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Study on the relationship between raw material characteristics of soybean protein concentrate and textured vegetable protein quality

Abstract

To investigate which protein properties affect the quality of high moisture textured vegetable protein (HTVP), we compared 12 commercial Soy Protein Concentrate (SPC). The comparison revealed that characteristics of different SPCs vary greatly, significant impact on the quality of extrusion products. There were significant differences in the degree of texturization, textural properties, and the sensory score of products. Statistical correlation analysis between SPC raw materials and products showed that the quality of products was significantly affected by the nitrogen solubility index (NSI), water holding capacity (WHC), emulsifying properties (EAI), foaming capacity (FC), gel strength, and sulfhydryl content of SPC raw materials. Our research provides a way to stabilize extrusion product quality.

Keywords:
soy protein concentrate; raw material characteristics; textured vegetable protein; quality properties

1 Introduction

High moisture extrusion cooking (moisture content ≥40%) started in the 1990s and was gradually developed based on low moisture extrusion technology. It is considered an emerging and promising technology to obtain fibrous structure similar to animal meat from plant protein (Kumar et al., 2017Kumar, P., Chatli, M., Mehta, N., Singh, P., Malav, O., & Verma, A. K. (2017). Meat analogues: health promising sustainable meat substitutes. Critical Reviews in Food Science and Nutrition, 57(5), 923-932. http://dx.doi.org/10.1080/10408398.2014.939739. PMid:25898027.
http://dx.doi.org/10.1080/10408398.2014....
; Zhang et al., 2022Zhang, Z., Zhang, L., He, S., Li, X., Jin, R., Liu, Q., Chen, S., & Sun, H. (2022). High-moisture extrusion technology application in the processing of textured plant protein meat analogues: a review. Food Reviews International, 1-36. http://dx.doi.org/10.1080/87559129.2021.2024223.
http://dx.doi.org/10.1080/87559129.2021....
). High moisture textured vegetable protein (HTVP) is an ideal substitute for animal meat due to its high protein, low saturated fat, no cholesterol, zero hormones, good digestibility, rich in essential amino acids, and direct edible advantages (Samard & Ryu, 2019Samard, S., & Ryu, G. H. (2019). Physicochemical and functional characteristics of plant protein‐based meat analogs. Journal of Food Processing and Preservation, 43(10), e14123. http://dx.doi.org/10.1111/jfpp.14123.
http://dx.doi.org/10.1111/jfpp.14123...
).

Soy ingredients are the most commonly used in meat analogues because of their characteristic functional properties, such as water-holding, fat-absorbing, gelling, and emulsifying capacities (Kyriakopoulou et al., 2019Kyriakopoulou, K., Dekkers, B., & van der Goot, A. J. (2019). Plant-based meat analogues. In C. M. Galanakis (Ed.), Sustainable meat production and processing (pp. 103-126). Cambridge: Academic Press. http://dx.doi.org/10.1016/B978-0-12-814874-7.00006-7.
http://dx.doi.org/10.1016/B978-0-12-8148...
). Soy protein concentrate (SPC) is a further purification of low-temperature defatted soybean meal. After removing the water-soluble or alcohol-soluble non-protein part, the product contains more than 65% protein. SPC-based meat analogues have been reported to be easier to extrude and more pronounced anisotropic than formulations based on soy protein isolate (SPI) under similar conditions (Chiang et al., 2019Chiang, J. H., Loveday, S. M., Hardacre, A. K. & Parker, M. E. (2019). Effects of soy protein to wheat gluten ratio on the physicochemical properties of extruded meat analogues. Food Structure, 19, 100102. https://doi.org/10.1016/j.foostr.2018.11.002.
https://doi.org/10.1016/j.foostr.2018.11...
). The result may be that the polysaccharides in SPC contribute to the formation of the dispersed phase (Wittek et al., 2021aWittek, P., Ellwanger, F., Karbstein, H. P., & Emin, M. A. (2021a). Morphology development and flow characteristics during high moisture extrusion of a plant-based meat analogue. Foods, 10(8), 10. http://dx.doi.org/10.3390/foods10081753. PMid:34441530.
http://dx.doi.org/10.3390/foods10081753...
). Furthermore, SPC, at low cost, is used as raw material to produce textured vegetable protein, which has good economic benefits (Wittek et al., 2021bWittek, P., Ellwanger, F., Karbstein, H. P., & Emin, M. A. (2021b). Morphology development and flow characteristics during high moisture extrusion of a plant-based meat analogue. Foods, 10(8), 1753. http://dx.doi.org/10.3390/foods10081753. PMid:34441530.
http://dx.doi.org/10.3390/foods10081753...
; Chajuss, 2001Chajuss, D. (2001). Soy protein concentrate: processing, properties, and prospects-soy protein concentrate is a widely used food ingredient that can be very profitable to manufacturers and users, as Daniel Chajuss of. Inform-International News on Fats Oils and Related Materials, 12, 1176-1182.).

