CN115708561B - Functional corn flour and efficient preparation method and application thereof - Google Patents
Functional corn flour and efficient preparation method and application thereof Download PDFInfo
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- CN115708561B CN115708561B CN202211478646.1A CN202211478646A CN115708561B CN 115708561 B CN115708561 B CN 115708561B CN 202211478646 A CN202211478646 A CN 202211478646A CN 115708561 B CN115708561 B CN 115708561B
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- procyanidine
- quercetin
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Landscapes
- Cereal-Derived Products (AREA)
Abstract
A functional corn flour and its high-efficiency preparation method and application are provided. The invention belongs to the field of functional food processing. The invention aims to solve the technical problems of low efficiency and serious loss of active ingredients in the existing wet pulverizing process. The method comprises the following steps: firstly, adding water into corn kernels to carry out microwave cooking; then carrying out enzymolysis on cellulase; adding alkaline protease and quercetin, and soaking under ultrasonic assistance; then dissolving procyanidine in water, spraying the procyanidine on the surface of soaked corn, and finally vacuum drying to obtain the functional corn powder. The corn flour is prepared by reasonably combining and linking through a plurality of technologies, so that the corn flour milling efficiency is effectively improved, the prepared corn flour is rich in two polyphenol bioactive components of quercetin and procyanidine, has high resistant starch content and antioxidant activity, the optimized process also improves the chemical stability and biological accessibility of the active components, and meanwhile, the corn flour has good gelatinization and anti-aging characteristics.
Description
Technical Field
The invention belongs to the field of functional food processing, and particularly relates to functional corn flour, and a high-efficiency preparation method and application thereof.
Background
Along with diversification of dietary structures of people and more hypertension, obesity, cardiovascular diseases and the like caused by refined main food and unreasonable dietary structures such as a large amount of meat and the like, the demands on coarse food grain foods such as corn and the like are more and more extensive. Corn is the first large grain crop in the world, is widely planted worldwide, has a annual corn yield of up to 2.7 hundred million tons in 2021 and is the second large corn producing country worldwide. The corn has high nutritive value and medicinal value, contains about 70% of starch and 8% -14% of protein, is rich in vitamins, carotenoid, dietary fiber and other nutritional ingredients, and has the effects of beautifying, improving eyesight, preventing coronary heart disease, hypertension and the like. The protein matrix in corn is a large and complex protein molecule formed by disulfide bonding of different protein subunits in gluten, prolamin is present in spheroids in the gluten matrix, and starch particles are embedded in the gluten matrix. Because of the unique combination mode of starch and protein in corn flour, the gelatinization of starch is limited in the food cooking process, so that the processing and eating quality of corn food are affected, and the development and utilization of corn as food raw material are not facilitated.
In the development of corn food products, grinding and milling are the basis of deep processing of corn kernels, and are also important pretreatment links. At present, the corn flour milling modes mainly comprise a dry method, a semi-dry method and a wet method, and different processing modes have important influence on the processing effect. The corn flour prepared by the dry method has good water absorption property, but has poor processing quality, coarse mouthfeel and easy aging; the corn flour prepared by adopting the wet method has less damaged starch and better processing characteristics and eating quality; the quality of the corn flour prepared by the semi-dry method is between that of the corn flour prepared by the dry method and that of the corn flour prepared by the wet method. However, the corn wet milling requires a long processing time, generally about 36-48 hours of soaking, which severely reduces milling efficiency and increases production cost.
Currently, corn steep processes have a variety of processes including fermentation, enzymatic and ultrasonic. The fermentation method has complex soaking process conditions and processes, and is not easy to control in production, so the method is not widely used in the corn flour milling production industry. The enzymatic soaking process shortens the soaking time, reduces the environmental pollution and improves the production efficiency. The proteases used at home and abroad mainly comprise acid protease, neutral protease, alkaline protease, bromelain, papain and the like. In addition, to break the cell wall structure in corn kernels, wall breaking enzymes such as cellulases, pectinases, xylanases and the like are also used. The soaking mode includes mixed soaking of acid and enzyme, stepwise soaking of wall breaking enzyme and protease, synchronous soaking after compounding of several proteases, etc. Ultrasound-assisted technology has become a routine technology for producing green, economical alternative foods and natural products. The ultrasonic cavitation shortens the soaking time, so that the protein in the corn is separated from the starch, the starch can be fully gelatinized in the processing process of the corn flour, and the protein can form a good network structure, so that the processing characteristics of the corn flour are improved, but in the wet soaking process, part of bioactive components in the corn can be lost along with soaking water, and the functional characteristics of the corn flour are reduced.
Disclosure of Invention
The invention aims to solve the technical problems of low efficiency and serious loss of active ingredients in the existing wet milling process, and provides functional corn flour, and a high-efficiency preparation method and application thereof.
