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CN117770317A - Preparation method of oat plant-based fermented milk for enriching polyphenol based on composite enzymolysis-fermentation combined technology - Google Patents

Preparation method of oat plant-based fermented milk for enriching polyphenol based on composite enzymolysis-fermentation combined technology Download PDF

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CN117770317A
CN117770317A CN202310708269.4A CN202310708269A CN117770317A CN 117770317 A CN117770317 A CN 117770317A CN 202310708269 A CN202310708269 A CN 202310708269A CN 117770317 A CN117770317 A CN 117770317A
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oat
fermentation
enzymolysis
milk
fermented milk
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王韧
何巧艳
罗小虎
李亚男
冯伟
王涛
张昊
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Ningbo University
Jiangnan University
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Ningbo University
Jiangnan University
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Abstract

The invention discloses a preparation method of oat plant-based fermented milk based on polyphenol enrichment by a composite enzymolysis-fermentation combined technology. The preparation method comprises the steps of soaking, cleaning and drying oat to obtain pretreated oat; baking, crushing and ball milling to obtain whole oat superfine powder; adding water into the whole wheat superfine powder to obtain oat milk, stirring, shearing at high speed, gelatinizing the oat milk completely dissolved, and adding enzyme for enzymolysis to obtain enzymolysis liquid; after enzyme deactivation, preparing and homogenizing to obtain a homogeneous liquid, sterilizing and cooling to obtain a uniform and stable system; finally, adding strains for inoculation, fermentation and after-ripening to obtain the oat plant-based fermented milk. The preparation method disclosed by the invention can effectively improve the polyphenol content of the system and the acidity of the product without adding an additional nitrogen source, and shortens the fermentation time.

Description

Preparation method of oat plant-based fermented milk for enriching polyphenol based on composite enzymolysis-fermentation combined technology
Technical Field
The invention relates to the technical field of fermented milk preparation, in particular to a preparation method of oat plant-based fermented milk based on polyphenol enrichment by a combined enzymolysis-fermentation technology.
Background
In recent years, consumer interest and research into vegetable dairy products has been growing due to the increasing negative impact of animal-derived products on human health, animal survival, and the environment. Dairy consumption is decreasing in many western countries, while sales of vegetable milk are increasing. Vegetable milk currently occupies 15% of the total milk industry, and other vegetable milk product substitutes are becoming more popular. Vegetable milk consumes less energy during production than animal milk, and emits less greenhouse gases per gram of protein, which makes them an attractive option for those desiring to reduce the carbon footprint.
The most common ingredient in plant-based fermented milk is soy, which is very suitable as a substrate for vegetable yoghurt substitutes due to the high protein content and beneficial amino acid composition of legumes, and in recent years other plant ingredients such as coconut and flaxseed have also been favored.
Oat is also called hulless oat, is one of grains with highest nutritive value, contains rich carbohydrate, balanced protein, unsaturated fatty acid, vitamin, mineral substance and other nutritive components, has the characteristics of three high and two low, has the medicinal and edible properties, and is recognized as functional food by FDA. However, the starch content is too high, so that the viscosity of the system is too high during gelatinization, and the consumer acceptability is reduced; meanwhile, as the raw materials used are whole oat, namely bran is reserved, the problem of poor solubility of oat milk can be solved.
At present, research on oat plant-based fermented milk mainly focuses on enzymolysis technology of oat starch and discusses influence of fermentation strains on oat fermented milk, and loss of key nutrition components is often caused by excessive processing in the processing process of oat products, so that the prepared oat milk has low polyphenol content and product acidity, long fermentation time and high cost.
Disclosure of Invention
In order to overcome the defects, the invention provides a preparation method of oat plant-based fermented milk based on the enrichment of polyphenol by a combined enzymolysis-fermentation technology. According to the invention, through research on enzyme and strain compounding and fermentation processes, the preparation method provided by the invention can effectively improve the polyphenol content of the system and the acidity of the product without adding an additional nitrogen source, and meanwhile, the fermentation time is shortened.
The technical scheme of the invention is as follows:
the preparation method of the oat plant-based fermented milk for enriching polyphenol based on the combined enzymolysis-fermentation technology comprises the following steps:
(1) Soaking, cleaning and drying oat to obtain pretreated oat;
(2) Baking, crushing and ball milling the pretreated oat to obtain whole oat superfine powder;
(3) Adding water into whole wheat superfine powder to obtain oat milk, stirring, shearing at high speed, gelatinizing the completely dissolved oat milk, and adding enzyme for enzymolysis to obtain enzymolysis solution;
(4) After enzyme deactivation, preparing and homogenizing to obtain a homogeneous liquid, sterilizing and cooling to obtain a uniform and stable system;
(5) Adding strain for inoculation, fermentation and after-ripening to obtain the oat plant-based fermented milk.
