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Article history: To improve tofu quality, the effects of using 0.12e0.18 g/100 mL citric acid, l-(-)-malic acid, or tartaric acid
Received 16 January 2017 as coagulation agents were investigated. The results showed that storage modulus, hardness, water-
Received in revised form holding capacity, non-freezable water content, and in-gel hydrophobic interactions were optimal
1 July 2017
when tofu was prepared with 0.14 g/100 mL organic acids, but deteriorated when more than 0.16 g/
Accepted 3 July 2017
Available online 4 July 2017
100 mL was used. Slower acidication during gelling in the presence of 0.14 g/100 mL organic acid
promoted proteineprotein hydrophobic interactions, resulting in a higher elastic modulus. Moreover, the
tofu products showed higher water-holding capacity and non-freezable water content, as well as more
Keywords:
Citric acid
compact gel microstructures. Notably, tofu prepared with citric or l-(-)-malic acid was of better quality
l-(-)-malic acid than tofu prepared with tartaric acid. Collectively, the results demonstrate that soft or rm tofu with
Tartaric acid desirable physicochemical properties can be prepared with organic acids if acidication is adequately
Tofu controlled during gelation.
Gelation 2017 Elsevier Ltd. All rights reserved.
http://dx.doi.org/10.1016/j.lwt.2017.07.005
0023-6438/ 2017 Elsevier Ltd. All rights reserved.
F.-H. Cao et al. / LWT - Food Science and Technology 85 (2017) 58e65 59
vegetables and are currently extensively used in the food industry analyzed on a TA-XT texture analyzer (Stable Micro System Co. Ltd.,
(Huang, Xu, Zhang, Xue, & Chen, 2007; DeNobili et al., 2016). Godalming, UK) according to Zhang et al. (2013). The samples were
Organic acids induce signicant gelation at concentrations suitable compressed twice to 50% deformation using a P/36R probe, with
for relatively slow acidication, a desirable property, as accelerated trigger force 5 g, pretest speed 5.0 mm/s, test speed 1.0 mm/s, and
acidication typically results in weaker gels (Jacob, No bel, Jaros, & post-test speed 10.0 mm/s. Hardness, springiness, cohesiveness,
Rohm, 2011). and gumminess were determined (Zhang et al., 2013).
The objective of this study was to investigate the effects of
organic acid (citric acid, l-(-)-malic acid, and tartaric acid) treat- 2.5. Determination of water-holding capacity
ments on the soymilk gelation process and tofu quality. The relative
contribution of various chemical interaction types, including ionic Approximately 3 g of each tofu sample was used to measure
bonds, hydrogen bonds, and hydrophobic interactions, to tofu water-holding capacity according to Hu et al. (2013), with some
properties was evaluated. In addition, the feasibility of organic acid modication. Samples were cut into 5 5 5-mm cubes, weighed
in the production of tofu with desirable physicochemical properties (Wt), and centrifuged at 5632 g for 20 min at 4 C in a CR21
was assessed. Texture, water-holding capacity, and water distri- centrifuge (Hitachi, Japan). Supernatants were discarded and re-
bution were assessed to measure gel strength, rigidity, and water sidual liquids were carefully removed with dry lter paper. The
state. pH, rheological properties, and in-gel chemical interactions removed water was weighed (Wr), and water-holding capacity was
were analyzed to investigate gel nucleation and formation. The gel calculated according to equation (1):
microstructure was evaluated to investigate its relationship with
gel macro-properties to allow quality control of organic acid- WHC Wt Wr =Wt 100 % (1)
induced tofu gel.
Fig. 2. Storage modulus (G0 ) in soymilk spiked with 0.14 g/100 mL citric acid (-), l-
(-)-malic acid (C) and tartaric acid (:) or 0.30 g/100 mL glucono-d-lactone (;) (a).
Maximum storage modulus (G'max) in soymilk spiked with 0.12e0.18 g/100 mL citric
acid (gray column), l-(-)-malic acid (white column), tartaric acid (hatched column) or
0.30 g/100 mL glucono-d-lactone (dark gray column), and incubated at 80 C for
30 min (b). Different letters on the top of bars indicate signicant differences
(P < 0.05).
Table 1
Texture of tofu prepared with organic acids.
