French scientist Louis Pasteur researched this issue during the 1850s, and the results of his experiments began the long road to elucidating the pathway of glycolysis.
His experiments showed that fermentation occurs by the action of living microorganisms, yeasts, and that yeast's glucose consumption decreased under aerobic conditions of fermentation, in comparison to anaerobic conditions (the Pasteur effect). Insight into the component steps of glycolysis were provided by the non-cellular fermentation experiments of Eduard Buchner during the 1890s.
The ability of boiled extract plus dialyzed juice to complete fermentation suggests that the cofactors were non-protein in character. In the 1920s Otto Meyerhof was able to link together some of the many individual pieces of glycolysis discovered by Buchner, Harden, and Young.
They further removed diphosphoglyceraldehyde as a possible intermediate in glycolysis. With all of these pieces available by the 1930s, Gustav Embden proposed a detailed, step-by-step outline of that pathway we now know as glycolysis.
By the 1940s, Meyerhof, Embden and many other biochemists had finally completed the puzzle of glycolysis.
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