Louis Pasteur: His Life and Labours

by René Vallery-Radot

ACETIC FERMENTATION.
THE MANUFACTURE OF VINEGAR.

Soon afterwards Pasteur came upon a most curious illustration of the 'fixation' of atmospheric oxygen by a microscopic organism—the transformation of wine into vinegar. As its name indicates, vinegar is nothing else than wine turned sour. Everybody has remarked that wine, left to itself, in circumstances which occur daily, is frequently transformed into vinegar. This is noticed more particularly when bottles, having been uncorked, are left in a half-empty condition. Sometimes, however, wine turns sour even in corked bottles. In this case we may be sure that the bottles have been standing upright, and that corks more or less defective have permitted the air to penetrate into the wine. The presence of air, in fact, is indispensable to the chemical act of transforming wine into vinegar. How does this air intervene? And what is the little microscopic creature which, in conjunction with the air, becomes the agent of this fermentation?

In a celebrated lecture given at Orleans at the request of the manufacturers of vinegar in that town, Pasteur, after having stated the two foregoing scientific questions, proceeded to examine the difference between wine and vinegar. What takes place in the fermentation of the juice of the grape which yields the wine? The sugar of this juice disappears, giving place to carbonic acid gas, which is exhaled during fermentation, and to alcohol, which remains in the fermented liquid. Formerly, chemists gave the name of 'spirit' to all volatile matters which could be collected from distillation. Now, when we distil wine and condense the vapour in a worm surrounded by cold water, we collect the spirit of wine at the extremity of the worm—this, when the water with which it is mixed during distillation is withdrawn from it, we designate by the name of alcohol. Vinegar contains no alcohol. When distilled it yields water and a spirit. But this spirit is acid, with a very pungent odour, and not inflammable like spirit of wine. Separated from the water which had accompanied it during the distillation, this spirit takes the name of acetic acid. This is the form in which it is used in smelling bottles—in those bottles of English salts the vapour of which is so penetrating.

In the formation of vinegar in contact with air the alcohol disappears, and is replaced by acetic acid. The air has thus given up something to the wine. Atmospheric air every one knows to be a mixture of nitrogen and oxygen, the nitrogen in the proportion of four-fifths of the total volume, and the oxygen of one-fifth. Well, in the transformation of wine into vinegar the nitrogen remains inactive. It is the oxygen alone which enters into combination with the alcohol. You ask for the proof of this? Take a bottle of wine turned sour, a bottle which at the same time is stopped hermetically; if the oxygen of the air contained in the bottle has combined with the alcohol, then, instead of air, there will be nothing in the bottle but nitrogen gas. Turn the bottle upside down and open it in a basin of water. The water of the basin will rush into the bottle to fill the partial vacuum created by the disappearance of the oxygen. The volume of water which enters the bottle is precisely equal to a fifth part of the total original volume of the air which the bottle contained at the time when it was closed. Moreover, it is easy to show that the gas which remains in the bottle has the properties of nitrogen gas. A lighted match is extinguished in it as if plunged into water, and a bird dies immediately in it of asphyxia.

If we confine our knowledge to what has gone before, it would seem that alcohol diluted with water and exposed to the air ought to furnish acetic acid. It is not so, however. Pure water alcoholised to the degree of ordinary wines may remain for whole years in contact with the air, without the least acetification. In this difference between natural wine and pure water alcoholised, and exposed to contact with air, we touch upon a vital point in the phenomena of fermentation. The celebrated theory of Liebig, which Pasteur was destined to overthrow, might be thus summed up:—If pure alcoholised water cannot become sour in contact with air, as is the case with wine, it is because the pure alcoholised water lacks the albuminoid substance which exists in the wine in a state of chemical alteration, and which is a ferment capable of causing the oxygen of the air to combine with the alcohol. And the proof, according to Liebig, that things act rigorously thus is, that if you add to the mixture of water and alcohol a little flour, or a little meat-juice, or even a minute quantity of any vegetable juice, the acetic fermentation arises, as if by compulsion. In other words, by the addition of a small quantity of any nitrogenised substance in process of alteration, you cause the union of the oxygen of the air with the alcohol.

There is doubtless always in the wine, when it turns sour, a necessary intermediary, producing the fixation of the oxygen of the air; since in no circumstances can pure alcohol, diluted to any degree whatever with pure water, transform itself into vinegar. But this necessary intermediary is not, as the German theory would have it, a dead albuminoid substance; it is a plant, and of all plants one of the simplest and most minute, which has been known from time immemorial under the name of flower of vinegar. This little fungus is invariably present on the surface of a wine which is being transformed into vinegar. Liebig was not ignorant of this, but he regarded it as a simple coincidence. Do we not know, said he, that whenever an infusion of organic matter is exposed to the air it becomes covered with a cryptogamic vegetation, or is invaded by a crowd of animalculæ? Is not vinegar a vegetable infusion? Vinegar affords a refuge to the flower of vinegar, just as it gives refuge to what are called the little eels of vinegar.

We can appreciate here the uncertainties of pure observation. The great art—and no one practised it better than Pasteur—consists in instituting decisive experiments which leave no room for an inexact interpretation of facts. These decisive proofs of the true part played by the little microscopic fungus, by this flower of vinegar, this mycoderma aceti, are thus formulated by Pasteur. It is but another example of the method which he used in alcoholic, lactic, and tartaric fermentations. The theories of Berzelius, of Mitscherlich, and of Liebig were destined again to receive the rudest shocks by the demonstration of these rigorous facts.

Let us place a little wine in a bottle, then hermetically seal it, and leave it to itself. In these conditions the wine becomes sour. But if we take the precaution of putting the bottle into hot water, so that the wine and the air in the bottle may be heated for some instants to a temperature of 60° Centigrade, and if, after cooling, we leave the bottle to itself, the wine in these conditions will never become transformed into vinegar. The heating, however, must have left intact the albuminoid or nitrogenous substances contained in the wine. These, then, cannot constitute the ferment of the vinegar. Can it be maintained that by heating the wine to 60° we have altered the albuminoid matter, which is, on this account, no longer able to act as a ferment, or, in other words, no longer able to determine the union of the oxygen of the air with the alcohol? This hypothesis falls to pieces before the following experiment. Open the bottle, blow into it with bellows, so that the once heated wine shall come into contact with ordinary air, and the acetification of the wine will take place.

But the master experiment is the following. We have seen that pure alcoholised water never turns sour unless some albuminoid matter is introduced into it. Pasteur saw that this albuminoid matter might be completely suppressed and replaced by saline crystallisable substances, alkaline and earthy phosphates, to which has been added a little phosphate of ammonia. In these conditions, especially if the alcoholised water be acidulated by small quantities of pure acetic acid, one actually sees the mycoderm developing, and the alcohol transforming itself into acetic acid. It is not possible to demonstrate in a more convincing manner that the albuminoid matters of the wine are not in this case the acetic ferment. These albuminoid matters, however, contribute to the acetic fermentation, but only as being an aliment to the mycoderma aceti, and notably a nitrogenous aliment. The true and only ferment of vinegar is the little fungus; it is the great agent of the phenomenon; it, indeed, accomplishes all.