One of the purposes of the invention is to provide a high-efficiency preparation method of functional corn flour, which comprises the following steps:
Step 1: adding water into corn kernels to carry out microwave cooking, and then grinding at a low speed to obtain a mixed solution;
Step 2: adjusting the pH value of the mixed solution to 5, and then adding cellulase for enzymolysis to obtain an enzymolysis solution;
Step 3: adjusting the pH value of the enzymolysis liquid to 9, then adding alkaline protease and quercetin, soaking under the assistance of ultrasound, and then filtering and draining to obtain modified corn particles;
Step 4: dissolving procyanidine in water to obtain procyanidine solution, spraying the procyanidine solution on the surface of modified corn particles, mechanically mixing, adjusting pH to neutrality when the water content of the modified corn particles reaches 35-40%, grinding at high speed, and vacuum drying to obtain functional corn powder.
Further defined, in step 1, the feed liquid ratio is 1:3.
Further defined, the power of the microwave cooking in the step 1 is 400-600W, and the time is 6-10min.
Further defined, the low-speed grinding in step 1 is carried out at a rotational speed of 2500-3000rpm.
Further limited, the addition amount of the cellulase in the step 2 is 0.5-1.5% of the mass of the mixed solution.
Further limited, the enzymolysis temperature in the step 2 is 40-60 ℃ and the enzymolysis time is 50-70min.
Further limited, the addition amount of the alkaline protease in the step 3 is 1-3% of the mass of the enzymolysis liquid.
Further limited, the concentration of quercetin in the enzymolysis liquid in the step 3 is 2-4mg/mL.
Further defined, the ultrasonic power in step 3 is 100-300W.
Further limited, the soaking temperature in the step 3 is 50-70 ℃ and the soaking time is 3-5h.
And (3) further limiting, namely refluxing the filtrate obtained by filtering after soaking in the step (3) into the enzymolysis liquid for recycling.
Further defined, the procyanidine solution concentration in step 4 is 4-6mg/mL.
Further defined, the high speed grinding in step 4 is performed at a rotational speed of 16000-17000rpm.
Further defined, the vacuum degree of the vacuum drying in the step 4 is 0.04-0.08MPa, and the temperature is 60-70 ℃.
The second object of the present invention is to provide a functional corn flour prepared by the above method.
The invention also provides the application of the functional corn flour prepared by the method, which is used for preparing functional corn food.
Compared with the prior art, the invention has the remarkable effects that:
The corn flour is prepared by reasonably combining and linking a plurality of technologies, the prepared corn flour is rich in two polyphenol bioactive components of quercetin and procyanidine, the optimized preparation process also improves the chemical stability and biological accessibility of the active components, and meanwhile, the corn flour has good gelatinization and ageing resistance and has the following specific advantages:
1) According to the invention, corn flour is prepared by reasonably combining and linking a plurality of technologies, corn kernels are firstly subjected to microwave cooking to enable the corn kernels to absorb water rapidly, so that corn cortex is softened and is partially destroyed, starch is pregelatinized, protein is partially denatured, subsequent enzymolysis of cellulase is facilitated, fibers in the corn cortex are further degraded, and partial insoluble dietary fibers are converted into soluble dietary fibers, so that subsequent soaking treatment is facilitated.
2) According to the invention, the pH of the enzymolysis liquid is adjusted to be alkaline, alkaline protease and quercetin are sequentially added, protease wet soaking is carried out under the assistance of ultrasonic cavitation, the acting force between starch and protein is effectively broken by ultrasonic cavitation, the starch-protein composite structure is destroyed, the separation of starch and protein is greatly promoted, the soaking time is shortened, the soaking efficiency is improved, in addition, the quercetin is added under the alkaline condition to promote the quercetin to form non-covalent and covalent complexes with starch and protein in corn respectively through interaction, the functionality of corn flour is greatly improved, and the raw material source guarantee can be provided for the development of novel functional corn foods. The soaking liquid can be recycled and used for soaking corn in the next batch, so that not only is the consumption of water and enzyme saved and the consumption of protease and quercetin saved, but also the loss of nutrient substances in the corn is reduced, and meanwhile, the corn soaking efficiency and the corn soaking effect are ensured. Greatly reduces the cost and realizes green and environment-friendly sustainable production.
3) According to the invention, the moisture of the drained modified corn particles is regulated by the procyanidine aqueous solution, and then the modified corn particles are prepared into powder by a semi-dry method, so that the defects of poor quality of dry powder preparation, long time consumption of wet powder preparation and the like are avoided, the good processing quality of corn powder is ensured while the high-efficiency powder preparation is realized, and the formation of starch-polyphenol compound is promoted by vacuum drying, so that the resistant starch content is improved.
4) The method shortens the soaking time of the traditional wet milling process from 36 hours to 5-6 hours, obviously improves the milling efficiency of corn, has the characteristics of green safety, short production time, high added value of products and the like, the prepared corn flour is rich in two polyphenol bioactive components of quercetin and procyanidine, improves the resistant starch content and antioxidant activity of the corn flour, and the chemical stability and biological accessibility of the active components are improved by a compound formed by starch and protein in corn, the quercetin and the procyanidine.
