APPuED MICROBIOLOGY, Jan. 1969, p. 10-13 Vol. 17, No.

I
Copyright © 1969 American Society for Microbiology Printed In U.S.A.

Fermentation of Philippine Vegetable Blends1

C. A. ORILLO, E. C. SISON, M. LUIS,2 AND C. S. PEDERSONI
College of Agriculture, University of the Philippines College, Laguna, Philippines
Received for publication 10 October 1968
Seven blends of Philippine vegetables, two of which contained soybeans and one
mongo bean sprouts, were prepared for fermentation and study of microbiological
and chemical changes. The fermentations were typical lactic acid bacterial fermenta-
tions, initiated by Leuconostoc mesenteroides and continued by Lactobacillus brevis,

Downloaded from http://aem.asm.org/ on July 18, 2020 by guest

Pediococcus cerevisiae, and L. plantarum. The combination of high acidity and
low pH resembled other vegetable fermentations, such as sauerkraut. The procedure
offers a method of preserving surplus vegetables, and, in addition, a method for in-
corporating and preserving the high-protein-containing soybeans.

Preservation of vegetables by fermentation
probably originated in Asia with methods that
have been in use for centuries. Yen tsai, paw tsay,
kimchi, sajur asin, mostasa, and nukamiso pickles
are among the better known preserved foods of
the Far East. In recent years, the Filipinos have
become increasingly aware of the value of vege-
tables to their diets. Although recently cucumber
pickling has become an important industry, the
Filipinos have not utilized fermentation as a
method of preserving other vegetables. They are
accustomed to eating sweet-sour foods, such as
"Atchara," a vegetable blend sweetened with
sugar and acidified with vinegar. The increase in
vegetable production, particularly in the moun-
tain province, has resulted in seasonal overpro-
duction which could be alleviated by preserving
vegetables by fermentation for later use.
Palo and Lapuz (2) isolated strains of bacteria
from fermenting mustard leaves, "mostasa,)
which were presumably strains of Leuconostoc
mesenteroides. Kim and Whang (1) reported that
this species played an important role in the fer-
mentation of kimchi. In view of the importance of
L. mesenteroides, Lactobacillus plantarum, and
other species in the production of sauerkraut of
superior quality (4) and the necessity for establish-
ing optimal environmental conditions for growth
of these species, it seemed desirable to determine
whether they also played a major role in all of the
I Approved by the Director of the New York State Agricul-
tural Experiment Station for publication as Journal Paper
No. 1630.
2 College of Home Economics, University of the Philippines,
Diliman.
' Visiting Professor, 1 July, 1965 to 31 August, 1967 from New
York State Agricultural Experiment Station, Cornell University,
Geneva, N. Y. 14456.
10
major vegetable blends that could be made in the
Far East.
MATERIALS AND METHODS
The vegetables used were purchased on the open
market. The vegetables of higher sugar content, such
as cabbage and singkamas, were included in the blends.
The edible roots of singkamas, a yam bean grown in
the Philippines, resemble small white turnips in ap-
pearance, but are white, crisp, and sweet. The blends
contained the percentages of trimmed vegetables
given in Table 1. The vegetables were washed, peeled
when necessary, cleaned, sliced or shredded, and then
mixed with 1.75 to 2.25% salt. The mixtures were
packed into jars of 1 to 3-gal-capacity. A bag-like
plastic cover was placed on the surface of the vege-
table, and sufficient water was placed in the bag to
contact the entire surface and to bring the vegetable
brine to a height level with the surface.
During fermentation, samples of brine were re-
moved at regular intervals for determination of pH,
total acidity, and salt content. With four blends,
samples of brine were plated for estimation of numbers
of bacteria, isolation of colonies, and study of the flora.
Plates were incubated for 2 days at room temperature,
which usually approximated 30 C. Colonies were
counted, and usually 25 colonies were isolated from
each plating. Later the cultures isolated were identified
by the method of Pederson and Albury (3). Repre-
sentative cultures were commonly studied further to
assure the validity of identification.
RESULTS AND DISCUSSION
With the uniform and comparatively high tem-
peratures of the Philippines, fermentations were
initiated rapidly and continued at a rapid rate.
Acidities (as lactic) were attained as follows: no.
1, 1.34% in 7 days; no. II, 1.72% in 11 days; no.
III, 1.60% in 8 days; no. IV, 1.75% with a pH
of 3.70 in 12 days; no. V, 1.83% with a pH of
3.78 in 7 days; no. VI, 1.75% with a pH of 3.70
VOL. 17, 1969 FERMENTATION OF PHILIPPINE VEGETABLE BLENDS 11
TABLE 1. Approximate percentages of vegetables used in each blend
Asparau Mustard Soybean Mongo
Blend no. Cabbage Singkamas Radishes Carrot Pepper beans green cooked banu

