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Effect of Calcium Sulfate Concentration in Soymilk on the Microstructure of

Firm Tofu and the Protein Constitutions in Tofu Whey

Article in Journal of Agricultural and Food Chemistry · November 2003

DOI: 10.1021/jf0342021 · Source: PubMed

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 Effect of Calcium Sulfate Concentration in Soymilk on the
Microstructure of Firm Tofu and the Protein Constitutions in
Tofu Whey
FUH-JUIN KAO, NAN-WEI SU, AND MIN-HSIUNG LEE*
Graduate Institute of Agricultural Chemistry, National Taiwan University, Taipei 10617, Taiwan

The effects of calcium sulfate concentration in soymilk on the microstructure of tofu and the
constitutions of protein in tofu whey were investigated. The firm tofu made with 0.4% CaSO4‚2H2O
was found to be most uniform and continuous in the microstructure. This tofu gave the maximal tofu
yield, maximal solid and protein recoveries, and the best water retention ability. The results of gel
electrophoresis and the ratio of amino nitrogen to total organic nitrogen indicated that the low molecular
weight proteins in tofu whey were at their lowest when the tofu was made with 0.4% CaSO4‚2H2O.
The SEM observations suggested that the tofu made with 0.4% CaSO4‚2H2O has the most uniform
and homogeneous microstructure and, consequently, can most efficiently retain soybean proteins
and water in the tofu gel.

