Application of perilla protein in preparation of sugar-controlled bread

Abstract

The invention provides an application of purple perilla protein in preparation of sugar-controlled bread, the purple perilla protein is obtained by deoiling and degreasing purple perilla seeds, and the sugar-controlled bread comprises the following raw materials in parts by mass: 80-95 parts of wheat flour and 5-20 parts of purple perilla protein powder. The sugar-controlled bread has the functions of reducing GI, reducing the release amount of reducing sugar, inhibiting the activity of alpha-glucosidase and alpha-amylase and reducing the content of RDS and SDS, and is beneficial to the control of postprandial blood sugar level.

Description

Technical Field

[0001] The present invention relates to the technical field of biomedicine, and in particular to an application of perilla protein in the preparation of sugar-controlled bread. Background Art

[0002] As people's living standards improve, high-sugar foods have become a major risk factor for obesity and diabetes. People consume 15% to 26% of their daily energy from sugar, and the World Health Organization recommends that daily sugar intake should be less than 10% of total energy intake. As people's health awareness gradually increases, the market demand for functional foods that reduce the glycemic index (GI) is rapidly increasing.

[0003] Perilla (Perilla Frutescens L.) is one of the plants that can be used as both medicine and food. The protein content of defatted perilla seed meal is as high as 40% to 65%. Perilla protein has the characteristics of anti-oxidation, immunomodulation, and improving metabolism, and can be used as an emerging plant protein resource for the development of functional foods. In the prior art, perilla soy sauce is prepared by low-salt solid-state fermentation using perilla meal as raw material. The results show that under the process conditions of 31% (mass fraction) replacement of perilla meal, 11% mass concentration of brine and 0.5% (mass fraction) addition of yeast, the soy sauce can meet the national first-class soy sauce standards, and the sensory quality is better than that of soy sauce without perilla meal. Summary of the invention

[0004] A first aspect of the present invention provides a use of perilla protein in preparing sugar-controlled bread or bread with a glycemic index (GI) lower than 85 or meal replacement bread.

[0005] The second aspect of the present invention also provides a use of perilla protein in preparing bread which reduces the release of reducing sugars, reduces the content of RDS and SDS, and inhibits the activity of α-glucosidase and α-amylase.

[0006] The third aspect of the present invention provides a sugar-controlled bread or a bread with a glycemic index (GI) of less than 85 or a meal replacement bread or a bread with reduced reducing sugar release, reduced RDS and SDS content, and inhibited activity of α-glucosidase and α-amylase.

[0007] The fourth aspect of the present invention provides a method for controlling postprandial blood sugar levels, which comprises eating controlled-sugar bread or bread with a glycemic index (GI) of less than 85 or meal replacement bread or bread with reduced reducing sugar release, reduced RDS and SDS content, and inhibited α-glucosidase and α-amylase activity instead of traditional bread.

[0008] The raw materials of the bread, sugar-controlled bread or meal replacement bread include wheat flour and perilla protein powder, wherein, by weight, they include:

[0009] Wheat flour: 80-95 parts, for example, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95 parts, etc., preferably 88-95 parts, more preferably 90-92 parts;

[0010] Perilla protein powder: 5-20 parts, for example 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20 parts, etc., preferably 5-12 parts, more preferably 8-10 parts.

[0011] Preferably, the raw materials of the bread, sugar-controlled bread or meal replacement bread also include yeast, salt, butter, egg liquid, xylitol and water.

[0012] In a specific embodiment of the present invention, the raw materials of the bread, sugar-controlled bread or meal replacement bread include, by weight:

[0013] Wheat flour: 80-95 parts, for example, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95 parts, etc., preferably 88-95 parts, more preferably 90-92 parts;

[0014] Perilla protein powder: 5-20 parts, for example 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20 parts, etc., preferably 5-12 parts, more preferably 8-10 parts;

[0015] Yeast: 1-2 parts, for example 1, 1.2, 1.4, 1.5, 1.8, 1.9 or 2 parts, etc., preferably 1.5-2 parts;

[0016] Salt: 0.5-1.5 parts, such as 0.5, 0.8, 0.9, 1.0, 1.2, 1.3 or 1.5, etc., preferably 0.8-1.2 parts;

[0017] Water: 30-50 parts, for example 30, 35, 40, 45 or 50 parts, preferably 35-45 parts;

[0018] Butter: 5-20 parts, such as 5, 10, 15 or 20 parts, etc., preferably 8-12 parts;

[0019] Xylitol: 5-10 parts, such as 5, 6, 7, 8, 9 or 10, etc., preferably 6-8 parts;

[0020] 10-30 parts of egg liquid, for example 10, 15, 20, 25 or 30, etc., preferably 15-25 parts.

[0021] The preparation method of the bread, sugar-controlled bread or meal replacement bread comprises:

[0022] Mix wheat flour and perilla protein powder to obtain mixed flour, then mix the mixed flour, yeast, salt, xylitol, egg liquid and water, then add butter and stir to form the dough. Seal the dough and ferment it, shape it and let it rise again to expand its volume to 1.5-3 times of its original volume. Sprinkle dry flour on the surface and bake it in the oven to obtain bread, sugar-controlled bread or meal replacement bread.

