The Possible Role of Perchlorates for Martian Life
The Possible Role of Perchlorates for Martian Life
(reprinted with permission from the publisher)
Journal of Astrobiology and Space Science Reviews, 1, 9-17, 2019

The Possible Role of Perchlorates for Martian Life

Joop M. Houtkooper, Ph.D.1, and Dirk Schulze-Makuch, Ph.D.2,
1 Center for Psychobiology and Behavioral Medicine, Justus-Liebig-University of Giessen, D-35394 Germany.
2School of Earth and Environmental Sciences, Washington State University, USA.


We previously suggested the presence of microbial organisms on Mars that use a mixture of hydrogen peroxide and water as an intracellular solvent as an adaptation to the harsh Martian surface environment (Houtkooper and Schulze-Makuch, 2007). These putative organisms would provide a consistent explanation of the observations by the Viking Biology Experiments, in contrast with a purely chemical explanation. The finding of surprisingly large amounts (0.4-0.6 wt%) of perchlorate salts in the Martian arctic soil (Hecht et al. 2009) by the Phoenix lander sheds a new light on these interpretations. An interesting property perchlorate salts share with hydrogen peroxide is their effectiveness as anti-freeze. Whereas the eutectic for H2O-H2O2 freezes at -56oC, the water-magnesium perchlorate eutectic is as low as -70oC. Therefore, Mg and Na perchlorate salts might well be employed as an antifreeze by hypothetical Martian microbes. The reason we still find H2O2 a more likely choice is the rapid evolution of O2 in the Gas Exchange (GEx) experiment, which is at variance with the stability of perchlorate under Viking's experimental conditions, and also the fact that H2O2 can be produced from only atmospheric constituents. At low temperatures on the Martian surface shortly before and after sunrise, the moisture level is close to saturation. This means any Mg-perchlorate would attract water. The apparent clumpiness of the soil, as well as what seemed drops of liquid water on one of Phoenix' struts, support that notion. The presence of perchlorates at a level of about 0.5% is surprising, because there is no straightforward chemical production mechanism for these compounds to account for the measured concentrations. The presence of perchlorates improves the habitability of the Martian surface, particularly for xerophilic organisms employing a mixture of hydrogen peroxide and water in their intracellular fluid.

Keywords: Astrobiology, Mars, Phoenix Lander, perchlorate salts, hydrogen peroxide-water hypothesis, anti-freeze, water activity

1. Introduction

The Phoenix Lander landed on Mars on 25 May 2008. During five months, the instruments on board explored the geology and climate of subpolar Mars and looked for the presence and dynamics of water ice, and explored if life could have ever arisen on Mars. Although the Phoenix mission was not a life detection mission per se, it looked for the presence of organic compounds and other evidence to support or discredit the notion of past or present life.

Therefore, the Phoenix Lander can be regarded as a successor mission to the Viking Landers of 30 years earlier. In 1976, these two spacecraft were the first two to successfully land on Mars. Their main purpose was to answer the question whether extant life on Mars could be detected. At that time, knowledge of the physical conditions on the Martian surface was scant, so that the designers of the Viking biology experiments had to work from what was known about life in dry and cold environments on Earth. Not surprisingly, the results of the Viking biology experiments were puzzling. No complete consensus about the meaning of the Viking results was reached among the researchers involved in the design of the experiments, although the majority opinion was that one or more highly oxidizing compounds in the Martian regolith were mainly responsible for the results (Klein, 1978, 1999). A different opinion was voiced by the researchers reponsible for the Labeled Release experiment (Levin and Straat, 1979, 1981; Levin, 1997, 2010), who favored a pro-life interpretation.