KEYWORDS: Firm tofu; microstructure; calcium sulfate; tricine SDS-PAGE

INTRODUCTION coagulant concentrations has been reported by many research

Tofu is one of the most popular food items in the traditional groups (1, 2, 5-13). However, no definite conclusion was
oriental diet (1). Because of its bland taste, the textural properties drawn. Because of the limited solubility of calcium sulfate, it
markedly influence its food quality and consumer acceptability was reported that a proper coagulant concentration for tofu
(2). Since the 1970s, the SEM and other analyses using making depends not only on coagulation temperature (7), stirring
instruments have been used to research tofu and have provided speed, and time (9, 11) but also on the stirring blade size, mixing
a more stereoscopic image of the fine structure of tofu. Saio tank dimension, and soybean variety (10). Various optimal
studied the relationship between fine structure and texture of concentrations from 9.5 to 58 mM (0.16-1.0%) CaSO4‚2H2O
tofu and concluded that the microstructure of tofu varied with in soymilk have been reported as appropriate for production of
the density of the gel network, the size of protein granules in tofu (1, 2, 5-13). However, very few reports concerning the
the network, and the size of coagulate prepared during the effects of coagulant concentration on the microstructure of tofu
coagulating process (3). All of them correlated with the hardness were reported. Additionally, the proteins in tofu whey are usually
of tofu. Furthermore, the fine structure of tofu is subject to regarded as the LMW proteins (14, 15); however, there were
change during subsequent processing, such as deep-fried tofu no direct evidences. In this study, we investigate the effects of
and frozen tofu (3). These processes lead to changes in the coagulant concentration on the microstructure of tofu, as well
texture of the tofu. deMan et al. (2) reported that the texture as the protein constitutions in tofu whey.
and microstructure of the soybean curd were greatly influenced
by the type of coagulant used. They found that curd produced MATERIALS AND METHODS
by GDL and CaSO4 gave a much finer and a more uniform Materials. Soybeans of the Ohio FG1 cultivar were obtained from
honeycomb-like structure than the curd prepared with CaCl2, a local agency. Food grade calcium sulfate dihydrate (Ako Kasei Co.,
MgCl2, and MgSO4 (2). Tsai et al. (1) considered that GDL Ltd., Hyogo, Japan) was used as the coagulant in the tofu production
and CaSO4 were the most suitable coagulants for tofu making and was obtained from Gemfont Corporation (Taipei, Taiwan).
and gave a higher yield than other calcium salt coagulants, but Tofu Making. Tofu was produced by the traditional method as
follows. Beans (300 g) were soaked in tap water (three times bean
the former was not suitable for Chinese style tofu due to its
weight) at room temperature for 9 h to bring the soaked beans weight
sour taste. to approximately 2.2 times their initial weight. Hydrated beans were
As with the type of coagulant, the concentration of the drained and ground with an amount of tap water to make the total weight
coagulant also affects the nature of tofu. The amount of of 3300 g in a grinder (Pineapple grinder, Great Yen Electric Food
coagulant added in tofu manufacturing is one of the critical Grinder Co., Ltd., Taipei, Taiwan) equipped with an automatic
control points, and it determines crucially the product’s texture, centrifugal filter to separate raw soymilk from the residue. This
flavor, and yield (4). The correlation of tofu properties and the corresponded to a water:soybean ratio of 10:1 by weight for extracting
solids from soybean into the raw soymilk and brought the total solid
* To whom correspondence should be addressed. Tel: +886-2-23630231 content of soymilk to approximately 6%. After adding 1 g of
ext. 2490. Fax: +886-2-23632714. E-mail: [email protected]. antifoaming agent (containing 90% glycerin fatty acid ester, 5% calcium
carbonate, 4.3% soybean phospholipids, and 0.7% silicone resin), the 20 min. The supernatant was collected and regarded as the soymilk
raw soymilk was heated to 95 °C and held at this temperature for 5 protein extracted. Tofu whey protein was extracted according to a
min. After the soymilk was cooled to about 73 °C, 50 mL of various method modified from Sorgentini and Wagner (20). Tofu whey was
concentrations of calcium sulfate solution was added to soymilk with centrifuged at 6000g for 20 min at 20 °C. The clarified supernatant
stirring at a speed of 250 rpm for 10 s and then incubated for 20 min was adjusted to pH 8.0 with 2 N NaOH, kept for 1 h at room
to form bean curd. The final CaSO4‚2H2O concentrations used in the temperature, and then centrifuged at 12 000g for 20 min at 20 °C.
soymilk were 0.20, 0.25, 0.30, 0.35, 0.40, 0.45, and 0.50% (w/v). The Polypeptides smaller than a molecular mass of 10 kDa in the supernatant
bean curd was then broken slightly and transferred into a muslin cloth- were removed by ultrafilteration (Centricon-10, Millipore Corp.,
lined stainless steel mold (13 × 13 × 5.5 cm3) and then pressed at Bedford, MA). The retentate was collected and regarded as tofu whey
21.8 g/cm2 for 10 min, 43.6 g/cm2 for 10 min, and 65.4 g/cm2 for 30 protein extract. The protein concentrations in soymilk protein and tofu
min. At the end of pressing, the cloth was removed and the weight of whey protein extract were both determined by the method of Bradford
tofu and the volume of drained whey were recorded. The tofu yield (21).
was expressed as grams of fresh tofu per 100 g of soybeans. The tofu Electrophoresis. TSDS-PAGE was performed according to the
was transferred into a plastic bag and stored at 4 °C overnight for texture method of Schägger and Gagow (22). The protein extract was adjusted
measurement. to 2 mg/mL with double-distilled water. An equal volume of the SDS
Determination of the Protein and Solid Contents and Their sample buffer containing 10% 2-mercaptoethanol was added and then
Recoveries. The protein contents in soybean, soymilk, tofu, and tofu was boiled for 5 min. An aliquot of sample solution containing about
whey were determined in triplicate by the micro-Kjeldahl method (16). 35 µg of proteins was loaded. The run was performed in 12%
The solid content of tofu was determined by drying 5 g of homogenized polyacrylamide gel in a Hoefer Mighty Small II SE-250 mini vertical
