In 1981, a paper was published by Weiss [286] that put the Clay hypothesis front and center among considerations on the problem of the origin of life. With phyllosilicates as models, the notion of replication, i.e., the spontaneous self-multiplication of an information carrier, can be proven to be a common property of some macromolecular systems. Mistakes in replication and feedback along with environmental effects may result in mutants with higher or lower replication rates, thereby facilitating evolution. Taking these findings into consideration, the question of whether or not chemical evolution resulted directly in the nucleic acid/protein system, i.e., the genetic basis shared among all living systems recognized thus far, has to be answered. It seems possible that as an initial step, a much simpler replicating system had developed: such a system may subsequently have experienced the evolution of replicating systems, producing the final nucleic acid/protein system. The principle of replication and self-multiplication is not restricted to the nucleic acid/protein system; it is a more general property of distinct macromolecular systems. More primitive forms of life or types of protolife, therefore, must also be discussed in the context of the origin of life. Perhaps the question of how nucleic acids and proteins were formed and aptly selected in the course of chemical evolution has been incorrectly phrased. Both might have developed in the course of the evolution of replicating systems. The highly expanding clay minerals are an excellent replicating system model; in the course of an immense number of replications, they might have experienced evolution and selection, forced on the system by the environment, in just the manner expected for the most primitive forms of protolife. The paper had many graphs and diagrams and made the case for the Clay hypothesis. It was well written and was very convincing, but no methods were described.

When the Armin Weiss paper was published, John Lewis and Hyman Hartman immediately thought that a Gordon Conference on the Origin of Life should be organized, so the Gordon Conference on the Origin of Life was begun in 1982 and has flourished since then. Lewis managed the planetary section, and Hartman coordinated the rest, concentrating on Clays and the Origin of Life with an emphasis on the Weiss paper. Hartman invited clay chemists that he knew and hoped that they would critically consider his claims. There was an added bonus for him in that he met Graham Cairns-Smith for the first time at this Gordon Conference. The conference was a great success, as the clay chemists were impressed with Armin Weiss, so Hartman proposed to meet in Glasgow at the University where Cairns-Smith was a lecturer. Having received funding from NASA, he met with other attendees in Glasgow for what was a set of talks that were to be written up for a book that would be published by Cambridge University Press. There were 26 speakers. This resulted in the book: Clay Minerals and the Origin of Life, edited by Cairns-Smith and Hartman [385].

The Glasgow meeting was held in 1983. It was thought that all of the chapters would be written by 1984. There were 25 authors of the book, and it was published in 1986, three years after the meeting. Why did this happen? It happened because there was a problem, and that was that the 26th speaker was Armin Weiss. The whole book was designed to culminate with Armin Weiss’ contribution to the book, this time providing the experimental details so that the experiments could be repeated in other laboratories. For two years, the editors asked for his chapter, and each time, another disaster occurred that prevented him from finishing his chapter. Finally, someone went to Munich and checked the references in his 1981 paper and found that they were irrelevant. Arrhenius et al. [287] were forced to send a letter to Angewendte Chemie indicating that there were troubling questions concerning Armin Weiss’s paper. In a paper published in 1981 in Angewandte Chemie, Professor Armin Weiss described observations with potential importance for the origin and early evolution of life [287]. This paper has been widely cited since it claimed experimental proof of clay mineral replication, mutational variability in replicating clay crystals, and specific catalytic effects. The results have been invoked in support of the idea that an inorganic evolution through natural selection preceded the evolution of organic life. The information presented in the 1981 Weiss paper is altogether inadequate for others to repeat the key experiments that are said to be “proof of replication”. A single reference in this section reads: “Unpublished results: published in part in G. Mai, Dissertation, Universitat München 1969; P. Brunner, ibid. 1978; S. Fritz, ibid. 1978”. Arrhenius et al. [287] were unable to find the first and third of these theses, while the second had no information that they deemed to have specific relevance to clay replication, nor were repeated attempts to obtain the experimental protocol from Professor Armin Weiss successful. Armin Weiss, like King Ludwig, had built castles that were based on his imagination, but Weiss’ castles were in his head and on paper. This was to be, in the context of a scientific project, the worst possible scenario: specifically, an experimental paper makes a central claim and then does not publish the methods in detail so that others can repeat the experiment. Hartman was furious that he had invested time and NASA money in what had turned out to be a fraud. The damage was done. Clays were no longer at the center of the discussion. The recovery would be long and painful.