A number of hypotheses have been put forward about the composition of the Martian soil and its apparent oxidizing properties: Evolution of O2 upon humidification was suggested to result from one or more reactive chemical species such as ozonides, superoxides, and peroxides (Oyama and Berdahl, 1977). Proposed inorganic oxidants include metal oxides such as Fe- and Ti-oxides (Quinn and Zent, 1999) and superoxide ions or radicals (Yen et al. 2000; Zent et al. 2006). Yet, a conclusive identification of the oxidizing compounds and their productive process is still lacking (Atreya et al. 2006; Bullock et al. 1994; Crawford et al. 2003; Horowitz, 1986; Levin, 2001; Ponnamperuma et al. 1977; Quinn and Zent, 1999; Tsapin et al. 2000; Yen et al. 2000).

Given the inconclusiveness of a purely chemical explanation, Houtkooper and Schulze-Makuch (2007) put forward the hydrogen peroxide-water hypothesis, entertaining the possibility that extant life on Mars is based on a mixture of hydrogen peroxide and water in their intracellular fluid. This interpretation would have a number of attractive properties and provide a viable alternative to the chemical explanation of many of the Viking results. The recent findings by the Phoenix Lander, especially that the Martian soil contains substantial amounts of perchlorate salts, shed new light on the possibilities for extant Martian life.

2. The H2O2-H2O Hypothesis

The H2O2-H2O hypothesis (Houtkooper and Schulze-Makuch, 2007) turns out to fit the results of the Viking Biology experiments remarkably well, in contrast to a purely chemical explanation (Schulze-Makuch et al. 2009). Particularly, the evolution of oxygen upon moisturizing the soil in the Gas Exchange (GEx) experiment (Oyama and Berdahl, 1977) and the lack of detectable organics (Biemann et al. 1977) are to be expected if the Martian soil contains organisms of which the intracellular fluid consists of a mixture of hydrogen peroxide and water, instead of only (salty) water as is the case in terrestrial life.

Why would organisms evolve to include hydrogen peroxide in their intracellular fluid? The physical and chemical properties of H2O2-H2O mixtures would be beneficial for life to cope with a harsh Martian near-surface environment. Any organisms on Mars would have been subject to evolutionary pressures, as the Martian surface became colder, dryer, and subject to higher radiation levels, to develop an adaptation to H2O2, similar to how life on Earth adapted to high concentration of free molecular oxygen about 2.5 billion year ago.

The Martian atmosphere contains small concentrations of H2O2 (Encrenaz et al. 2004; 2008). These are in the 10s of ppb range, and may be created by triboelectricity in dust devils (Atreya et al. 2006), but are not in sufficient quantity to explain the Viking results.

H2O2 shows superior properties as an antifreeze, a source of energy and oxygen, and as a scavenger of water vapor.

The H2O2-H2O eutectic mixture (61.2% H2O2) freezes at -56.5oC and tends to supercool below that temperature (Giguère and Secco, 1954). The mixture is hygroscopic because of its low water activity and is also a source of oxygen and energy. The biological compatibility of hydrogen peroxide has to be considered even though H2O2 is mostly known as a desinfectant. H2O2 is used for a variety of purposes, such as a messenger molecule in the biochemistry of Earth's organisms (Rhee, 2006). H2O2 is produced in the peroxisomes of most eukaryotic cells (De Duve, 1969) and the microbe Acetobacter peroxidans uses it in its metabolism (Tanenbaum, 1956). While H2O2 is often employed in defense mechanisms, most organisms which come into contact with H2O2 protect themselves with scavenging enzymes such as catalase. The most eye-catching use of H2O2 is in insects such as the bombardier beetle Brachinus crepitans, which produces a 25% solution of H2O2 to produce steam explosions in a chitinous chamber in its posterior, which can be directed at a pursuing predator (Eisner, 2003).

Ambient conditions on Mars are often deemed adverse to the existence of life. Mars is considered to be too cold and too dry, the soil too oxidizing, the atmosphere too thin, the solar radiation containing too much UV and there is much hard radiation from solar and cosmic origin because of the lack of a magnetosphere. However, some Earth organisms show a remarkable resistance to UV and to various types of irradiation, and adaptations to oxidative stresses and irradiation involve many of the same resistance responses in microorganisms.