Protein as the skeleton material of textured vegetable protein, protein intrinsic physicochemical properties, such as particle size, nitrogen solubility index (NSI), 11S and 7S content, and functional properties (hydration ability and active surface properties) by influencing the heat transfer rate, specific mechanical energy, and water or other protein interaction and so on, further affect the quality of extrusion products (Beniwal et al., 2021Beniwal, A. S., Singh, J., Kaur, L., Hardacre, A., & Singh, H. (2021). Meat analogs: Protein restructuring during thermomechanical processing. Comprehensive Reviews in Food Science and Food Safety, 20(2), 1221-1249. http://dx.doi.org/10.1111/1541-4337.12721. PMid:33590609.
http://dx.doi.org/10.1111/1541-4337.1272...
; Zhang et al., 2022Zhang, Z., Zhang, L., He, S., Li, X., Jin, R., Liu, Q., Chen, S., & Sun, H. (2022). High-moisture extrusion technology application in the processing of textured plant protein meat analogues: a review. Food Reviews International, 1-36. http://dx.doi.org/10.1080/87559129.2021.2024223.
http://dx.doi.org/10.1080/87559129.2021....
). During extrusion cooking, larger particles typically require a large input of mechanical energy (Singh & Koksel, 2021Singh, R. & Koksel, F. (2021). Effects of particle size distribution and processing conditions on the techno-functional properties of extruded soybean meal. LWT, 152, 112321. https://doi.org/10.1016/j.lwt.2021.112321.
https://doi.org/10.1016/j.lwt.2021.11232...
; Onwulata & Konstance, 2006Onwulata, C., & Konstance, R. (2006). Extruded corn meal and whey protein concentrate: effect of particle size. Journal of Food Processing and Preservation, 30(4), 475-487. http://dx.doi.org/10.1111/j.1745-4549.2005.00082.x.
http://dx.doi.org/10.1111/j.1745-4549.20...
). Wang et al. (2012)Wang, X., Zhu, X., & Yang, S. (2012). Effect of soybean protein properties on its fibrosis products by high moisture extrusion. Science and Technology of Food Industry, 33, 100-104. concluded that the extruded products with higher NSI, smaller particle size, and higher 11S globulin content had better fibrous structure. Furthermore, Liu et al. (2022)Liu, Y., Huang, Z. H., Hu, Z. X., Yu, Z., & An, H. Z. (2022). Texture and rehydration properties of texturised soy protein: analysis based on soybean 7S and 11S proteins. International Journal of Food Science & Technology, 58(1), 323-333. https://doi.org/10.1111/ijfs.15787.
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added 11S and 7S to the raw materials and concluded that a higher 11S protein content resulted in a brighter color, better texture properties, and a higher degree of texturization. Existing studies primarily explore products extruded from types of protein raw material (Schreuders et al., 2019Schreuders, F. K., Dekkers, B. L., Bodnár, I., Erni, P., Boom, R. M., & van der Goot, A. J. (2019). Comparing structuring potential of pea and soy protein with gluten for meat analogue preparation. Journal of Food Engineering, 261, 32-39. http://dx.doi.org/10.1016/j.jfoodeng.2019.04.022.
http://dx.doi.org/10.1016/j.jfoodeng.201...
; Palanisamy et al., 2019Palanisamy, M., Töpfl, S., Berger, R. G., & Hertel, C. (2019). Physico-chemical and nutritional properties of meat analogues based on Spirulina/lupin protein mixtures. European Food Research and Technology, 245(9), 1889-1898. http://dx.doi.org/10.1007/s00217-019-03298-w.
http://dx.doi.org/10.1007/s00217-019-032...
), extrusion parameters (Ferawati et al., 2021Ferawati, F., Zahari, I., Barman, M., Hefni, M., Ahlström, C., Witthöft, C., & Östbring, K. (2021). High-moisture meat analogues produced from yellow pea and faba bean protein isolates/concentrate: effect of raw material composition and extrusion parameters on texture properties. Foods, 10(4), 843. http://dx.doi.org/10.3390/foods10040843. PMid:33924424.
http://dx.doi.org/10.3390/foods10040843...
; Pietsch et al., 2019Pietsch, V. L., Werner, R., Karbstein, H. P., & Emin, M. A. (2019). High moisture extrusion of wheat gluten: Relationship between process parameters, protein polymerization, and final product characteristics. Journal of Food Engineering, 259, 3-11. http://dx.doi.org/10.1016/j.jfoodeng.2019.04.006.
http://dx.doi.org/10.1016/j.jfoodeng.201...
), system parameters (Chen et al., 2010Chen, F. L., Wei, Y. M., Zhang, B., & Ojokoh, A. O. (2010). System parameters and product properties response of soybean protein extruded at wide moisture range. Journal of Food Engineering, 96(2), 208-213. http://dx.doi.org/10.1016/j.jfoodeng.2009.07.014.
http://dx.doi.org/10.1016/j.jfoodeng.200...
; Zhang et al., 2015Zhang, B., Zhang, Y., Dreisoerner, J., & Wei, Y. (2015). The effects of screw configuration on the screw fill degree and special mechanical energy in twin-screw extruder for high-moisture texturised defatted soybean meal. Journal of Food Engineering, 157, 77-83. http://dx.doi.org/10.1016/j.jfoodeng.2015.02.019.
http://dx.doi.org/10.1016/j.jfoodeng.201...
) and additives (Chen et al., 2021Chen, Y., Liang, Y., Liu, H., Wang, X., & Wang, J. (2021). Extrusion of wheat gluten-peanut oil complexes and their rheological characteristics. Food Chemistry, 364, 130435. http://dx.doi.org/10.1016/j.foodchem.2021.130435. PMid:34175619.
http://dx.doi.org/10.1016/j.foodchem.202...
; Dou et al., 2022Dou, W., Zhang, X., Zhao, Y., Zhang, Y., Jiang, L., & Sui, X. (2022). High moisture extrusion cooking on soy proteins: Importance influence of gums on promoting the fiber formation. Food Research International, 156, 111189. http://dx.doi.org/10.1016/j.foodres.2022.111189. PMid:35651098.
http://dx.doi.org/10.1016/j.foodres.2022...
) etc. However, the relationship between the characteristics of protein raw materials and the quality of extruded products has not been studied deeply. So far, it is not clear which properties of protein raw materials are critical to the production of products.