[0023] Preferably, the fermentation conditions are: temperature 25-35° C., relative humidity 80-90%, and fermentation time 0.5-2 h.

[0024] Preferably, the baking temperature is 170-190° C., and the baking time is 10-30 min.

[0025] Preferably, stir for 20-40 minutes after adding the butter.

[0026] The mass content of perilla protein in the perilla protein powder is 40%-65%, such as 40%, 45%, 50%, 55%, 60% or 65%, preferably 45%-55%.

[0027] The perilla protein is obtained by deoiling and defatting perilla seeds.

[0028] The preparation method of the perilla protein powder comprises:

[0029] The perilla seeds are deoiled to obtain perilla cake, the perilla cake is defatted with an organic solvent, and then the organic solvent is removed, and then dried and crushed to obtain perilla protein powder.

[0030] Preferably, the organic solvent is ethanol.

[0031] Preferably, the defatting temperature is 60-80° C., the defatting time is 2-4 hours, and the mass volume ratio of the perilla cake to the organic solvent is 1:3.

[0032] The GI value of the bread, sugar-controlled bread or meal replacement bread is lower than 85, for example, lower than 85, 84, 83, 82, 81, 80, 75, 70, 65, etc., preferably lower than 70.

[0033] The beneficial effects of the present invention are as follows: the bread containing perilla protein has high moisture content, good water retention, and improved texture characteristics, recovery and sensory scores. It also has the functions of lowering GI, reducing reducing sugar release, lowering RDS and SDS contents, and inhibiting the activity of α-glucosidase and α-amylase, which is beneficial to the control of postprandial blood sugar levels. BRIEF DESCRIPTION OF THE DRAWINGS

[0034] The embodiments of the present invention are described in detail below with reference to the accompanying drawings, wherein:

[0035] Figure 1 : Effects of different amounts of perilla protein powder on bread color, where C was the control group.

[0036] Figure 2 :The inhibitory activity of perilla protein powder bread on α-glucosidase. Different lowercase letters in the figure indicate significant differences (P<0.05).

[0037] Figure 3 :The inhibitory activity of perilla protein powder bread on α-amylase. Different lowercase letters in the figure indicate significant differences (P<0.05).

[0038] Figure 4 : Reducing sugar content of bread with added perilla protein powder during the in vitro digestion process. Different lowercase letters in the figure indicate that the effect of the amount of perilla protein powder added on the reducing sugar content at the same digestion time was significantly different (P<0.05); different uppercase letters indicate that the effect of digestion time on the reducing sugar content at the same amount of perilla protein powder added was significantly different (P<0.05).

[0039] Figure 5 : Sensory scoring results of bread with added perilla protein powder. Different lowercase letters in the figure indicate significant differences (P<0.05).

[0040] Figure 6 :Effects of different addition amounts of hemp (H group) protein powder and perilla (P group) protein powder on the moisture content of bread. Different lowercase letters in the figure indicate significant differences (P<0.05).

[0041] Figure 7 : Comparison results of response, different lowercase letters in the figure indicate significant differences (P<0.05). DETAILED DESCRIPTION

[0042] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, not all. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative work are within the scope of protection of the present invention.

[0043] Materials, instruments and data processing used in the examples:

[0044] Hemp protein powder is obtained by deoiling and defatting hemp seeds, specifically by the same method as in Example 1, with a protein content of 50%; wheat flour, Hebei Jinshahe Noodle Group Co., Ltd.; yeast powder, Angel Yeast Co., Ltd.; butter, Inner Mongolia Mengniu Cheese Co., Ltd.; xylitol, Shandong Futian Pharmaceutical Co., Ltd.; refined edible salt, Jiangsu Ruifeng Salt Co., Ltd.; fresh eggs, Beijing Xiangfeng Trading Co., Ltd.

[0045] Glucose AR, Tianjin Hengxing Chemical Reagent Co., Ltd.; pepsin USP, pancreatin BR, Shanghai McLean Biochemical Technology Co., Ltd.; 3,5-Dinitrosalicylic acid (DNS) AR, Tianjin Damao Chemical Reagent Factory; p-Nitrophenyl-α-D-glucopyranoside (pNPG) AR, sodium carbonate AR, Shanghai McLean Biochemical Technology Co., Ltd.; α-glucosidase AR, α-amylase AR, soluble starch AR, Shanghai Aladdin Biochemical Technology Co., Ltd.