The physical properties of mixtures of H2O2 and H2O mean that the low temperatures and the low water content of the atmosphere are conditions that allow the organisms with such mixtures in their intracellular fluid not to freeze and to be able to scavenge water vapor from the atmosphere. While H2O2-H2O based life seems to be possible under Martian ambient conditions, H2O2 as a cellular component has its disadvantages as well: H2O2 is a reactive compound, although less so at lower temperatures, and the organisms may need an active stabilizing mechanism to keep the organic cellular components intact. The organisms would also need a mechanism to produce their own H2O2. For this, the H2O2-producing bacterium Lactococcus lactis and Brachinus crepitans provide an analog.

The H2O2-H2O hypothesis implies that the addition of water vapor at a relatively high temperature could only be withstood by the organisms for a short time, as they perished due to hyperhydration (Houtkooper and Schulze-Makuch, 2007). Thus, the evolution of oxygen in the GEx experiment is explained by the high oxidative content of the organisms. The lack of detected organics by the Viking GCMS could be the result of autooxidation of the organisms as these were gradually heated. Especially heating together with perchlorate salts would result in combustion to CO2 and H2O of any organics present. And indeed, the production of CO2 was detected by the Viking GCMS (Navarro-Gonzalez et al. 2006).

3. Results from the Phoenix Lander

Major results of the Phoenix Lander mission are the presence of water ice (Smith et al. 2009) with an ice table being within a few cm of the Martian surface in the subpolar region and the presence of calcium carbonate (Boynton et al. 2009). In addition, the Phoenix mission characterized the soluble chemistry of the Martian soil, revealing the alkalinity of the soil with a pH of 7.7 +/- 0.5 and the detection of surprisingly large amounts (0.4-0.6 wt %) of perchlorate salts in the Martian subpolar soil (Hecht et al. 2009). The presence of perchlorates was primarily detected by the Wet Chemistry Laboratory of the MECA package on board the Phoenix Lander (Kounaves et al. 2009), and was also confirmed by the Thermal and Evolved-Gas Analyzer (TEGA).

The Wet Chemistry Laboratory (WCL) detected the perchlorates by using an "ionspecific" electrode for different anions, apart from ClO4-, also for NO3-. However, the presence of a significant concentration of ClO4- overwhelms the response for NO3-, so that the biologically significant nitrate anion could not be detected (West et al. 1999; Kounaves et al. 2009). The detection of perchlorates may also shed new light on the interpretation of the Viking biology experiments.

4. The Role of Perchlorates: Anti-freeze and Water Activity

As an analog to Mars, perchlorate salts have also been detected on Earth in hyperarid soils (Fig. 1). For instance, Ericksen found that the soils of the Atacama desert contain 0.03-0.6 wt% perchlorate (Ericksen, 1981). Perchlorate is thought to be produced by atmospheric electricity or photooxidation (Miller et al. 2006). For Mars, the scarcity of atmospheric oxygen could be a problem.

Fig. 1: Top: Halite crusts in the very dry core of the Atacama desert in Chile, which are colonized by cyanobacteria (inset) that live within the rocks and take advantage of the hygroscopic properties of the mineral to obtain liquid water from the atmosphere. Any organisms organisms utilizing a water-hydrogen peroxide or perchlorate solution would be hygroscopic as well and could in principle attract water from the Martian atmosphere. Bottom: Chloride-bearing deposits on Mars (bright) which could have similar properties as the salt crusts in the Atacama desert and could provide a habitable niche for well-adapted microorganisms. Top picture and inset are courtesy of Jacek Wierzchos from the Institute of Natural Resources, CSIC, Spain. Bottom picture is credit to NASA/JPL/University of Arizona.

