Process for Utilizing Moulds or Microscopic Fungi for Extracting from Residues, particularly those arising from the Treatment of Amylaceous and Sugary Substances, "Alcohol and Mucedines capable of being used in Panification, Distillation, Saccharification, and the like.
Year: 1897

page_text: Date of Application, 15th Jan., 1897
1007
4000
PROVISIONAL SPECIFICATION.
À Process for Utilizing Moulds or Microscopic Fungi for Extracting
from Residues, particularly those arising from the Treatment of
Amylaceous and Sugary Substances, "Alcohol and Mucedines
capable of being used in Panification, Distillation, Saccharification,
and the like.
We, AUGUSTE COLLETTE, the Younger, and AUGUSTE BOIDIN, both of Séchin,
Departement du Nord, in the Republic of France, Distillers, do hereby declare the
nature of this invention to be as follows:
This invention relates to a process for the utilisation of amylaceous or sugary
substances and likewise of a portion of the organic and mineral substances con-
tained in residues of any description whatsoever, by employing, for converting
them into alcohol, moulds possessing a certain diastasic activity and a fairly high
degree of fermentability.
The moulds which we employ are preferably such as are not pathogenetic in
order that they may be utilised in the manufacture of bread or in brewing beer
and also in the saccharification of the musts or worts of cereals.
In order to simplify the description, we will take by way of example the utilisa-
tion of the hydrates of carbon contained in the malt residuum of distillation, by a
mould discovered in Indo-China by Doctor A. Calmette and named by him
Amylomyces Rouxii.
According to the researches of Maerker and of Schultze a hectolitre of residuum
of the manufacture of yeast contains 2 kilogrammes 600 grammes of fermentable
products; sugar, dextrine and soluble starch, 720 grammes of albuminoid
substances and 250 grammes of mineral substances.
If after having sterilised a medium of this nature we introduce into it a trace of
amylomyces, this mucor develops; it first of all transforms the soluble starch and
the dextrine into glucose and finally into alcohol. At the same time the density
of the liquid diminishes; when it rests constant it. is passed through a sieve in
order to collect the mycelium formed and is distilled to recuperate the alcohol.
If it is aërated considerably a large quantity of mycelium is obtained. If, on the
other hand, the supply of air is restricted, the weight of mycelium is less but the
quantity of alcohol obtained is greater. In an ordinary commercial operation we
have obtained 1 kilogramme of pressed amylomyces and a litre of phlegm at
100° C. per hectolitre of residuary distillation liquors treated.
The operation can be carried out either with malt residuum clarified by decanta-
tion or filtered by being heated under pressure or else with thick or cloudy malt
residuum. The mycelium obtained undergoes a washing process in order to free it
from the fermentable substances in which it is immersed, it is then vigorously
compressed. In this state it keeps much longer than compressed yeast; in order
that it may keep still longer it may be submitted to a process of desiccation;

page_text: residues is obtained without the addition of acids or of malt; this saccharification
takes place spontaneously owing to the action of the diastases secreted by the
mucedine which was set or sown.
3. If a mucedine is selected which has sufficient diastasic power, the final attenu-
ation (that is to say the quantity of sugar per litre which is not fermented) _is
perfect and is invariably less than the attenuation obtained by the use of yeast. In
fact it frequently happens that with musts saccharified by amylomyces only traces
of sugar, which are not capable of being directly determined, are found. This
result can never be obtained by the simultaneous use of the most attenuating
yeasts and of malt.
4. Not only is alcohol produced in sufficient quantity for the extraction of that
'product to be remunerative, but a mould is also obtained which, either in a fresh
condition, or desiccated and compressed into tablets or reduced to powder, is capable
of being used as a substitute for malt or for the preparation of diastases or as a
leaven for bakery or finally as yeast.
5. Certain moulds, and in particular Amylomyce Rouxii, the chlamydomucor
orizoe of Java, and the aspergillus orizce of Japan resist for longer than yeasts to
desiccation. After being kept for three years in a dry state, amylomyce commences
to work in a few hours.
6. The use in bakeries of moulds prepared according to the aseptic methods of
Mr. Pasteur prevents all diseases which might be caused by the employment of raw
Chinese yeast.
In fact, Chinese yeast, according to the results of the examination made by
Dr. Calmette, always contains in addition to amylomyces, a large number of
aspergillus and other microscopic organisms some of which under certain conditions
may become pathogenetic for human beings and for animals.
