Life in the Lab -- Proof of Concept (Long) Part 1
Tue, 11 May 1999 08:28:07 EDT

In this second part I will describe the research that provides the basis for
the consensus among biologists that life has been synthesized in the lab.
The references I will provide will by no means be complete, but they should
be enough to not only verify the veracity of my claim, but to also provide
anyone the basis for doing their own more extensive literature search. Also,
as I explained in the first part, while the significance this breakthrough
has for abiogenesis in general or the origin of life on earth is not yet
fully determined, it does serve as proof of concept that life can be derived
abiotically from simpler chemicals.

As I also pointed out in the first part, however, I have none of the types of
references that would be most convincing to certain key people. I wish I
did, to be perfectly frank. It would have been wonderful to have been able
to report that _Nature_, _Science_ and _Scientific American_ all ran news
reports and major articles describing how scientists in some lab had
announced that they had synthesized life, or to provide references to _New
York Times_, _Newsweek_ and _Time_ articles with screaming headlines like
"ARTIFICIAL LIFE MADE IN LAB". Neither can I provide the kinds of references
that Brian asked for, even though I looked for them. Outside of
popularizations, as far as I know no biologist or chemist has claimed in
textbooks, reference books or journal articles that life has been synthesized
in the lab, even though that is the general consensus. The reason of course
is very simple: most scientists are cautious about making bold claims and
even more cautious about accepting them. What I have found, however, are
books and articles like the one authored by Fox that Brian quoted from, that
seem to endorse the consensus without actually stating it or claiming that it
is true. If one reads between the lines the endorcement is obvious, but none
make any explicit claims.

For example, Albert L. Lehninger describes Fox's work in the 1970 edition of
his textbook _Biochemistry_ on pages 782 to 785. Though Lehninger never
quite states that they are alive, it is clear from his theoretical discussion
that even as early as the late Sixties Fox's proteinoid microspheres were
being recognized as possibly being alive. I was taught about Fox's
protocells in my undergraduate biochemistry classes in the early Eighties; my
professors accepted that they were at least technically alive. Stephen F.
Mason, a chemist, reported in his book _Chemical Evolution_ [Oxford:Clarendon
Press (1991)] that Fox's proteinoids could form microspheres that had
"proto-cellular attributes" (pg. 244) such as reproduction and enzymatic
activity (pg. 251); his discussion is matter-of-fact and distinctly
uncritical, though he does not himself claim that the microspheres are
genuinely alive. In his book _Evolution and the Myth of Creationism_ Timothy
M. Berra (a biologist) all but agrees with Fox that his microspheres are
alive (pg. 75). And so on. Again, no one that I could find actually comes
right out and says that Fox synthesized living cells in his lab, but nearly
all who discuss it appear to accept it.

So why do I claim that there is a consensus among biologists that life has
been synthesized in the lab, if I cannot produce even one citation that
supports such a claim? That in and of itself is an interesting story. I
shall try to be brief.

Early last year, I was approached by Sheldon Gottlieb, a retired biologist
formerly of the University of Southern Alabama, to join his team that was
working on writing a response to _The Creation Hypothesis_, edited by J. P.
Moreland [Downers Grove, IL:InterVarsity Press (1994)]. He specifically
wanted me to critique the chapter "Information and the Origin of Life" by
Walter L. Bradley (an engineer and materials scientist) and Charles B.
Thaxton (a chemist). After reading the chapter I declined, not because I
found their arguments unassailable (they had in fact made a number of
fundamental biochemical errors), but because their position was ultimately
based on information theory, something I knew nothing about. I informed Dr.
Gottlieb of this and included the statement, "Even if in the next three
months [the time before his writing deadline] I could manage to create life
in the lab, it would be no serious objection to their basic argument." He
responded, in part, to say that Fox had in fact succeeded in making life in
the lab.

At the time I was familiar with Fox's work and I had read some of his early
papers, but all I remembered were his proteinoids. So I told Dr. Gottlieb
that Fox's proteinoids were simply random polymers of amino acids without
catalytic activity and that they did not constitute living organisms. He
responded by reminding me that they formed microspheres, then informing me
that the microspheres had catalytic activity, that they could reproduce, they
could convert light into usable chemical energy and they could react to
external stimuli. He concluded by saying that the general consensus among
biologists was that Fox's microspheres were alive and could be considered
true protocells.

