Fermented Soy
Foods and solid or semisolid. They are made by
Sauce
Lewis B. Lockwood^
Allan K. Smith
Soybeans, in the form of soy sauce
and soybean cheese, paste, sprouts,
milk, and curd, have been an impor-
tant source of protein in the diet of the
Chinese and Japanese for centuries. In
Asia the whole bean is not ordinarily
eaten. The people favor mostly fer-
mented soy products or other modifi-
cations—sprouts, curd, or milk—in
which the characteristic flavor and
shape of the beans are lost. Only soy
sauce and monosodium glutamate have
found much favor in Western countries.
Asiatic people live largely on a vege-
table diet. It is estimated that in China
meat and eggs make up less than 3
percent of the food of the peasants,
compared to 21 percent among Ameri-
cans. About 95 percent of the protein
eaten is of vegetable origin. Much of
it comes from soybeans, w^hich form
about 20 percent of the basic diet in
Northern China.
Soy sauce, shoyu in Japanese, is the
most popular use made of the soybean.
It is a dark-brown liquid, very salty and
sharp in flavor. Its odor suggests cooked
beef. It accentuates the flavor of vege-
tables and meat. Sweetening and thick-
ening agents and spices may be added
to give the sauce variety in flavor.
Soybean paste is a semisolid, mushy
food. Its flavor is like that of soy sauce.
In Japan it is know'n as miso. It is used
as a relish for rice, in soups, and to
add flavor to vegetables.
Soybean cheeses of the Orient are
fermenting the soybean curd. Soybean
cheeses are unlike American and Euro-
pean cheeses in flavor and appearance;
it is too bad that the name cheese has
been applied to them. Some of the soy-
bean cheeses have a flavor resembling
soy sauce. Others are quite different.
Many are too salty for the American
taste.
The Chinese made soy sauce in an-
cient times as a household industry.
Descriptions of the process are found in
books written more than 1,500 years
ago. It has remained largely a family
art; even now^ some manufacturers
point wdth pride to the fact that their
factories have been operated as family
enterprises for five centuries.
Soy sauce is manufactured by two
basic processes. One involves a fermen-
tation technique and the other a chem-
ical method. A third procedure, which
is thought to have some advantages, is
a combination of the two. The third
method is still in the developmental
stage. The products of the difTerent
methods difler somewhat in taste and
odor; the fermentation product is the
most acceptable. Sometimes the prod-
ucts of tw^o basic processes are blended.
The fermentation method is a mixed
fermentation by three micro-organ-
isms: A mold, Aspergillus oryzae; a
bacterium, Lactohacillus delbruckii;
and a yeast, Zygosaccharomyces soja,
Z. major y or a yeast closely related to
Haiisenula anómala. In the traditional
Chinese method, the manufacturer
adds a prepared mold culture, a koji,
to the soybeans, but relies on chance
inoculation for the bacterium and
yeast. Modern methods include the use
of pure-culture inocula of all the micro-
organisms needed in the fermentation.
The materials are soybeans, wheat
or other starchy grains or flour, and
salt. First, the beans are washed and
357
358 1950-19 5 1 YEARBOOK
then soaked in water for 12 to 24 hours,
depending on the temperature or sea-
son of the year. Longer soaking is
needed in winter or when the tempera-
ture is low. The soaking finished, the
beans are drained of excess water and
cooked with steam under 10 pounds
pressure. The cooking period covers
several hours.
The second step includes the addi-
tion of soft wheat, which has been
cleaned, roasted, and cracked or very
coarsely ground. It is mixed with the
cooked soybeans in the ratio of about
3 pounds of w^heat (initial weight) to
10 pounds of cooked soybeans (initial
weight).
In the third step, the mixture of
beans and w^heat is inoculated with a
culture of one or more strains of the
mold Aspergillus oryzae. The inocula-
tion cultures are made by growing the
mold on steamed polished rice. An
ounce of rice will make enough koji to
inoculate 10 pounds of the soybean-
parched wheat mash. In modern fac-
tories, cultures of the necessary yeast
and bacteria are added at this point.
A good yeast growth before the mold
growth becomes apparent is believed to
result in a sauce of superior quality. In
some modern plants the order of add-
ing the micro-organisms is changed;
the yeast is. added to the steamed beans
about a day before mixing them with
the parched w^heat^ and the yeast starts
to grow before the mold gets under
way. The procedure is said to give very
good results.
After inoculation, the mash is spread
in a layer 3 inches deep in wooden
trays or baskets about 4 inches deep.