READINESS OF FILTRATION.
Persons who are conversant with the methods of distilling cereals are well
acquainted with the difficulties experienced in separating the solid from the liquid
part of the malt residuum. Our process of fermentation of the malt residuuni by
means of moulds entirely obviates these difficulties.
If, for example, we are treating the malt residuum by amylomyces, as soon as
the blue coloration produced by the iodine upon the soluble starch as a result of
the saccharification has disappeared, the filtration of the liquid becomes very simple
and extremely hard cakes can then be obtained by means of the filter-press. These
cakes are of greater value than those which could be obtained by submitting to the
hydraulic press the drainings of ordinary malt residues, because the amylomyces
which has been reproduced has fixed a large quantity of soluble nitrogeneous
substances. In this manner the liquid is to some extent deprived of its nutritive
substances for the benefit of what is insoluble, which is an advantage if clear
residuary distillation liquors cannot be used.
If on the other hand this latter is to be used the cloudy malt résiduum is sub-
mitted to distillation after fermentation by our moulds, the fecula which remains
which has not been converted into starch at the time of the treatment and which
consequently was not susceptible of being saccharified, is dissolved during the
passage to the columns; at the same time a portion of the nitrogeneous substances
contained in the mycelium of the mould used, passes in solution in such a manner
that the residuary distillation liquors in this case preserves practically the same
degree of richness in nutritive elements and also remains as advantageous as
residuary distillation liquor which has not been treated, for being used again in
the manufacture of yeast by the usual methods.
MANUFACTURE OF THE MOULDS IN A STATE OF PURITY.
Our process also permits of the manufacture in a very economical manner of the
moulds in a pure state. In fact, owing to the facility with which the filtration of
the malt residuum can be effected as a result of the treatment by our moulds, it
is only necessary to filter this malt residuum aseptically in order to obtain a' straw,

page_text: cake containing a large quantity of mycelium; this latter may be utilised either in
a fresh condition or in a dry state for the manufacture of alcohol by the Chinese
method. In order to dry it a current of air heated to 30° to 35° C. or any other
suitable method of desiccation may be employed; we employ with great success the
process of low temperature desiccation (10° to 12° C.) by means of the anhydrous
fecula described in the Specification of Patent No. 23224 of 1893.
MANUFACTURE OF PURE YEASTS.
We can likewise set or sow in our malt residuum or sterile residuary distillation
liquor, at the same time as the amylomyces, one or more kinds of pure yeast and
thus obtain pure yeast at an extremely low cost.
The amylomyces which is mixed with it does not in any way interfere with the
alcoholic fermentation, the yeast acting more quickly and at a lower temperature
than the amylomyces. It is also possible to obtain pure yeast free from all foreign
substances and from every other micro-organism by employing for the manufacture
of pure yeast the liquid fermented by the amylomyces and by employing it before
distillation. This separation of the liquid can be effected either by means of a
filter-press, the filtered liquid being again sterilised, or by allowing the malt
residuum to remain in the vat when the fermentation excited by the mucedine is
terminated, the upper part is decanted and this becomes absolutely limpid and it is
then placed in an aseptic vat; or finally by arranging in the midst of the liquid a
sterilisable filter of any appropriate description, a clear liquid may be obtained
during the continuance of the fermentation. The liquid obtained by one or other
of these methods may be employed in the usual process of fermentation as source of
diastase.
EMPLOYMENT OF ANTISEPTICS.
Instead of making use of aseptic apparatus, open vats may be used equally well,
use being made (in spite of their defects) of antiseptic methods.
The antiseptic method of fermentation which have been so far patented, have,
according to the information in our possession, only been protected for the special
case of fermentation by means of yeasts.
We apply to working by means of moulds, these methods and also the antiseptic
methods hitherto known and employed.
We have found that the moulds can work in the presence of antiseptic substances
which alcoholic yeasts will not support at first without losing their reproductive
power. It is therefore within the bounds of probability that, even without
acclimatation of the moulds to the antiseptics, satisfactory results may be obtained.
In this case, instead of exciting the fermentation of a few hundreds of hectolitres
of malt residues by a single ball containing one or two grammes of moulds it will
be preferable to previously make a small leaven the volume of which would be
increased little by little until the volume of a fermenting vat is reached, whereupon
a second vat might be put into fermentation by syphon action, and so on.
Dated this 15th day of January 1897.