Further, in step (1), the oat is whole oat kernel (oat kernel with bran); the temperature of the drying is 105 ℃ and the time is 20-40min.
Further, in the step (1), the drying refers to drying until the surface is dry and no scorched smell is generated.
Further, in the step (2), the frequency of the ball milling is 4000Hz and the time is 15min.
Further, in the step (2), the baking is by infrared baking; the crushing is that a universal crusher is used for grinding for 3 times, and the powder is sieved by a 40-mesh sieve.
Further, in the step (2), the baking temperature is 170-190 ℃, the time is 10-20min, and the baking is performed once every 2-4 min.
Further, in the step (3), the volume ratio of the Quan Maichao micro powder to the water is 1:5-8; the temperature of the water is 50-55 ℃, preferably the volume ratio of the Quan Maichao micro powder to the water is 1:6, preparing a base material; the temperature of the water was 55 ℃.
Further, in the step (3), the stirring speed is 300-500r/min and the stirring time is 10-20min; the high-speed shearing rotating speed is 4000-7000r/min, and the shearing time is 10-20min; preferably, the high-speed shearing rotating speed is 6000rpm, and the shearing time is 10-15min; the gelatinization temperature is 80-90deg.C, and the time is 10-20min.
Further, in step (3), the enzyme comprises one or more of an alpha-amylase, a beta-amylase, a cellulase, an alkaline protease; the addition amount E/S of each enzyme (the percentage of enzyme powder and a substrate is the gelatinized liquid before enzyme addition) is 0.1% -2%; the enzymes were all purchased from Jiangsu Bo Bioproducts Co., ltd; the enzymolysis temperature is 55-65 ℃.
Further, in the step (3), the specific process of adding enzyme for enzymolysis comprises the steps of adding alpha-amylase, beta-amylase and cellulase for reaction at 55-65 ℃ for 90-120min, adjusting the temperature to 55-65 ℃ and the pH value to 8, and adding alkaline protease for reaction; the reaction time is 30-60min.
Further, in the step (4), 1% -7% of sucrose is added to the preparation; homogenizing for 3 times at 20-30deg.C and 30-40 MPa; the sterilization is carried out for 10min at 95 ℃; the cooling refers to cooling to 43-48 ℃.
Further, in the step (5), the strains are lactobacillus acidophilus and lactobacillus casei, and the volume ratio of the two strains is 1:1; the concentration of the strains is 1 multiplied by 10 8 CFU/mL; the total inoculation amount of the strain is 1% (v/v), namely, 1% of activated bacterial liquid is added into the culture substrate; the temperature of the fermentation is 37 ℃, and when the pH value of the fermentation liquid is less than or equal to 4.5 and the acidity is more than or equal to 30 DEG T, the fermentation is finished, and after-ripening is carried out; the post-ripening temperature is 4 ℃ and the time is 12-24 hours.
An oat plant-based fermented milk prepared by the method.
The beneficial technical effects of the invention are as follows:
the invention releases the polyphenol in a combined state through the synergism of fermentation and specific enzymes by an enzymolysis-fermentation combined technology, thereby improving the polyphenol content and enriching the polyphenol in the product, and enhancing the antioxidant activity; the enzymolysis technology effectively improves the acid production rate of the lactobacillus, thereby improving the acidity and shortening the fermentation time.
The nitrogen source required in the fermentation process is completely provided by oat protein in whole oat, and the nitrogen source is not added additionally, so that the production cost is saved.
According to the invention, ball milling processing is carried out on whole oat, the obtained superfine crushed oat flour is not filtered, the grain utilization rate is improved, meanwhile, the coarse sense of the grain is not generated, the finished product has good tissue state, the taste is smooth and fine, and the unique oat flavor is provided.
The invention realizes that the whole oat milk is used as a fermentation substrate, and the whole oat milk is used for fermentation, does not contain raw cow milk, and meets the requirements of a plurality of consumers, such as consumers allergic to dairy products.
The invention hydrolyzes alpha-1, 4 glycosidic bond in starch by alpha-amylase to generate micromolecular sugar, and can break ester bond combined with polyphenol in the hydrolysis process, so that the polyphenol combined with starch is released; the pullulanase hydrolyzes starch, so that the product contains more free amylose molecules, thereby maximally utilizing starch raw materials, promoting the action of beta-amylase and being beneficial to forming oligosaccharide and maltose as main monosaccharides; the whole oat milk is subjected to enzymolysis by using cellulase, so that cellulose and hemicellulose in bran can be effectively subjected to enzymolysis, and combined phenolic acid, particularly ferulic acid, in the bran can be released; the alkaline protease can decompose proteins in the oat fermented milk into small-molecule polypeptides, amino acids and the like, so that the release of polyphenol substances combined with the proteins is promoted; the protease treatment is also beneficial to the release of more free amino acids and small peptide substances from the whole oat milk, and the substances are a high-efficiency nitrogen source and are more beneficial to the growth and propagation of lactic acid bacteria and the fermentation and acid production.