[0046] XMTD-205C digital display constant temperature magnetic stirring water bath, Hunan Frontier Technology Co., Ltd.; ZX-9146MBE electric heating blast drying oven, Shanghai Boxun Industrial Co., Ltd.; ZYCGF-20 ultrapure water preparation system, Sichuan Excellence Water Treatment Equipment Co., Ltd.; DHS-10A moisture meter, Hangzhou Jingfei Instrument Technology Co., Ltd.; SC-10 colorimeter, Shenzhen Sanenshi Technology Co., Ltd.; TA.XTC texture analyzer, Shanghai Baosheng Industrial Development Co., Ltd.; FX10 fermentation box, Guangdong Henglian Food Machinery Co., Ltd.; MG38CB-AA oven, Guangdong Midea Microwave Electric Manufacturing Co., Ltd.; SJJ-B10W5 dough mixer, Xiaoxiong Electric Co., Ltd.; P4 UV-visible spectrophotometer, Shanghai Meipuda Instrument Co., Ltd.

[0047] All quantitative data in this application were measured three times, and the measured data results were expressed as mean ± standard deviation (X ± SD). IBM SPSS Statistics 26 and Microsoft Excel 2019 were used to organize and statistically analyze the data. P < 0.05 was considered significant and expressed by the Alphabet Mark Method. Origin 2021 was used for drawing.

[0048] Example 1: Preparation of Perilla Protein Powder

[0049] Perilla seeds are cold pressed to remove oil to obtain perilla cake, which is defatted for 3 hours at an ethanol concentration of 95%, a solid-liquid ratio of 1:3, and a temperature of 70°C, and repeated twice. The ethanol is removed, the mixture is air-dried, and ultrafinely ground to obtain perilla protein, the defatting rate of which can reach 98%, and the protein content of the perilla protein is 50%.

[0050] Example 2: Preparation of Perilla Protein Sugar-Controlled Bread

[0051] According to the formula in Table 1, accurately weigh various raw materials, first mix wheat flour and perilla protein powder, then pour the mixed flour, yeast, salt, xylitol, egg liquid and water into the dough mixer and mix evenly, finally add butter and stir quickly for 30 minutes. Ferment the dough in a fermentation box at a temperature of 30°C and a relative humidity of 85% for 1 hour. Divide the fermented dough into 50±1g / piece, manually shape and plate it, and put it back into the fermentation box to ferment for 1 hour, so that its volume expands to about twice its original size. Sprinkle dry flour on the surface of the fermented dough and put it into the oven. The baking temperature is 180°C and the baking time is 15 minutes to obtain bread.

[0052] Table 1: Bread ingredients

[0053] Ingredients(g) Control group P5 Group P10 Group P15 Group P20 Group wheat flour 100 95 90 85 80 Perilla protein powder 0 5 10 15 20 Yeast powder 1.7 1.7 1.7 1.7 1.7 Refined edible salt 1 1 1 1 1 water 40 40 40 40 40 butter 10 10 10 10 10 Xylitol 7 7 7 7 7 Egg liquid 20 20 20 20 20

[0054] Example 3: Determination of bread crumb color

[0055] 1. Determination of bread crumb color

[0056] Determination of L content of bread crumbs by colorimeter * (brightness), a * (redness) and b * (yellowness) value, analyze the color change of bread, and calculate the whiteness index (WI).

[0057]

[0058] Note: WI——whiteness index;

[0059] L * ——brightness value;

[0060] a * ——Redness value;

[0061] b * ——Yellowness value.

[0062] 2. Results

[0063] Color is one of the important indicators of bread quality. As the amount of perilla protein powder added increases, the color of the bread crumb gradually darkens (see Figure 1), Table 2 shows the color index results of the bread core. The bread without perilla protein powder has the highest brightness. As the amount of perilla protein powder added increases, the brightness L * The value gradually decreased. Compared with the control group, the bread with perilla protein powder had a * The redness index and whiteness index WI value were significantly reduced (P < 0.05), indicating that the addition of perilla protein powder will affect the color of the bread crumb. The more the amount added, the darker the color of the bread crumb. The WI value is positively correlated with L*. In terms of color tone, the addition of perilla protein powder leads to a * Value and yellowness b * The values ​​gradually increased. Compared with the control group, the P5 group * The value did not change significantly (P>0.05). * The value changed significantly (P<0.05). In principle, the color of bread gradually deepened after baking, which may be due to the Maillard reaction between amino acids and reducing sugars during the baking stage of bread, resulting in the addition of perilla protein powder to affect the color of the bread core.

[0064] Table 2: Effect of perilla protein powder on bread color

[0065] Group <![CDATA[L * ]]> <![CDATA[a * ]]> <![CDATA[b * ]]> WI Control group <![CDATA[66.98±0.59 a ]]> <![CDATA[1.46±0.17 e ]]> <![CDATA[17.53±0.15 c ]]> <![CDATA[62.59±0.60 a <!-- 4 -->]]> P5 Group <![CDATA[59.20±0.57 b ]]> <![CDATA[2.70±0.11 d ]]> <![CDATA[17.64±0.26 c ]]> <![CDATA[55.47±0.53 b ]]> P10 Group <![CDATA[54.62±1.06 c ]]> <![CDATA[3.35±0.18 c ]]> <![CDATA[18.89±0.52 b ]]> <![CDATA[50.73±1.17 c ]]> P15 Group <![CDATA[48.35±0.22 d ]]> <![CDATA[4.97±0.06 b ]]> <![CDATA[19.15±0.55 b ]]> <![CDATA[44.69±0.23 d ]]> P20 Group <![CDATA[47.50±0.19 d ]]> <![CDATA[5.98±0.02 a ]]> <![CDATA[19.95±0.03 a ]]> <![CDATA[43.52±0.17 d ]]>

[0066] Among them, different lowercase letters in the same column indicate significant differences (P<0.05).