HASELTINE, LAKE & Co.,
45, Southampton Buildings, W.C., Agents for the Applicants.
COMPLETE SPECIFICATION.
A Process for Utilizing Moulds or Microscopic Fungi for Extracting
from Residues, particularly those arising from the Treatment of
Amylaceous and Sugary Substances, Alcohol and Mucedines
capable of being used in Panification, Distillation, Saccharification,
and the like.
We, AUGUSTE COLLETTE, the Younger, and AUGUSTE BOIDIN, both of Séchin,
Departement du Nord, in the Republic of France, Distillers, do hereby declare the

page_text: nature of this invention and in what manner the same is to be performed, to be
particularly described and ascertained in and by the following statement:-
This invention relates to a process for the utilization of amylaceous or sugary
substances and likewise of a portion of the organic and mineral substances con-
tained in residues of any description whatsoever, by employing, for converting
them into alcohol, moulds possessing a certain diastasic activity and a fairly high
degree of fermentability.
The moulds which we employ are preferably such as are not pathogenetic in
order that they may be utilized in the manufacture of bread or in brewing beer
and also in the saccharification of the musts or worts of cereals.
In order to simplify the description, we will take by way of example the utilisa-
tion of the carbohydrates contained in the malt residuum of distillation, by a
mould discovered in Indo-China by Doctor A. Calmette and named by him
Amylomyces Rouxii.
According to the researches of Maerker and of Schultze a hectolitre of residuum
of the manufacture of yeast contains 2 kilogrammes 600 grammes of fermentable
products, sugar, dextrine and soluble starch, 720 grammes of albuminoid
substances and 250 grammes of mineral substances.
If after having sterilised a medium of this nature we introduce into it a trace of
amylomyces, this mucor develops; it first of all transforms the soluble starch and
the dextrine into glucose and finally into alcohol. At the same time the density
of the liquid diminishes; when it rests constant it is passed through a sieve in
order to collect the mycelium formed and is distilled to recuperato the alcohol.
If it is aerated considerably a large quantity of mycelium is obtained. If, on the
other hand, the supply of air is restricted, the weight of mycelium is less but the
quantity of alcohol obtained is greater. In order to obtain in an industrial opera-
tion 1 kilogramme of pressed amylomyces and a litre of phlegm at 100° of the Gay
Lussac alcoholmeter, that is to say, of theoretically pure phlegm per hectolitre of
residuary distillation liquors treated, I proceed as follows:-
A trace of amylomyces is sown in a hectolitre of yeast-manufactory residuary
distillation liquor, it is acrated for several hours the liquor being then left to
ferment for several days whereby a fermented must is obtained which no longer
contains either starch or sugar and containing one litre of alcohol at 100° and one
kilo of amylomyces in a pressed condition.
The operation can be carried out either with malt residuum clarified by decanta-
tion or filtered by being heated under pressure or else with thick or cloudy malt
residuum. The mycelium obtained undergoes a washing process in order to free it
from the fermentable substances in which it is immersed, it is then vigorously
compressed. In this state it keeps much longer than compressed yeast; in order
that it may keep still longer it may be submitted to a process of desiccation.
PRODUCTION OF SPORES.
For the purpose of producing spores a sterilisable receptacle is filled with large
straws or an inert body of any suitable description in order to increase the extent
of surface and to facilitate the access of the air to every part of the
receptacle; the liquid to be treated is introduced into this receptacle, sterilisation is
then effected and after cooling has taken place the liquid is caused to run off from
below whilst avoiding the ingress of any un-filtered air. The liquid to be treated
need not be a decoction of an amylaceous or sugary substance, thus, for example,
the yeast of beer, the price of which has fallen to such an extent that it frequently
forms only a waste product which the brewer would rather be without, may be
utilized. In this case it is preferable to prepare a kind of broth by boiling either
in an open vessel or under pressure and either with or without the addition of acid.
Although not containing any reducing sugar, this broth gives an abundant supply
of mould.
In the same way the washing waters of yeast, the residuary liquors of beetroot
distillation and so on, may be employed, these being brought to the most suitable'
degree of acidity for each mould.
.

page_text: is obtained which archougn or very small weight is yet
extremely active and which may be employed either as a saccharifying agent or as
yeast.
EMPLOYMENT IN BAKERIES.
