Improvements in and in the Production of Diastatic and Alcoholic Ferments, and the Utilization of the Bye-products.
Year: 1894

page_text: Date of Application, 11th Sept., 1894-Accepted, 17th Nov., 1994
CUMI LEID SIMITICATION.
Improvements in and in the Production of Diastatic and Alcoholic
Ferments, and the Utilization of the Bye-products.
I, JOKICHI TAKAMINE, of 2111 North Jefferson Avenue, Peoria, County of
Peoria, State of Illinois, United States of America, Chemist, do hereby declare
the 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:-
The object of this invention is to prepare and make diastatic enzyme, Taka-Koji
diastase, a soluble ferment in a concentrated form which possesses the power of
transforming starch into sugar for use in various industries, in an economical and
practical manner, and by a process not heretofore practised; also to prepare and
manufacture Taka-Koji ferment for use in the arts, such as alcoholic fermentation,
beer-brewing, yeast-making, etc. in an active appropriate, efficient and concentrated
condition; also an alcoholic ferment mash for developing active alcoholic ferment
cells, possessing fermenting properties similar to, but more effective than that
substance commonly known as yeast mash, for use in the arts in a more effective
condition than heretofore produced.
My invention is based upon the utilization of the property possessed by certain
fungi during their growth upon proper media, of producing diastatic and fermenting
properties. For the purpose of my invention I use the microscopic fungus
eurotium oryza with the best results, but other mould fungi belonging to the
genus aspergillis and to the genera mucor and penecilium may also be used. I
prefer to use as the raw material upon which to grow the fungus, comminuted or
broken grains of the cereals, such as wheat, corn, oats, etc. and preferably the
comminuted or broken grains from which the greater part of the starchy matter
has been removed, leaving the broken or comminuted parts in the form of brans,
shorts, middlings, etc, and particularly wheat, bran. I may use other substances
which supply the necessary ingredients for the growth of the fungi, such as slop,
solid portions from alcoholic distillation, grain from beer brewing, oil cake, brans,
and slop from glucose and starch factories, cloth crash or other fabric coated with
an artificial compound. I prefer to use the branny part of the grain because it is
of a loose and coarse nature, thus affording a large surface for the growth of the
fungus and a ready access of air, one of the necessary conditions of its growth..
Being rich in albuminoids and nitrogenous matter, they form a nutritious soil upon
which to sow and propagate the fungus growth.
They contain a large percentage of woody fibre which makes their use especially
beneficial in the process of extraction, acting as a filter. They are cheap and
abundant at all seasons of the year.
In describing my invention I shall apply the term "Taka-Koji" to a mass such
as above indicated, upon and throughout which the specified fungus has been
developed and grown to the desired stage under proper conditions of temperature
and manipulation, see the Specification of Letters Patent granted to me No. 17374
of 1891.
I have discovered that during the growth of the fungus under the manipulations
and conditions above described, an abundant supply of enzyme is produced.
The enzyme is extracted by percolating water, or water mixed with alcohol,
through the Taka-Koji, or by steeping the Taka-Koji in the liquid. By repeatedly
treating fresh quantities of Taka-Koji with the same liquid, the strength of the
liquid may be increased. The enzyme contained in the solution may be precipi-
tated therefrom by the addition of from one-and-a-half to three times its volume
of 90 % absolute alcohol and agitating the same. The precipitate is in the form

page_text: of solid matter, consisting mainly of diastatic enzyme and of traces of mineral
matter. The precipitate is separated from the supernatant liquid by decantation
or filtration, or wholly by filtration. It may then be washed with alcohol
containing about 85% absolute alcohol, and dried slowly at a low temperature in
a dry room, or it may be dehydrated by strong alcohol, by which latter means
enzyme can at once be obtained in a powdery form. It may be further purified
by redissolving in water and reprecipitated, by washing with alcohol, and drying.
The product thus obtained is an amorphous dry mass or powder of almost white
or light, yellowish-brown color. It is readily soluble in water, and possesses the
power of transforming gelatinized starch into sugar. By concentrating the
extract as above described, the quantity of alcohol required in precipitating or
dehydrating or washing the same is economized. The product may be further
concentrated by evaporation at a temperature not to exceed 50° C. so as to form a
thick syrup or semi-solid in which form it can be preserved without change before
precipitating with alcohol.
The article obtained as above described, while possessing diastatic property in
a bigh degree, lacks the characteristic of giving blue coloration when treated with
tincture of guiacum mixed with hydrogen peroxide, as in the case of malt diastase.
In the case of unsteamed bran, however, this coloration is given in a slight degree,
since the raw cereals, of which the bran is a portion, originally contain a small
portion of malt diastase, which is necessarily extracted in the case of unsteamed
bran simultaneously with the enzyme.