[0067] Example 4: Inhibitory activity against α-glucosidase

[0068] 1. Experimental methods

[0069] A certain amount of bread crumbs (the perilla protein powder bread prepared in Example 2 and the wheat flour control bread) were weighed, and PBS buffer with pH = 7 was ultrasonicated at 37°C for 1 h at a ratio of 1:10, then filtered and centrifuged, and the supernatant was taken to prepare sample solutions with concentrations of 0, 12.5, 25, 50, and 100 mg / mL, respectively.

[0070] 200 μL of wheat flour control bread and perilla protein powder bread with different mass concentrations (0, 12.5, 25, 50, 100 mg / mL) were mixed with 200 μL 1U / mL α-glucosidase solution and 100 μL pNPG, respectively, reacted in a 37°C water bath for 30 minutes, and then 200 μL 0.1M Na2CO3 was added to measure the absorbance at 400 nm. The inhibition rate of bread on α-glucosidase (I1) was calculated by the formula.

[0071] The calculation formula of enzyme activity inhibition rate is:

[0072]

[0073] Where: A1 is the absorbance of the sample;

[0074] A2 is the absorbance of the solution without α-glucosidase;

[0075] A3 is the absorbance of the solution without sample.

[0076] 2. Results

[0077] The mass concentration (mg / mL) is the horizontal axis and the inhibition rate (%) is the vertical axis. Figure 2 As shown. Compared with the control bread, the perilla protein powder bread has an inhibitory effect on α-glucosidase, and the product solution concentration shows a more obvious dose-dependent relationship in the range of (12.5-100) mg / mL. As the dose increases, the inhibitory effect is better. Within the experimental concentration range, the inhibitory effect of perilla protein powder bread on α-glucosidase is significantly better than that of the control group. When the mass concentration is 100 mg / mL, the inhibition rate of perilla protein powder bread on α-glucosidase is 70.92% higher than that of the control bread, indicating that it has a certain in vitro hypoglycemic effect.

[0078] Example 5: Inhibitory activity against α-amylase

[0079] 1. Experimental methods

[0080] A certain amount of bread crumbs (the perilla protein powder bread prepared in Example 2 and the wheat flour control bread) were weighed, and PBS buffer with pH = 7 was ultrasonicated at 37°C for 1 h at a ratio of 1:10, then filtered and centrifuged, and the supernatant was taken to prepare sample solutions with concentrations of 0, 12.5, 25, 50, and 100 mg / mL, respectively.

[0081] Take 100 μL of the prepared sample solution of each concentration, add 200 μL of 1% soluble starch solution and 100 μL of α-amylase solution, mix well and place in a 37°C water bath for 10 minutes. After the water bath, add 1 mL of DNS color developer to terminate the reaction, boil in a boiling water bath for 10 minutes, dilute with ice distilled water after the water bath, and measure the absorbance at 520 nm. The inhibition rate (I2) of perilla protein powder bread on α-amylase was calculated by the formula.

[0082] The calculation formula of enzyme activity inhibition rate is as follows:

[0083]

[0084] Where: A1 is the absorbance of the sample;

[0085] A2 is the absorbance of the solution without α-glucosidase;

[0086] A3 is the absorbance of the solution without sample;

[0087] A4 is a blank control.

[0088] 2. Results

[0089] like Figure 3 As shown in the figure, compared with the control bread, the inhibition rate of α-amylase by perilla protein powder bread increased significantly with the increase of mass concentration. At 100 mg / mL, the inhibition rate of P10 on α-amylase was 31.64%, which was 21.74% higher than that of the control bread. This may be because as the protein concentration increases, the protein and starch in the extract form a complex system, which reduces the contact between starch and digestive enzymes and effectively inhibits the activity of α-amylase, thereby increasing its inhibition rate and playing a role in inhibiting the increase of blood sugar after a meal.

[0090] Example 6: Reducing sugar content

[0091] 1. Experimental methods

[0092] 1) Simulated gastric digestion conditions: Take 1g of bread, add 30mL of distilled water, adjust the pH value of the solution to 2.5, then add 1mL of 10% pepsin solution (mass fraction, prepared with a mass concentration of 0.05mol / L HCl), and put it into a water bath for oscillation reaction (130r / min, 37℃, 30min). Simulated intestinal digestion: Adjust the pH value with 2mL of 1mol / L NaHCO3 solution and 5mL of 0.1mol / L phosphate buffer (PBS buffer). Add 100μL of amyloglucosidase solution and 5mL of 2.5% pancreatic enzyme solution (mass fraction, prepared with a mass concentration of 0.1mol / L PBS buffer), add distilled water to 55mL, and slowly stir and digest at 37℃ for 120min. At 0 (no amyloglucosidase solution and pancreatin solution were added), 20, 60, 90 and 120 min, 0.5 mL of the sample was taken, 2 mL of anhydrous ethanol was added to terminate the reaction, and the reducing sugar content was determined by DNS colorimetry.