We have found that if 1 to 2 per cent. of mycelium is incorporated with a paste
or dough it rises or works in a few hours. After baking, the bread obtained is
exactly similar to that obtained when the best yeasts from cereals or from beer are
used.
In order to make use of our moulds in the bakery or the making of
pastry, all
that is necessary is to dilute them with either warm water, sugared water or a
vegetable decoction containing either sugar or starch.
With amylomyce can also be made what in England is made with yeast in
preparing what is called a "fruit"; rice, potatoes or other farinaceous substances
are cooked and mashed, then when the mash has cooled down to 35° to 40° C.
it is sprinkled with the mould, the whole is placed in an earthenware vessel which
is kept in a warm place. It is not necessary, as in the English "fruit," to add flour.
The saccharification and fermentation commence immediately and it is only
necessary to wait a few hours in order to obtain a semi-solid mass, a few kilo-
grammes of which are sufficient to produce in one or two hours the quantity of gas
necessary to the raising of 100 kilogrammes of flour.
By this method bread of perfect whiteness and exquisite taste is obtained; the
bread does not turn sour and keeps much longer than that made with leaven of
sour dough. It is not possible to obtain the same degree of working with raw
Chinese yeast which besides imparts to the bread a musty taste.
This process will be found extremely useful in hot countries because our moulds
are ready for fermentation at the temperature of such countries which is almost
always too high for yeasts to be employed.
Our process possesses the following advantages.
1. It allows of the employment of carbohydrates and a portion of the
albuminoid and mineral substances of all residues which are not sufficiently rich
in these products for their extraction by means of the processes ordinarily employed
to be remunerative. This result is obtained by turning to account the property
which these moulds possess of utilizing in a manner which is almost perfect the
least traces of nutritive elements which are placed within reach of their action.
2. The saccharification of the starch and of the dextrine contained in these
residues is obtained without the addition of acids or of malt; this saccharification
takes place spontaneously owing to the action of the diastases secreted by the
mudecine which was set or sown.
3. If a mucedine is selected which has sufficient diastasic power, the final attenu-
ation (that is to say the quantity of sugar per litre which is not fermented) is
perfect and is invariably less than the attenuation obtained by the use of yeast. In
fact it frequently happens that with musts saccharified by amylomyces only traces
of sugar, which are not capable of being directly determined, are found. This
result can never be obtained by the simultaneous use of the most attenuating
yeasts and of malt.
4. Not only is alcohol produced in sufficient quantity for the extraction of that
product to be remunerative but a mould is also obtained which either in a fresh
condition, or desiccated and compressed into tablets or reduced to powder, is capable
of being used as a substitute for malt or for the preparation of diastases or as a
leaven for bakery or finally as yeast.
5. Certain moulds, and in particular Amylomyce Rouxii, the chlamydomucor
orizoe of Java, and the aspergillus orizoe of Japan resist far longer than yeasts to

page_text: desiccation. After being kept for three years in a dry state, amylomyce commences
to work in a few hours.
6. The use in bakeries of moulds prepared according to the aseptic methods of
M. Pasteur prevents all diseases which might be caused by the employment of raw
Chinese yeast.
In fact, Chinese yeast, according to the results of the examination made by
Dr. Calmette always contains in addition to amylomyces, a large number of
aspergillus and other microscopic organisms some of which under certain conditions
may become pathogenetic for human beings and for animals.
READINESS OF FILTRATION.
Persons who are conversant with the methods of distilling cereals are well
acquainted with the difficulties experienced in separating the solid from the liquid
part of the malt residuum. Our process of fermentation of the malt residuum by
means of moulds entirely obviates these difficulties.
If, for example, we are treating the malt residuum by amylomyces, as soon as
the blue coloration produced by the iodine upon the soluble starch as a result of
the saccharification has disappeared, the filtration of the liquid becomes very simple
and extremely hard cakes can then be obtained by means of the filter press. These
cakes are of greater value than those which could be obtained by submitting to the
hydraulic press the drainings of ordinary malt residues, because the amylomyces
which has been reproduced has fixed a large quantity of soluble nitrogeneous
substances. In this manner the liquid is to some extent deprived of its nutritive
substances for the benefit of what is insoluble, which is an advantage if clear
residuary distillation liquors cannot be used.