The alcoholic mother liquid contains a small quantity of unprecipitated enzyme
which may be used again in admixture with fresh alcohol for precipitating fresh
enzyme solution. The use of the mother liquid may be repeated until its efficiency
is impaired by the accumulation of various substances, then the alcohol contained
therein may be recovered by distillation. After such distillation, the residue,
containing a considerable quantity of sugars, may be fermented or distilled for
alcohol, or used for other purposes. The bran remaining after the extraction with
water of the enzyme may be pressed and dried for cattle food, or it may be used
over again as a base upon which to grow fresh quantities of the fungus.
I have discovered that enzyme may be precipitated from a watery extract from
a mixture in about equal proportions of Taka-Koji and fresh raw material, such
as brans, shorts, etc. I have also discovered that satisfactory results may be
obtained by first obtaining a watery extract from Taka-Koji, then a watery extract
from the raw materials, brans, shorts, etc. mixing the two extracts and then
precipitating the enzyme. I may also obtain a precipitate from the Taka-Koji and
then a precipitate from the raw material, brans, shorts, etc. and mix the two
-precipitates.
Taka-Koji possesses two distinct properties, namely, diastatic property, or property
of converting starch into sugar; and fermenting property. It is exceedingly
important from an economical standpoint to separate these two properties, as well
from the Taka-Koji, as from each other, a separation that has never before been
successfully accomplished on an economical scale, so that when a conversion is
desired the ferment property need not be wasted, and when fermentation is desired,
only the ferment property of the Taka-Koji may be employed without waste of the
diastatic property. The ferment portion may be directly separated from the dried
Taka-Koji, in the form of a fine powder, by sifting the Taka-Koji. This powder
comprises the bloom of the fungus and possesses fermenting properties in a
remarkable degree. The residue left after silting contains the diastatic property,
and may be used as a diastatic agent. These two properties may be extracted
together from the Taka-Koji by soaking or steeping the same in water, thoroughly
stirring and then pressing the mass, the diastatic property dissolving in the water,
and the ferment part, insoluble in water, becoming detached by attrition and
remaining suspended in the liquid. The soaking or steeping, separating and pressing
operations may be repeated as often as may be desired, in order to effect a thorough
separation. The ferment may be separated from this solution by decantation or

page_text: filtration. It is, in the form of a fine dry powder, composed of young, immature
spores of the mycelial fungus, the color varying with the different fungi employed.
It consists of microscopical, spherical cells, possessing the property of being
transformed into alcoholic ferment cells when submerged in sugar solution under
suitable conditions.
The residue of Taka-Koji, after separation therefrom of the diastase and ferment,
may be used over again several times to grow a fresh supply of fungi, either as a
raw material, or by mixing it with fresh material. When wheat bran is used, the
residue can thus be used for the propagation or production of four or five crops of
fungi, without the addition of fresh material, or of any nutritious or fertilizing
substances, and with the addition of only about 10% of fresh bran, each time, the
original residue may be used to grow more than 10 crops of the fungi.
I have discovered that a mixture of Taka-Koji and the raw material, as above
specified for making Taka-Koji, namely, comminuted or broken grains of cereals,
preferably from which the greater starchy matter has been removed, and
particularly wheat brans, shorts, middlings, etc. which I shall designate generically
by the word brans, forms a converting agent for converting starch into sugar.
A mixture of Taka-Koji diastase and braus produces good results, as does also an
extract from brans and either Taka-Koji or Taka-Koji diastase, or an extract from
a mixture of brans and Taka-Koji. The starch material to be converted is first
cooked in water in the usual way, and then the converting agent, as above
specified, is added. The residue left after extracting the enzyme, diastase and
fermenting properties from Taka-Koji, constitutes a rich cattle food, inasmuch as
during the process described to which Taka-Koji is subjected, the carb-hydrate
constituents of the original raw material are eliminated, leaving the residuum rich
in protein, the property that gives cattle food its richness.
To prepare alcoholic ferment, I mix brans of the different cereals, including
shorts, middlings, etc., or a mixture of the same with from 3% to 10% by weight
of Taka-Koji, as above described, or Taka-Koji diastase corresponding to from 3%
to 10% by weight of Taka-Koji, thus enriching the mash with the nitrogenous
ingredients or ferment cells and with comparatively a small amount of saccharine
inatter, in order to induce or cause the nitrogenous matter of the brans, etc. to change
into a more assimilable condition for the multiplication of ferment cells, and also
for the conversion into sugar of any starchy matter contained in the same.
