ACTURE
Year: 1899

page_text: Impr
We, LA SOCIÉTÉ ANONYME DES MANUFACTURES DE PRODUITS CHIMIQUES DU
NORD, of Lille (Nord), France, and EUGENE BOULLANGER, Chemist, of the Pasteur
Institute, Lille aforesaid, do hereby declare the nature of this invention to be
as follows:-
The principal processes for the commercial manufacture of lactic acid are at
present based on the conversion of milk sugar or of glucose into lactic acid by
the lactic ferment in presence of bicarbonate of soda or of carbonate of lime.
The lactic ferment produces a splitting up of the molecule of glucose into two
molecules of lactic acid. There may be employed for the purpose either whey
which is fermented with old cheese, or, as proposed by Jacquemin, a saccharine
wort prepared with commercial glucose or with grain saccharified by diastase of
malt, or with such fort of grain mixed with wort of commercial glucose; which
worts being then, according to Jacquemin's process, sown with a lactic ferment
isolated in the pure state by bacteriological methods.
If as in many processes, the operation is not conducted in an aseptic medium,
100 kilogrammes of glucose, which should theoretically yield 100 kilogrammes of
lactic acid, will only produce from 60 to 70 kilogrammes of said acid, hence
there are considerable losses due to the invasion and competition of foreign
microbe germs which consume the lactate or produce other. acids at the expense
of the sugar; moreover the production is very irregular.
If on the other hand, the operation be performed by sowing with a pure lactic
ferment a sterile saccharine wort resulting from the saccharification of amyla-
ceous matters, by means of malt or by acids, the vield is still (referred to one
hundred of starch) much below the theoretical yield, as several inconveniences
are encountered, viz. :-
(1) The unavoidable formation of a greater or less quantity of dextrine which
is attacked much more slowly and difficultly by the lactic ferment, hence a
diminution of yield;
(2) To this loss is to be added the losses of starch resulting from the malting
or partial caramelization of the sugar if acids be employed:
(3) The lactic ferment gives a maximum yield when the richness in sugar
does not exceed from 3% to 4%. in more concentrated worts the vield is less
and fermentation becomes much slower; now this concentration of 3% to 4%

page_text: must be considerably exceeded in practice to avoid a very large expense for
fuel for sterilizing and facilitating the extraction of the acid product.
Lastly, if to partially obviate these objections worts prepared from commercial
glucose he employed the last objection relative to the concentration of the wort
still remains and moreover the cost of the acid produced will be increased and
the more so as one is obliged in this case to add to the malt very costly nutri-
tive nitrogenous matters (peptones) to enable the lactic ferment to propagate.
Commercial glucose further always contains dextrine, so that the objection above
indicated in the case of worts from grain still obtains in the present case.
The present invention relates to a process for the commercial manufacture of
lactic acid in which the whole of the above objections are avoided and a much
larger vield consequently obtained.
We have found on certain plants mucedines which possess the property both
of energetically saccharifying starch and of producing at same time at the
expense of sugars, considerable quantities of lactic acid. One of these mucedines
which we have more particularly studied up to the present, belongs to the group
of mucors and to it we have given the name of lactomyces on account of its
distinguishing properties. We have found that other mucedines are equally
capable of furnishing lactic acid in variable quantities at the expense of starch
and at the expense of sugars. By the cultivation of these mucedines under
favourable conditions and such that they will be compelled to propagate in the
midst of the liquid mass, without forming aerial fructifications, there is obtained,
commencing with starch, a saccharification and an integral lactic fermentation
of this substance. The yield is consequently considerably augmented.
Our process is based on the employment of these saccharifying lactomyces
either alone or associated with lactic ferments of known kind, or the employment
of ordinary saccharifying mucedines together with lactic ferments or the lacto-
myces before referred to, for the production of lactic acid by means of amylaceous
worts by simultaneous saccharification and lactic fermentation.