If on the other hand this latter is to be used the cloudy malt residuum is sub-
mitted to distillation after fermentation by our moulds, the fecula which remains
which has not been converted into starch at the time of the treatment and which
consequently was not susceptible of being saccharified, is dissolved during the
passage to the column; at the same time a portion of the nitrogeneous substances
contained in the mycelium of the mould used, passes in solution in such a manner
that the residuary distillation liquors in this case preserves practically the same
degree of richness in nutritive elements and also remains as advantageous as
residuary distillation liquor which has not been heated, for being used again in
the manufacture of yeast by the usual methods.
MANUFACTURE OF THE MOULDS IN A STATE OF PURITY.
Our process also permits of the manufacture in a very economical manner of the
moulds in a pure state. In fact, owing to the facility with which the filtration of
the malt residuum can be effected as a result of the treatment by our moulds, it
is only necessary to filter this malt residuum aseptically in order to obtain a straw
cake containing a large quantity of mycelium; this latter may be utilised either in
a fresh condition or in a dry state for the manufacture of alcohol by the Chinese
method. In order to dry it a current of air heated to 30° to 35° C. or any other
suitable method of desiccation may be employed; we employ with great success the
process of low temperature desiccation (10° to 12° 0.) by means of the anhydrous
fecula described in the Specification of Patent No. 23224 of 1893.
MANUFACTURE OF PURE YEASTS.
We can likewise set or sow in our malt residuum or sterile residuary distillation
liquor, at the same time as the amylomyces, one or more kinds of pure yeast and
thus obtain pure yeast at an extremely low cost.
The amylomyces which is mixed with it does not in any way interfere with the
alcoholic fermentation, the yeast acting more quickly and at a lower temperature
than the amylomyces. It is also possible to obtain pure yeast free from all foreign
substances and from every other micro-organism by employing for the manufacture
of pure yeast the liquid fermented by the amylomyces and by employing it before
distillation. This separation of the liquid can be effected either by means of a

page_text: filter-press, the filtered liquid being again sterilised, or by allowing the malt
residuum to remain in the vat when the fermentation excited by the mucedine is
terminated, the upper part is decanted and this becomes absolutely limpid and it is
then placed in an aseptic vat; or finally by arranging in the midst of the liquid a
sterilisable filter of any appropriate description, a clear liquid may be obtained
during the continuance of the fermentation. The liquid obtained by one or other
of these methods may be employed in the usual process of fermentation as source of
diastase.
EMPLOYMENT OF ANTISEPTICS.
Instead of making use of aseptic apparatus, open vats may be used equally well,
use being made (in spite of their defects) of antiseptic methods.
The antiseptic methods of fermentation which have been so far patented,. nave
according to the information in our possession, only been protected for the special
case of fermentation by means of yeasts.
We apply to working by means of moulds these methods and also the antiseptic
methods hitherto known and employed.
We have found that the moulds can work in the presence of antiseptic substances
which alcoholic yeasts will not support at first without losing their reproductive
power. It is therefore within the bounds of probability that, even without
acclimatation of the moulds to the antiseptics, satisfactory results may be obtained.
In this case, instead of exciting the fermentation of a few hundreds of hectolitres
of malt residues by a single ball containing one or two grammes of moulds it will
be preferable to previously make a small leaven the volume of which would be
increased little by little until the volume of a fermenting vat is reached, whereupon
a second vat might be put into fermentation by syphon action, and so on.
Having now particularly described and ascertained the nature of our said in-
vention and in what manner the same is to be performed, we declare that what we
claim is: :―
1. The process above described for the utilisation of moulds for extracting from
all residues and from those arising from the treatment of amylaceous and sugary
substances in particular, of alcohol and mucedines susceptible of use in panification,
distillation, saccharification and so forth, to which we give the names amyl-leavens
and amyl-yeasts.
2. The application of saccharifying moulds for obtaining malt residues which may
be readily filtered and for increasing the weight and the richness in nitrogeneous
substances of the insoluble portion.
3. The economical manufacture of moulds in the form of malt residues reduced to
· powder or compressed into tablets.
4. The simultaneous manufacture of moulds and of yeasts in a pure state and at
a. very low cost.
5. The application to the fermentation by means of moulds of the antiseptics
employed in fermentation by means of saccharomyces.
6. The use of malt residues fermented and filtered before distillation in order
to be re-introduced in the usual methods with a view to utilising the diastases
secreted by the saccharifying mould.
Dated this 14th day of October 1897.
HASELTINE, LAKE & Co.,
45, Southampton Buildings, London, W.C., Agents for the Applicants.