Therefore, this study aims to establish possible connections between SPC raw material characteristics and the quality of HTVP, to solve the phenomenon of unstable product quality. A wide range of tests (physicochemical and functional properties, etc.) were performed to characterize SPC. Furthermore, extruded product indexes were measured. The relationship between SPC raw material characteristics and extruded product quality was explored by cluster analysis and correlation analysis, which provided a theoretical basis for the selection of raw material and control of product quality in the processing of plant protein texturization.

2 Materials

Twelve commercial SPC were used, obtained from different batches from six manufacturers (Yihai Kerry, Yuwang, Vanderfor, Yuxin, Jiahua, and DuPont) in China, marked SPC 1-12. SPI contains, on a dry basis, 94.24% protein, 5.99% moisture, 4.64% ash, and 0.39% fat. Wheat gluten (WG) contains about 87.66% protein, 5.32% moisture, 2.00% ash, and 2.11% fat. SPI and WG were purchased from Yihai Kerry Food Industry Co., LTD. (Henan, China).

2.1 Raw material characteristics of SPC

Chemical composition of SPC

The moisture, ash, and fat in the samples were determined according to the method of AACC (American Association of Cereal Chemists, 1999aAmerican Association of Cereal Chemists - AACC. (1999a). Ash—basic method. Saint Paul: AACC., bAmerican Association of Cereal Chemists - AACC. (1999b). Calculation of percent moisture. Saint Paul: AACC., cAmerican Association of Cereal Chemists - AACC. (1999c). Crude fat in wheat, corn, and soy flour, feeds, and mixed feeds. Saint Paul: AACC.). The crude protein content of the samples was determined by the Kjeldahl method (N × 6.25) (National Health and Family Planning Commission of the People Republic of China, 2016National Health and Family Planning Commission of the People Republic of China. State Administration of Food and Drug Administration. (2016). National food safety standard -- determination of protein in foods. Beijing: National Health and Family Planning Commission of the People Republic of China.).

Nitrogen solubility index

Nitrogen solubility index (NSI) reference to AACC - 46 -23 (American Association of Cereal Chemists, 1999dAmerican Association of Cereal Chemists - AACC. (1999d). Nitrogen solubility index. Saint Paul: AACC.).

pH

The SPC was prepared into 8% slurries with distilled water pH 7.0 as a dispersant. The pH of the solution was determined using a pH meter (HI2210, HANNA, Italy).

Water and oil holding capacity (WHC and OHC)

The determination of WHC or OHC was performed according to Zahari et al. (2021)Zahari, I., Ferawati, F., Purhagen, J. K., Rayner, M., Ahlstrom, C., Helstad, A., & Ostbring, K. (2021). Development and characterization of extrudates based on rapeseed and pea protein blends using high-moisture extrusion cooking. Foods, 10(10), 2397. http://dx.doi.org/10.3390/foods10102397. PMid:34681446.
http://dx.doi.org/10.3390/foods10102397...
. 3 g SPC was accurately weighed into a centrifuge tube, 17 mL distilled water or soybean oil was added, shaken for 1 min, then stood for 15 min, centrifuged at 5 000 g for 10 min, and the supernatant was poured out and weighed. The WHC and OHC were calculated by the following formula (Equation 1).

W H C or O H C (%) = M 3 - M 2 - M 1 M 1 × 100 % (1)

Where M1 = the mass of the SPC, M2 = the mass of the empty centrifuge tube, and M3 = the total weight of SPC and centrifuge tube after the supernatant was decanted.

Emulsifying activity index (EAI) and emulsifying stability index (ESI)

EAI and ESI were determined according to the method of Ahmed et al. (2018)Ahmed, J., Al-Ruwaih, N., Mulla, M., & Rahman, M. H. (2018). Effect of high pressure treatment on functional, rheological and structural properties of kidney bean protein isolate. LWT, 91, 191-197. http://dx.doi.org/10.1016/j.lwt.2018.01.054.
http://dx.doi.org/10.1016/j.lwt.2018.01....
and Wani et al. (2015)Wani, I. A., Sogi, D. S., Shivhare, U. S., & Gill, B. S. (2015). Physico-chemical and functional properties of native and hydrolyzed kidney bean (Phaseolus vulgaris L.) protein isolates. Food Research International, 76, 11-18. http://dx.doi.org/10.1016/j.foodres.2014.08.027.
http://dx.doi.org/10.1016/j.foodres.2014...
. The SPC was prepared into 1 g/100 mL of slurries, and the solution was stirred for 30 minutes, centrifuged at 3,000 g for 15 min, 5 mL soybean oil was added to 15 mL supernatant, and homogenized for 1 min at 10,000 rpm. At 0 min and 10 min, the absorbance was measured at 500 nm. The EAI and ESI were calculated by the following formula (Equations 2-3).

E A I ( m 2 / g ) = 2 × 2.303 × A 0 × N θ × L × C × 10000 (2)
E S I ( min - 1 ) = A 0 × 10 A 0 - A 10 (3)

Where A0 and A10 are the absorbance values measured at 0 min and 10 min; N = the dilution ratio (100); θ = the oil volume fraction of the emulsion (0.25); L = the optical path (0.01); C = the initial protein concentration (g/mL).