The trays are stacked so that air will
circulate freely over the beans. During
the fermentation stage, the mold grows
throughout the mash and gives off
considerable heat. The temperature of
the material may reach 40° C. (104°
F.) or higher, if it is not controlled.
This phase of the process lasts about 3
days. A thin white surface growth of
mold appears, and turns yellowish as
spore formation begins. The brine fer-
mentation, or second phase of the
OF AGRICULTURE
fermentation process, is then started.
If it is started too soon, the supply of
the mold enzymes, which are respon-
sible for certain necessary chemical re-
actions, will be inadequate. If the start
is delayed too long, many spores will
cover the surface of the beans and may
contribute undesirable flavors to the
sauce.
This molded soybean-wheat mash is
placed in deep vessels and barely cov-
ered with brine made with 22-percent
salt solution. In the old Chinese fac-
tories, 50-gallon earthenware vessels
are used. These are set in the open and
are covered only during rain. Many of
the old Chinese manufacturers believe
that sunlight, even moonlight, afTects
the flavor of the sauce.
The beans are stirred daily for the
first few^ weeks, then weekly until the
end of the fermentation period. In
modern Japanese and Chinese facto-
ries the aging process is carried out in
concrete vats of about 5,000-gallon
capacity. Air is blown through the bean
mash to stir and mix it every 2 or 3
days in the beginning, but after several
weeks the material is aerated for 30
minutes once a week. Gas is given on'
during the first 2 weeks of the brine
fermentation.
After 3 months—preferably after a
year or longer—the mash is pressed to
remove the liquid. This is considered
the best grade of soy sauce. A second
grade is made by suspending the press
cake in 18- to 20-percent brine and
pressing. A third (occasionally a fourth,
or even a fifth) grade is prepared by
further extraction of the press cake in
like manner. Each extraction gives a
product of weaker flavor and hence of
less commercial value than the pre-
ceding one. The sauces are pasteurized
at about 65° C. (149° F.). If higher
pasteurization temperatures are used,
a cloudy sauce results because of the
precipitation of partly degraded pro-
teins. Alum is then added as a floccu-
lating agent, and the%auce is filtered.
The salt content of the sauce, 18 to
20 percent, prevents the growth of most
micro-organisms. Sodium salicylate or
FERMENTED SOY FOODS
^'-naphthol may be added as preserva-
tive, but that is not neeessary if the sah
content is high enough. Cloves, cinna-
mon, nutmegs, pepper, ginger, and
other spices enhance the flavor and are
believed to prevent spoilage. They are
added only to the lower-grade prod-
ucts. Caramel may be added to darken
the sauce. Licorice or maltose may be
added to sweeten it.
The chemical changes in the pro-
duction of soy sauce are complex and
interrelated. Wheat servies as a carbo-
hydrate source for the growth of the
micro-organisms. The mold undoubt-
edly supplies the enzymes necessary to
convert the starch to sugar, which, in
turn, is acted upon by all three micro-
organisms. The mold and yeast pro-
duce some alcohol from the sugar. The
bacteria produce lactic acid and other
organic acids. Esters, such as ethyl
acetate, are also formed by interaction
of th(i alcohols and organic acid. They
account for much of the aroma and
flavor of the sauce. Other important
flavor constituents are amino acids or
salts of amino acids. Monosodium glu-
tamate is the most important of these.
The amino acids are produc(^d by enzy-
matic decomposition of the proteins
of the soybeans and wheat. Most of the
protein degradation and carbohydrate
fermentation occurs during the first 2
v/eeks of the brine fermentation. After
this time, the flavor matures by very
slow reactions, which involve the for-
mation of esters and the* splitting of
clextrins.
The chemical com]30sition of soy
sauce with a specific gravity, at 15° C,
of 1.19 to 1.20 is (in parts per 1,000
parts of soy sauce) : Total solids, 250;
sodium chloride, 130 to 150; total
nitrogen, 6 to 13 ; protein nitrogen, 0.8 ;
amino nitrogen, 3 to 6; volatile acids
(as acetic), 8.0 to 40.0; nonvolatile
acids (as lactic), 50; sugar (as glu-
cose), 20 to 70; and dextrin, 8.