tofu in an oven at 105 °C until constant weight was obtained. Protein gel electrophoresis unit at a 135 V constant voltage. After the
recovery in tofu was expressed as the amount of protein in tofu divided electrophoresis, the gel was stained in a solution of methanol, acetic
by the amount of protein in raw soybeans multiplied by 100% on a acid, and water (5:1:4) containing 0.25% Coomassie Brilliant Blue
dry weight basis. The same calculation was applied to tofu solid R-250. The Pharmacia LMW kit (R-lactalbumin, 144 000; trypsin
recovery. inhibitor, 20 100; carbonic anhydrase, 30 000; ovalbumin, 43 000;
Determination of WRA of Tofu. WRA was determined by a bovine serum albumin, 67 000; and phosphorylase b, 94 000) was
modification of the water holding capacity method (WHC) of Puppo employed as a standard to determine the molecular masses of the
and Añón (17). About 5 g (w1) of tofu was placed on a cotton cloth extracted proteins. Protein quantification of the electrophoresis gels was
membrane maintained in the middle position of a 250 mL centrifuge carried out by direct densitometric scanning. The gels were scanned
tube (62 mm × 120 mm). The sample weight was recorded after and analyzed by a Kodak EDAS 290 scanner equipped with Kodak
centrifugation at 120g for 5 min at 15 °C (w2) and subsequently heated Digital Science 1D 3.5 image analysis software.
to a constant weight (w3) at 105 °C. The WRA of tofu was calculated Determination of FN, Ammonium Nitrogen, and AN. FN and
as the following equation: ammonium nitrogen in the soymilk and tofu whey were determined
by titration according to the Chinese National Standards method (23).
WRA ) ((w2 - w0)/(w1 - w0)) × 100% To determine the content of FN, 5 g of soymilk or clarified tofu whey
was diluted to 250 mL with distilled water and adjusted to pH 8.5
Determination of Textural Properties. Textural properties of tofu with 0.1 N NaOH. Forty milliliters of sample solution was mixed with
were measured by a texture analyzer (model TA-XT2, Stable Micro an equal volume of 37% formaldehyde solution (previously adjusted
systems, Haslemere, Surrey, U.K.). A 5 kg load cell was used with the to pH 8.5) and then titrated to pH 8.5 with 0.1 N NaOH. To determine
crosshead controlled at 1.5 mm/s. A cylindrical plunger with a diameter the content of ammonium nitrogen, 10 g of soymilk or clarified tofu
of 35 mm was used to compress the tofu cakes (35 mm diameter × 22 whey was mixed with 25 mL of 40% NaOH and distilled for 5 min. A
mm height) to 50% deformation. Hardness, cohesiveness, gumminess, conical flask containing 40 mL of 4% boric acid was placed under the
springiness, and chewiness were calculated using the textural profile condenser outlet to receive the distillate. The ammonium borate solution
analysis curve (18). Hardness was defined as the height of the force formed was titrated to pH 8.5 with 0.1 N NaOH. The amount of FN
peak on the first compression cycle, which was the force necessary to and ammonium nitrogen was expressed as g of nitrogen/100 g of
attain a given deformation. The ratio of the positive force areas under sample. AN was determined by subtraction of the ammonium nitrogen
the second and the first compression was defined as cohesiveness. from FN.
Gumminess was defined as the product of hardness and cohesiveness. Data Analysis. All of the results were analyzed by ANOVA using
The springiness was the degree to which the tofu returned to its original the general linear model (24). Duncan’s multiple range test was used
shape after it had been decompressed and was expressed as the to determine differences among the samples. Significant levels were
horizontal distance (mm) between the point when the second curve defined as probabilities of 0.05 or less. All processing treatments were
started and the point when the second curve reached a peak. Chewiness in triplicate.
was defined as the product of gumminess and springiness.
SEM. A JEOL JSM-6300 scanning electron microscope (JEOL USA, RESULTS AND DISCUSSION
INC., Peabody, MA) was used to examine the fine structure of tofu
Effect of CaSO4‚2H2O Concentration in Soymilk on
coagulated with different CaSO4‚2H2O concentrations. The procedure
used for sample preparation was that of deMan et al. (2) with some Microstructure of Firm Tofu. A better understanding of the
modifications. A small piece of each tofu sample was fixed at room changes in the microstructure of tofu that corresponded to the
temperature with 2.5% glutaraldehyde in 0.1 M phosphate buffer (pH coagulation conditions would provide a more rational basis to
7.1) for 2 h. After five washings with 0.1 M phosphate buffer (pH 7.1) determine the suitable concentration of coagulant. In this study,
at 10 min intervals, it was then postfixed with 1% osmium tetroxide in the SEM images of the tofu prepared with different CaSO4‚
the same buffer for 90 min at room temperature. The fixed sample 2H2O concentrations showed clearly different fine structures
was rinsed five times with phosphate buffer at 10 min intervals. The (Figure 1). The network of tofu obtained with 0.2% CaSO4‚
sample was frozen in liquid nitrogen and freeze-dried. The dried sample 2H2O revealed a coarse and discontinuous structure with many
was finally mounted on an aluminum stub and sputter-coated with gold/ fragments and large pores (Figure 1A). On increasing the
palladium (60/40). The observations were carried out at 20 kV.
CaSO4‚2H2O concentration to 0.3%, there was a trend toward
Preparation of Soymilk Protein and Tofu Whey Protein. The
preparation of soymilk protein was based on a method modified from decreasing pore size and increasing regularity and uniformity
Cai and Chang (19). Fifty milligrams of freeze-dried soymilk was (Figure 1B). The microstructure of tofu with 0.4% CaSO4‚2H2O
dispersed in 1 mL of buffer (86 mM Tris-90 mM glycine-4 mM Na2‚ showed the most continuous and uniform honeycomb-like
EDTA, pH 8.0). The mixture was sonicated at 60 °C for 90 min to structure (Figure 1C). This structure was very uniform with
extract the proteins. The extract was then centrifuged at 12 000g for smaller holes than those prepared with 0.2 and 0.3% CaSO4‚
Figure 1. SEM images of freeze-dried tofu prepared with different concentrations of CaSO4‚2H2O in soymilk; (A) 0.2, (B) 0.3, (C) 0.4, and (D) 0.5%.