Drawings
FIG. 1 is a bar graph showing the starch-polyphenol recombination rate in functional corn flour prepared in examples 1-3 and comparative examples 1-3;
FIG. 2 is an infrared spectrum of a starch-polyphenol complex in functional corn flour prepared in examples 1-3 and comparative examples 1-3;
FIG. 3 is a bar graph showing the free amino and free thiol content of the protein-polyphenol complexes of functional corn flour prepared in examples 1-3 and comparative examples 1-3;
FIG. 4 is a schematic diagram showing sodium dodecyl sulfate-polyacrylamide gel electrophoresis of protein-polyphenol complexes in functional corn flour prepared in examples 1-3 and comparative examples 1-3;
FIG. 5 is a bar graph of the resistant starch content and antioxidant activity of the functional corn meal prepared in examples 1-3 and comparative examples 1-3 after digestion;
FIG. 6 is a bar graph of the bioavailability of the active ingredient after digestion of the functional corn meal prepared in examples 1-3 and comparative examples 1-3;
FIG. 7 is a bar graph showing chemical stability of active ingredients in functional corn flour prepared in examples 1-3 and comparative examples 1-3 under 65℃heat treatment conditions;
FIG. 8 is a bar graph showing chemical stability of active ingredients in functional corn flour prepared in examples 1-3 and comparative examples 1-3 under heat treatment conditions of 85 ℃;
FIG. 9 is a bar graph showing chemical stability of active ingredients in functional corn flour prepared in examples 1-3 and comparative examples 1-3 under heat treatment conditions at 100deg.C.
Detailed Description
The present invention will be described in further detail with reference to the following examples in order to make the objects, technical solutions and advantages of the present invention more apparent. It should be understood that the specific embodiments described herein are for purposes of illustration only and are not intended to limit the scope of the invention.
The experimental methods used in the following examples are conventional methods unless otherwise specified. The materials, reagents, methods and apparatus used, without any particular description, are those conventional in the art and are commercially available to those skilled in the art.
The terms "comprising," "including," "having," "containing," or any other variation thereof, as used in the following embodiments, are intended to cover a non-exclusive inclusion. For example, a composition, step, method, article, or apparatus that comprises a list of elements is not necessarily limited to only those elements but may include other elements not expressly listed or inherent to such composition, step, method, article, or apparatus.
When an equivalent, concentration, or other value or parameter is expressed as a range, preferred range, or a range bounded by a list of upper preferable values and lower preferable values, this is to be understood as specifically disclosing all ranges formed from any pair of any upper range limit or preferred value and any lower range limit or preferred value, regardless of whether ranges are separately disclosed. For example, when ranges of "1 to 5" are disclosed, the described ranges should be construed to include ranges of "1 to 4", "1 to 3", "1 to 2 and 4 to 5", "1 to 3 and 5", and the like. When a numerical range is described herein, unless otherwise indicated, the range is intended to include its endpoints and all integers and fractions within the range. In the description and claims of the application, the range limitations may be combined and/or interchanged, if not otherwise specified, including all the sub-ranges subsumed therein.
The indefinite articles "a" and "an" preceding an element or component of the invention are not limited to the requirement (i.e. the number of occurrences) of the element or component. Thus, the use of "a" or "an" should be interpreted as including one or at least one, and the singular reference of an element or component includes the plural reference unless the amount clearly dictates otherwise.
Example 1: the efficient preparation method of the functional corn flour comprises the following steps:
step 1: adding water into corn kernels (feed-liquid ratio is 1:3), steaming at 500W for 8min by microwave, and grinding at 2800rpm for 20s to obtain mixed solution;
Step 2: adjusting the pH value of the mixed solution to 5, then adding cellulase accounting for 1% of the mass of the mixed solution, and carrying out enzymolysis for 60min at 50 ℃ to obtain an enzymolysis solution;
Step 3: adjusting the pH value of the enzymolysis liquid to 9, then adding alkaline protease and quercetin which account for 2% of the mass of the enzymolysis liquid, wherein the concentration of the quercetin in the enzymolysis liquid is 3mg/mL, soaking for 4 hours under the assistance of 200W ultrasonic at 60 ℃, then filtering, refluxing the obtained filtrate into the enzymolysis liquid for recycling, and draining the obtained precipitate to obtain modified corn particles;
Step 4: dissolving procyanidine in water to obtain procyanidine solution with concentration of 5mg/mL, spraying the procyanidine solution on the surface of modified corn particles, mechanically mixing, adjusting pH to neutrality when the water content of the modified corn particles reaches 38%, grinding at 16800rpm at high speed, vacuum drying wet corn flour under vacuum degree of 0.06MPa and temperature of 65 ℃, sieving with 100 mesh sieve to obtain functional corn flour, and measuring to obtain corn flour with quercetin content of 9mg/g and procyanidine content of 5mg/g.