What he said sounded fantastic to me, so I simply responded by advising him
not to do a biological refutation of the chapter, but to get someone who knew
information theory, preferably a biological theorist. He thanked me for my
advice and we have not been in touch since. However, what he said about Fox
intrigued me, so I began to pursue it. I started by contacting all the
biologists I knew to ask them what they thought of Fox's work. I have to
admit that most of them were even less familiar with it than I was, but among
those who did know of it they agreed that Fox had synthesized life in the
lab. Every one of them, without dissent. That in and of itself made me
leery, so I broadened my scope to include textbook authors and college
professor that I did not know. This time nearly everyone whom I contacted
was familiar with Fox's work. There were a handfull that were unimpressed
and preferred the theories advanced by those who advocated the primacy of
nucleic acids over amino acids as the intitial abiotic carriers of
information, or the metabolic theories of Morowitz, but the majority accepted
virtually without reservation that Fox's microspheres were living protocells.

At this point most of you may be expecting me to reproduce a list of these
people to satisfy Brian's request, but I will not for two reasons. First of
all, most of these people refused to allow me to do so, saying that they do
not want to get involved in a debate with creationists. Secondly, however,
these were personal correspondences, not publications, so it would be
impossible for anyone to independently verify the truth of my claims without
contacting my correspondents directly. I want to keep these people friendly,
and I'm afraid I would alienate them if they each received a dozen requests
for verification.

Besides, as I explained in an earlier post, what matters is not how many
people claim that life has been synthesized in the lab, but whether their
claim can be supported by the scientific evidence. Hence late last year I
started doing an intense literature search for all papers on protocells,
proteinoid microspheres and Fox's work. It is by no means finished, and I
have only begun to read all this literature, but what I have so far
discovered convinces me that the consensus among biologists is correct.

To start off, let me give a list of books and review articles, then I'll move
on to the critical references. First the books:

Sidney Fox. _The Emergence of Life: Darwinian Evolution From the Inside_.
New York:Basic Books (1988) ISBN: 0465019250

Sidney W. Fox and Klaus Dose. _Molecular Evolution and the Origin of Life_,
Revised Edition. New York:M. Dekker (1977) ISBN: 0824766199

Duane L. Rohlfing and A. I. Oparin, editors. _Molecular Evolution:
Prebiological and Biological_. New York:Plenum Press (1972)

Sidney W. Fox, editor. _The Origins of Prebiological Systems and Their
Molecular Matrices_. New York:Academic Press (1965)

The first is the one Brian quoted from in his post; I have not seen it, but
from what I have heard of it, it sounds like a popularization. As such a
more scientifically rigorous book (that I have seen) would be the second one,
of which there is also an earlier edition (1972). It contains a general
discussion of the question of whether protonoid microsphere protcells are
actually a primitive synthetic organism or simply represent plausible
evolutionary precursor (pg. 250-2). The general conclusion was, first of
all, "that proteinoid microspheres are *not* contemporary organisms [emphasis
in original]." Secondly: "The essential defensible statement is that the
proteinoid microsphere is a plausible, or the most plausible, model for the
evolutionary precusor of the contemporary cell." Then later they said, "A
perhaps fuller assertion of the same kind is one that a proliferative (page
214) protoorganism has been synthesized in the laboratory." Fox and Dose
were confident that the experiments that had been done up to that time
"demonstrated that some of the evolutionary gap between the primordial and
contemporary cell has been bridged." In the summary of their chapter
"Interpreting Experiments: Proteinoid Microsystems" (pg. 241-67) they state:
"The unit of contemporary life is the cell. The unit of primitive life, by
back-extrapolation, was the protocell." And in a flowchart of the principle
stages of molecular evolution and the origin of life (pg. 350) they label the
formation of protoproteinous cells as the beginning of primitive life (in the
same chart they label the development of internal synthesis of coded
macromolecules as the onset of contemporary cellular life). All of this
would indicate that Fox and Dose accept that proteinoid microsphere
protocells are living systems, even if they qualify that acceptance with such
terms as "proto-" and "primitive". From this general discussion, however,
they use those terms to avoid the mistaken impression that protocells are
indistinguishable from modern contemporary cells.