Foaming capacity (FC) and foaming stability (FS)

FC and FS were determined according to the procedure used by Yust et al. (2010)Yust, M. D. M., Pedroche, J., Millán-Linares, M. D. C., Alcaide-Hidalgo, J. M., & Millán, F. (2010). Improvement of functional properties of chickpea proteins by hydrolysis with immobilised Alcalase. Food Chemistry, 122(4), 1212-1217. http://dx.doi.org/10.1016/j.foodchem.2010.03.121.
http://dx.doi.org/10.1016/j.foodchem.201...
. Slurries of 1 g/100 mL were prepared with distilled water, and the sample was stirred for 30 min to ensure complete dissolution. After centrifugation at 3,000 g for 10 min, 25 mL protein solution was homogenized at 10,000 rpm for 1 min. The volume of the foam was measured after homogenization and standing for 25 min. The FC and FS were calculated by the following formula (Equations 4-5).

F C ( % ) = V 1 V 0 × 100 % (4)
F S ( % ) = V t V 1 × 100 % (5)

Where V0 = the initial volume of protein solution, V1 = the foam volume after homogenization for 1 min, and Vt = the foam volume after standing for 25 min.

RVA viscosity

The viscosity of the proteins was measured on a Rapid Visco Analyzer (RVA-TM, Perten, Sweden). SPC was prepared in 10% slurries, and rapidly stirred at 960 rpm for 15 s and 160 rpm for 10 min. The temperature was constant at 25 °C, and the value was recorded at 5 min.

Gel strength

The determination was performed according to Jin et al. (2013)Jin, I.-H., Kim, J.-E., Seo, J.-H., & Lee, S.-P. (2013). Physicochemical properties of soy protein isolate gels emulsified with various oils using a microbial transglutaminase. Food Science and Biotechnology, 22(S1), 129-136. http://dx.doi.org/10.1007/s10068-013-0058-0.
http://dx.doi.org/10.1007/s10068-013-005...
. Preparation of gel: The SPC was made into 10% slurries, homogenized at 10,000 rpm with a high speed homogenizer for 5 s, and 20 mL was placed in a beaker, covered with plastic wrap, and heated in a water bath at 90 °C for 35 min. After removal, it was cooled for 20 min and placed in a refrigerator at 4 °C for 24 h. Gel strength was measured on a physical property tester (Ta. XT Plus, Stable Micro-systems, UK) with a P/0.5 probe at a speed of 1 mm/s.

Sulfhydryl content

Ellman's reagent was used to determine the content of free sulfhydryl groups in the samples. Take 100 mg of SPC, add 4.7 g guanidine hydrochloride, and then dissolve in 10 mL of buffer. Take 1 mL of the prepared solution, add 4 mL of urea-guanidine hydrochloride solution and 0.05 mL of Ellman's reagent, stand still for 30 min, then measure the absorbance value at the wavelength of 412 nm (Equation 6).

S H ( μ m o l / g ) = 73.53 × A 412 × D C (6)

Where A412 = the absorbance value of the sample at 412 nm; D = the dilution factor; C = the sample concentration (mg(dry base)/mL).

11S/7S

SDS-PAGE was performed according to the method of Yadav et al. (2010)Yadav, M. P., Parris, N., Johnston, D. B., Onwulata, C. I., & Hicks, K. B. (2010). Corn fiber gum and milk protein conjugates with improved emulsion stability. Carbohydrate Polymers, 81(2), 476-483. http://dx.doi.org/10.1016/j.carbpol.2010.03.003.
http://dx.doi.org/10.1016/j.carbpol.2010...
.

2.2 High moisture extrusion cooking

A CLEXTRAL Ev025 (Firminy, France) type twin screw extruder was adopted with a screw diameter of 25.5 mm and an aspect ratio of 24. The barrel contains six temperature control areas, which are heated by an electric heating cylinder and cooled by circulating water.

The formula of extruder ingredients includes SPI, SPC, and WG in a ratio of 3: 4: 3 and 5% corn starch. The mixtures were fed into the extruder at a constant speed of 3.4 kg/h. Based on preliminary experiments and operational stability of the extruder, water entered the extruder at a constant speed of 3.0 L/h, the screw speed was 280 rpm, and the temperatures of extruder barrel were kept at 30, 90, 120, 140, 150, and 160 °C from the first zone to the sixth zone, respectively. After the extrusion material was stable, it was collected, packed with a vacuum seal, and stored at -18 °C for further experiments.

2.3 Color

The color difference meter (CR-400, Konica Minolta, Japan) was used for determination, and L* and ∆E* values were recorded.

2.4 Texture properties

The texture properties of the extrudates were evaluated using a TA.XT plus Texture Analyzer (Stable Micro Systems, UK). The shape samples (2 cm × 2 cm × 0.5 cm) were compressed to 75% of the original thickness with a P/36R probe at a speed of 2mm /s, and the hardness, springiness, and chewiness were recorded. Samples of the same shape were cut using a knife blade to 75% of their original thickness at the same speed, and the cutting strength was recorded.

2.5 Sensory evaluation

Ten professional sensory evaluators were invited to evaluate 12 HTVPs. The sensory evaluation criteria are shown in Table 1.

Table 1
Sensory evaluation criteria of HTVP.

2.6 Statistical analysis

All experiments were performed at least three times, and the results were expressed as mean ± standard deviation (SD). Analysis of variance (ANOVA) and Duncan's multiple interval test were used to compare the results. Origin 2021 was used for mapping.