Example 2: the efficient preparation method of the functional corn flour comprises the following steps:
Step 1: adding water into corn kernels (feed-liquid ratio is 1:3), carrying out microwave cooking for 9min at 450W, and grinding for 20s at 2800rpm to obtain mixed liquor;
Step 2: adjusting the pH value of the mixed solution to 5, then adding cellulase accounting for 0.5% of the mass of the mixed solution, and carrying out enzymolysis for 65min at 55 ℃ to obtain an enzymolysis solution;
Step 3: adjusting the pH value of the enzymolysis liquid to 9, then adding alkaline protease and quercetin which account for 1.5% of the mass of the enzymolysis liquid, wherein the concentration of the quercetin in the enzymolysis liquid is 2.5mg/mL, soaking for 3.5h under the assistance of 250W ultrasonic and at 65 ℃, then filtering, refluxing the obtained filtrate into the enzymolysis liquid for recycling, and draining the obtained precipitate to obtain modified corn particles;
Step 4: dissolving procyanidine in water to obtain procyanidine solution with concentration of 5.5mg/mL, spraying the procyanidine solution on the surface of modified corn particles, mechanically mixing, adjusting pH to neutrality when the water content of the modified corn particles reaches 38%, grinding at 16800rpm at high speed, vacuum drying wet corn flour under vacuum degree of 0.05MPa and temperature of 70 ℃, sieving with 100 mesh sieve to obtain functional corn flour, and measuring to obtain corn flour with quercetin content of 7.2mg/g and procyanidine content of 5.5mg/g.
Example 3: the efficient preparation method of the functional corn flour comprises the following steps:
Step 1: adding water into corn kernels (the feed-liquid ratio is 1:3), carrying out microwave cooking for 7min at 550W, and grinding for 20s at 2800rpm to obtain mixed liquor;
Step 2: adjusting the pH value of the mixed solution to 5, then adding cellulase accounting for 1.5% of the mass of the mixed solution, and carrying out enzymolysis for 55min at 50 ℃ to obtain an enzymolysis solution;
step 3: adjusting the pH value of the enzymolysis liquid to 9, then adding alkaline protease and quercetin which account for 2.5% of the mass of the enzymolysis liquid, wherein the concentration of the quercetin in the enzymolysis liquid is 3.5mg/mL, soaking for 4.5 hours under the assistance of 150W ultrasonic and at 55 ℃, then filtering, refluxing the obtained filtrate into the enzymolysis liquid for recycling, and draining the obtained precipitate to obtain modified corn particles;
Step 4: dissolving procyanidine in water to obtain procyanidine solution with concentration of 4.5mg/mL, spraying the procyanidine solution on the surface of modified corn particles, mechanically mixing, adjusting pH to neutrality when the water content of the modified corn particles reaches 38%, grinding at 16800rpm at high speed, vacuum drying wet corn flour under vacuum degree of 0.07MPa and temperature of 60 ℃, sieving with 100 mesh sieve to obtain functional corn flour, and measuring to obtain corn flour with quercetin content of 8.4mg/g and procyanidine content of 4.5mg/g.
Comparative example 1: corn flour is prepared by a dry method. The specific process is as follows: peeling and degerming the screened corn seeds by using a high-efficiency peeling pulverizer to obtain corn grits, adding quercetin and procyanidine, pulverizing in a high-speed universal pulverizer to obtain corn flour, sieving with a 100-mesh sieve, wherein the final content of quercetin and procyanidine in dry corn flour is 9mg/g and 5mg/g respectively.
Comparative example 2: corn flour is prepared by a semi-dry method. The specific process is as follows: the corn grits are prepared by peeling and degerming the screened corn seeds by using a high-efficiency peeling powder making machine, distilled water containing quercetin and procyanidine is added into the corn grits, the moisture of the corn grits is adjusted to 38%, then the corn grits are crushed in a high-speed universal crusher to obtain corn flour, the corn flour is put into a drying box and dried at 40 ℃ and then is sieved by a 100-mesh sieve, and the final contents of the quercetin and procyanidine in the semi-dry corn flour are 9mg/g and 5mg/g respectively.
Comparative example 3: the corn flour is prepared by adopting a wet method. The specific process is as follows: putting the screened corn kernels into a beaker, adding distilled water, wherein the mass ratio of corn to distilled water is 1:3, adding quercetin and procyanidine, soaking for 36 hours, putting into a colloid mill for grinding, filtering corn steep liquor obtained after grinding to remove most of water to obtain wet corn flour, putting into a drying oven, drying at 40 ℃, sieving with a 100-mesh sieve, and respectively obtaining the final content of the quercetin and procyanidine in the wet corn flour of 9mg/g and 5mg/g.
Detection test:
Test method (one)
1. Corn starch-polyphenol recombination rate determination
Mixing corn flour added with polyphenol with water according to a ratio of 1:10, adding 1% neutral protease for enzymolysis for 1h to remove protein, centrifuging, and taking starch precipitate. The starch pellet was then made into a 10% dispersion, which was gelatinized by heating at 95℃for 10min, 1mL of the gelatinized sample was added to 5mL of distilled water, vortexed for 2min and centrifuged at 4000rpm for 15min. mu.L of the supernatant was added to 15mL of distilled water and 2mL of iodine solution, and the tube was turned about 10 times. The absorbance of the solution was measured at 620nm using a spectrophotometer. The absorbance of corn flour without added polyphenol was used as a blank. The starch-polyphenol recombination rate is calculated from the following formula:
Wherein: CI is the recombination rate; a 0 is absorbance of corn flour without polyphenol; a 1 is absorbance of corn flour added with polyphenol.