Hartman met Gerry Soffen in Cambridge, MA, who, at that time, was a project scientist for NASA’s Viking program of Mars landers, the first successful mission to operate unmanned experiments on the Martian surface. As such, he supervised all scientific experiments performed by both landers, managing more than 70 scientists around the USA. Hartman told him about the Clay hypothesis, and he suggested collaborating with a group working at NASA Ames led by Jim Lawless (smectites) and Sherwood Chang (kaolinites). They were working on clays, especially on their interaction with peptides. Hartman joined Jim Lawless’ group and spent three summers there (1978, 1979, and 1980). He spent three summers adding coenzymes, especially pyridoxal phosphate (vitamin B6), to clays. He began with the serine + pyridoxal phosphate reaction. The enzyme is called serine dehydratase (Scheme 12).

The enzyme is composed of a (protein) apoenzyme + a coenzyme (pyridoxal phosphate). Could the clay substitute for the protein (apoenzyme) (Scheme 13)?

The clay used was Wyoming bentonite—a montmorillonitic clay that was developed from the alteration of volcanic ash in seawater. The pyridoxal phosphate was added to the Mt in a buffered solution, and the reaction was monitored at various temperatures. The experiment worked beautifully—the Mt did substitute for the protein. Then, Hartman ran a control, which entailed the removal of the Mt and addition of the supernatant to serine and pyridoxal phosphate, and it worked as well. He learned that E.E. Snell, a biochemist, had found that serine underwent this reaction using pyridoxal phosphate and metal ions such as Cu²⁺ and Al³⁺. Serine underwent rapid transamination by pyridoxal phosphate in an Al³⁺- or Cu²⁺-catalyzed reaction. There was no need for clay. The bentonite was not a pure compound, and there were ions in the supernatant not attached to the clay. While Hartman was conducting these experiments, Max Mortland visited his laboratory. He was intrigued by these experiments. Mortland decided to focus on enzyme transaminase with the apoenzyme as a protein and pyridoxal phosphate as a coenzyme. Hartman decided not to publish this paper, as Snell had already published his results [386].

Mortland [387] showed that glutamic acid was selectively deaminated through a mixture of pyridoxal phosphate (PLP) and Cu²⁺-smectite. Ammonia was formed together with α-ketoglutaric acid (Scheme 14).

This reaction was far more active with Cu²⁺-smectite compared to when the Cu²⁺ cation was simply in solution. Likewise, Cu²⁺-kaolinite, vermiculite, and faujasites X and Y showed some activity, though at a significantly lower rate, indicating that their activity was probably confined to external surfaces only, whereas that of the smectites involved catalysis in the interlayer spaces. Following the addition of a strong ligand (o-phenanthroline) to the reaction, the reaction rate dropped, clearly indicating that coordination of the reacting species with Cu²⁺ on the smectite was essential. The suggested reaction pathway consists first of the formation of a Schiff base between the amino acid and pyridoxal phosphate, followed by complexation with Cu²⁺ at the smectite surface. Next, hydrolysis of the complex to NH₃ and the α-keto acid occurs, or transamination takes place to form pyridoxamine phosphate, which is subsequently deaminated by Cu²⁺-smectite. The pyridoxal phosphate-Cu²⁺-smectite behaves like a pseudo enzyme, with the clay structure replacing the apoenzyme. Mortland thanked Drs. H. Hartman of the Massachusetts Institute of Technology and J. G. Lawless of the Ames Research Center (NASA), Moffett Field, CA, for sharing their results on pyridoxal phosphate–clay interactions with serine. The reaction mechanisms in both the glutamic and serine cases with pyridoxal phosphate and Cu²⁺ were identical. Mortland’s paper then stimulated others to explore the catalytic properties of clay minerals with coenzymes and metal ions.