I 48 25 25
II 36 19 24 5 8 6
III 32 31 35
IV 36 27 10 15 10
V 32 21 10 4 31
VI 39 21 11 9 18
VII 38 20 11 9 20

Downloaded from http://aem.asm.org/ on July 18, 2020 by guest

TABLE 2. Microbiological and chemical changes during the fermentation
of vegetable blend with soybean, blend no. V
Estimated no. of each species of bacteria X 106
Total Total-- - -_ __
Time Temp Salt acid pH count
X 106 Aerobic Leuconostoc Lacto,-
bacillus
PeIc
occus
Laco-
bacillus
Sp mcsenteroides brevis cercisca~e plaxbarum
hr C %
3 30 0 5.90 7 6.3 0.7
10 2.77 0.17 5.10 470 470
21 30 2.23 0.49 4.64 3,700 3,700
27 32 2.34 0.65 4.50 2,700 2,600 100
34 33 2.25 0.77 4.30 1,200 864 46 144 144
45 31 2.49 0.99 4.15 1,630 68 204 340 1,020
51 31 2.23 1.14 4.09 2,100 84 336 336 1,343
60 2.40 1.24 3.94 1,540 124 62 1,354
68 2.55 1.32 3.94 1,660 330 1,330
76 31 2.23 1.41 3.90 2,500 900 1,600
94 31 2.27 1.50 3.86 1,320 50 1,270
119 30 2.30 1.51 3.86 1,340 60 1,280
143 30 2.34 1.63 3.75 570 250 320
165 30 2.23 1.83 3.78 64 6 58
213 2.30 1.85 3.80 171 64 107
285 29 2.34 1.86 3.78 90 35 55

in 5 days; and no. VII, 1.74% with a pH of 353.
in 7 days.
Blends no. I and III resembled sauerkraut in
both appearance and flavor. The varying shades
of green of mustard leaves, beans, and peppers,
combined with the white of the other vegetables,
cabbage, singkamas, and radish, in blend no. II
presented the least attractive appearance of all
blends and also had the least appealing flavor.
In contrast, blend no. IV was most attractive in
color and flavor. The red and green colors of the
peppers and with the carrot-red hue of the carrots
blended well with the white vegetables. These
vegetables were included also in blends V, VI,
and VII. Although the soybeans in blends V and
VI were very noticeable, the mongo bean sprouts
in blend VII were barely discernible.
After fermentation was completed, the blends
were diluted with one-half their volume of water
to reduce the acidity to about 1.2%. They
were then heated to 165 F (74 C), packed into
heated glass jars at this temperature, sealed, and
cooled.
Microbiological and chemical changes. The
microbiological and chemical changes that oc-
curred during fermentation were quite similar to
those that occur in the sauerkraut fermentation
(3). The detailed microbiological and chemical
changes that occurred in blend no. V (Table 2)
show the rapid rise in acidity with a corresponding
reduction of pH. L. mesenteroides was entirely
responsible for the early stages of this fermenta-
tion as it was in the other blends studied. A maxi-
mal count of 3,700 X 106 was attained in 21 hr.
The count dropped to 2,700 X 106 during the
next 6 hr, and a marked change in flora occurred
during the next 7 hr, when an acidity of 0.77%
was attained. This change in flora occurred at a
12 ORILLO ET AL. APPL. MICROBIOL.