2. Infrared spectroscopic determination of corn starch-polyphenol complexes
The change in the structure and functional groups of the starch-polyphenol complex in the corn flour was measured and analyzed using an infrared spectrometer. Mixing functional corn flour with water at a ratio of 1:10, adding 1% neutral protease for enzymolysis for 1h to remove protein, centrifuging, collecting starch precipitate, and lyophilizing. The lyophilized sample was mixed with potassium bromide and ground, pressed into small discs. The scanning range of the wave number is 500-4000 cm -1, the resolution in the scanning process is 4cm -1, and air is used as background for scanning, so that an infrared spectrum graph is finally obtained.
3. Determination of free amino and free thiol groups of zein-polyphenol complexes
Mixing functional corn flour with water according to a ratio of 1:10, adding 1% of alpha-amylase for enzymolysis for 2 hours to remove starch, centrifuging, taking protein precipitate, freeze-drying, and measuring free amino and free sulfhydryl of a freeze-dried sample.
The free amino content was determined by the o-phthalaldehyde method (OPA). 80mg of OPA was dissolved in 2mL of 95% ethanol, and mixed with 50mL of 10mM sodium tetraborate buffer (pH 9.7), 5mL of 20% (w/w) SDS, and 200. Mu.L of beta-mercaptoethanol, and the mixture was diluted with distilled water to 100mL to prepare an OPA reagent. 200. Mu.L of the sample solution (2 mg/mL) was reacted with 4mL of OPA reagent at room temperature for 5min, and then absorbance at 340nm (A 340) was measured using an ultraviolet-visible spectrophotometer, using distilled water instead of the sample as a blank, and its free amino content was analyzed according to A 340.
The free thiol content was determined using the Ellman reagent method. A4 mg/mL solution of DTNB, the Ellman reagent, was prepared in Tris-Gly buffer (0.086 mol/LTris, 0.09mol/L glycine, 0.04mol/L EDTA, 8mol/L urea, pH 8). 15mg of the sample was dissolved in 5mL of Tris-glycine buffer, 50. Mu.L of Ellman reagent was added, the reaction was carried out at 25℃for 1 hour, 5,000Xg was centrifuged for 10 minutes, the supernatant was taken and absorbance (A 412) was measured at 412nm, and the free thiol content was analyzed according to A 412 using an equivalent amount of distilled water instead of the sample as a blank.
4. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of zein-polyphenol complexes
Mixing functional corn flour with water at a ratio of 1:10, adding 1% alpha-amylase for enzymolysis for 2 hours to remove starch, centrifuging, collecting protein precipitate, and lyophilizing. The concentrations of the separation gel and the concentration gel are 12% and 4%, respectively, and the concentration of the protein in the sample is 2mg/mL. The sample was dissolved in buffer (pH 6.80.5M Tris-HCl and glycerol, SDS, beta-mercaptoethanol and bromophenol) and then heated at 95℃for 5min to a loading of 15uL. After electrophoresis, the protein was stained with coomassie brilliant blue R250. SDS-PAGE gel electrophoresis was performed using Image Lab Software 3.0.0 software.
5. In vitro digestion experiments
500Mg corn meal samples were immersed in 50mL simulated gastric fluid (2 g NaCl and 7mL37% HCl in 1000mL distilled water with pH adjusted to 1.2) containing 0.1% pepsin. The mixture was then thermostatically shaken at 100rpm for 2h at 37℃to simulate gastric digestion. Subsequently, the gastric digested sample was transferred to simulated intestinal fluid (6.8 g KH 2PO4 in 250mL distilled water, mixed with 190mL 0.2M NaOH, and the volume was adjusted to 1000mL with distilled water, pH was adjusted to 7.5) containing 1% pancreatin, 0.5% amyloglucosidase, and 0.5% bile salts, and the intestinal digestion was simulated by shaking under the same conditions for another 4 hours. The sample was placed in an ice bath for 30min to terminate the digestion reaction.
6. Determination of resistant starch content
Starch is classified into three categories based on its bioavailability after digestion: rapidly Digestible Starch (RDS) refers to starch that is rapidly digested and absorbed in the small intestine within 20 minutes; slowly Digestible Starch (SDS) refers to starch that is fully absorbed in the small intestine within 20-120min but at a relatively slow rate of absorption; resistant Starch (RS) refers to starch that is not digested and absorbed by the human small intestine within 120 minutes. Glucose content (FG) was measured by the DNS method. The calculation formula is as follows:
Wherein: FG is the amount of free glucose (mg) contained prior to enzymatic hydrolysis; g 20 is the glucose content (mg) at 20min of enzymatic hydrolysis; g 120 is the glucose content (mg) at 120min of enzymatic hydrolysis; TS is the total starch content (mg) of the system.