[0093] The reducing sugar concentration is expressed as milligrams of reducing sugar per gram of food, and the calculation formula is as follows:

[0094]

[0095] Note: W——reducing sugar content (mg / g, mass fraction);

[0096] c——reducing sugar concentration of sample solution (mg / mL, mass concentration);

[0097] v——sample volume (mL);

[0098] 2) Preparation of standard curve and determination of reducing sugar

[0099] Accurately weigh 0.1g of glucose to prepare a glucose standard solution with a mass concentration of 1.0mg / mL. Take 0, 0.2, 0.4, 0.6, 0.8, and 1.0mL of the glucose standard solution in 10mL stoppered test tubes, add distilled water to 1mL, and then accurately add 0.75mL of DNS reagent to mix evenly, heat in a boiling water bath for 5min to develop a color reaction, cool in running water, add 3.0mL of distilled water to each, shake well, and measure the absorbance value A at a wavelength of 540nm 540 . Draw a standard curve with glucose content (mg) as the horizontal axis and absorbance as the vertical axis.

[0100] Determination of reducing sugar in samples: Take 1 mL of the supernatant obtained in step 1), add 0.75 mL of DNS reagent, heat in a boiling water bath for 5 min, cool in running water, add 3.0 mL of distilled water, shake well, and measure the absorbance at a wavelength of 540 nm. Calculate the corresponding reducing sugar concentration based on the standard curve.

[0101] 2. Results

[0102] The reducing sugar content results are shown in Figure 4 As shown in the figure, within the first 20 minutes, the reducing sugar content increased rapidly, the reducing sugar content of the control group ranged from 353.85 mg / g to 749.96 mg / g, and the reducing sugar content of the bread with perilla protein powder added ranged from 207.35 mg / g to 586.49 mg / g. The reducing sugar release rate slowed down within 20 to 120 minutes. During the entire digestion process, the reducing sugar release of the perilla protein group bread was significantly lower than that of the control group bread at the same time point. With the increase of the amount of perilla protein powder added, the reducing sugar release showed a downward trend, indicating that the addition of perilla protein powder helps control blood sugar levels. This may be due to the addition of perilla protein powder, which forms a protein-starch complex system with starch, inhibiting the contact between α-amylase and starch, thereby reducing starch digestion.

[0103] Example 7: Sensory evaluation of bread

[0104] 1. Sensory evaluation method

[0105] Invite 30 sensory assessors to conduct sensory assessment of bread. The assessors are required to rinse their mouths with water before and after the assessment. The specific assessment criteria are shown in Table 3, with a total of 100 points.

[0106] Table 3: Evaluation criteria

[0107]

[0108] 2. Results

[0109] As the amount of perilla protein powder added increased, the sensory score of the bread showed a trend of first increasing and then decreasing (see Figure 5). The sensory scores of the P5 and P10 groups were high, above 80, and the P10 group had the highest sensory score. This was due to the special aroma of the perilla protein powder in the bread, and the delicate and elastic structure of the bread. However, with the further addition of perilla protein powder, the sensory score showed a downward trend. This is because as the amount of perilla protein powder added increases, the fermentation process of yeast and the formation of gluten may be hindered, causing the bread's structure to become compact, the taste to become hard, and difficult to chew and swallow.

[0110] Example 8: Preparation of comparative bread

[0111] According to the formula in Table 4, accurately weigh various raw materials, first mix wheat flour and hemp protein powder, then pour the mixed flour, yeast, salt, xylitol, egg liquid and water into the dough mixer and mix evenly, finally add butter and stir quickly for 30 minutes. Ferment the dough in a fermentation box at a temperature of 30°C and a relative humidity of 85% for 1 hour. Divide the fermented dough into 50±1g / piece, manually shape and plate it, and put it back into the fermentation box to rise for 1 hour, so that its volume expands to about twice its original size. Sprinkle dry flour on the surface of the proofed dough and put it into the oven. The baking temperature is 180°C and the baking time is 15 minutes to obtain bread.

[0112] Table 4: Bread ingredients

[0113] Ingredients(g) H5 Group H10 Group H15 Group H20 Group wheat flour 95 90 85 80 Hemp protein powder 5 10 15 20 Yeast powder 1.7 1.7 1.7 1.7 Refined edible salt 1 1 1 1 water 40 40 40 40 butter 10 10 10 10 Xylitol 7 7 7 7 Egg liquid 20 20 20 20

[0114] Example 9: Determination of moisture content and water retention of bread

[0115] 1. Determination of moisture content and water retention

[0116] 1) Determination of moisture content

[0117] GB 5009.3-2016 “National Food Safety Standard Determination of Water in Food” was adopted and the direct drying method was used for determination.