I have discovered that a mash made by using Taka-Koji, and especially
Taka-Koji diastase, as above described, mixed with brans produces a medium for
developing ferment cells far superior to any other, and the ferment cells thus
developed possess greater activity and fermenting power than any other ferment
cells. To the mixture specified, is added from four to eight times its weight of
water and the mixture heated to about 65° C. After remaining at that temperature
from 15 to 30 minutes, the temperature is raised to the boiling point and maintained
at that temperature from 10 to 20 minutes. The mash is then cooled down to
about 60° C., and the same quantity of Taka-Koji, or Taka-Koji diastase, is added
in order to convert the remaining starch. The liquid portion is then separated
from the solid material and is rapidly heated to the boiling point and maintained at
that point from 10 to 20 minutes in order to sterilize the same. Any insoluble
matter remaining in the liquid is allowed to settle, or is filtered out, and the clear
liquid quickly cooled to a temperature of from 12 to 35° C. when it is ready
for the introduction of ferment cells for development and multiplication. I prefer
to employ the ferment cells known as Moto, prepared as described in ny said
former Specification, and which are hereinafter called for distinction Taka-Moto.
There is no special requirement or proportion as to the amount of the clear
liquid or the ferment. A large amount of Taka-Moto will complete the develop-
ment and multiplication of the ferment cells soine what earlier than a small amount,
but the proper proportion to be added will be readily understood by those skilled
in the art.
After adding the Taka-Moto, the mash is allowed to stand from 12 to 60 hours

page_text: until the ferment cells of the Taka-Moto settle to the bottom and there
further development and multiplication ceases, leaving the liquid bright and clear.
The ferment cells which have settled to the bottom are separated, washed with
water and made into a pasty mass, or pressed into solid form, or otherwise treated
and applied to various uses in the arts, such as bread-inaking, brewing, etc.
small percentage of alcohol remaining in the supernatant liquid may be recovered
by distillation.
The
Having now particularly described and ascertained the nature of my inven-
tion, and in what manner the same is to be performed, I declare that what I
claim is.
1. The process of making Taka-Koji diastase and Taka-Koji ferment, which
consists in treating Taka-Koji with water, whereby the diastatic property of the
Taka-Koji is dissolved and extracted, leaving behind the ferment cells or portion;
as and for the purpose set forth.
2. As a composition of matter, Taka-Koji diastase in the form of a liquid
solution, formed by percolating water through Taka-Koji, which possesses diastatic
property; as and for the purpose specified.
3. As a composition of matter, Taka-Koji ferment in the form of a fine, dry
powder, comprising the ferment cells of Taka-Koji, which possesses fermenting
properties; as and for the purpose set forth.
4. The process of making Taka-Koji diastase and Taka-Koji ferment, which
consists in treating Taka-Koji with water, whereby the diastatic property possessed
thereby dissolves and passes into solution and the ferment properties are detached
and held in suspension in the solution, and finally separating from the solution the
ferment cells held suspended therein, by filtration; all substantially as and for
the purpose described.
5. The process of preparing and making diastatic enzyme, which consists in
treating Taka-Koji with water whereby the soluble enzyme is extracted therefrom,
then precipitating with alcohol, washing the precipitated soluble matter with alcohol,
and drying; all substantially as described.
6. The process of preparing and making diastatic enzyme, which consists in
extracting from Taka-Koji the soluble matter contained therein, then precipitating
said solid matter, then washing said precipitate, and finally drying; as and for
the purpose set forth.
7. The process of preparing and making diastatic enzyme, which consists in
first extracting with water the soluble matter contained in Taka-Koji, next
extracting with water the soluble matter contained in the raw material, brans, etc.
then thoroughly mixing the two extracts, then precipitating with alcohol the solid
matter contained in said mixture of extracts, and finally drying said precipitates;
all substantially as and for the purpose set forth.
8. The process of preparing and making diastatic enzyme, which consists in first
extracting with water the soluble matter contained in Taka-Koji, then precipitating
the solid matter contained in said extract, then drying said precipitate, next
extracting with water the soluble matter contained in a mass of the proportion
specified of fresh, raw material, comminuted or broken grains of cereals, brans,
shorts, middlings, etc. next precipitating from said last-mentioned extract the
solid matter contained therein, then drying said precipitate, and finally mixing the
two precipitates and washing with alcohol; all substantially as and for the purpose
specified.
9. The process of preparing and making diastatic enzyme in a crude form
which consists in extracting the soluble matter contained in Taka-Koji with water,
concentrating the extract by repeatedly treating fresh quantities of said mass
therewith, and finally evaporating the extract; all substantially as described.
10. The process of preparing and making diastatic enzyme in a crude form which
consists in mixing Taka-Koji and brans, then extracting with water from this
mixture the soluble matter contained therein, concentrating the solution by repeatedly

page_text: treating fresh quantities of said mixture therewith and evaporating, substantially as
and for the purpose set forth.
11. The process of preparing and making diastatic enzyme in a crude form, which
consists in first extracting with water the soluble matter contained in a mass of
Taka-Koji, next, extracting with water the soluble matter contained in a mass of
brans, next, thoroughly mixing the two extracts thus obtained, and, finally,
concentrating the mixture by evaporation; as and for the purpose set forth.