IN THE MANUFACTURE OF SOY
SAUCE by the chemical method, soy-
bean meal is first extracted with water,
w^hich has been adjusted with acid to
AND SAUCE 359
the isoelectric point of the protein in
the meal. The isoelectric w^ash removes
soluble carbohydrates and nitrogen
compounds and raises the protein con-
tent of the meal above 65 percent. The
meal is then cooked with 17 percent
hydrochloric acid under a steam pres-
sure of 30 pounds to the square inch
for 15 to 18 hours to reduce the protein,
to amino acids. In place of the soy-
bean meal, soybean-protein curd may
be used as the starting material, and
constant-boihng hydrochloric acid used
in the cooking process. After hydrolysis
of the protein, the acid is neutralized
with sodium hydroxide or sodium
carbonate, and the resultant solution is
marketed as chemical soy sauce. Some-
times, after incomplete neutralization,
a crop of crystals of monosodium gluta-
mate is removed. The remaining amino
acids arc neutralized, and th(^ concen-
tration is adjusted to a suitable amino-
nitrogen content. Such a sauce is poorer
in glutamic acid but richer in other
amino acids.
In the soy sauce prepared by the
chemical method, the ])rotein hydrol-
ysis is more complete than in the fer-
mented product. The chemically
produced material is essentially a solu-
tion of salt and amino acids. Some of
the flavor and odor constituents of the
fermented sauce (such as peptides,
alcohols, esters, and nonnitrogenous
organic acids) are not found in the
chemically manufactured sauce.
Recent attempts to combine the con-
venience, economy, and speed of the
chemical method with the desirable
flavor characteristics obtained by the
fermentation process involve the par-
tial hydrolysis by acid treatment, fol-
lowed by neutralization of the acid.
The material is then fermented for
about a month by the organisms used in
the fermentation method.
Miso is the most important soybean
food product of Japan. It is prepared in
three varieties—white, red, and black.
Soybeans, wheat or wheat flour, and
rice are used to make white miso in
the proportions of 2: 1: 1. The soy-
beans arc soaked in water about 20
36o 1950-1951 YEARBOOK
hours, drained, and cooked for 2 hours
in a pressure cooker under 10 pounds
steam pressure. Soybean meai can be
substituted for th(î soybeans. When
wheat flour is used it is made into
a dough with water, and baker's yeast
is added. The dough is allowed to rise
for a day. It is then molded into loaves
about 2 inches square, and steamed for
1 hour at atmospheric pressure. Pol-
ished rice is soaked 2 hours in water,
drained, anci steamed in a collander.
The loaves of dough arc broken and
mixed thoroughly with the steamed
rice and soybeans. The mixtures is inoc-
ulated with a rice culture of soy-sauce
mold, Aspergillus oryzae. Shallow pans
arc filled with the paste to a depth of 2
inches, and left at room temperature
until the surface is covered with a white
mold growth. If the mold grow^th con-
tinues until green spores are formed,
the miso will be of poor quality. The
moldy paste is put in a deep vessel, and
7 volumes of salt water added for each
10 volumes of miso paste. The brine
contains 7 ounces of salt per gallon.
The fermentation time in the salt mash
will vary from 1 to 4 weeks, depending
on the temperature. At the end of the
fermentation period, the mash is
steamed and is ready for use without
further treatment. The salt content of
miso is much lower than that of soy
sauce. Consequently, miso is much
more subject to spoilage by micro-
organisms. The paste is usually con-
sumed within a month or two after
manufacture. Spices are commonly
added to miso shortly before it is
served.
Red miso is made from a combina-
tion of soybeans with rice and wheat or
barley, but the relative quantity of soy-
beans is greater in the red miso than
in the white. More salt is also used in
its preparation.
Black miso is made entirely from soy-
beans. The brine-fermentation stage of
the process is 1 to 3 years, although an
inferior-quality product may be pre-
pared in a 2- to 3-month brine fermen-
tation. The salt content of black miso
approximates that of soy sauce.
OF AGRICULTURE
Protein hydrolysis in miso manu-
facture is less complete than in soy-
sauce preparation. The protein content
of miso is about 15 percent. The mois-
ture content is about 50 percent.
Soybean cheeses are made by fer-
menting the curd obtained from soy-
bean milk. To make the curd, the soy-
beans are soaked in water 6 or 7 hours
in summer, or up to 24 hours in w'inter.
If the soaking period is too long, a
good, firm curd will not be formed
later in the process. The raw, soaked
beans are ground with cold water, then
pressed to remove the milk. More
^vater may be added to complete the
protein extraction. Ordinarily, about
10 pounds of water is used for each
pound of soybeans. The curd is usually
precipitated from the milk, which has
been heated almost to the boiling point,
by the addition of calcium or magne-
sium sulfates. The type of salt, the
hydrogen-ion concentration, and the
temperature of the milk at the time
of precipitation markedly influence the
physical properties and taste of the
curd. Too much magnesium sulfate
results in a bitter curd, and the bitter-
ness is retained in the finished cheese.