[0118] 2) Determination of water retention

[0119] The water loss rate was used as an indicator of water retention. The bread was weighed immediately after it was baked and then measured every 24 hours. Samples were taken for a total of 5 days to evaluate the water retention performance of bread stored at room temperature (23±2)℃.

[0120]

[0121] Note: Y——water loss rate (%, mass fraction); M——original mass of bread (g);

[0122] m——the mass of bread after storage (g).

[0123] 2. Results

[0124] The moisture content directly affects the quality, aging and chewing difficulty of bread. If the moisture content is too low, the bread will be of low quality, easy to age, and difficult to chew and swallow. Figure 6 It can be seen that with the increase in the amount of perilla protein powder or hemp protein powder added, the moisture content of both types of bread increased. The groups of bread with added perilla protein powder had higher moisture content than the groups of bread with added hemp protein powder. Specifically, when the addition amount of perilla protein powder was 5% to 20%, the moisture content of the bread was significantly increased compared with the control (P<0.05), and the moisture content increased in the range of 28.71% to 36.04%. When the addition amount of hemp protein powder was 5%, the moisture content of the bread did not change significantly. This shows that perilla protein has good water retention and increases the water absorption rate of the dough. This may be due to the higher dietary fiber content in the bread fortified with perilla protein powder.

[0125] The water loss rate of the bread with added perilla protein powder was further measured, and the results are shown in Table 5. Within 5 days of storage, the amount of perilla protein powder added was significantly correlated with the water retention of bread (P<0.05), and with the increase in the amount of perilla protein powder added, the water loss rate gradually decreased, that is, the water retention was enhanced. In addition, compared with the bread in the control group, the bread with added perilla protein powder always had a lower water loss rate. Specifically, the effect on the bread was greater when placed for 1 day, and the water loss rate of bread with different amounts of perilla protein powder added decreased by 27.40% to 33.36%; it tended to be flat from 2 to 5 days.

[0126] Table 5: Effect of different amounts of perilla protein powder added on the water loss rate of bread (%)

[0127] Group 1d 2d 3d 4d 5d Control group <![CDATA[14.60±0.07 aD ]]> <![CDATA[15.20±0.05 aC ]]> <![CDATA[15.87±0.08 aB ]]> <![CDATA[15.93±0.11 aB ]]> <![CDATA[16.37±0.30 aA ]]> P5 <![CDATA[10.60±0.07 bC ]]> <![CDATA[13.08±0.55 bAB ]]> <![CDATA[13.87±0.28 bAB ]]> <![CDATA[14.01±0.40 bAB ]]> <![CDATA[14.87±0.19 bA ]]> P10 <![CDATA[10.40±0.29 cD ]]> <![CDATA[12.20±0.32 cBC ]]> <![CDATA[12.80±0.32 cBC ]]> <![CDATA[13.53±0.40 cAB ]]> <![CDATA[14.13±0.23 cA ]]> P15 <![CDATA[10.33±0.16 cD ]]> <![CDATA[12.18±0.44 cC ]]> <![CDATA[12.73±0.32 cBC ]]> <![CDATA[13.33±0.28 dAB ]]> <![CDATA[13.87±0.11 dA ]]> P20 <![CDATA[9.73±0.22 dD ]]> <![CDATA[11.73±0.11 dC ]]> <![CDATA[12.27±0.19 dBC ]]> <![CDATA[13.06±0.12 eAB ]]> <![CDATA[13.76±0.10 eA ]]>

[0128] Among them, different lowercase letters on the right shoulders in the same column indicate significant differences (P<0.05), and different uppercase letters on the right shoulders in the same row indicate significant differences (P<0.05).

[0129] Example 10: RDS and SDS content

[0130] 1. Experimental methods

[0131] 1) Simulated gastric digestion conditions: Take 1g of bread, add 30mL of distilled water, adjust the pH value of the solution to 2.5, then add 1mL of 10% pepsin solution (mass fraction, prepared with a mass concentration of 0.05mol / L HCl), and put it into a water bath for oscillation reaction (130r / min, 37℃, 30min). Simulated intestinal digestion: Adjust the pH value with 2mL of 1mol / L NaHCO3 solution and 5mL of 0.1mol / L phosphate buffer (PBS buffer). Add 100μL of amyloglucosidase solution and 5mL of 2.5% pancreatic enzyme solution (mass fraction, prepared with a mass concentration of 0.1mol / L PBS buffer), add distilled water to 55mL, and slowly stir and digest at 37℃ for 120min. At 0 (no amyloglucosidase solution and pancreatin solution were added), 20, 60, 90 and 120 min, 0.5 mL of the sample was taken, 2 mL of anhydrous ethanol was added to terminate the reaction, and the reducing sugar content was determined by DNS colorimetry.