In addition, when oat is used as a substrate, the lactobacillus casei has an advantage over other strains in increasing the total polyphenol content, and possibly, during fermentation, enzymes secreted by the lactobacillus casei hydrolyze oat polysaccharide, protein or chemical bonds of cell walls and polyphenol, so that a large amount of free polyphenol is released; meanwhile, the metabolite possibly weakens the ether bond action of the combined polyphenol compound and the cell wall, so that the combined polyphenol compound is easier to release, and then the cellulase can well release the combined polyphenol to the free polyphenol, so that the free polyphenol in the oat plant-based fermented milk is greatly increased; it is also possible that it produces new polyphenolic compounds or that it enzymatically degrades macromolecular polyphenolic compounds, such that the free polyphenol content increases after fermentation.
According to the invention, when oat is used as a substrate, lactobacillus acidophilus is more suitable for growth than other strains, so that the two strains are subjected to compound fermentation, the total polyphenol content in the oat plant-based fermented milk is increased, the acidity is increased, and the fermentation time is greatly shortened.
Detailed Description
The present invention will be specifically described with reference to examples.
In the following description, numerous specific details are set forth in order to provide a thorough understanding of the present invention, but the present invention may be practiced in other ways other than those described herein, and persons skilled in the art will readily appreciate that the present invention is not limited to the specific embodiments disclosed below.
Further, reference herein to "one embodiment" or "an embodiment" means that a particular feature, structure, or characteristic can be included in at least one implementation of the invention. The appearances of the phrase "in one embodiment" in various places in the specification are not necessarily all referring to the same embodiment, nor are separate or alternative embodiments mutually exclusive of other embodiments.
The polyphenols in oat can be combined with polysaccharide, protein and cell wall components in a combined state, and the combined state polyphenols can be released through fermentation and hydrolysis of corresponding enzymes. Prior studies have shown that polyphenols in starch granules can interact with amylose to form clathrate inclusions and can bind to the side chains of amylopectin, the amorphous regions of starch granules.
The invention provides a preparation method of oat plant-based fermented milk for enriching polyphenol based on a composite enzymolysis-fermentation combined technology, which comprises the following steps:
(1) Soaking, cleaning and drying oat to obtain pretreated oat;
(2) Baking, crushing and ball milling the pretreated oat to obtain whole oat superfine powder;
(3) Adding water into whole wheat superfine powder to obtain oat milk, stirring, shearing at high speed, gelatinizing the completely dissolved oat milk, and adding enzyme for enzymolysis to obtain enzymolysis solution;
(4) After enzyme deactivation, preparing and homogenizing to obtain a homogeneous liquid, sterilizing and cooling to obtain a uniform and stable system;
(5) Adding strain for inoculation, fermentation and after-ripening to obtain the oat plant-based fermented milk.
In one embodiment of the present invention, in step (1), the oat is whole oat kernel (oat kernel with bran); the temperature of the drying is 105 ℃ and the time is 20min, 30min or 40min.
In one embodiment of the present invention, in step (1), the drying refers to drying to the surface without scorched smell.
In one embodiment of the present invention, in the step (2), the frequency of the ball milling is 4000Hz for 15min.
In one embodiment of the invention, in step (2), the baking is by infrared baking; the grinding is carried out by grinding for 3 times by a universal grinder, sieving by a 40-mesh sieve, and the grinding times are not limited to 3 times.
In one embodiment of the present invention, in the step (2), the baking temperature is 170 ℃, 180 ℃, 185 ℃ or 190 ℃, and the time is 10min, 15min or 20min, and the baking is performed once every 2min, 3min or 4 min.
In one embodiment of the present invention, in the step (3), the volume ratio of the Quan Maichao micro powder to the water is 1:5, a step of; 1:6 or 1:8, 8; the temperature of the water is 50-55 ℃, preferably the volume ratio of the Quan Maichao micro powder to the water is 1:6, preparing a base material; the temperature of the water was 55 ℃.
In one embodiment of the present invention, in the step (3), the stirring speed is 300-500r/min for 10min, 15min or 20min; the high-speed shearing rotating speed is 4000r/min, 6000r/min or 7000r/min, and the shearing time is 10min, 15min or 20min; preferably, the high shear rotational speed is 6000rpm; the gelatinization temperature is 80 ℃, 85 ℃ or 90 ℃ and the time is 10min, 15min or 20min.