The third and the fourth books are worth a look if as much for a historical
perspective as for scientific information. The third is a volume
commemorating the sixtieth birthday of Sidney W. Fox, the fourth contains
proceedings of the second of a series of meetings on the origin of life.
These books should be available through interlibrary loan if they are not in
your local library system.

The review articles (including letters and "editorials") follow. (Some of
these I haven't read yet, so I cannot vouch for their relevence.)

Fox SW. "My Scientific Discussions of Evolution for the Pope and His
Scientists." _The Harbinger_ 1997 May-June; 15(15):1. Accessible at
<> [In this
paper, Fox openly claims that he indeed created a living cellular system in
his lab, though again he is careful to explain that it is not a modern
cellular system.]

Fox SW. "Thermal synthesis of amino acids and the origin of life."
_Geochimica et Cosmochimica Acta_ 1996; 59:1213-4

Pappelis A, Fox SW. "Thermal peptides as the initial genetic system." In
Chela-Flores J and Raulin F, eds, _Chemical Evolution: Physics of the Origin
and Evolution of Life_. Dordrecht:Kluwer Academic Publishers (1996) pg.

Fox SW, Bahn PR, Pappelis A, Yu B. "Experimental retracement of terrestrial
origin of an excitable cell: Was it predictable?" In Chela-Flores J and
Raulin F, eds, _Chemical Evolution: Physics of the Origin and Evolution of
Life_. Dordrecht:Kluwer Academic Publishers (1996) pg. 21-32

Pappelis A, Fox SW. "Domain protolife: protocells and metaprotocells within
thermal protein matrices." In Ponnamperuma C and Chela-Flores J, eds,
_Chemical Evolution: Structure and Model of the First Cell_.
Dordrecht:Kluwer Academic Publishers (1995) pg. 129-32

Fox SW, Bahn PR, Dose K, Harada K, Hsu L, Ishima Y, Jungck J, Kendrick J,
Krampitz G, Lacey JC Jr, Matsuno K, Melius P, Middlebrook M, Nakashima T,
Pappelis A, Pol A, Rohlfing DH, Vegotsky A, Waehneldt TV, Wax H, Yu B.
"Experimental retracement of the origins of the protocell: It was also a
protoneuron." In Ponnamperuma C and Chela-Flores J, eds, _Chemical
Evolution: Structure and Model of the First Cell_. Dordrecht:Kluwer
Academic Publishers (1995) pg. 17-36

Pappelis A, Fox SW. "Domain protolife: The protocell theory." In Poglazov
BF, Kurganov BI, Kritsky MS and Gladilin KL, eds, _Evolutionary Biochemistry
and Related Areas Physicochemical Biology_. Moscow:Bach Institute of
Biochemistry and ANKO (1995) pg. 151-9

Pappelis A, Fox SW, Papagiannis MD. "Protocell and metaprotocells of the
Protolife Kingdom." _Illinois State Academy of Science_ 1993; 86(Suppl.):57

Fox SW. "Synthesis of life in the lab? Defining a protoliving system."
_Quarterly Review of Biology_ 1991 June; 66(2):181-5 [In this paper, Fox
again argues that proteinoid microsphere protocells are representatives of
"(_proto_) living organisms" (pg. 182) and he continues to use the qualifiers
"proto-" and "primitive". However, he also more freely refers to his
protocells as "organisms" and "cells" without qualifiers, and he asserts that
(pg. 181): "The 'synthesis' of an organism in the lab holds special
significance as an affirmation of cell analysis....Such verification is an
extension to cellular science from organic which synthesis of
bioorganic compounds is a traditional confirmation of the analysis by organic
chemists of the chemical structure of a natural compound." This is also (to
my knowledge) the first time he specifies the criteria he will use to assert
that his protocells are alive: metabolism, cellularity, reproduction and
growth, and response to external stimuli. He even quotes critics who admit
that (except for the lack of a genetic system) they would willingly accept
that his protocells are alive because they demonstrate metabolism, growth and
reproduction. Even so, he still does not come right out and claim that he
synthesized life in the lab. I therefore believe that this paper represents
a transition in his thinking based on the accumulating evidence from his more
cautious position in 1977 to his bolder claims in 1997.]