2H2O, and this was similar to that obtained with CaSO4‚1/2H2O Table 1. Effect of Calcium Sulfate Concentration in Soymilk on
by deMan et al. (2). When the CaSO4‚2H2O concentration was Moisture Content, WRA, Tofu Yield, Solid Recovery, and Protein
increased to 0.5%, the network of tofu became too porous and Recovery of Firm Tofua
too compact (Figure 1D) and not as uniform and smooth as CaSO4‚ moisture tofu yield solid protein
that obtained with 0.4% CaSO4‚2H2O. 2H2O content WRAb (g tofu/ recovery recovery
Effect of CaSO4‚2H2O Concentration on Yield, WRA, (%, w/v) (%) (%) 100 g soybean) (%) (%)
Protein, and Solid Recoveries of Firm Tofu. In this study, 0.20 80.2 a 67.5 c 205 e 45.9 c 62.6 e
we investigated the effects of CaSO4‚2H2O concentrations, from 0.25 80.2 a 67.7 c 230 d 49.7 b 67.0 cd
0.2 to 0.5%, on the microstructure and physical properties and 0.30 80.9 a 70.3 b 237 cd 51.2 b 67.9 bcd
0.35 80.8 a 75.6 a 243 bc 51.9 b 68.4 bc
found that 0.4% CaSO4‚2H2O in soymilk produced tofu with a 0.40 80.5 a 77.2 a 253 a 55.2 a 70.9 a
maximum tofu yield (253 g tofu/100 g soybean), maximum 0.45 80.7 a 76.9 a 247 ab 53.7 ab 69.5 ab
protein recovery (70.9%), maximum solid recovery (55.2%), 0.50 81.0 a 76.0 a 240 bc 51.5 b 68.3 bc
and a maximum WRA (77.2%). All four parameters increased
significantly as the CaSO4‚2H2O concentration increased from a Mean scores bearing the same letters among the same column are not
0.2 to 0.4% but decreased as the CaSO4‚2H2O concentration significantly different (p < 0.05). b WRA of tofu.
increased from 0.4 to 0.5% (Table 1). At the lower calcium
sulfate concentrations, the interaction of the protein molecules and other soluble substances. These events would seem to
with calcium ion was not strong enough to form a firm tofu account for the lower yield of tofu (240 g tofu/100 g soybean),
gel. There were many discontinuous fragments and large holes lower protein recovery (68.3%), and lower solid recovery
in the microstructures of tofu made with 0.2 or 0.3% CaSO4‚ (51.5%) at this concentration. At the optimum coagulant
2H2O (Figure 1A,B). The exclusion of noncoagulated proteins concentration (0.4% CaSO4‚2H2O), a uniform, continuous tofu
and other soluble solids with the water during the pressing step network was formed (Figure 1C), and it was able to trap more
resulted in a lower tofu yield and the lower protein and solid water and soluble substances. Therefore, it could obtain the
recoveries (Table 1). On the contrary, at a high concentration highest tofu yield, protein and solid recoveries, and WRA (Table
of 0.5% CaSO4‚2H2O, too many cross-linkages were formed 1). These observations were in agreement with the findings of
and resulted in a too compact and too porous structure (Figure Puppo and Añón (17), who showed that a protein gel with a
1D). This structure was less uniform and less homogeneous than homogeneous and fine structure gave high WRA, when
that with 0.4% CaSO4‚2H2O. This, in turn, resulted in an compared to the gel with a nonhomogeneous structure, that had
increased syneresis and induced the loss of water, whey proteins, a high degree of syneresis.
Table 2. Effect of Calcium Sulfate Concentration in Soymilk on the
Textural Characteristics of Firm Tofua
CaSO4‚
2H2O hardness springiness gumminess chewiness
(%, w/v) (kg) (mm) cohesiveness (kg) (kg mm)
0.20 0.40 b 7.84c 0.35 d 0.14 c 1.11 d
0.25 0.45 b 8.70 b 0.39 c 0.18 c 1.52 c
0.30 0.66 a 9.74 a 0.40 bc 0.26 b 2.57 b
0.35 0.68 a 9.76 a 0.41 b 0.28 b 2.72 b
0.40 0.71 a 9.76 a 0.45 a 0.32 a 3.11 a
0.45 0.73 a 9.84 a 0.45 a 0.33 a 3.21 a
0.50 0.74 a 10.32 a 0.45 a 0.33 a 3.39 a