3 Results and discussion

3.1 Analysis of SPC raw material characteristics

The chemical composition of SPC 1-12 is summarized in Table 2. The moisture and ash content of the 12 SPCs follows the General Standard for Soy Protein Products CXS 175-1989 (Food and Agriculture Organization, 2019Food and Agriculture Organization - FAO. (2019). CXS 175-1989 general standard for soy protein products. Codex Committee on Vegetable Protein. Rome: FAO.) for SPC moisture content (≤10.0%) and ash content (≤8.0). With the exception of SPC 5, crude protein content is just under 65%, and all others met the crude protein content standard (65%~90%).

Table 2
Chemical composition of SPC.

The raw material characteristics of protein are the basis and guarantee of producing extruded plant protein. The characteristics of SPC 1-12 raw materials are shown in Table 3. There are significant differences in the characteristics of SPC from different manufacturers and different batches of the same manufacturer, which may be related to the soybean varieties (Min et al., 2005Min, S., Yu, Y., & Martin, S. St. (2005). Effect of soybean varieties and growing locations on the physical and chemical properties of soymilk and tofu. Food and Chemical Toxicology, 70(1), C8-C21. https://doi.org/10.1111/j.1365-2621.2005.tb09026.x.
https://doi.org/10.1111/j.1365-2621.2005...
), production process (Preece et al., 2017Preece, K. E., Hooshyar, N., & Zuidam, N. J. (2017). Whole soybean protein extraction processes: a review. Innovative Food Science & Emerging Technologies, 43, 163-172. http://dx.doi.org/10.1016/j.ifset.2017.07.024.
http://dx.doi.org/10.1016/j.ifset.2017.0...
), and storage conditions (Ziegler et al., 2018Ziegler, V., Ferreira, C. D., Hoffmann, J. F., de Oliveira, M., & Elias, M. C. (2018). Effects of moisture and temperature during grain storage on the functional properties and isoflavone profile of soy protein concentrate. Food Chemistry, 242, 37-44. http://dx.doi.org/10.1016/j.foodchem.2017.09.034. PMid:29037703.
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) used by the manufacturer. All SPCs used in this study were prepared using the alcohol method. The NSI of SPC produced by the alcohol method is generally lower (<15%). Still, the NSI difference in the measurement results is very significant, which is mainly caused by the different degrees of protein denaturation caused by process conditions, such as leaching concentration (Peng et al., 2021Peng, Y., Kyriakopoulou, K., Ndiaye, M., Bianeis, M., Keppler, J. K., & van der Goot, A. J. (2021). Characteristics of soy protein prepared using an aqueous ethanol washing process. Foods, 10(9), 10. http://dx.doi.org/10.3390/foods10092222. PMid:34574332.
http://dx.doi.org/10.3390/foods10092222...
), drying methods (Ghribi et al., 2015Ghribi, A. M., Gafsi, I. M., Blecker, C., Danthine, S., Attia, H., & Besbes, S. (2015). Effect of drying methods on physico-chemical and functional properties of chickpea protein concentrates. Journal of Food Engineering, 165, 179-188. http://dx.doi.org/10.1016/j.jfoodeng.2015.06.021.
http://dx.doi.org/10.1016/j.jfoodeng.201...
) and different modification methods (Sui et al., 2021Sui, X., Zhang, T., & Jiang, L. (2021). Soy protein: Molecular structure revisited and recent advances in processing technologies. Annual Review of Food Science and Technology, 12(1), 119-147. http://dx.doi.org/10.1146/annurev-food-062220-104405. PMid:33317319.
http://dx.doi.org/10.1146/annurev-food-0...
; Moreno et al., 2020Moreno, H. M., Tovar, C. A., Domínguez-Timón, F., Cano-Báez, J., Díaz, M. T., Pedrosa, M. M., & Borderías, A. J., (2020). Gelation of commercial pea protein isolate: effect of microbial transglutaminase and thermal processing. Food Science and Technology (Campinas), 40(4), 800-809. http://dx.doi.org/10.1590/fst.19519.
http://dx.doi.org/10.1590/fst.19519...
). Studies had found that the protein solubility of ethanol-treated lentil protein isolate was below 10% when 35~55% ethanol was applied and increased to approximately 30% when the water/ethanol ratio increased from 55 to 75% (Chang et al., 2019Chang, C., Stone, A. K., Green, R., & Nickerson, M. T. (2019). Reduction of off-flavours and the impact on the functionalities of lentil protein isolate by acetone, ethanol, and isopropanol treatments. Food Chemistry, 277, 84-95. http://dx.doi.org/10.1016/j.foodchem.2018.10.022. PMid:30502215.
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). There are significant differences in WHC, OHC, and EAI among different SPCs. The higher WHC may be due to the lower loss of soluble protein and the higher polar amino acids, while OHC is attributed to protein hydrophobicity and the binding performance of non-polar amino acid side chains to fat (Ghribi et al., 2015Ghribi, A. M., Gafsi, I. M., Blecker, C., Danthine, S., Attia, H., & Besbes, S. (2015). Effect of drying methods on physico-chemical and functional properties of chickpea protein concentrates. Journal of Food Engineering, 165, 179-188. http://dx.doi.org/10.1016/j.jfoodeng.2015.06.021.
http://dx.doi.org/10.1016/j.jfoodeng.201...
). EAI refers to the ability of protein to quickly adsorb on the oil-water interface, and ESI is to evaluate the stability of the protein staying at the water-oil interface for a period of time (Yao et al., 2022Yao, G., Guo, Y., Cheng, T., Wang, Z., Li, B., Xia, C., Jiang, J., Zhang, Y., Guo, Z., & Zhao, H. (2022). Effect of γ-irradiation on the physicochemical and functional properties of rice protein. Food Science and Technology (Campinas), 42, e12422. http://dx.doi.org/10.1590/fst.12422.
http://dx.doi.org/10.1590/fst.12422...
). There is no significant difference in ESI among SPCs. There are significant differences in FC, divided into two levels. SPC 2, 4, 5, 7, 8, 9, and 11 are the ones with higher FC, and SPC 1, 6, and 10 are the ones with lower FC. Differences in FS are also very significant. SPC 2, 5, 8, and 9 are free of foam, and SPC 6 and 10 are relatively stable at 40%. The foaming ability of protein mainly depends on its soluble part. Generally, there is a strong positive correlation between FC and NSI. At the same time, it can be seen that the data of FC and FS data show opposite trends. The viscosity can be divided into three apparent levels. 19.00~22.00 cP is the lowest batch, 164.50~397.50 cP is the middle batch, and 1 093.50~2 188.50 cP is the highest batch. The three clusters of raw materials have a very significant difference. There are significant differences in sulfhydryl content, composition, and gel strength among SPCs. Sulfhydryl oxidation generates new disulfide bonds to organize protein molecules in a more orderly manner and form a more stable network structure (Bruneel et al., 2011Bruneel, C., Lagrain, B., Brijs, K., & Delcour, J. A. (2011). Redox agents and N-ethylmaleimide affect the extractability of gluten proteins during fresh pasta processing. Food Chemistry, 127(3), 905-911. http://dx.doi.org/10.1016/j.foodchem.2011.01.048. PMid:25214077.
http://dx.doi.org/10.1016/j.foodchem.201...
). Therefore, free sulfhydryl groups have a great impact on the formation of gel and protein cross-linking. The ratio of 7S and 11S is different in different soybean varieties, which has an important impact on the emulsification (Cheng et al., 2006Cheng, C. L., Shi, Y. G., Zhu, X. Q., & Jiang, L. Z. (2006). The effect of soybean variety on emulsifying function of soubean protein. Science and Technology of Cereals, Oils and Foods, 14, 25-27.) and gel ability (Shi et al., 2005Shi, Y. G., Cheng, C. L., Zhu, X. Q., & Jiang, L. Z. (2005). The effect of soybean variety on gelling function of SPI. Journal of the Chinese Cereals and Oils Association, 20, 58-60.) of soybean protein. 11S globulin contains more disulfide bonds and sulfhydryl groups, so when the proportion of 11S globulin increases, high gel strength. In conclusion, SPC raw material characteristics of different manufacturers and different batches produced by the same manufacturer are significantly different.