The process essentially comprises the following operations:-
(1) The wort is prepared by heating amylaceous matters such as grain, pota-
toes, starch, etc. In order to prevent the thickening of the starch by cooling, it
is liquefied with the aid of a small quantity of malt, or by means of a little aci:
or by any other means usually employed. To the wort is then added chalk or
other substance capable of saturating the lactic acid as fast as it is formed, and
it is then heated in a closed vessel, to a sufficient temperature to destroy all the
germs which the mass of liquid may contain.
(2) The sterile wort is then distributed aseptically in closed rats which are
also sterilized and provided with means for enabling the impregnation with lac-
tomyces in a pure condition and for promoting its propagation in the mass of
liquid by a suitable agitation and maintaining the temperature at about 35° C.
(3) When the lactomyces is sufficiently propagated and the saccharification of
the wort is in progress, the conversion into lactic acid of the sugar formed may
be facilitated by adding to the wort a very small quantity of pure seed of lactic
ferment. This latter lives perfectly in symbiosis with the lactomyces, and the
whole of the sugar as fast as it is formed by the mucedines at the expense of the
starch, is both converted into lactic acid by the lactomyces and by the lactic
ferment.
(4) When the saccharification and formation of lactate of lime are considered
as terminated (which can be easily ascertained by taking a sample of the fer-
mented liquid) the heated wort is decanted and run into special vats in which
the lactate of lime crystallizes. It is only necessary to decompose it by means
of a regulated quantity of sulphuric acid in order to obtain lactic acid.
The progress of the operation is similar when an ordinary saccharifying muce-
dine is employed in lieu of lactomyces, but in this case it is indispensable to
Sow with an ordinary lactic ferment in considerable quantity and to slightly
prolong the duration of the fermentation. The lactomyces may also be employed
{

page_text: either alone or associated with lactic ferments in saccharine worts prepared by
means of glucose or commercial maltose or in worts obtained by mixing these
saccharine worts with an ordinary amylaceous wort.
The following are the advantages of this improved process:-
(1) The process permits of avoiding all the expense attending either the
employment of large quantities of malt for saccharifying starch (loss of starch
by malting, hand labour, etc.,) or the employment of acids (loss of sugar by
caramelization) as well as the indispensable subsequent operations such as satura-
tion of the acid, etc.
(2) It is possible to employ amylaceous matters for the manufacture of lactic
acid while avoiding the losses which have been hitherto inevitably incurred when
employing these substances, such as the formation of dextrine, etc., as before
indicated.
(3) The worts may be raised to a high degree of concentration because the
lactic acid is produced as fast as the sugar is formed, so that the sugar is never
at a sufficient degree of concentration to interfere with the ferment, retard the
fermentation, and lower the yield as was previously pointed out.
(4) The duration of the fermentation is reduced in that the process enables a
malt considerably more concentrated than that employed in other processes of
manufacture to be fermented in an almost equal space of time.
(5) The extraction of the lactic acid from the worts is proportionately less
costly by reason of this very concentration and of their greater richness in
lactates.
(6) In consequence of all the advantages above indicated the process gives
a yield in lactic acid much higher than that obtained by any other known
processes.
A. M. & WM. CLARK,
Chartered Patent Agents, 53, Chancery Lane, London.
Improvements in the Manufacture of Lactic Acid.
We, LA SOCIETE ANONYME DES MANUFACTURES DE PRODUITS CHIMIQUES DU
NORD, of Lille (Nord), France, and EUGÈNE BOULLANGER, Chemist, of the Pasteur
Institute, Lille aforesaid, do hereby declare the nature of this invention and in
what manner the same is to be performed to be particularly described and ascer-
tained in and by the following statement:-
The principal processes for the commercial manufacture of lactic acid are at
present based on the conversion of milk sugar or of glucose into lactic acid by
the lactic ferment in presence of bicarbonate of soda or of carbonate of limě.
The lactic ferment produces a splitting up of the molecule of glucose into two
molecules of lactic acid. There may be employed for the purpose either whey
which is fermented with old cheese, or, as proposed by Jacquemin, a saccharine
wort prepared with commercial glucose or with grain saccharified by diastase of
malt, or with such wort of grain mixed with wort of commercial glucose; said
worts being then, according to Jacquemin's process sown with a lactic ferment
isolated in the pure state by bacteriological methods.