7. In vitro antioxidant Activity assay
DPPH free radical scavenging ability was used to evaluate antioxidant activity of corn meal samples after digestion. Samples were collected after completion of simulated gastrointestinal digestion, 1.0mL of 0.5mg/mL sample was mixed with 2.0mL of 0.2mM DPPH-ethanol solution, reacted in the dark for 30min, and absorbance was measured at 517nm after the reaction. The DPPH distilled water solution is used as a blank. The DPPH radical scavenging capacity was calculated as follows:
8. Determination of the biological availability of active ingredients
After the simulated gastrointestinal digestion of the corn meal sample is completed, 4000g is centrifuged at 4℃for 40min. The supernatant collected represents the "mixed micelle" portion of intestinal fluid, which contains the bioavailable active ingredient. Extracting quercetin and procyanidine in the mixed micelle with absolute ethanol, and centrifuging at 4000g for 20min after vortex. And then measuring the absorbance of the supernatant by an ultraviolet spectrophotometer at the wavelength of 373nm and 546nm respectively, and taking the absorbance into a standard curve to obtain the content of quercetin and procyanidine, wherein the content is used for calculating the biological accessibility of the active ingredients after the simulated gastrointestinal digestion. The bioavailability is expressed as the ratio of the amount of active ingredient in the micelle to the amount of active ingredient in the sample prior to simulated digestion.
9. Determination of chemical stability of active ingredient
And (3) respectively placing the functional corn flour samples in water baths at 65, 85 and 100 ℃ for heating treatment for 2 hours, and then respectively measuring the content of quercetin and procyanidine in the corn flour subjected to heat treatment at different temperatures. The retention of the active ingredient is expressed as the ratio of the active ingredient content in the sample after heat treatment to the initial active ingredient content in the sample before heat treatment.
10. Corn flour gelatinization property determination
3.5G of corn meal sample and 25mL of distilled water were added to an aluminum box of a Rapid Viscosimeter (RVA), the rotating paddles were calibrated first, then the sample was thoroughly stirred with water using the rotating paddles and then mounted on the RVA meter for testing. The sample was kept at 50℃for 1min, then heated to 95℃at a rate of 4℃per min, kept at 95℃for 5min, and then cooled to 50℃at the same rate and kept for 5min, to form a corn paste and the sample was analyzed for gelatinization properties.
11. Corn meal aging characteristic determination
Placing the gelatinized corn flour sample in a small-sized culture dish, sealing with a preservative film, refrigerating at 4 ℃ for 7 days to enable the sample to form gel, and testing the texture of the gelatinized sample. The TPA assay mode was selected using a cylindrical metal probe P/0.5. Test conditions: the pre-test rate was 1.0mm/s, the test rate was 2.0mm/s, the post-test rate was 2.0mm/s, the test distance was 10.0mm, the compression degree was 40%, the interval between two compressions was 2s, the trigger force was 5g, and the aged hardness (g) of the sample was analyzed.
(II) results and analysis
1. Corn starch-polyphenol complex characterization
The starch-polyphenol complexing ratio refers to the degree of complexing of starch with polyphenol. FIG. 1 is a bar graph of the starch-polyphenol recombination rate in functional corn flour. The corn flour of comparative example 1 has very low starch-polyphenol complexing ratio, and the starch and polyphenol cannot be effectively contacted due to the fact that the dry milling does not have water, so that the complexing ratio is low. The corn flours of comparative example 2 and comparative example 3 gradually increase the starch-polyphenol complexation rate, which may be related to the water participation and longer reaction times. The 3 examples corn flour significantly increased the starch-polyphenol complexation rate compared to 3 comparative examples, and example 1 had the greatest starch-polyphenol complexation rate, which greatly promoted the interaction and binding of starch and polyphenol.
The infrared absorption spectrum forms essentially a transition of rotational-vibrational energy level, and the absorption peak of the molecule and the intensity and position of the peak can be determined according to the vibration condition of the molecule. FIG. 2 is an infrared spectrum of a starch-polyphenol complex in functional corn flour. All samples had a broad band between 3000-3500cm -1 due to the stretching vibration and absorption of hydrogen bonding groups (O-H). The peak at 2927cm -1 is due to the antisymmetric stretching vibration of CH 2 and the band at 1649cm -1 is due to COO-stretching vibration. In addition, there are many absorption peaks at 500-1800cm -1, which are mostly caused by vibrations of some double bond carbons, ester bonds and ether bonds. All absorption peak intensities of the starch-polyphenol complexes in the corn flour of comparative examples 1-3 were lower, while examples 1-3 significantly increased the infrared spectrum absorption peak intensities of the starch-polyphenol complexes, especially the starch-polyphenol complex of example 1 was the strongest. Notably, the absorption peak becomes broader and the peak intensity is greater in the wavelength range of 3000-3500cm -1, indicating that starch is non-covalently bound to polyphenols via hydrogen bonds. The starch-polyphenol complexes of example 1 have stronger absorption peaks at wavelengths 576, 930, 1020 and 1155cm -1, indicating that stronger interactions of starch with polyphenols result in changes in starch structure. In addition, the complexing of starch with polyphenols did not produce new absorption peaks, indicating that no new covalent bonds were formed. Thus, the starch in the functional corn flour forms a non-covalent complex with the polyphenol through non-covalent interactions.