Fox SW. "Molecular selection and natural selection." _Quarterly Review of
Biology_ 1986 September; 61(3):375-86

Fox SW. "Self-sequencing of amino acids and origins of polyfunctional
protocells." _Orig Life_ 1984; 14(1-4):485-8 [This article discusses the
information content of proteinoid microsphere protocells.]

Fox SW, Nakashima T, Przyblyski A, Syren RM. "The updated experimental
proteinoid model." _International Journal of Quantum Chemistry_ 1982; QBS

Fox SW. "How many theories for the origin of (proto)life?" in Srinivasan R,
ed., _Biomolecular Structure, Conformation, Function and Evolution_.
Oxford:Pergamon Press (1981) pg. 643-6

Fox SW. "A model for protocellular coordination of nucleic acid and protein
synthesis." in Kageyama M, Nakamura K, Oshima T, Uchida T, eds., _Science
and Scientists_. Tokyo:Japan Science Society Press (1981) pg. 39-45

Fox SW. "From inanimate matter to living systems." _American Biology
Teacher_ 1981; 43(3):127-35, 140

Matsuno K. "Material self-assembly as a physiochemical process."
_BioSystems_ 1981; 13:237-41

Fox SW. "Life from an orderly cosmos." _Naturwissenschaften_ 1980 December;
67(12):576-81 [This article ties in the nonrandomness of thermal proteinoids
with the order seen in the universe and discusses how this is a reflection of
the second law of thermaldynamics "on a cosmic scale".]

Fox SW. "Metabolic microspheres: origins and evolution."
_Naturwissenschaften_ 1980 August; 67(8):378-83 [This article discusses the
catalytic activity of proteinoid microspheres as a model for the evolution of

Fox SW, Nakashima T. "The assembly and properties of protobiological
structures: the beginnings of cellular peptide synthesis." _Biosystems_
1980; 12(3-4):155-66 [The research that forms the basis for this proposal is
in Nakashima T, Fox SW. "Synthesis of peptides from amino acids and ATP with
lysine-rich proteinoid." _Journal of Molecular Evolution_ 1980 May;

Fox SW. "The origins of behavior in macromolecules and protocells."
_Comparative Biochemistry and Physiology_ 1980; 67B:423-36

Fox SW. "Organic microstructures and terrestrial protocells."
_Naturwissenschaften_ 1977 July; 64(7):380-1

Fox SW. "Bioorganic chemistry and the emergence of the first cell." In van
Tamelan EE, ed., _Bioorganic Chemistry_. New York:Academic Press (1977) pg.

Fox SW. "The evolutionary significance of phase-separated microsystems."
_Orig Life_ 1976 January; 7(1):49-68 [This article discusses how proteinoid
microspheres can serve as a models for the cell and the protocell.]

Fox SW> "Stereomolecular interactions and microsystems in experimental
protobiogenesis." _BioSystems_ 1975 July; 7(10):22-36

Fox SW. "Looking forward to the present." _Biosystems_ 1975 March;

Fox SW. "Origins of biological information and the genetic code." _Mol Cell
Biochem_ 1974 April 15; 3(2):129-42

Fox SW, Jungck JR, Nakashima T. "From proteinoid microsphere to contemporary
cell: formation of internucleotide and peptide bonds by proteinoid
particles." _Orig Life_ 1974 January-April; 5(1):227-37

Fox SW. "The proteinoid theory of the origin of life and competing ideas."
_American Biology Teacher_ 1974; 36:161-72, 181

Fox SW. "Origin of the cell: experiments and premises."
_Naturwissenschaften_ 1973 August; 60(8):359-68

Fox SW. "Molecular evolution to the first cells." _Pure Appl Chem_ 1973;

Hsu LL, Brooke S, Fox SW. "Conjugation of proteinoid microspheres: a model
of primordial communication." _Current Models in Biology_ 1971; 4:12-25