If as in many processes, the operation is not conducted in an aseptic medium,
100 kilogrammes of glucose, which should theoretically yield 100 kilogrammes of
lactic acid, will only produce from 60 to 70 kilogrammes of said acid, hence

page_text: there are considerable losses due to the invasion and competition of foreign
microbe germs which consume the lactate or produce other acids at the expense
of the sugar; moreover the production is very irregular.
If on the other hand, the operation be performed by sowing with a pure lactic
ferment a sterilized saccharine wort resulting from the saccharification of amyla-
ceous matters, by means of malt or by acids, the yield (referred to one hundred
of starch) is still much below the theoretical yield, as several inconveniences
are encountered, viz. :-
(1) The unavoidable formation of a greater or less quantity of dextrine which
is attacked much more slowly and difficultly by the lactic ferment, hence a
diminution of yield;
(2) To this loss is to be added the losses of starch resulting from the malting
or partial caramelization of the sugar if acids be employed.
(3) The lactic ferment gives a maximum yield when the richness in sugar
does not exceed from 3% to 4%, in more concentrated worts the yield is less
and fermentation becomes much slower; now this concentration of 3% to 4%
must be considerably exceeded in practice to avoid a very large expense for
fuel for sterilizing and facilitating the extraction of the acid product.
Lastly, if to partially obviate these objections worts prepared from commercial
glucose be employed the last objection relative to the concentration of the wort
still remains and moreover the cost of the acid produced will be increased and
the more so as one is obliged in this case to add to the malt very costly nutri-
tive nitrogenous matters (peptones) to enable the lactic ferment to propagate.
Furthermore, commercial glucose always contains dextrine, so that the objection
above indicated in the case of worts from grain still obtains in the present case.
The present invention relates to a process for the commercial manufacture of
lactic acid in which the whole of the above objections are avoided and a much
larger yield consequently obtained.
The following is the method of obtaining and identifying the special ferments
employed in carrying out this process.
We have found on certain plants mucedines which possess the property both
of energetically saccharifying starch and of producing at same time at the
expense of sugars, considerable quantities of lactic acid. One of these mucedines
which we have more particularly studied up to the present, belongs to the group
of mucors and to it we have given the name of lactomyces on account of its
distinguishing properties.
Our lactomyces belong to a group of mucedines well known with regard to
morphological properties and called mucors, a complete description of which may
be found in any book relating to botany. The characteristic of these mucedines
is a ramified mycelium on which grow very soon, under propitious circum-
stances some long and slender skins ending in a ball called a sporangium, inside
of-which are contained the spores.
All mucors and especially our lactomyces possess the same morphological
characteristics and are to be distinguished only by their physiological proper-
ties. This is a constant fact in bacteriology: it often happens that several
species which are identical in aspect are however widely different with regard to
physiological properties. The one for instance produces alcohol and the other
citric acid, both at the expense of sugar. And so it happens with our lactomyces
which in shape are like any known mucor: but they are characterized by the
fact that they produce lactic acid when brought into contact with sugar or starch,
whilst any other mucor would give alcohol, water or carbolic acid, consequently
the physiological properties of the species and those properties only are important
as the characteristics of form or shape may be modified according to the condi-
tions of the growth and besides are often identical among species which are in
fact quite different. Anyone who has studied bacteriology is aware of this fact.
We have discovered that these mucedines can, for example, be developed by
placing fragments of the leaves of Rumex Acetosella in a nutritive medium, but

page_text: it is not to be understood from this that they are nowhere else to be found,
as microbes as a rule are too abundant in nature and undergo too much mixing
and carrying through the medium of the air and animals for it to be possible
to assign to them any special or fixed location.
In order to isolate the lactomyces of Rumex Acetosella, we place the fragments
of the leaves in the following nutritive medium :-
Yeast water obtained by boiling beer yeast with water
Glucose
.
100 grammes.