Values are the mean SD from triplicates. Different letters within a column indicate signicant differences (P < 0.05). CA, citric acid; MA, l-(-)-malic acid; TA, tartaric acid; GDL,
glucono-d-lactone.
F.-H. Cao et al. / LWT - Food Science and Technology 85 (2017) 58e65 63
Fig. 5. Total water (Wt) (a) and non-freezable water (Wnf) (b) in tofu prepared with
0.12e0.18 g/100 mL citric acid (gray column), l-(-)-malic acid (white column), or tar-
taric acid (hatched column), and pressed at 8 g/cm2 for 30 min. Different letters on the
top of bars indicate signicant differences (P < 0.05).
non-covalent interactions, including ionic bonds, hydrogen bonds, correlated with Wnf values, and thus, may be further interpreted in
and hydrophobic interactions (Tay & Perera, 2010; Wang et al., terms of hardness, water-holding capacity, and G'.
2015). These contributions of these interactions in the acid-cured
tofu are summarized in Fig. 6; hydrophobic interactions contrib- 3.6. Microstructure of organic acid-induced tofu
uted to gelation and gel structure more signicantly than hydrogen
bonds or ionic bonds. Several studies have indicated that hydro- Changes in the physicochemical properties of organic acid-
phobic interactions occur after charge neutralization (Guo & Ono, induced tofu corresponded with different microstructures (Lee &
2010; Kohyama & Nishinari, 2002). Hydrogen bonds and hydro- Kuo, 2011; Wang et al., 2017). Scanning electron micrographs of
phobic interactions are the main contributors to gel-like structure tofu coagulated with organic acids are presented in Fig. 7. A
(Wang et al., 2015). Moreover, Wang et al. (2017) suggested that honeycomb-like structure with varying size of pores were observed
ionic and hydrogen bonds are generally disrupted by heating to in tofu prepared with various organic acids at different concen-
90 C, highlighting the relatively low contribution of these bonds to trations. The surface charge of soy protein were screened gradually
chemical interactions in tofu prepared at 80 C. In addition, acidi- by protons, the hydrophobic interactions among proteins were
cation may also have interfered with the formation of ionic and favored, resulting in the lamentous and porous network (Lee &
hydrogen bonds. Kuo, 2011; Liu et al., 2013). Tofu prepared with lower coagulant
Hydrophobic interactions contributed 6.07 g/L, 5.40 g/L, and concentrations (0.12e0.14 g/100 mL) exhibited with denser
5.15 g/L in tofu coagulated with 0.14 g/100 mL citric acid, l-(-)-malic honeycomb-like topography than that prepared with higher acid
acid and tartaric acid, respectively. In comparison, hydrophobic coagulant concentrations (0.16e0.18 g/100 mL). In addition, tofu
interactions contributed 6.18 g/L in tofu prepared with 0.30 g/ prepared with GDL exhibited a rmer structure with small pores, in
100 mL GDL, likely as a result of increased total protein in the gel accordance with its higher springiness and hardness (Table 1).
matrix as well as moderated acidication. Remarkably, these values Tofu prepared with citric acid formed a higher number of small
Fig. 7. Scanning electron micrographs of tofu prepared with (A) citric acid, (B) l-(-)-malic acid, and (C) tartaric acid at (a) 0.12 g/100 mL, (b) 0.14 g/100 mL, (c) 0.16 g/100 mL, and (d)
0.18 g/100 mL. Tofu prepared with (D) 0.30 g/100 mL glucono-d-lactone was used as a reference. Scale bars, 100 mm.
F.-H. Cao et al. / LWT - Food Science and Technology 85 (2017) 58e65 65
pores (Fig. 7Aa-Ad) than that prepared with l-(-)-malic (Fig. 7Ba- Food Science, 70, C258eC262.
Hayashi, N., Hayakawa, I., & Fujio, Y. (2010). Hydration of heat-treated soy protein
Bd) or tartaric acid (Fig. 7Ca-Cd), which was in accordance with its
isolate and its effect on the molten ow properties at an elevated temperature.
higher storage modulus and gel hardness. This might have resulted International Journal of Food Science & Technology, 27, 565e571.
from differences in acidication rate and pH. Indeed, the small Huang, C., Xu, T., Zhang, Y., Xue, Y., & Chen, G. (2007). Application of electrodialysis
pores in the former appear to have contributed to higher water- to the production of organic acids: State-of-the-art and recent developments.
