Effective ingredients from marine biotechnology

Biotechnology encompasses the use of microorganisms to come up with novel active ingredients that fulfil two of the demands that are leading trends in the cosmetic industry: natural and sustainable. Besides, complex molecules can be obtained, which otherwise would be impossible due to technical or economic limitations. Our approach is to take advantage of biotechnology to develop cosmetic ingredients which are naturally occurring in non-genetically modified organisms, through sustainable production while preserving the environment, since there is no harvesting nor extracting from nature. Intertidal rocky shores are highly dynamic systems that are exposed to a combination of harsh factors, such as wave action, thermal and desiccation stress, UV exposure and nutrient depletion. The relative frequency of these fluctuations poses both physical and biochemical challenges to microorganisms that live in such environments.1 As a consequence, these intertidal inhabitants have developed several activities to protect themselves. Marine bacterial strains with different physiological and biochemical characteristics can produce exopolysaccharides (EPS) to protect themselves against the environmental stressors. Many of these EPSs present unique properties, and the search for new EPS-producing microorganisms is still promising.3 Organisms that inhabit intertidal areas suffer from desiccation when the tide recedes. The removal of water from cells, the storage of cells in the air-dried state, and the rewetting of air-dried cells impose physiological constraints that few organisms can tolerate. The removal of water molecules can cause mild, moderate, severe or extreme water deficit, depending on the quantity removed.2 Desiccation appears to induce the production of copious amounts of microbial EPS, presumably of a highly hygroscopic nature. Hyadisine (INCI name: Pseudoalteromonas Exopolysaccharides) is an exopolysaccharide obtained through biotechnology by fermentation of a marine bacterial strain that belongs to the genus Pseudoalteromonas sp. This bacterium was collected from a colony of mussels in an intertidal area in the Douarnenez bay. This bay is located in the Finistère breakwater (Brittany department, France) and covers a distance of more than 600 kilometres of coast that is some the most wild, with the spikiest reefs, not only in France, but also in the world. This coastline is declining fast as a result of sea erosion, especially during heavy storms coinciding with high tides, because of the winds blowing from the West. During high tides, these mussels are covered with water and gradually become exposed to desiccation and other drastic environmental changes at low tide. The organisms inhabiting this area must have had to develop some kind of protection against these stressors and in all probability marine exopolysaccharides play a precious role in it. Glucuronic acid is one of the main monomers of pseudoalteromonas exopolysaccharides. As hyaluronic acid (HA) is rich in the same monosaccharide, they were thought to have similar cosmetic properties.4 Skin contains about 50% of the total HA in a given organism. HA is produced mainly by fibroblasts and keratinocytes and thanks to its water retention capacity is able to maintain the extracellular space and facilitate the transport of ion solutes and nutrients.4 The levels of HA in the skin naturally decrease with each passing year, resulting in dermal dehydration and rhytide formation (wrinkles), a process accelerated by free radicals.5 Hyaluronic acid is extremely hydrophilic and biochemically retains water: hydrogen bonding occurs between adjacent carboxyl and N-acetyl groups to the extent that it retains up to 1000 times its weight in water. In the superficial epidermis, it acts as a humectant contributing to moisture content, and it decreases transepidermal water loss. Once absorbed into deeper dermis, it increases the water retention leading to a plumping effect in the skin.5 However, intertidal areas are not the only relevant sources for cosmetic researchers; we should also look into extreme cold environments. In fact, in recent years many microbiologists have focused their attention on the identification of Antarctic bacterial isolates. For almost six months a year, Antarctica does not receive solar energy, and when it does, it is not quantitatively comparable to that of other latitudes. This phenomenon gives rise to unique ecosystems, where evolution and biological adaptation processes have been developed without human intervention.6 Temperatures may sink to –40°C or lower in the winter months, and in contrast to the rather limited biodiversity of plants and animals, the microbial variety has been shown to be surprisingly diverse. Barren soil, rock landscapes, and numerous lakes at the edges of the continent, harbour a range of prokaryotes, which indicate that the extremely low temperatures are no obstacle to microbial colonisation.7 The sea pack ice and coastal attached ice around Antarctica has been found to contain abundant populations of bacteria. Approximately 30 new species of Antarctic bacteria have been described and many of them have not been found elsewhere in the world.8 Extremophiles are single-celled, prokaryotic organisms that thrive in extreme conditions, which would otherwise be toxic to life. They have adapted to their environments by optimising their metabolic processes and are believed to be some of the oldest forms of life on Earth. One type of extremophile are low-temperatureadapted microorganisms, which grow at temperatures around 0°C.9 Psychrotrophs are cold-loving extremophiles adroitly adapted to these environmental conditions and produce extracellular compounds that may present interesting properties for the cosmetic industry.9 Psychrotrophic species appear to be as common in sea ice as in the underlying seawater, with Pseudoalteromonas strains the most frequently isolated psychrotrophs.10 Antarctic Pseudoalteromonas strains protect themselves against cold environments by producing high amounts of extracellular polymeric substances, which could enhance bacterial growth and survival in these extreme conditions. When an organism cools below the freezing point of its tissue fluids, ice may form, which is usually lethal for cells. At lower tissue temperatures (<0°C), skin water freezing may start; ice-like water increases while decreasing useful water available for normal physiological processes.9 Cryoprotective products can modify the crystal’s morphology or even avoid their formation and so any damage to the lipidic bilayers. Extracellular polymeric substances produced by cold-loving extremophiles may present cryoprotective properties, which could help them to protect these organisms from freezing. Antarcticine (INCI name: Pseudoalteromonas Ferment Extract) is a glycoprotein obtained through biotechnology from a marine cold-loving extremophilic strain. During a scientific expedition to the Antarctic continent, the bacterial strain Pseudoalteromonas Antarctica NF3 was isolated from sludge collected from the base of a glacier located in the region of Inlet Admiralty Bay (King George Island, South Shetland Island). It was observed that the psychrotrophic bacteria produced large amounts of an extrapolymeric substance, which was mainly composed of proteins and sugars. This extrapolymeric substance had to provide some kind of protection to Pseudoalteromonas Antarctica NF3 so that it was able to thrive in such extreme cold environmental conditions. Therefore, it was thought that it could present potential benefits in cosmetics, which were confirmed by performing some in vitro and in vivo assays on it.

Dynamic vapour sorption (DVS) profile

The water retention profile of pseudoalteromonas exopolysaccharides was compared to hyaluronic acid (MW 106 Da) by the Dynamic Vapour Sorption technique (DVS). Experiments were performed in a TGA Q5000 SA and the obtained values were analysed by the software Universal Analysis 2000 version 4.5. Pseudoalteromonas exopolysaccharides retained more moisture and faster than HA, proving that they retain 12.7% more water at 95% relative humidity than the known moisturising agent.