2. Zein-polyphenol complex characterization
Polyphenols are susceptible to oxidation to form quinones under aerobic alkaline conditions and when contacted with proteins, are susceptible to further covalent complexing reactions by attack of nucleophilic groups (amino and sulfhydryl groups, etc.) on the proteins, which results in reduced free amino and free sulfhydryl content of the proteins. FIG. 3 is a bar graph of the free amino and free thiol content of a protein-polyphenol complex in functional corn flour. The protein-polyphenol complexes in the 3 comparative corn flours had higher free amino groups and free thiol groups. The wet milling process increases the contact area of the corn protein and the polyphenol in a water system, accelerates the composite reaction rate, and leads the corn powder prepared in the comparative example 3 to obviously reduce the free amino and the free sulfhydryl of the protein-polyphenol compound. Examples 1-3 further reduced the free amino and free thiol groups of the protein-polyphenol complex as compared to comparative examples 1-3. SDS was used in the determination of free amino groups, which disrupt non-covalent bonds, so that a decrease in free amino groups demonstrated covalent binding of the protein to polyphenols; since 8mol/L urea, which can inhibit the conversion of the thiol into disulfide, was used in the measurement of the free thiol, the decrease in the free thiol also demonstrated covalent binding of the polyphenol to the free thiol of the protein. The quinone formed by oxidation of polyphenols can react not only with free amino groups to form C-N covalent bonds, but also with thiol groups of cysteines to form C-S covalent bonds. Furthermore, the protein-polyphenol complexes in the corn flour of example 1 had the lowest free amino groups and free thiol groups, indicating that more protein-polyphenol covalent bonds were formed in the corn flour of example 1.
Covalent binding between protein and polyphenol can be verified by SDS-PAGE. FIG. 4 is a SDS-PAGE diagram of protein-polyphenol complexes in functional corn flour. Marker is the standard protein molecular weight, and the movement distance of protein molecules in electrophoresis is mainly determined by the relative molecular weight of the protein. The protein-polyphenol complexes of all sample corn flour appear as high molecular weight aggregates at greater than 140 kDa. Since SDS-PAGE electrophoresis uses a loading buffer solution containing SDS and beta-mercaptoethanol to break the noncovalent interactions and covalent disulfide bonds in the protein-polyphenol complex, the presence of this high molecular weight aggregate band indicates that C-N and/or C-S covalent bonds are formed between the protein and the polyphenol, thereby producing the protein-polyphenol covalent complex. The weak high molecular weight bands of the protein-polyphenol complexes in the corn flours of comparative examples 1-3, while the darker high molecular weight aggregate bands of examples 1-3, demonstrate that the corn flours prepared in examples effectively increase covalent bonds between protein and polyphenol. In addition, the high molecular weight bands of the protein-polyphenol complex in the corn flour of example 1 are darkest, indicating that the covalent bonds between the protein and the polyphenol are the greatest, and the covalent bonds between the protein and the polyphenol are the most obvious, forming a more stable protein-polyphenol covalent complex, which has positive effects on the improvement of polyphenol stability and bioavailability and the protection of antioxidant activity.
3. Resistant starch content, antioxidant activity and active ingredient bioavailability
Fig. 5 is a bar graph of resistant starch content and antioxidant activity after functional corn meal digestion, and fig. 6 is a bar graph of active ingredient bioavailability after functional corn meal digestion.
As can be seen from fig. 5, the resistant starch content of the 3 comparative corn flours is relatively low, while the resistant starch content of the 3 example corn flours is significantly increased, and example 1 has the highest resistant starch content. The resistant starch has various physiological functions of reducing blood sugar, preventing colon cancer, obesity and the like, and has important functions of regulating blood sugar, blood fat and digestion. In addition, the DPPH radical scavenging ability of the corn meal of all examples was significantly higher than that of all comparative examples, especially example 1 had the highest DPPH radical scavenging ability and exhibited the strongest antioxidant activity.
As can be seen from fig. 6, quercetin and procyanidins in the comparative example have lower bioavailability, both below 35%, and procyanidins have higher bioavailability than quercetin. All examples improved the bioavailability of the active ingredient compared to the comparative examples, whether quercetin or procyanidin, and example 1 had the highest bioavailability, which may be related to its improved chemical stability of the active ingredient. Higher bioavailability results in higher bioavailability, indicating that the body has higher absorption efficiency of nutrients or active ingredients. In addition, all the examples change the bioavailable mode of the active ingredient from higher procyanidin bioavailability to higher quercetin bioavailability, so that the more expensive quercetin (200 yuan/g) is easier to be absorbed by the organism than the procyanidin (150 yuan/g) with relatively low price, and the added value of the product can be effectively improved. Therefore, the corn flour prepared by the invention has higher resistant starch content, stronger antioxidant activity and higher biological accessibility, and can be applied to the development of starch-based functional foods.