[0132] The reducing sugar concentration is expressed as milligrams of reducing sugar per gram of food, and the calculation formula is as follows:

[0133]

[0134] Note: W——reducing sugar content (mg / g, mass fraction);

[0135] c——reducing sugar concentration of sample solution (mg / mL, mass concentration);

[0136] v——sample volume (mL);

[0137] TS——total mass of sample (g);

[0138] 0.9 – conversion factor;

[0139] RDS – rapidly digestible starch (%, mass fraction);

[0140] SDS——slowly digestible starch (%, mass fraction);

[0141] G0——mass of free reducing sugar before enzymatic hydrolysis (mg);

[0142] G 20 ——Mass of reducing sugar in the supernatant after 20 min digestion (mg);

[0143] G 120 ——Mass of reducing sugar in the supernatant after 120 min of digestion (mg).

[0144] Among them, starch that is quickly digested and absorbed within the first 20 minutes in the small intestine is called rapidly digestible starch (RDS), and starch that is slowly digested within 20 to 120 minutes in the small intestine is called slowly digestible starch (SDS).

[0145] 2) Preparation of standard curve and determination of reducing sugar

[0146] Accurately weigh 0.1g of glucose to prepare a glucose standard solution with a mass concentration of 1.0mg / mL. Take 0, 0.2, 0.4, 0.6, 0.8, and 1.0mL of the glucose standard solution in 10mL stoppered test tubes, add distilled water to 1mL, and then accurately add 0.75mL of DNS reagent to mix evenly, heat in a boiling water bath for 5min to develop a color reaction, cool in running water, add 3.0mL of distilled water to each, shake well, and measure the absorbance value A at a wavelength of 540nm 540 . Draw a standard curve with glucose content (mg) as the horizontal axis and absorbance as the vertical axis.

[0147] Determination of reducing sugar in samples: Take 1 mL of the supernatant obtained in step 1), add 0.75 mL of DNS reagent, heat in a boiling water bath for 5 min, cool in running water, add 3.0 mL of distilled water, shake well, and measure the absorbance at a wavelength of 540 nm. Calculate the corresponding reducing sugar concentration based on the standard curve.

[0148] 2. Results

[0149] The trends of RDS and SDS contents in breads of hemp protein powder group and perilla protein powder group are shown in Table 6. With the increase of the addition amount, the RDS and SDS contents were significantly lower than those in the control bread (P<0.05). The RDS and SDS of hemp protein powder bread in group H10 and perilla protein powder bread in group P10 were reduced by 36.34%, 17.36% and 48.09%, 31.46% respectively compared with the control, indicating that the addition of perilla protein powder is more conducive to inhibiting starch digestion in bread than hemp protein powder. Under the same addition amount, the RDS content of bread in the perilla protein powder group was lower than that of bread in the hemp protein powder group, indicating that the addition of perilla protein in bread is more conducive to controlling the extent of postprandial blood sugar increase. The reduction in RDS and SDS contents may be due to the fact that the hydroxyl groups in the molecular structure of the added protein can bind to starch through non-covalent interactions (hydrogen bonds, electrostatics, hydrophobic interactions, van der Waals forces, etc.), thereby affecting the digestion of bread.

[0150] Table 6: RDS and SDS content of bread

[0151]

[0152] Among them, different lowercase letters on the right shoulder of the same column in the hemp protein powder group indicate significant differences (P<0.05), and different lowercase letters on the right shoulder of the same column in the perilla protein powder group indicate significant differences (P<0.05).

[0153] Example 11: GI analysis

[0154] 1. The calculation formulas for hydrolysis index (HI) and glycemic index (GI) are as follows:

[0155]

[0156] Note: HI——Starch hydrolysis index (%, mass fraction);

[0157] AUC1——Area under the starch hydrolysis curve of the sample, cm 2 ;

[0158] AUC0——Area under the starch hydrolysis curve of white bread, cm 2

[0159] GI = 39.71 + 0.549 × HI 120

[0160] Note: GI - glycemic index (%, mass fraction);

[0161] HI 120 ——HI value of the sample after 120 minutes of hydrolysis.

[0162] 2. Results

[0163] GI usually reflects the ability of a food to cause the blood sugar level of the human body to rise. Generally speaking, when GI <55, the food can be considered as a low GI food, when GI is between 55 and 70, the food is a medium GI food, and when GI>70, the food is a high GI food. The GI values ​​of bread in the hemp protein powder group and the perilla protein powder group are shown in Table 7. After adding different amounts of hemp protein powder and perilla protein powder, the GI values ​​of the bread were significantly lower than those of the control group. The GI value decreased with the increase in the amount of protein added. Among them, the GI values ​​of the H10 and P10 groups decreased by 19.05% and 26.31% respectively compared with the control group. Bread in the perilla protein powder group is a medium glycemic index food, which can effectively control blood sugar levels and can be selected by diabetics.

[0164] Table 7: GI analysis of bread

[0165]

[0166] Among them, different lowercase letters on the right shoulder of the same column in the hemp protein powder group indicate significant differences (P<0.05), and different lowercase letters on the right shoulder of the same column in the perilla protein powder group indicate significant differences (P<0.05).

