UK, and so I was delighted to be offered a place on the JIC rotation programme. I am keen to encourage and help build relations between JIC and Vietnam. I was very happy to see the JIC is involved in joint work with Vietnam to sequence the genomes of different varieties of rice" said Tung, who left Vietnam at the age of seventeen for a bioscience career in the UK.

The researchers have also recently received a grant from the BBSRC to continue investigating the complexities of the regulation of antibiotic biosynthetic pathways, focussing on SimR and two other antibiotic-responsive transcription factors encoded in the simocyclinone biosynthetic cluster. This will establish the roles that the antibiotic plays in regulating self-resistance and its own biosynthesis. With the ever-growing problem of resistance, this kind of fundamental research is vital.

Provided by Norwich BioScience Institutes