This may be observed to a lesser
extent when other chValent ions are
used as precipitating agents. The curd
is pressed to a consistency suitable for
cutting into small cubes.
Sufu is a light-gray soft cheese made
by the fermentation of cubes of curd
1 to 1/2 inches thick. The cubes are
dipped in dry salt, placed in a shallow-
pan, and inoculated with a culture of
the mold Mucor sujii. After 3 days at
a temperature of about 25° G. (77°
F. ), they are covered with a white mold
growth, which turns gray by the fifth
day as sporangia arc formed. If the
curd is too wet or too dense, an am-
moniacal odor may be noticed at this
time. Such curds make cheese of in-
ferior quality. When the mold com-
pletely covers the cubes and has started
to turn gray, the moldy curd is im-
mersed in brine, to which a culture of
wine yeast has been added. The final
salt content should be 12 to 15 percent.
FERMENTED SOY FOODS AND
The brine fermentation is continued in
a scaled vessel for 3 or 4 months.
Red sufu is a red cheese. It is pre-
pared in much the way that sufu is,
except that the curd cubes and red rice
arc placed in alternate layers in deep
vessels when the brine fermentation is
started. The red rice is a Chinese prod-
uct made by growing the mold Monas-
cus rub er in the grains of polished
white rice until the entire grain is per-
meated with the coloring matter. The
red rice contributes a pleasant taste
and aroma to the red sufu cheese.
Chee-fan is a brownish soft cheese
made from soy-milk curd cut into cubes
about one-half inch square. The curd
obtained from 70 parts of soybeans is
salted with 20 parts of salt^ the cubes
are inoculated with the Mucor, and a
week is allowed for mold development.
Both the Mucor and a blue Aspergillus
appear to play a part in this fermenta-
tion. The week-old moldy cubes are
covered with Shaohing wine, and a
rice or wheat-flour culture of the mold
and wine yeast is added. The cheese is
allowed to age in wine in an earthern-
ware vessel for about a year. Shaohing
wine is a yellow wine made from rice.
Tsue-fan is prepared in the same
manner as chee-fan, except that the
quantity of curd is greater in propor-
tion to the salt and wine than in chee-
fan.The cubes arc about one-half inch
on each side. They are boiled in water,
cooled, and partly dried before inocu-
lation or salting. This cheese is aged in
rice wine for 6 months.
Hon-fan is a red cheese prepared like
red sufu, except that the cheese is aged
in soy sauce instead of brine.
SAUCE 361
The principal flavor constituents of
the soy cheese are a combination of the
amino acids derived by the hydrolysis
of protein and fats. The red rice and
its fermentation products, along with
the rice wine, contribute sugars, esters,
alcohols, and organic acids to the flavor
and odor of these foods.
IN THE United States, soy sauce is
manufactured by both the fermentative
and chemical methods. Sauce blended
from the products of both processes has
been marketed. Soy sauce is the prin-
cipal constituent of the popular meat
sauces of the Worcestershire type. The
use of soy sauce without added condi-
ments is not popular in this country^
except in the so-called Chinese foods.
The recent introduction on a broad
scale into American markets of mono-
sodium glutamate as a seasoning agent
indicates the wide field for expansion
of the soy-sauce market in the food
industries.
LEWIS B. LOGKWOOD^ a native of
Indiana, joined the Department of
Agriculture in 1931. From 1940 until
his resignation in July 1950, he con-
ducted research on bacterial oxidations
and organic acid production at the
Northern Regional Research Labora-
tory.
ALLAN K. SMITI-I received his doc-
tor's degree in chemistry in 1926 from
Columbia University. He is senior
chemist at the Northern Laboratory,
where he is in charge of the protein
properties, isolation, and food use sec-
tion. Dr. Smith has worked in the De-
partjnent of Agriculture since 1937.

THE FARM BOY of a half century ago was familiar with the custom of occa-
sionally washing or rinsing with oatmeal water the inside of the glazed clay
jug in which he carried fresh w^ater from the well to the harvest hands in the
fields. It w^as believed that the water in w^hich oatmeal had been soaked had
some cleansing or purifying quality that kept the jug sweet and clean. Now we
find oat flour used in foods, on food packages, and on women's skin, all for the
purpose of preserving sweetness.—T. R. Stanton, Bureau of Plant Industry,
Soils, and Agricultural Engineering.