[0167] Example 12: Determination of bread texture

[0168] 1. Determination of bread texture characteristics

[0169] The texture characteristics of bread were determined using a texture analyzer. The method was as follows: the bread crumbs were cut into small cubes of 36 mm × 36 mm × 25 mm, and then the hardness, elasticity, resilience and cohesion of the bread were measured using a P36R probe. The parameters were set as follows: pre-test speed 1.7 mm / s, test speed 1.7 mm / s, post-test speed 1.7 mm / s, displacement 6.25 mm, trigger force 5 g, and compression time 5 s.

[0170] 2. Results

[0171] The results of bread texture test showed that the addition of hemp protein powder and perilla protein powder had no significant effect on the elasticity and cohesion of the bread, but had a significant effect on the hardness and resilience. The effects of adding hemp protein powder and perilla protein powder on hardness are shown in Table 8.

[0172] Table 8: Hardness comparison results

[0173]

[0174] Among them, different lowercase letters on the right shoulder of the same column in the hemp protein powder group indicate significant differences (P<0.05), and different lowercase letters on the right shoulder of the same column in the perilla protein powder group indicate significant differences (P<0.05).

[0175] Adding protein can promote the growth of yeast, thus increasing the gas production rate in the bread and making the texture of the bread containing protein hydrolysate softer. However, the results in Table 8 show that although the bread hardness of the P5 group, P10 group, H5 group, and H10 group were significantly lower than that of the control group (P<0.05), the bread hardness increased significantly with further increase in the amount of addition (P<0.05). Among them, under the premise of the same amount of addition, the bread prepared with perilla protein powder was significantly better than the bread prepared with hemp protein powder. Compared with the bread in the control group, the bread hardness of the P5 group, P10 group, H5 group, and H10 group was significantly lower than that of the control group (P<0.05). The hardness of the bread prepared with hemp protein powder was reduced by 35.87% and 7.69%, respectively, and the hardness of the bread prepared with perilla protein was reduced by 57.39% and 43.05%, respectively. Therefore, the P5 group, P10 group, H5 group, and H10 group were selected to further verify the resilience. Results are shown in Figure 7 . Figure 7The results showed that the recovery of the H5 group with added hemp protein powder showed a downward trend, while the recovery of the P5 and P10 groups with added perilla protein powder increased.

[0176] The preferred embodiments of the present invention are described in detail above. However, the present invention is not limited to the specific details in the above embodiments. Within the technical concept of the present invention, a variety of simple modifications can be made to the technical solution of the present invention, and these simple modifications all belong to the protection scope of the present invention.

[0177] It should also be noted that the various specific technical features described in the above specific embodiments can be combined in any suitable manner without contradiction. In order to avoid unnecessary repetition, the present invention will not further describe various possible combinations.

Claims

1. Application of perilla protein in the preparation of sugar-controlled bread, wherein the perilla protein is obtained by deoiling and defatting perilla seeds, and the raw materials of the sugar-controlled bread include wheat flour and perilla protein powder, wherein: Calculated by weight, the invention comprises 80-95 parts of wheat flour and 5-20 parts of perilla protein powder.

2. The use according to claim 1, characterized in that: The raw materials of the controlled sugar bread also include yeast, salt, butter, egg liquid, xylitol and water.

3. The use according to claim 1, characterized in that: The raw materials of the sugar-controlled bread include, by weight: Wheat flour: 80-95 parts; Perilla protein powder: 5-20 portions; Yeast: 1-2 parts; Salt: 0.5-1.5 parts; Water: 30-50 parts; Butter: 5-20 portions; Xylitol: 5-10 parts; 10-30 portions of egg liquid.

4. The use according to claim 1, characterized in that: The preparation method of the sugar-controlled bread comprises: Mix wheat flour and perilla protein powder to obtain mixed flour, then mix the mixed flour, yeast, salt, xylitol, egg liquid and water, then add butter and stir to obtain the dough. Seal the dough for fermentation, shape it and let it rise again to expand its volume to 1.5-3 times of its original volume, sprinkle dry flour on the surface, and bake in the oven to obtain the controlled sugar bread.

5. The use according to claim 4, characterized in that: The fermentation conditions are: temperature 25-35° C., relative humidity 80-90%, and fermentation time 0.5-2 h.

6. The use according to claim 4, characterized in that: The baking temperature is 170-190° C., and the baking time is 10-30 min.

7. The use according to claim 1, characterized in that: The mass content of perilla protein in the perilla protein powder is 40%-65%.

8. The use according to claim 1 or 7, characterized in that: The preparation method of the perilla protein powder comprises: The perilla seeds are deoiled to obtain perilla cake, the perilla cake is defatted with an organic solvent, and then the organic solvent is removed and then dried and crushed to obtain perilla protein powder.

9. The use according to claim 8, characterized in that: The degreasing temperature is 60-80° C., the degreasing time is 2-4 hours, and the mass volume ratio of the perilla cake to the organic solvent is 1:

3.

10. The use according to claim 1, characterized in that: The GI value of the sugar-controlled bread is lower than 85.

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