An interesting property perchlorate salts share with hydrogen peroxide is their effectiveness as anti-freeze. Whereas the eutectic for H2O-H2O2 freezes at -56oC, the water- magnesium perchlorate eutectic temperature is as low as -70oC. Therefore, as Mg and Na are prevalent cations in the Martian soil, their perchlorate salts might well be employed as an antifreeze by hypothetical Martian microbes. Although perchlorates are known as powerful oxidants, there is no perchlorate known to release oxygen upon moisturizing at the Viking experimental temperature of about 10oC as was observed during the GEx experiments of the Viking mission. Thus, we still consider H2O2 a more likely choice for the agent causing the rapid evolution of O2 in the GEx experiment. In addition, H2O2 has the advantage that it can be produced from atmospheric constituents only. What could be another role of perchlorates in addition to allow liquidity at lower temperatures? Like some terrestrial plants living in extremely arid conditions, Martian organisms may employ perchlorate salts to scavenge water from the atmosphere. Both H2O2-H2O mixtures and concentrated solutions of perchlorate salts have very low water activity (down to approximately 0.5 at the respective eutectics). The presence of perchlorate salts on their surface would enable organisms to pump water into their cells at relatively modest energy cost.

5. Discussion

Xerophilic organisms on Earth are often also halophilic, occurring in hypersaline solutions (Grant, 2004). Organisms on Earth evolved a number of organic (e.g., glycerol) and ionic (KCl) solutes to obtain osmotic balance with the environment. The adaptation to H2O2 as a solute which is compatible with the cell content, would for Martian organisms enable growth at very low water activity, comparable to that in concentrated perchlorate solutions. The diurnal temperature cycle on the Martian surface is concomitant with a water saturation cycle in the local atmosphere. At low temperatures shortly before and after sunrise, the moisture level is close to saturation. For example, ground fogs were observed by the Phoenix lander. This means any Mg-perchlorate would attract water. The clumpiness of the soil, which posed such a problem for getting a proper sample into the TEGA instrument of Phoenix (Fig. 2), as well as what seemed drops of liquid water on one of Phoenix' struts (Fig. 3), support that notion.

Fig. 2: NASA's Phoenix Lander's Surface Stereo Imager took this image on Sol 14 (8 June 2008). It shows two trenches dug by Phoenix's Robotic Arm. Soil from the right trench, informally called "Baby Bear", was delivered to Phoenix's Thermal and Evolved-Gas Analyzer, or TEGA, on Sol 12 (6 June 2008). The following several sols included repeated attempts to shake the screen over TEGA's oven number 4 to get fine soil particles through the screen and into the oven for analysis. When analyzed, a sample from "Baby Bear" showed an oxygen release consistent with perchlorate on Sol 25 (20 June 2008).

Fig. 3: Robot Arm Camera images of a strut on Sols 8, 31 and 44. The two spheroids enclosed by the circle appear to merge with each other. The resulting spheroid moves to the right, that is downslope. For a discussion, see Renno et al. (2009).

The fact that the upper few cm of the soil is crusted (Smith et al. 2009), together with the nightly precipitation of water ice crystals (Whiteway et al. 2009) has also been found in a parallel study of the migration of moisture in the soil of the hyperarid core of the Atacama desert (Davis et al. 2009). The occurrence of substantial amounts of perchlorate salts, both in the Atacama and at the Phoenix landing site, raises the question of the mechanisms producing perchlorate in both places. The presence of nitrates could not be ascertained on Mars, but for the Atacama a correlational study of the presence of perchlorates, nitrates and biomass might elucidate possible mechanisms.

The presence of perchlorates at the Phoenix landing site, at a level of about 0.5% is surprising, because there is no straightforward chemical production mechanism for these compounds to account for the measured concentrations. The production of perchlorates by putative organisms is therefore an alternative possibility - this could be a case of organisms modifying their environment to serve their needs. The presence of perchlorates improves the habitability of the Martian surface, particularly for xerophilic organisms employing a mixture of hydrogen peroxide and water in their intracellular fluid.


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