Journal of Membrane Science, 288, 1e12.
holding capacity and hardness. The random and coarse structure Hu, H., Fan, X., Zhou, Z., Xu, X., Fan, G., Wang, L., et al. (2013). Acid-induced gelation
of tofu prepared with 0.16 or 0.18 g/100 mL acid indicates the lower behavior of soybean protein isolate with high intensity ultrasonic pre-treat-
quality of these gels. ments. Ultrasonics Sonochemistry, 20, 187e195.
Jacob, M., No bel, S., Jaros, D., & Rohm, H. (2011). Physical properties of acid milk
gels: Acidication rate signicantly interacts with cross-linking and heat
4. Conclusion treatment of milk. Food Hydrocolloids, 25, 928e934.
Kamizake, N. K. K., Silva, L. C. P., & Prudencio, S. H. (2016). Effect of soybean aging on
the quality of soymilk, rmness of tofu and optimum coagulant concentration.
The use of different types and concentrations of organic acids as Food Chemistry, 190, 90e96.
coagulant resulted in textural and microstructural differences in Kohyama, K., & Nishinari, K. (2002). Rheological studies on the gelation process of
tofu. Rheological properties, pH, and in-gel chemical interactions soybean 7 s and 11 s proteins in the presence of glucono-d-lactone. Journal of
Agricultural & Food Chemistry, 41, 8e14.
suggested that accelerated acidication reduces the amount of time Kohyama, K., Sano, Y., & Doi, E. (1995). Rheological characteristics and gelation
available for ordered aggregation of soy proteins, thus lowering mechanism of tofu (soybean curd). Journal of Agricultural & Food Chemistry, 43,
tofu quality. 1808e1812.
Kuiper, B. J. H., van Koningsveld, G. A., Alting, A. C., Driehuis, F., Gruppen, H., et al.
Tofu acidied with 0.14 g/100 mL organic acids exhibited
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interactions. On the other hand, tofu prepared with 0.12 and 0.16 g/ Li, M., Chen, F., Yang, B., Lai, S., Yang, H., Liu, K., et al. (2015). Preparation of organic
100 mL citric acid were comparable in hardness, although the tofu using organic compatible magnesium chloride incorporated with poly-
former was more elastic. Similar trends were observed for tofu saccharide coagulants. Food Chemistry, 167, 168.
Liu, H. H., Chien, J. T., & Kuo, M. I. (2013). Ultra high pressure homogenized soy our
prepared with 0.12 and 0.16 g/100 mL L-(-)-malic acid. However, for tofu making. Food Hydrocolloids, 32, 278e285.
tofu prepared with tartaric acid had comparatively poorer physical Malaki Nik, A., Alexander, M., Poysa, V., Woodrow, L., & Corredig, M. (2011). Effect of
properties and less robust chemical interactions. Finally, tofu pre- soy protein subunit composition on the rheological properties of soymilk
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pared with 0.18 g/100 mL organic acid was of low quality, as indi- Meng, L., Chen, F., Bao, Y., Lai, S., Yang, H., Liu, K., et al. (2015). Preparation of organic
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Qin, X. S., Luo, S. Z., Cai, J., Zhong, X. Y., Jiang, S. T., Zhao, Y. Y., et al. (2016). Trans-
tively, the data indicated that the quality of tofu prepared with glutaminase-induced gelation properties of soy protein isolate and wheat
organic acids is comparable to that prepared with GDL. Our study gluten mixtures with high intensity ultrasonic pretreatment. Ultrasonics Sono-
revealed that different types tofu (soft or rm) can be produced in a chemistry, 31, 590e597.
Rui, X., Fu, Y., Zhang, Q., Li, W., Zare, F., Chen, X., et al. (2016). A comparison study of
straightforward manner by using suitable types and concentrations
bioaccessibility of soy protein gel induced by magnesiumchloride, glucono-d-
of organic acids. lactone and microbial transglutaminase. LWT - Food Science and Technology, 71,
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Anhui Province of China (No. 1301031031 and 15czz03096). salt-extracted pea protein gels. Food Hydrocolloids, 28, 325e332.
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