TABLE 3. Fermentation of vegetable blend containing soybean, blend no. VI

Estimated no. of each species of bacteria X 106
count plate
Time pH Total acid Salt Total - ____________ -actobacillu_
X 106 Leuconosloc
mesenteroides
Lactobacillus
brevis
Laclobacillus-
plantarum
hr
0 1.69 0.135
9 0.1 2.49 7.20 7.2
21 0.59 2.09 1,410 1,410
27 4.50 0.76 1.90 900 900
34 4.30 0.80 2.02 800 544 96 160
45 4.10 0.98 1.90 610 80 50 48&
3.92 1,020 43 934

Downloaded from http://aem.asm.org/ on July 18, 2020 by guest

52 1.13 1.90 43
58 3.85 1.18 1.84 700 28 672
68 3.79 1.44 1.90 1,000 1,000
73 3.78 1.70 1.90
82 3.72 1.75 1.90 260 260
97 3.69 1.75 1.90 470 470
116 3.70 1.75 1.90 224 224
164 3.73 1.78 1.90 121 121
212 3.75 1.80 1.90 15 15

lower acidity than that which occurs in sauerkraut
fermentation. Lactobacillus plantarum became the
predominant florum. L. brevis and Pediococcus
cereviseae played minor roles; the acid-tolerant
L. brevis persisted in spite of the high acidity
attained.
In one of the vegetable fermentations without
soybeans, e.g., blend no. IV, and the second
fermentation with soybeans, blend no. VI (Table
3), P. cereviseae was never isolated at any stage;
Lactobacillus brevis played a less important role
than in blend V. In blend no. IV, strains of strep-
tococci and micrococci were isolated from the
second plating when the acidity was low, e.g.,
0.26%, but they were entirely absent thereafter.
The soybeans were added to vegetable blends
no. V and IV to increase the protein content.
Protein analysis of the heat-processed sample of
blend no. IV yielded 8.80% protein (N X 6.25)
calculated on the dry basis. The soybean sample,
blend no. V, yielded 34.44% protein on the dry
basis. Although this may seem abnormally high
when only 32% by weight of soybean had been
added, soybeans even after cooking contain a
high percentage of total solids, whereas the other
vegetables are low in total solids. Blend no. VI, in
which only 18% soybeans were added, contained
23.0% protein.
Because of the high-protein content of blend V,
it was assumed that the buffer content of the
brine may have increased to such an extent that
insufficient acid would be produced to lower the
pH for effective preservation. Actually equilib-
rium between the soybean and the other vege-
table was not attained rapidly, and the soybean
protein apparently remained within the bean.
Therefore, the buffering effect of the fermenting
brine was not affected greatly. A pH below 4.00-
was attained at an acidity level of 1.0 to 1.2%.
The blend of carrots and red peppers contrasted
well to the white cabbages, singkamas, and
radishes, providing an attractive and acceptable
appearance. In general, the Americans who were
accustomed to eating sauerkraut found the flavor
of the blends acceptable. The processed blends
were too sour for the majority of the Filipinos;
however, when they were sweetened with 10 to
15% sugar or blended with crushed pineapple,
the products were relished by them. They com-
pared them to their "Atchara." The soybeans
acquired a somewhat chewy, peanut-like flavor.
There may be a chemical change responsible for
development of this character. The mongo beans
in blend VII were barely discernible.
Technically, the low oxidation-reduction po-
tential attained in lactic acid fermentation systems
is effectual in retaining the light color and bright
appearance of the vegetables. Low pH or high
acidity delays enzyme action and inhibits growth
of other microorganisms.
Preservation of mixed vegetables, such as those
seasonally available, can be readily adapted to
small-scale operations using almost any blend of
vegetables as is done in other Asian countries.
After fermentation, the vegetable blends may
be preserved by heating the blend to 165 F (74 C),
filling into containers, and hermetically sealing
the containers at a temperature of 165 F.
VOL. 17, 1969 FERMENTATION OF PHILIPPINE VEGETABLE BLENDS 13
LITERATURE CITED the synthesis of the gum and its production in coconut water.
Philippine J. Sci. 83:327-357.
1. Kim, Ho-Sik, and Kyn-Chan Whang. 1959. Microbiological 3. Pederson, C. S., and M. N. Albury. 1954. The influence of salt
studies on kimches. Part I. Isolation and identification of and temperature on the microflora of sauerkraut fermenta-
tion. Food Technol. 8(1):1-5.
anaerobic bacteria. Bull. Sci. Res. Inst. (Seoul, Korea) 4:56. 4. Pederson, C. S., and M. N. Albury. 1969. The sauerkraut fer-
2. Palo, M. A., and M. M. Lapuz, 1955. On a new gum-forming mentation. N.Y. State Agr. Expt. Sta. (Geneva, N.Y.)
streptococcus, with studies on the optimal conditions for Bull. 824.

Downloaded from http://aem.asm.org/ on July 18, 2020 by guest