a Mean scores bearing the same letters among the same column are not

significantly different (p < 0.05).

Tofu Texture. The increase of CaSO4‚2H2O concentration
from 0.2 to 0.4% resulted in a steady increase in the hardness,
springiness, cohesiveness, gumminess, and chewiness of tofu
(Table 2). Texture is considered to be a direct consequence of
microstructure as determined by chemical composition and the
physical forces involved (25). A more dense and compact Figure 2. TSDS−PAGE profile of proteins of soymilk and tofu whey
structure maintained by the Ca-protein bridges and hydrogen obtained during tofu making with different CaSO4‚2H2O concentrations
bonds might account for the higher hardness and higher in soymilk. Lane 1, soymilk; lane 2, 0.2%; lane 3, 0.3%; lane 4, 0.4%;
springiness of the tofu at higher concentrations of CaSO4‚2H2O. lane 5, 0.5%; and lane 6, protein molecular mass markers.
Similar results were reported by Shih et al. (9). Scaefer and
Table 3. Effect of Calcium Sulfate Concentration during Tofu Making
Love (26) also reported a significant correlation between calcium
on TN and AN of Tofu Wheya
concentration and hardness (r ) 0.73) and elasticity (r ) 0.83)
for tofu. When the concentration of the coagulant was increased TNb ANb AN/TONb
from 0.2 to 0.4%, cohesiveness increased significantly but sample (%) (%) (%)
remained constant at concentrations between 0.4 and 0.5% soymilk 0.47 a 0.004 e 0.93 e
(Table 2). Wang and Hesseltine (7) found that cohesiveness tofu whey CaSO4‚
increased with increasing CaSO4‚2H2O concentration from 10 2H2O (%, w/v)
to 20 mM (0.17-0.34%) and remained the same when beyond 0.20 0.14 b 0.053 a 43.1 c
0.30 0.08 c 0.040 b 66.7 a
the range. Tofu with a less intensive network gave less 0.40 0.04 e 0.013 d 39.4 d
cohesiveness. Because gumminess was defined as the product 0.50 0.06 d 0.027 c 57.5 b
of hardness and cohesiveness and chewiness was the product
of hardness, springiness, and cohesiveness, they all had similar a Mean scores bearing the same letters among the same column are not