The catalytic activity of smectites in numerous chemical reactions led various researchers to study their possible involvement in the origin of life [385,388,389]. Smectite-catalyzed amino acid polymerization has been shown. These initial results led researchers to think about clays playing the role of enzymes in biological systems. Several researchers, in particular, Mortland [387], Boyd and Mortland [390,391], Siffert and Naidja [392], and Naidja and Siffert [393], studied the catalytic activity of smectites in some simple biochemical reactions. The research by Naidja and Siffert [393] was aimed at learning more about the role of clay minerals as catalysts in certain biochemical reactions. They looked at the oxidative decarboxylation reaction of isocitric acid, which forms the third step of the Krebs cycle. The isocitric acid oxidative decarboxylation reaction was studied with and without homoionic Na⁺, Mn²⁺-, and Cu²⁺-Mt present. The catalytic activity of the Mt was a function of the nature of the interlayer exchangeable cation. Isocitric acid was converted to α-ketoglutaric acid in the presence of Na⁺-Mt, where Na⁺ does not produce a complex with the isocitrate anion (Scheme 15).

Instead, with Na⁺-Mt, the isocitrate anion was oxidized and converted to the α-ketoglutarate anion. The interlayer exchangeable Na⁺ ions first permitted the intercalation of isocitrate in the interlayer spaces, which did not result in the formation of complexes with isocitrate. Divalent cations (Mn²⁺ and Cu²⁺) were removed from the interlayer spaces, which resulted in the formation of “external complexes” with the isocitrate anions. However, the reaction rate was far lower than with the enzymatic system (isocitrate dehydrogenase enzyme and nicotinamide adenine dinucleotide phosphate coenzyme). The outstanding transition metal involved in the origin of life is iron, especially Fe²⁺ and Fe³⁺ in the iron clays found on Mars and carbonaceous chondrites. This was clear to Hartman when he convinced Richard Frankel at the Magnet Laboratory (MIT) to investigate the coenzyme flavomononucleotide (FMN) and Fe-clay.

Mortland et al. [394] observed that the adsorption isotherms and UV–visible and Mossbauer spectroscopic data suggested specific interactions between FMN and Fe³⁺-smectite. The highest adsorbed FMN amount was determined to be 0.3 mmole/g Fe³⁺-smectite, yielding a 1:1 molar proportion of Fe³⁺ to FMN. This pointed to the presence of a Fe³⁺–FMN complex on the smectite surface. Other homoionic smectites (Cu²⁺, Zn²⁺, and Ca²⁺) showed less adsorption and less obvious specific interaction. It is likely that the observed interaction takes place between the Fe³⁺ on the smectite and the phosphate group on the sugar moiety of FMN. Since this study, the emphasis has been on transition-state metals rather than coenzymes. However, the behavior of the transition-metal ions in and on the clay lattice is significantly different from their behavior in solution.

