A Non-Enzymatic Gluconeogenesis and Fatty Acid Cycle

Project Information

NonEnzymatic

Grant agreement ID: 896219

DOI

10.3030/896219

Project closed

EC signature date 9 April 2020

Start date 1 September 2020

End date 31 August 2022

Funded under

• EXCELLENT SCIENCE - Marie Skłodowska-Curie Actions

Total cost

€ 196 707,84

EU contribution

€ 196 707,84

Coordinated by

UNIVERSITE DE STRASBOURG
France

Periodic Reporting for period 1 - NonEnzymatic (A Non-Enzymatic Gluconeogenesis and Fatty Acid Cycle)

Reporting period: 2020-09-01 to 2022-08-31

Summary of the context and overall objectives of the project

It is generally accepted that the emergence of life occurred gradually, by the natural flow of molecules in metabolism like reaction pathways, which gradually developed complexity over time. In this scenario, key pathways like the rTCA would be present and occurring without the need for enzymes, but rather catalysts like metals (homogenous and heterogenous) and when new catalysts for any one reaction were formed, they would further accelerate their own synthesis (autocatalysis). Other pathways may have their roots in a prebiotic and enzyme free world as well. The overall objective of the project was to investigate gluconeogenesis and the fatty acid cycle to evaluate if an enzyme free reaction could be plausible. Furthermore, we investigated the robustness of these catalysts in the presence of common catalyst poisons and modulators which would also be present in a prebiotic context.
Since becoming self-aware, certain questions have plagued humanity, one of which is the question of the emergence of life. The project here further expands the hypothesis that life began from a bottom-up approach, where metabolism was the precursor to complexity. The overall project would solidify that not only the Wood–Ljungdahl and rTCA cycles are prebiotic in nature, but also sugar and fatty acid synthesis.

Work performed from the beginning of the project to the end of the period covered by the report and main results achieved so far

A meta-analysis of the possible homogenous and heterogenous catalysts which could have been present in a prebiotic context was performed and certain minerals and metals were purchased. New analytical methods using various equipment like NMR, GCMS, HPLC-ELSD, and SFC were developed for the analysis of small and fleeting intermediates using standards synthesized by FFS providers. Many reactions in various pH, temperature, and pressure-controlled environments were conducted to observed key intermediates in gluconeogenesis were performed. Many reactions were also conducted to either synthesize (anabolism) or degrade (catabolism) short-long chain fatty acids. In the case of gluconeogenesis, promising results were obtained for many of the reactions, investigations into which are still ongoing by others in the host lab. In the case of fatty acid synthesis, all reactions failed, suggesting that this pathway may have come later in the emergence of life. These catalysts however were highly active in other pathways, such as the rTCA cycle, and the reduction of NAD+. The reduction of this molecule was further investigated with an in-depth look into catalyst modulators which has been a long-standing critique of metal catalysts in prebiotic chemistry.
The work was discussed at an early stage at the conference, Molecular Origins of Life, and is now in the stage of being submitted for publication. Furthermore, other work in organic chemistry methods has been published in Chemical Science.

Progress beyond the state of the art and expected potential impact (including the socio-economic impact and the wider societal implications of the project so far)

The negative results in the fatty acid pathway have allowed us to turn our focus to some more plausible prebiotic metabolic pathways, such as amino acid biosynthesis. These form the basis
of enzymes and therefore their presence in higher concentration may be crucial for these other pathways to emerge.
Based on the discovery of catalyst modulators, we now have a clearer imagine into how catalysts behave in the presence of the complex mixture of molecules present in prebiotic conditions. Very likely the reactions which eventually led to life were not as clean as a test tube in a lab, but rather with a plethora of small molecules ready to latch onto catalysts, and this understanding is paramount.

Modulating reactivity