Table 3
Raw material characteristics of SPC.

3.2 Color and functional properties analysis

Color L* is lightness value. The larger L*, the brighter the sample. ∆E* indicates the degree of difference between each value and the standard whiteboard. Table 4 shows that there are significant differences in color between products. L*, ∆E* coefficient of variation is 1%, 2%, and the coefficient of variation is small (<15%). The color datas for 12 HTVP products did not vary much, suggesting that SPC raw materials have little impact on the color of the products. Therefore, the color index of product quality can be considered a secondary factor in selecting raw materials.

Table 4
color and functional properties of HTVP.

WHC and OHC are related to the taste and flavor of products, reflecting the adsorption and retention capacity of HTVP products for water or oil, which depends on protein composition, protein denaturation, and the degree of interaction with water and oil (Samard & Ryu, 2019Samard, S., & Ryu, G. H. (2019). Physicochemical and functional characteristics of plant protein‐based meat analogs. Journal of Food Processing and Preservation, 43(10), e14123. http://dx.doi.org/10.1111/jfpp.14123.
http://dx.doi.org/10.1111/jfpp.14123...
). Table 3 shows significant differences in WHC and OHC among products, with the maximum WHC being 289.89% and the minimum WHC being 222.61%. The maximum OHC was 109.94%, and the minimum OHC was 90.72%. After the extrusion of high temperature, high pressure, and high shear, the molecular structure of proteins from different raw materials is destroyed and assembled to form different structures. However, the variation coefficient of WHC and OHC of the product was small (<15%), indicating that different SPC had little influence on the WHC and OHC of the product.

3.3 Product texture properties and sensory analysis

The texture properties of HTVP play an important role in people's acceptability and repeat purchases. In Figure 1, the hardness, springiness, and chewiness range from 18 145.40 to 29 551.09, 0.85 to 0.97, and 13 706.35 to 24 497.62, and the coefficient of variation is 16.10%, 3.72%, and 22.39%, respectively. Through the significance analysis and coefficient of variation of HTVP texture characteristics, it can be concluded that there are significant differences in the hardness and chewiness of HTVP. Combined with the sensory score, people's acceptance is relatively high when the hardness is kept at about 20 000 g, and the chewiness is maintained at 15 000 g. If the hardness and chewiness are too high or too low, it will reduce the taste and decrease the approval of people.

Figure 1
Textural properties of HTVP.