In 1980, Hartman went to MIT, where he joined John Lewis in the Earth Atmosphere and Planetary Sciences. He had received a grant from NASA to work in the field of Clays and the Origin of Life and began working with John Lewis on carbonaceous chondrites. Becker and Epstein [239] performed solvent extractions with CCl₄ and CH₃OH on the carbonaceous chondrites Murray, Murchison, Orgueil, and Renazzo. Around 2–10% of the total carbon in these meteorites could be extracted using normal methods, mostly in CH₃OH. The extracts from Renazzo exhibited isotopic ratios indicating that they consisted primarily of terrestrial organic matter, with smaller amounts from indigenous organics. The CH₃OH-soluble organic matter from Murchison and both untreated and HF-treated Murray showed δ¹³C values between approximately +5 and +10‰ and δ¹³N values between approximately +90 and +100‰, both of which are substantially higher than those determined for the bulk meteorites. Likewise, the Orgueil CH₃OH extract exhibited a δ¹⁵N value considerably higher than that of the residual organic matter. δD values between +300 and +500‰ were observed for the CH₃OH-soluble organic matter. These results for C, H, and N isotopes make it extremely doubtful that the CH₃OH-soluble components were derived from, or simply related to, the insoluble organic polymer present in these meteorites. Organic matter soluble in CCl₄ contained almost no N and had lower δ¹³C and δD values compared to the CH₃OH-soluble compounds. Either large isotopic fractionations for C and H occurred between different soluble organic compounds, or the less polar compounds were in part of terrestrial origin. They concluded that the soluble organic material (amino acids, etc.) was probably introduced to the meteorites, along with high-δ¹³C carbonates, during a hydrothermal event that produced hydrous silicates in CI1 and CM2 meteorites. Hartman et al. [249] indicated that organic compounds found in carbonaceous chondrites could be divided into three different fractions. The first fraction, which is insoluble in chloroform and methanol, has a portion that is of interstellar origin. The remaining two fractions, which are chloroform-soluble hydrocarbons and methanol-soluble polar organics, were assumed to have been formed on a planetoid body. They suggested that the polar organic compounds, i.e., amino acids, were formed near the surface of the chondrite through the radiolysis of hydrocarbons and ammonium carbonate in a liquid water environment. Certain hydrocarbons could have been produced through a Fischer–Tropsch mechanism inside the chondrite body. The ferrous ion was thought to function as a protection against reverse reactions. The concurrent formation of Fe-rich clays and polar organics may be an indication of events associated with the origin of life on Earth. During the outgassing of the early Earth, events similar to those observed in the carbonaceous chondrites must also have taken place. In particular, in recent years, a new model for the origin of life has been proposed. It emphasizes the role of liquid water, Fe-rich clays, and ultraviolet light in the production of biologically significant molecules. Thus, although the radiation involved is not ultraviolet, carbonaceous chondrites may provide invaluable indications of the early steps that led to life on pre-Archean Earth.

In the summers that Hartman spent at NASA Ames, discussions were centered on how Viking was a failed mission, so he told the exobiology division what he thought about Mars and the Viking Mission, which was that they were looking only for organic molecules and completely ignoring the clays on Mars. They had set the laboratories of Sherwood Chang and Jim Lawless to study clays and paid no attention to what was happening in those laboratories. His ideas were published as an abstract at a meeting on Exobiology and Future Mars Missions [395]. To detect life in the Martian soil, the two Viking landers contained tests that were developed to search for respiration and photosynthesis. These two experiments (labeled release, LR, and pyrolytic release, PR) searching for life in the Martian soils provided positive results. However, no organic molecules were found in the soils of Mars. The explanation provided was that the inorganic compounds present in the Martian soil caused these results. The inorganic composition of the Martian soil was best modeled with a mixture consisting of 60–80% clay, iron oxide, and quartz, together with soluble salts such as halite (NaCl). The minerals most effective in replicating the PR and LR tests were Fe-rich clays. One theory considers clays to be the first organisms able to replicate, mutate, and catalyze and, therefore, to evolve. Clays were formed as a result of the weathering of rocks by liquid water. The distribution of ions, e.g., of Al, Mg, and Fe, was thought to act in a manner similar to the sequence of bases in DNA. The information was stored in the distribution of these ions in the octahedral and tetrahedral sites in the clay layers, and they might, similar to RNA and DNA, replicate. When the clays replicated, each clay layer would act as a template for the next layer. The ion substitutions in one clay layer would result in a complementary or similar pattern in the next clay layer formed on its surface. It was hypothesized that on the surface of replicating Fe-rich clays, CO₂ might react under light to form organic acids, e.g., formic or oxalic acid. If Mars had liquid water during a warm period in its past, clay formation would have been abundant, something that has since been proven to be the case. These clays would have been able to replicate and evolve until the liquid water disappeared as a result of the cooling of Mars. His suggestion was that they should study the Fe-rich clays of Mars. Since that time, Fe-rich clays have become central to the missions to Mars and to the origin of life on Mars.

