The history of Buchner's discovery is of great interest [Gruber, [1908]; Hahn, [1908]]. As early as 1893 Hans and Eduard Buchner found that the cells of even the smallest micro-organism could be broken by being ground with sand [Buchner, E. and H., and Hahn, [1903], p. 20], and in 1896 the same process was applied by these two investigators to yeast, with the object of obtaining a preparation for therapeutic purposes. Difficulties arose in the separation of the cell contents from the ground-up mixture of cell membranes, unbroken cells, and sand, but these were overcome by carrying out the suggestion of Martin Hahn (at that time assistant to Hans Buchner) that kieselguhr should be added and the liquid squeezed out by means of a hydraulic press [Buchner, E. and H., and Hahn, [1903], p. 58]. The yeast-juice thus obtained was, in the first instance, employed for animal experiments, but underwent change very rapidly. The ordinary antiseptics were found to be unsuitable, and hence sugar was added as a preservative, and it was the marked action of the juice upon this added cane sugar that drew Eduard Buchner's attention to the fact that fermentation was proceeding in the absence of yeast-cells.
As in the case of so many discoveries, the new phenomenon was brought to light, apparently by chance, as the result of an investigation directed to quite other ends, but fortunately fell under the eye of an observer possessed of the genius which enabled him to realise its importance and give to it the true interpretation.
In his first papers [[1897, 1], [2]; [1898]], Buchner established the following facts: (1) yeast-juice free from cells is capable of producing the alcoholic fermentation of glucose, fructose, cane sugar, and maltose; (2) the fermenting power of the juice is neither destroyed by the addition of chloroform, benzene, or sodium arsenite [Hans Buchner, [1897]], by filtration through a Berkefeld filter, by evaporation to dryness at 30° to 35°, nor by precipitation with alcohol; (3) the fermenting power is completely destroyed when the liquid is heated to 50°. [p019]
From these facts he drew the conclusion "that the production of alcoholic fermentation does not require so complicated an apparatus as the yeast cell, and that the fermentative power of yeast-juice is due to the presence of a dissolved substance". To this active substance he gave the name of zymase.
Buchner's discovery was not received without some hesitation. A number of investigators prepared yeast-juice, but failed to obtain an active product [Will, [1897]; Delbrück, [1897]; Martin and Chapman, [1898]; Reynolds Green, [1897]; Lintner, [1899]]. A more accurate knowledge of the necessary conditions and of the properties of yeast-juice, however, led to more successful results [Will, [1898]; Reynolds Green, [1898]; Lange, [1898]], and it was soon established that, given suitable yeast, an active preparation could be readily procured by Buchner's method. Criticism was then directed to the effect of the admitted presence of a certain number of micro-organisms in yeast-juice [Stavenhagen, [1897]], but Buchner [Buchner and Rapp, [1897]] was able to show by experiments in the presence of antiseptics and with juice filtered through a Chamberland candle that the fermentation was not due to living organisms of any kind.
The most weighty criticism of Buchner's conclusion consisted in an attempt to show that the properties of yeast-juice might be due to the presence, suspended in it, of fragments of living protoplasm, which, although severed from their original surroundings in the cell, might retain for some time the power of producing alcoholic fermentation. This, it will be seen, was an endeavour to extend Nägeli's theory to include in it the newly discovered fact.
In favour of this view were adduced the similarity between the effects of many antiseptics on living yeast and on the juice, the ephemeral nature of the fermenting agent present in the juice, the effect of dilution with water, and the phenomenon of autofermentation which is exhibited by the juice in the absence of added sugar [Abeles, [1898]; v. Kupffer, [1897]; v. Voit, [1897]; Wehmer, [1898]; Neumeister, [1897]; Macfadyen, Morris, and Rowland, [1900]; Bokorny, [1906]; Fischer, [1903]; Beijerinck, [1897], [1900]; Wroblewski, [1899], [1901]].
A brief general description of the actual properties of yeast-juice and of the phenomena of fermentation by its means is sufficient to show the great improbability of this view.
The juice prepared by Buchner's method forms a somewhat viscous opalescent brownish-yellow liquid, which is usually faintly acid in reaction [compare Ahrens, [1900]] and almost optically inactive. It has a specific gravity of 1·03 to 1·06, contains 8·5 to 14 per cent. [p020] of dissolved solids, and leaves an ash amounting to 1·4 to 2 per cent. About 0·7 to 1·7 per cent. of nitrogen is present, nearly all in the form of protein, which coagulates to a thick white mass when the juice is heated.