10 grammes,
The leaves which are cut up with scissors which have been passed through a
flame, are placed in sterilized tubes containing 5 c. c. of the above described nutri-
tive medium which are then placed in a stove at 28° until the development of
the mucedines which are separated by the usual methods of bacteriology.
We have also found that other mucedines are equally capable of furnishing
lactic acid in variable quantities at the expense of starch and at the expense
of sugars. By the cultivation of these mucedines under favourable conditions
and such that they will be compelled to propagate in the midst of the liquid
mass, without forming aerial fructifications, there is obtained, commencing with
starch, a saccharification and an integral lactic fermentation of this substance.
The yield is consequently considerably augmented. The species thus developed
are then isolated in a pure state by bacteriological methods.
In order to obtain a culture of the lactomyces a fragment of potato is steri-
lized in a test tube closed by a plug of cotton at a temperature of 120° C. and
upon this sterilized fragment of potato are deposited with the aid of a platinum
wire which has been passed through a flame, a few spores of a precedent pure
culture of lactomyces, the precautions usual in bacteriology being taken to avoid
all, contamination by foreign microbes. The tube is then placed in a stove
heated to 30° C. and at the end of 48 hours, a culture is obtained which may
itself serve as seed for the obtainment of another culture.
Cultures of lactic ferments are obtained in the same manner except that instead
of potato, sugared gelatine is used at the surface of which the ferment develops
very rapidly.
Or we may proceed as follows:--In order to obtain spores of lactomyces neces-
sary for sowing a vat we proceed in the following manner:--In a flask of about,
500 c. c capacity are placed 20 grammes of rice and 5 c. c of water and after
letting swell for from four to five hours the rice is caused by agitation to deposit
upon the interior surface of the flask. The flask is then closed by a plug of cotton
and sterilized for twenty minutes in a digester at 120°.
We then separately prepare in a test tube containing 5 c. c. of sterilized beer
wort, a culture of lactomyces, for which purpose it suffices to sow in it a fragment
of a previous culture. The culture thus obtained is then added to the flask con-
taining the 20 grammes of sterilized rice, after which the flask is placed in a
stove at 28° and at the end of three or four days, it will be entirely covered
with lactomyces in full fructification.
A microscopical examination suffices to show whether the culture of lactomyces
or of lactic ferment is pure.
Our process is based on the employment of these saccharifying lactomyces
either alone or associated with lactic ferments of known kind, or the employment
of ordinary saccharifying mucedines together with lactic ferments or the lacto-
myces before referred to, for the production of lactic acid by means of amyla-
ceous worts by simultaneous saccharification and lactic fermentation.
The process essentially comprises the following operations:-
(1) The wort is prepared by heating amylaceous matters such as grain, pota-
toes, starch, etc. In order to prevent the thickening of the starch by cooling,
it is liquefied with the aid of a small quantity of malt, or by means of a little
acid or by any other means usually employed. To the wort is then added chalk
or other substance capable of saturating the lactic acid in proportion as it is
formed, and it is then heated in a closed vessel to a sufficient temperature to

page_text: destroy all the germs which the mass of liquid may contain. For example for
the purposes of lactic fermentation a wort is employed averaging 15 kilo-
grammes of maize per hectolitre of water, but this proportion would however
vary according to the amount of starch contained in the grain and the desired
duration of the fermentation. Thus the proportion of grain may be reduced if
the fermentation is desired to be effected in a very short time, or it may be
increased if it be desired to treat a more concentrated wort and prolong the
period of fermentation. To this wort is added carbonate of lime in the propor-
tion of 100 kilogrammes of carbonate for every 180 kilogrammes of total glucose
present as determined by a test of the wort.
(2) The sterile wort is then distributed aseptically in closed vats which are
also sterilised and provided with means for enabling the sowing with lactomyces
in a pure condition and for promoting its propagation in the mass of liquid by
a suitable agitation and maintaining the temperature at about 35° C. The
sowing of the sterile wort is effected with a flask of 500 cubic centimetres of
wort containing a pure culture of lactomyces or lactic ferment for a vat of a
capacity of from 200 to 500 hectolitres.