The degree of texturization is an important index used to evaluate the quality of HTVP products. For example, Zhang et al. (2018a)Zhang, J., Liu, L., Zhu, S., & Wang, Q. (2018a). Texturisation behaviour of peanut-soy bean/wheat protein mixtures during high moisture extrusion cooking. International Journal of Food Science & Technology, 53(11), 2535-2541. http://dx.doi.org/10.1111/ijfs.13847.
http://dx.doi.org/10.1111/ijfs.13847...
used it to describe the degree of fibrosis of products. The higher the value of texturization degree, the better the fiber structure. In Figure 2, The texturization degree of the 12 HTVPs ranges from 1.28 to 2.06, and the coefficient of variation is 16.32%. The range of a batch with a reasonable degree of texturization is about 2.00, mainly No.5, No.8, and No.9. However, Osen et al. (2014)Osen, R., Toelstede, S., Wild, F., Eisner, P., & Schweiggert-Weisz, U. (2014). High moisture extrusion cooking of pea protein isolates: raw material characteristics, extruder responses, and texture properties. Journal of Food Engineering, 127, 67-74. http://dx.doi.org/10.1016/j.jfoodeng.2013.11.023.
http://dx.doi.org/10.1016/j.jfoodeng.201...
concluded that when the cooking temperature exceeded the denaturation temperature of the protein, the functional characteristics of the protein played a small role in the process of fiber formation. This is inconsistent with the experimental results, possibly due to the different extrusion materials used and the significant differences in the quality of SPC raw materials measured in this experiment.

Figure 2
Texturization degree and sensory of HTVP.

Sensory evaluation has been used as a valid tool for assessing the quality of food products, bringing predictive results on the acceptance of a product by consumers (Priulli et al., 2021Priulli, É., Pereira, C. M. T., Oliveira, A. C., & Pires, C. R. F. (2021). Use of the quantitative descriptive analysis for sensory assessment of tapiocas with adding waste from soy processing. Food Science and Technology (Campinas), 42, e08621. https://doi.org/10.1590/fst.08621.
https://doi.org/10.1590/fst.08621...
). For TVP to replace meat in the daily diet, these products need to be accepted by the public in terms of their overall preferences. In addition, sensory assessment combined with instrumental measures (e.g. texture and colour) can also better guide product processing and recipe optimization, helping to improve the end product (Fiorentini et al., 2020Fiorentini, M., Kinchla, A. J., & Nolden, A. A. (2020). Role of sensory evaluation in consumer acceptance of plant-based meat analogs and meat extenders: a scoping review. Foods, 9(9), 1334. http://dx.doi.org/10.3390/foods9091334. PMid:32971743.
http://dx.doi.org/10.3390/foods9091334...
). Sensory scores for the 12 HTVPs are shown in Figure 2. The sensory evaluation of HTVP shows that No.2 has the highest score. No.2 HTVP, showing a light yellow color, has a flat surface without burrs, good taste, flavor, and fiber structure, so the overall score is the highest. The lowest score is No.1, which was low because of its poor taste, smell and fiber structure, dark color, hard taste and lack of acceptance.

3.4 Cluster analysis of HTVP quality

To find the optimal HTVP and the most suitable SPC raw material characteristics, the quality of HTVP is clustered. The results are shown in Figure 3, Table 5, and Table 6.

Figure 3
Pedigree of HTVP quality characteristics cluster analysis.
Table 5
Quality characteristics of HTVP after clustering.
Table 6
Characteristics of SPC raw materials of different classes after clustering.

The 12 products were divided into three categories and the clustered HTTPP quality characteristics are shown in Table 5. The WHC, OHC, and sensory scores of the first category are the highest among the three types. The second category has the highest degree of texturization, followed by the first. The hardness and chewiness of the third group are the highest. According to the data presented above, springiness and color are not significantly different among the three groups, and the hardness and chewiness keep at the appropriate level. Comprehensive analysis showed that the first type of product had an excellent WHC, OHC, and the highest sensory score, indicating the best acceptance. Although the degree of texturization of the first type is inferior to the second type of products, the difference is slight. So the first type of sample can be used as the optimal product. According to products 2, 7, 4, and 9 included in the first category, the optimal range of the HTVP protein quality index can be selected as follows: WHC is 252.58%~289.89%, OHC is 93.47%~104.38%, texturization degree is 1.46~2.00, hardness is 18 145.40 g~20 536.49 g, springiness is 85.00%~87.00%, and chewiness is 13 706.35 g~15 277.18 g.

The characteristics of raw materials corresponding to the three categories of HTVP products obtained by cluster analysis are shown in Table 6. The functional characteristics of SPC raw materials corresponding to the first category of products with the best product quality performance are the highest, such as NSI, WHC, OHC, EAI, FC, and gel strength. Therefore, it can be concluded that the functional characteristics of SPC play an essential role in promoting the quality of HTVP. In addition, moisture, ash, crude protein, fat content, and color of the three kinds of HTVP related raw materials do not have noticeable differences. To sum up, when the quality of HTVP is better, the required range of SPC raw material characteristics is as follows: NSI is 50.44%~70.56%, pH is 6.99~7.84, WHC is 425.62%~492.90%, OHC is 118.00%~150.00%, EAI is 8.82~10.27%, ESI is 14.70%~21.47%. FC is 140.54%~150.82%, FS is 10.34%~23.55%, viscosity is 397.50 CP~856.29 cP, gel strength is 82.00 g~144.76 g, sulfhydryl content is 3.62 μmol/g~5.31 μmol/g, the ratio of 11S to 7S is 1.52~2.49.