Fox SW. "Self-ordered polymers and propagative cell-like systems."
_Naturwissenschaften_ 1969 January; 56(1):1-9

Fox SW. "Spontaneous generation, the origin of life, and self assembly."
_Current Models in Biology_ 1968 November-December; 2(5):235-40

Now something I'm sure you've noticed is that Fox is the sole author on the
vast majority of these papers and a coauthor on all of them save one. The
reason, however, is quite simple. Of all the candidates for synthesized life
in the lab, including self-replicating molecules and enzymatic RNA molecules,
the one with the strongest case are the proteinoid microspheres. Other
people have investigated them, and I will discuss their work shortly, but Fox
made the major effort; in fact, he devoted virtually the whole of the last 25
years of his career to trying to understand them and their implications for
life in general and abiogenesis in particular. As the acknowledged expert,
it should be no surprise that he wrote the majority of the papers dealing
with them.

What might be a surprise is why so few have followed up on his research. The
reason may involve a combination of three factors. The first is that Fox and
his colleagues seemed to investigate them so thoroughly that many researchers
may feel that there is little new to discover. There is also the fact that
certain experiments need only be replicated so many times before everyone
comes to believe the results are real and so stop replicating them simply to
verify their reality. Since the general biological consensus is that
proteinoid microsphere protocells are alive, there is little need for anyone
to continue to verify that. Instead, most of the people who investigate them
do so to try to find medical or industrial uses for them, not to understand
them or abiogenesis.

The second factor is that, while most of his colleagues appear to have
accepted that his protocells were alive, as Brian has pointed out most of
them did not accept that they had much of anything to do with abiogenesis or
the origin of life on earth. They were investigating their own ideas, so
they were rivals -- friendly, cooperative rivals who valued and respected
each others' work, but rivals nonetheless. As a result, they probably
convinced succeeding generations of biologists that Fox's protocells were
little more than an interesting sideline or even a complete deadend. Fox
himself has pointed this out, with regard to the RNA-first and DNA-first
concepts. And why would anyone investigate a deadend when they could pursue
more fruitful avenues? (In point of fact, I do not believe that Fox's
protocells are a deadend, but that is a different topic from the one we are
currently discussing. Besides, more of the current generation of biologists
appear to be abandoning the genetic-code-first concepts for the protein-first
concept of thermal proteinoids.)

The third factor is somewhat like the second. Nearly all of Fox's colleagues
are dead now, and unfortunately with them went their own concepts, for much
the same reasons as for Fox's protocells. Their critique of each others'
work as having little or no relevence to abiogenesis may have worked too
well, so that most contemporary biologists have largely rejected their
specific concepts in favor of more exotic ideas. However, it is interesting
that whereas many seem now to be concentrating on protocells composed solely
of amphiphilic molecules, they still accept the fundamental precepts
established by Fox: the first life must be cellular; information in the form
of proteins and metabolism preceeded a genetic code; and the key biomolecules
must be capable of self-assembly.

This should not, however, be misconstrued to believe that no one is currently
investigating proteinoid microsphere protocells in terms of abiogenesis. At
Southern Illinois University in Carbondale is the Thermal Protein Study Group
( headed by Aristotel Pappelis and Donald Ugent.
Also, Duane L. Rohlfing of the University of Southern Carolina continues to
do research in proteinoids. The names of additional researchers can be found


which is a publication and abstract listing for the 1994 American Chemical
Society Symposium on the Origin of Life. They include such scientists as

C. Steven Sikes of the University of Southern Alabama;
K- Harada of Shoin Women's University, Japan;
Klaus Dose of Gutenberg University, Germany;
Paul Melius of Auburn University, Alabama;
Koichiro Matsuno of the Nagaoka University of Technology, Japan;
Peter R. Bahn of Bahn Biotechnology Co.;
R.M. Ottenbrite of the Virginia Commonwealth University;
Alexander T. Pol of the College of Engineering, University of Miami;
R. Rosen of Dalhousie University, Halifax, Nova Scotia; and
Etsuo Kokufuta of the Institute of Applied Biochemistry, University of
Tsukuba, Japan

I will post the third and last part when I know this one has propogated.