4. Chemical stability of the active ingredient
Figures 7-9 are bar graphs of chemical stability of active ingredients in functional corn flour under heat treatment conditions of 65 ℃, 85 ℃ and 100 ℃. It can be seen that the retention of the active ingredient was lower in all of the comparative examples under the conditions of 3 heat treatment temperatures, and the retention of the active ingredient was further decreased as the heat treatment temperature was increased. The retention of quercetin and procyanidins in the comparative examples at 65, 85 and 100 ℃ was 47% -53%, 37% -43% and 23% -34%, respectively. The retention of active ingredient in all examples was significantly improved at 3 heat treatment temperatures compared to the comparative examples, reaching 78% -82%, 69% -73% and 58% -64%, respectively. The best retention of quercetin and procyanidins in example 1 exhibited the best chemical stability against high temperatures. As the quercetin and procyanidin molecules form physical and chemical interactions with starch and protein in corn flour by specific groups, the structural stability of the quercetin and procyanidin molecules is improved, and degradation is slowed down under the influence of external environment, so that the retention rate of the quercetin and procyanidin molecules after heat treatment is improved. Therefore, the functional corn flour prepared by the invention can ensure that active ingredients of quercetin and procyanidine in the functional corn flour have good chemical stability in the hot processing process.
5. Gelatinization characteristics and aging characteristics
Table 1 shows gelatinization parameters and aged hardness of functional corn flour. There were differences in the gelatinization properties of the 3 comparative examples, but not so much. Comparative example 3 (wet corn flour) has a higher gelatinized viscosity than comparative example 1 (dry corn flour) and comparative example 2 (semi-dry corn flour), probably due to the smaller particle size of the corn particles by wet processing and the higher integrity of the starch particles, which makes the corn flour easier to gelatinize. However, wet-process corn flour has a too long soaking time and a low milling efficiency. The milling process (5-6 h) of all the embodiments obviously reduces the soaking time of the traditional wet milling process (36 h) and improves the corn milling efficiency. The gelatinization characteristics of the 3 examples of corn flour are all better than those of comparative example 3, especially the corn flour of example 1 has the highest gelatinization viscosity and exhibits the optimal gelatinization characteristics. Furthermore, all of the example corn flours had lower aged hardness than all of the comparative examples, and the example 1 corn flour had the lowest aged hardness, exhibiting significant anti-aging characteristics. Therefore, the functional corn flour prepared by the invention has good processing quality.
TABLE 1 gelatinization parameters and aged hardness of functional corn flour
Different letters in the same column indicate significant differences (p < 0.05).
In the foregoing, the present invention is merely preferred embodiments, which are based on different implementations of the overall concept of the invention, and the protection scope of the invention is not limited thereto, and any changes or substitutions easily come within the technical scope of the present invention as those skilled in the art should not fall within the protection scope of the present invention. Therefore, the protection scope of the present invention should be subject to the protection scope of the claims.
Claims (5)
1. The efficient preparation method of the functional corn flour is characterized by comprising the following steps of:
step 1: adding water into corn kernels to carry out microwave cooking, and then grinding at a low speed to obtain a mixed solution; the feed liquid ratio is 1:3, the power of microwave cooking is 400-600W, the time is 6-10 min, and the rotating speed of low-speed grinding is 2500-3000 rpm;
Step 2: adjusting the pH value of the mixed solution to 5, and then adding cellulase for enzymolysis, wherein the addition amount of the cellulase is 0.5-1.5% of the mass of the mixed solution, so as to obtain an enzymolysis solution; the enzymolysis temperature is 40-60 ℃ and the enzymolysis time is 50-70 min;
Step 3: adjusting the pH value of the enzymolysis liquid to 9, then adding alkaline protease and quercetin, wherein the addition amount of the alkaline protease is 1-3% of the mass of the enzymolysis liquid, the concentration of the quercetin in the enzymolysis liquid is 2-4 mg/mL, soaking under the assistance of ultrasound, and then filtering and draining to obtain modified corn particles; the ultrasonic power is 100-300W, the soaking temperature is 50-70 ℃ and the time is 3-5 h;
Step 4: dissolving procyanidine in water to obtain procyanidine solution with concentration of 4-6 mg/mL, spraying the procyanidine solution on the surface of modified corn particles, mechanically mixing, adjusting pH to neutrality when the water content of the modified corn particles reaches 35-40%, grinding at high speed at 16000-17000 rpm, and vacuum drying to obtain functional corn powder.
2. The method of claim 1, wherein the filtrate obtained by filtering after soaking in step 3 is refluxed to the enzymatic hydrolysate for recycling.
3. The method according to claim 1, wherein the vacuum degree of the vacuum drying in the step 4 is 0.04-0.08 MPa and the temperature is 60-70 ℃.
4. A functional corn flour made by the process of any one of claims 1-3.
5. A functional corn flour made by the method of any one of claims 1-3 for use in preparing a functional corn food.
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