3.5 Correlation analysis

The correlation analysis results between SPC raw materials and HTVP quality indexes are shown in Figure 4. The influence of raw materials on products is mainly discussed in this part. The WHC of the product is positively correlated with NSI, OHC, EAI, and negatively correlated with ESI of the SPC raw material. The water holding capacity of proteins is directly proportional to their water binding capacity, so WHC is positively correlated with NSI. The NSI, WHC, OHC, and EAI of SPC raw materials have significant correlation. The OHC of the product is positively correlated with pH, WHC, OHC, EAI, FC, viscosity, gel strength, 11S/7S, and negatively correlated with the sulfhydryl content of raw materials. The hardness and chewiness of HTVP are negatively correlated with NSI, WHC, OHC, EAI, FC, and gel strength of raw materials. The chewiness is also negatively correlated with pH and 11s/7s of raw materials. The springiness is negatively correlated with the OHC and FC of raw materials. The higher the NSI value, the lower the degree of protein denaturation and the better the texture properties of extruded products (Zhang et al., 2017Zhang, B., Zhang, J. C., & Wei, Y. M. (2017). The effect of raw physicochemical properties on the quality of extruded texturized vegetable protein. Journal of Chinese Institute of Food Science and Technology, 17, 155-163.). In addition, the stronger the binding capacity of the protein raw material with water or oil, the softer the product.

Figure 4
Correlation analysis between SPC raw material and HTVP quality indexes.

The texture degree of HTVP is positively correlated with pH, WHC, OHC, EAI, FC, viscosity, gel strength, and 11S/7S of raw materials. It is negatively correlated with the content of the sulfhydryl group. Increasing the pH value of raw materials appropriately can improve the organizational quality of products. Alkaline conditions promoted the formation of gluten network and induced a more fibrous cross section and the regular surface of gluten extrudate (Li et al., 2018Li, T., Guo, X.-N., Zhu, K.-X., & Zhou, H.-M. (2018). Effects of alkali on protein polymerization and textural characteristics of textured wheat protein. Food Chemistry, 239, 579-587. http://dx.doi.org/10.1016/j.foodchem.2017.06.155. PMid:28873608.
http://dx.doi.org/10.1016/j.foodchem.201...
). The viscosity is the internal friction force when the material flows. The greater the viscosity value, the slower the flow speed of the material in the barrel, and the more orderly fiber structure may be formed in the cooling mold (Chen et al., 2010Chen, F. L., Wei, Y. M., Zhang, B., & Ojokoh, A. O. (2010). System parameters and product properties response of soybean protein extruded at wide moisture range. Journal of Food Engineering, 96(2), 208-213. http://dx.doi.org/10.1016/j.jfoodeng.2009.07.014.
http://dx.doi.org/10.1016/j.jfoodeng.200...
). Proteins with poor gel ability, such as peanut protein, are difficult to form rich fibrous structure during high moisture extrusion (Zhang et al., 2018bZhang, J., Liu, L., Zhu, S., & Wang, Q. (2018b). Texturisation behaviour of peanut-soy bean/wheat protein mixtures during high moisture extrusion cooking. International Journal of Food Science & Technology, 53(11), 2535-2541. http://dx.doi.org/10.1111/ijfs.13847.
http://dx.doi.org/10.1111/ijfs.13847...
). Soybean 11S protein has a higher content of sulfur amino acids, and the tensile and shear strength of heat-induced gels are higher than 7S protein. It may be easier to promote the formation of fiber structure through covalently cross-linking during extrusion, thus improving the texture degree of HTVP (Liu et al., 2022Liu, Y., Huang, Z. H., Hu, Z. X., Yu, Z., & An, H. Z. (2022). Texture and rehydration properties of texturised soy protein: analysis based on soybean 7S and 11S proteins. International Journal of Food Science & Technology, 58(1), 323-333. https://doi.org/10.1111/ijfs.15787.
https://doi.org/10.1111/ijfs.15787...
). The oxidation reaction that occurs during extrusion reduces the content of sulfhydryl group, which is conducive to the formation of disulfide bonds, and the protein rearranged to form new chemical cross-linking in the die (Peng et al., 2022Peng, H., Zhang, J., Wang, S., Qi, M., Yue, M., Zhang, S., Song, J., Wang, C., Zhang, D., Wang, X., & Ma, C. (2022). High moisture extrusion of pea protein: Effect of l-cysteine on product properties and the process forming a fibrous structure. Food Hydrocolloids, 129, 107633. http://dx.doi.org/10.1016/j.foodhyd.2022.107633.
http://dx.doi.org/10.1016/j.foodhyd.2022...
). The sensory evaluation of HTVP is positively correlated with the NSI, pH, WHC, OHC, EAI, and FC of SPC. According to the above, we can try to produce better quality extrusion products by changing some characteristics of SPC raw materials.

4 Conclusion

In conclusion, we conclude that the characteristics of SPC raw materials produced by different manufacturers and different batches of the same manufacturer vary significantly. SPC was extruded with other raw materials to prepare HTVP, and it was found that there were significant differences in the quality of the various products, in which the texture properties, degree of texturization, and sensory score were significantly different. At the same time, WHC, OHC, and color coefficient of variation were small, and the differences between the data were slight. According to the correlation analysis results of SPC and HTVP quality, the SPC raw material characteristics that have a very significant impact on product quality are NSI, WHC, EAI, FC, gel strength, and sulfhydryl content.

Acknowledgements

This work was supported by the National Key Research and Development Program of China [2022YFD2101403].

  • Practical Application: The phenomenon of product quality instability often arises in the process of producing high moisture textured vegetable protein, which is closely related to the raw materials of soy protein concentrate used. It is useful to explore the relationship between the raw material characteristics of soy protein concentrate and the quality of high moisture textured vegetable protein products to address product instability in industry.

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Publication Dates

  • Publication in this collection
    13 Feb 2023
  • Date of issue
    2023

History

  • Received
    02 Nov 2022
  • Accepted
    22 Dec 2022
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