In 1988, Hartman moved back to the University of California, Berkeley, where he met Garrison Sposito in the Soil Science Department. They began to study smectites using an atomic force microscope (AFM). The only path that he could envision at the time was to explore new experimental techniques such as Mossbauer spectroscopy and atomic force microscopy. For this purpose, Hartman et al. [396] purified Mt samples of Crook County and Silver Hill Illite and prepared the Na forms, which were subsequently studied under 80% relative humidity with an atomic force microscope. Direct imaging distinctly showed the hexagonal rings of basal oxygen atoms of the SiO₄ tetrahedra in the tetrahedral sheet in Mt. AFM might provide useful information at the molecular-scale resolution of clay mineral surfaces such as Mt containing adsorbed organic molecules. This period was followed by several years in which he was not involved in any research related to clay minerals and the origin of life.

Hartman returned to the clay world and decided to replicate clay minerals. It began when he spent the summer of 2005 with Dennis Eberl, who was with the USGS, Boulder, Colorado. They were motivated to explore the possibility that oxalic acid could catalyze the formation of clays from a gel. This was again inspired by the work of Becker and Epstein [239], who showed the coupling of clay formation on carbonaceous chondrites with dicarboxylic acids, amino acids, and other water-soluble organic chemicals. This experiment was run, and the results were analyzed by a group of clay chemists led by Hojatollah Vali (McGill University). The research was carried out by Dirk Schumann, a graduate student, which resulted in his PhD thesis and a paper in Astrobiology [154]. The possible role of clay minerals in the abiotic origin of life is the topic of ongoing debate that started several decades ago. The main focus is on the clay minerals detected in a class of meteorites called carbonaceous chondrites. These clay minerals were formed through aqueous alteration of anhydrous minerals, e.g., olivine and orthopyroxene, that are frequently found in the chondrules. Furthermore, a strong correlation exists between the existence of these clay minerals and the occurrence of polar organic molecules. Laboratory experiments have proven that at low temperature and ambient pressure, polar organic molecules, e.g., the oxalate ion observed in meteorites, are able to catalyze the formation of clay minerals. In this particular investigation, it was proven that oxalate was a strong catalyst in the formation of saponite, an Al- and Mg-rich trioctahedral clay mineral, from a silicate gel at a temperature of only 60 °C and ambient pressure. High-resolution transmission electron microscopy (TEM) of the synthetic saponite intercalated with octadecylammonium cations showed the presence of 2:1 layer structures with variable negative layer charge. The formation of these differently charged 2:1 layers within the saponite particles most likely took place independently. If polar organic molecules, such as oxalate, are able to catalyze the crystallization of clay minerals, which then can promote the formation of clay microenvironments and offer a large number of adsorption sites on the clay surfaces for other organic molecules existing in the solution, the interaction between the adsorbed molecules could result in polymerization, resulting in more complex organic molecules, such as RNA from nucleotides on early Earth. The major findings of this paper were twofold: (1) Oxalic acid was a catalyst of clay formation. This was suggested by B. Siffert in a chapter in Clay Minerals and the Origin of Life entitled “The role of Organic Complexing Agents” [397]. He stated, “A number of successful attempts to synthesize aluminum phyllosilicates at ambient temperature have been reported. In most cases organic aluminum complexes were used”. (2) The crystallization of these differently charged saponites most likely occurred independently. The fact that saponites with variable charge formed from the same gel has consequences for our interpretation of how life originated, as these 2:1 clay minerals most likely replicate via a process of template-catalyzed polymerization and transmit the charge distribution from one layer to the next layer. The reason that the second finding was so interesting was that Hartman had discussed his publication on the formation of clay minerals in soils with Isaac Barshad in the Soil Department at the University of California, Berkeley. He pointed out that his experience with clay formation in soils was that where conditions for clay formation were the same, the clays formed a diverse assemblage of different clays. The reason for his observation was that clay formation is catalyzed by seed clays [398].