12. As a composition of matter, diastatic enzyme in a crude form, obtained by
extracting with water the soluble matter contained in a mass of Taka-Koji, and
which has been repeatedly used with fresh quantities of said mass, and then
evaporated to the form of a syrupy liquid or semi-solid of a yellowish brown
to dark-brown color, of a pleasing mushroom flavor, possessing the power of
converting hydrolized starch into sugar, and differing from starch converting
enzyme of germinating grains in not giving the characteristic chemical reaction
of the starch converting enzyme of germinating grain; substantially as described.
13. As a composition of matter, diastatic enzyme in a crude form, composed of
a mixture of extracts from brans and from Taka-Koji, said mixture of extracts
concentrated to a syrupy liquid of semi-solid form, of light-brown or dark-brown
color, of mushroom flavor, possessing the power of converting starch into sugar and
not giving the characteristic reaction of the starch converting enzyme of germinating
grains except in a slight degree, due to the existence of a small quantity of starch
converting enzyme originally present in the raw grain used in admixture from
which the watery extract has been prepared; substantially as and for the purpose
set forth.
14. As a composition of matter, diastatic enzyme, that is, soluble ferment in
the form of a solid of light-brown or yellowish-white color, possessing the power of
converting hydrolized starch into sugar aud giving a milky-white coloration when
treated with a mixture of solution of hydrogen peroxide and tincture of guiacum ;
substantially as described.
15. As a composition of matter, diastatic enzyme in the form of a solid, of
yellowish-white or light-brown color, soluble in water, possessing the power of
converting hydrolized starch into sugar to a degree at least equal to the
starch converting power of the diastase of germinating grain, and giving
a faint
blue coloration when treated with a mixture of hydrogen peroxide and tincture
of guiacum, obtained by precipitating with alcohol from the admixed extract of the
soluble matters of Taka-Koji and of brans; all substantially as and for the purpose
described.
16. As a composition of matter, diastatic enzyme in the form of a solid, of a
yellowish-white or light-brown color, soluble in water, possessing the power of
converting hydrolized starch into sugar, composed of a mixture of the precipitate
obtained from a watery solution or extract of a mass of brans and Taka-Koji ; all
substantially as and for the purpose described.
17. The process of developing and cultivating alcoholic ferment cells, which
consists in heating gradually to the boiling point, a mixture of the bran of the
cereals, Taka-Koji or Taka-Koji diastase, and water in the proportions specified,
adding after cooling a portion of Taka-Koji or Taka-Koji diastase equal to that
first introduced into the mixture, separating and sterilizing the liquid portion, and
adding Taka-Moto to the clarified liquid for development and multiplication of the
ferment cells under the temperature and manipulation specified; all substantially as
described.
18. The process of cultivating and developing alcoholic ferment cells, which
consists in heating gradually to the boiling point, a mixture of the bran of cereals,
Taka-Koji or Taka-Koji diastase, and water in the proportions specified, adding after
cooling a portion of Take-Koji or Taka-Koji diastase equal to that first introduced
into the mixture, separating and sterilizing the liquid portion, adding Taka-Moto
to the clarified liquid, for development and multiplication of the ferment cells under
the temperature and manipulation specified, drawing off the clarified liquid, washing

page_text: the ferment cells with water and forming the same into a pasty or semi-solid mass;
all substantially as described.
19. The process of preparing a mash for the cultivation and development of
alcoholic ferment cells, which consists in heating gradually to the boiling point, &
mixture of the bran of cereals, Taka-Koji or Taka-Koji diastase, and water in the
proportions specified, adding after cooling a portion of Taka-Koji or Taka-Koji
diastase equal to that first introduced into the mixture, separating the liquid portion
from the mixture, and sterilizing the same, allowing the same to rest and cool; all
substantially as described.
20. The method of converting starchy matter into sugar which consists in
cooking the starch material in water, then introducing into and mingling with it a
mixture or composition consisting of brans and Taka-Koji, or brans and Taka-
Koji diastase, or a mixture of the watery extract of brans and. Taka-Koji diastase,
or a watery extract from a mixture of brans and Taka-Koji; as and for the purpose
set forth.
21. As a converting agent for converting starchy matter into sugar a mixture of
bran and Taka-Koji, or brans and Taka-Koji diastase, or a watery extract from a
mixture of brans and Taka-Koji diastase, or a watery extract from a mixture of
brans and Taka-Koji; as and for the purpose specified.
22. A cattle food comprising the residue of Taka-Koji after treating the same to
extract therefrom the enzyme, diastase and ferment portions; as and for the purpose
set forth.
11
11 1
A
T
1004
Chicago, Ills., U.S.A.
JOKICHI TAKAMINE.
W. Lloyd Wise,
46, Lincoln's Inn Fields, London, W.C., Agent for the Applicant.