trends (Table 2). significantly different (p < 0.05). b TON, subtraction of ammonium nitrogen from
Effect of CaSO4‚2H2O Concentration on the Constitution TN.
of Tofu Whey Proteins. The effect of CaSO4‚2H2O concentra-
tions on the constitution of tofu whey proteins was analyzed The increase of CaSO4 concentration from 0.2 to 0.4% in
by TSDS-PAGE (Figure 2), a method that had been shown to tofu making resulted in a decrease in TN and AN in tofu whey
have an improved resolution for LMW proteins (22). The and an increase in TN and AN when beyond 0.4% CaSO4‚2H2O
TSDS-PAGE pattern of soymilk protein showed that the protein (0.04 and 0.013%, respectively). The results supported the
bands with molecular masses in the range of 16-40 and 42- previous finding that tofu with a homogeneous and uniform
80 kDa were the two major proteins of soybean, i.e., 11S network gave the highest protein recovery at the optimum
(glycinin) and 7S (β-conglycinin) globulins. Glycinin is com- coagulant concentration (Table 1). Among the four tofu whey
posed of acidic (34-45 kDa) and basic (17-20 kDa) polypep- samples, the percentages of LMW proteins (Figure 2, lane 4)
tide chains paired by a disulfide bond (27, 28). β-Conglycinin and the ratio of AN to TON (Table 3) were the lowest in tofu
consists of R, R′, and β subunits, wherein the molecular masses whey obtained from the soymilk with 0.4% CaSO4‚2H2O (29.0
of R and R′ subunits are reported to be in the range of 57-83 and 39.4%, respectively). This suggested that a homogeneous
kDa, while that of the β subunit is 42-53 kDa (29). network can more efficiently retain the LMW proteins in tofu
As compared with soymilk protein composition, the tofu whey matrixes.
protein contained higher amounts of LMW proteins with a A possible gelation mechanism for firm tofu in the presence
molecular mass below 16 kDa (Figure 2). The protein bands of CaSO4 is schematically represented in Figure 3. It has been
on the gel were densitometrically scanned, and the relative ratio suggested that a Ca2+ gel is induced in the denatured protein
of bands was estimated. The percentages of LMW proteins in solution to form a filamentous gel structure (30-35). Heating
soymilk and tofu whey that were discharged from the pressed a soybean protein solution causes the proteins to unfold and
tofu made with 0.2, 0.3, 0.4, and 0.5% CaSO4‚2H2O were 1.8, form protein filaments due to the formation of disulfide bonds
41.1, 81.0, 29.0, and 53.7%, respectively. A similar trend was and hydrophobic interactions. A filamentous gel structure is
found in the ratio of AN to TON in soymilk and tofu whey formed when CaSO4 is added, and this reduces the electrostatic
(Table 3), with the values of 0.93, 43.1, 66.7, 39.4, and 57.5%, repulsion between the charged filaments, causing them to
respectively. These results confirmed the suggestion that the aggregate with the Ca2+ ions acting as bridges between the
proteins remaining in tofu whey were mainly LMW proteins adjacent charged carboxylic groups on neighboring protein
(15). molecules. At the end of pressing, the ratio of tofu whey volume
 Figure 4. Representation (highly schematic) of likely states of constitution
of protein filaments and LMW proteins in the tofu network and in the tofu
whey obtained with different concentrations of CaSO4‚2H2O in soymilk;
(O) representing HMW protein; (b) representing LMW protein. The eight
Figure 3. Gelation mechanism of firm tofu in the presence of CaSO4; states were as follows: (A) 0.2% (tofu network), (B) 0.3% (tofu network),
(O) representing HMW protein; (b) representing LMW protein. Each (C) 0.4% (tofu network), (D) 0.5% (tofu network), (E) 0.2% (tofu whey),
square area represents a relative volume. (F) 0.3% (tofu whey), (G) 0.4% (tofu whey), and (H) 0.5% (tofu whey).
The left-hand square areas represent the relative volumes of tofu, and
to firm tofu volume was in the range of 2.4-3.2. An increase the right-hand square areas represent the relative volumes of tofu whey.
in the CaSO4 concentration can promote gelation of the protein
filaments. At a low CaSO4 concentration (0.2% CaSO4‚2H2O), and consequently, resulted in the maximal tofu yield, maximal
the protein filaments failed to form a firm tofu gel (Figure 4A), protein and solid recoveries, and the best WRA.
with many individual protein filaments and LMW proteins
dispersed in tofu whey (Figure 4E). When the CaSO4‚2H2O ABBREVIATIONS USED
concentration was increased to 0.3%, the extent of aggregation SEM, scanning electron microscopy; GDL, glucono-δ-lac-
of the protein filaments in tofu gel increased remarkably. tone; LMW, low molecular weight; HMW, high molecular
However, the gel was still not homogeneous and exhibited a weight; WRA, water retention ability; TSDS-PAGE, tricine
fragmentary network with a relatively low retention of water sodium dodecyl sulfate-polyacrylamide gel electrophoresis; TN,
and LMW proteins (Figure 4B) and resulted in a maximum total nitrogen; FN, formol nitrogen; AN, amino nitrogen; TON,
percentage of LMW proteins dispersed in tofu whey (Figure total organic nitrogen; ANOVA, analysis of variance.
4F). As the CaSO4‚2H2O concentration further increased to the
optimum concentration (0.4% CaSO4‚2H2O), the LMW proteins ACKNOWLEDGMENT
were well-dispersed in the spaces (pores) of the homogeneous
network structure (Figure 4C) with only a few protein filaments We are grateful to Miss Chien Lin for technical assistance with
and LMW proteins dispersed into tofu whey (Figure 4G). As picture drawing.
the CaSO4 concentration was further increased to 0.5%, the
ability of the gel to retain water and LMW proteins adversely LITERATURE CITED
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