(3) When the lactomyces is sufficiently propagated and the saccharification of
the wort is in progress, the conversion into lactic acid of the sugar formed may
be facilitated by adding to the wort a very small quantity of pure seed of lactic
ferment. This latter lives perfectly in symbiosis with the lactomyces, and the
whole of the sugar in proportion as it is formed by the mucedines at the expense
of the starch is converted into lactic acid by both the lactomyces and the lactic
ferment.
(4) When the saccharification and formation of lactates of lime are considered
as terminated (which can be easily ascertained by taking a sample of the fer-
mented liquid) the heated wort is decanted and run into special vats in which
the lactate of lime crystallizes. It is only necessary to decompose it by means
of sulphuric acid in the proportion of about 34 kilogrammes of acid to 100 kilo-
grammes of lactate in order to obtain lactic acid.
7.
The progress of the operation is similar when an ordinary saccharifying muce-
dine is employed in lieu of lactomyces, but in this case it is indispensable to
sow with an ordinary lactic ferment in considerable quantity and to slightly pro-
long the duration of the fermentation.
The mode of sowing the ferments is as follows:-At the upper part of the
fermenting vat is a pipe connection to which is attached an indiarubber tube
closed by an iron stopper. When it is desired to sow the lactomyces or lactic
ferment, the orifice of the indiarubber tube closed by the stopper is exposed
to the flame of a spirit lamp, after which the stopper is removed and the neck
of the flask containing the 500 cubic centimetres of culture is introduced into
the end of the tube and its contents poured off into the mass. The flask is then
withdrawn and the tube then again immediately re-stoppered, after having first
passed the stopper through a flame.
The mode of determining the state of development of the lactomyces is as
follows:-
The fermenting vat has a second pipe connection similar to the one above
referred to through which, after removing the stopper, a sample may be taken.
The lactomyces is considered to be sufficiently developed when on microscopic-
ally examination of this sample, there is found in each field of the microscope
a mycelian filament of the mucedine.
The lactomyces may also be employed either alone or associated with lactic
ferments in saccharine worts prepared by means of glucose or commercial maltose
in worts obtained by mixing these saccharine worts with an ordinary amylaceous
wort
The following are the advantages of this improved process:-
(1) The process permits of avoiding all the expense attending either the
employment of large quantities of malt for saccharifying starch (loss of starch

page_text: by malting hand labour etc.) or the employment of acids (loss of sugar by cara-
melization) as well as the indispensable subsequent operations, such as saturation
of the acid etc.
(2) It is possible to employ amylaceous matters for the manufacture of lactic
acid while avoiding the losses which have been hitherto inevitably incurred when
employing these substances, such as the formation of dextrine, etc., as before
indicated.
(3) The worts may be raised to a high degree of concentration because the
lactic acid is produced as fast as the sugar is formed, so that the sugar is never
at a sufficient degree of concentration to interfere with the ferment, retard the
fermentation, and lower the yield as was previously pointed out.
(4) The duration of the fermentation is reduced in that the process enables
a malt considerably more concentrated than that employed in other processes of
manufacture, to be fermented in an almost equal space of time.
(5) The extraction of the lactic acid from the worts is proportionately less
costly by reason of this very concentration and of their greater richness in
lactates.
(6) In consequence of all the advantages above indicated the process gives a
yield in lactic acid much larger than that obtained by any other known processes.
Having now particularly described and ascertained the nature of the said
invention and in what manner the same is to be performed, we declare that what
we claim is:-
A process for the commercial manufacture of lactic acid by means of amy-
laceous and saccharine matters, comprising the series of operations hereinbefore
described and based:-
1st. On the use of lactic and saccharifying mucedines, such as our so-called
lactomyces, alone or associated with ordinary lactic ferments:
2nd. On the use of ordinary saccharifying mucedines associated with lactic
ferments or with our so-called lactomyces, under the conditions herein indicated
for the manufacture of lactic acid by simultaneous lactic saccharification and
fermentation.
1
Vanah 1000
A. M. & WM. CLARK,
Chartered Patent Agents, 53, Chancery Lane, London.