The synthesis of clays in the years 2005–2006 took 3 months at 60 °C. This was not going to allow a lot of experiments to be performed, so Hartman searched for a simpler and shorter synthesis at temperatures lower than 100 °C. Then came the reason that he became the editor for Clays and the Origin of Life in the journal Life. He came across the paper by Kloprogge et al. [130] on the synthesis of smectite clay minerals, and on page 533, he read the following paragraph:

“Following a new approach, Vogels et al. (1995) synthesized saponites at 90 °C from a Si-Al gel and a solution containing urea and an M²⁺-nitrate [M²⁺ = Zn, Mg, Ni, Co] in only a few (5–20) h. Precipitation from homogeneous solution was induced by the slow hydrolysis of the urea, which resulted in a homogeneous release of hydroxyl ions. The octahedral divalent metal had a large influence on the characteristics of the saponite product, such as stacking order, surface area, and pore volume. With Mg the nucleation and growth were rather slow compared to saponites containing Co or Zn. After 20 h with Mg some gel was still present and even after 2 d almost no stacking was observed by transmission electron microscopy (TEM) in agreement with the absence of the (001) reflection in the XRD pattern. The results with different divalent cations indicated that stacking increased in the order Ni = Mg < Co < Zn. Both the surface area and pore volume increased as Zn < Co < Ni < Mg. The A1 distribution over the tetrahedral, octahedral, and interlayer sites was influenced by increasing the initial Si/A1 ratio in the starting gel from 5.7 to 39. Adjusting both the octahedral-sheet and tetrahedral-sheet composition by the choice of divalent metals or combinations of metals and the Si/A1 ratio offers the possibility to control properties like surface area, pore volume, and acidity” [399]. Hartman immediately began to synthesize Zn-clays using the method of Vogels et al. [399] in Roger Summon’s laboratory at MIT, and it worked. Hartman modified it by adding oxalic acid and NaOH, and this worked as well. He then contacted Marcelo Guzman at the University of Kentucky, and they began a collaboration with his graduate student Ruixin Zhou. The synthetic Zn-clays were characterized by Chris Matocha and Hojatollah Vali and his group (McGill University). The role of primordial metabolic networks, e.g., the reverse tricarboxylic acid (rTCA) cycle, and the coevolution of clay mineral catalysts in the origin of life is still not properly understood. Although prebiotic reactions from the rTCA cycle have been achieved through photochemistry on semiconductor minerals [400], the formation of clay minerals has been shown to be catalyzed by oxalate at low temperature and ambient pressure [154]. Zhou et al. [156] reported on the succinate-catalyzed crystallization of sauconite as a model for clay minerals using a photoproduced intermediate from central metabolism as an example. In addition, they showed that seeding induced nucleation at low temperatures, speeding up the crystallization process. Their results indicate that the coevolution of clay minerals and early metabolites on the Earth’s surface might have been accelerated by sunlight-induced photochemistry, which played a major role in the intricate interactions between rock surfaces and life at a geological timescale. The most important finding is that seeding increased crystallization. The catalytic power of the synthesized clay was validated by replicating the synthesis at 70 °C after adding a single (macroscopic) sauconite particle obtained from the 90 °C synthesized product to the starting gel. The fact that a single sauconite particle acted as a seed crystal to produce a larger amount of sauconite provides an excellent example of the self-catalytic power of clay minerals with respect to their nucleation and crystallization. The surface of the seed particle is thought to enable heterogeneous nucleation at lower temperature via a reduction in the activation energy for crystallization. The detected acceleration in crystallization by seeding does not merely depend on random events but is the result of clay surface interactions with chemical species present in the starting gel at 70 °C. The seed particle interaction with soluble free and complexed ions moving freely in the gel creates intermolecular forces that are required to produce the crystal lattice. The addition of a seed particle to the gel offers a pathway to direct a process that is then independent of random interactions.