In one embodiment of the invention, in step (3), the enzyme comprises one or more of an alpha-amylase, a beta-amylase, a cellulase, an alkaline protease; the addition amount E/S of the enzyme (the percentage of enzyme powder and a substrate is the gelatinized liquid before enzyme addition) is 0.15%, 0.5%, 0.6%, 1%, 1.5% or 2%; the enzymes were all purchased from Jiangsu Bo Bioproducts Co., ltd; the specific process of adding enzyme for enzymolysis comprises adding alpha-amylase, beta-amylase and cellulase, reacting at 55deg.C, 60deg.C or 65deg.C for 90min, 100min, 110min or 120min, adjusting temperature to 55deg.C, 58 deg.C, 60 deg.C or 65deg.C, and adding alkaline protease to react for 30min, 45min or 60min.
In one embodiment of the invention, in step (4), the formulating is adding 1%, 3%, 5% or 7% sucrose; the homogenizing condition is 20 ℃,25 ℃ or 30 ℃, 30MPa, 35MPa or 40MPa, homogenizing for 3 times; the sterilization is carried out for 10min at 95 ℃; the cooling means cooling to 43 ℃, 45 ℃ or 48 ℃.
In one embodiment of the invention, in the step (5), the strains are lactobacillus acidophilus (accession number: CICC 6089) and lactobacillus casei (accession number: CICC 6117), and the inoculation volume ratio of the two strains is 1:1; the concentration of the strains is 1 multiplied by 10 8 CFU/mL; the total inoculation amount of the strain is 1% (v/v), namely, 1% of activated bacterial liquid is added into the culture substrate; the temperature of the fermentation is 37 ℃, and when the pH value of the fermentation liquid is less than or equal to 4.5 and the acidity is more than or equal to 30 DEG T, the fermentation is finished, and after-ripening is carried out; the post-ripening temperature is 4 ℃ and the time is 12 hours, 16 hours, 20 hours or 24 hours.
An oat plant-based fermented milk prepared by the method.
The present invention will be further illustrated by the following specific examples and comparative examples.
Example 1:
the oat plant-based fermented milk enriched in polyphenol by the combined enzymolysis-fermentation technology comprises the following steps:
(1) Carefully selecting whole oat grains with good quality, soaking for half an hour, repeatedly cleaning for 3 times, placing in an oven, and drying at 105 ℃ for 30min to obtain pretreated oat grains.
(2) Baking the obtained pretreated oat grains for 10min at 180 ℃ by infrared rays, and turning over for one time after 2min to ensure that the pretreated oat grains have rich baking fragrance; coarsely crushing the roasted seeds for 3 times by using a universal crusher, sieving by using a 40-mesh sieve, and grinding for 15min by using a ball mill (the ball milling frequency is 4000 Hz) to obtain the whole oat superfine powder.
(3) Adding 6 times of 55 ℃ deionized water into the whole oat superfine powder, stirring the oat milk for 20min at the rotating speed of 500rpm/min, shearing at the high speed of 6000rpm for 15min, and gelatinizing the oat milk completely sheared at 85 ℃ for 15min. And then carrying out enzymolysis, namely adding 1% of alpha-amylase, 0.6% of beta-amylase and 0.5% of cellulase into the gelatinized oat milk, wherein the enzymolysis temperature is 55 ℃, the time is 120min, the pH is regulated to 8 by sodium hydroxide, the temperature is regulated to 60 ℃, and 0.15% of alkaline protease is added for heating reaction for 30min to obtain an enzymolysis liquid.
(4) After enzyme deactivation, 7% sucrose (mass fraction of sucrose in oat milk) was added to oat milk. Homogenizing after uniformly stirring, wherein the homogenizing condition is 25 ℃, 35MPa, and homogenizing for 3 times; sterilizing at 95deg.C for 10min, and rapidly cooling to 45deg.C to obtain uniform stable system.
(5) Inoculating strain, fermenting, measuring pH and acid value at the whole point, and after fermentation, finishing the fermentation when the pH value of the fermentation liquid is less than or equal to 4.5 and the acidity is more than or equal to 30 DEG T, and obtaining the oat plant-based fermented milk. The fermentation strain is lactobacillus casei and lactobacillus acidophilus, and the mixing ratio is 1:1, the concentration of the strains is 1 multiplied by 10 8 CFU/mL; the total inoculum size of the strain is 1% (v/v), the fermentation temperature is 37 ℃, and the post-maturation temperature is4℃for 18h. The experimental results of the physicochemical indexes related to the oat plant-based fermented milk are shown in table 1 below.
Example 2:
the oat plant-based fermented milk enriched in polyphenol by the combined enzymolysis-fermentation technology comprises the following steps:
(1) Carefully selecting whole oat grains with good quality, soaking for half an hour, repeatedly cleaning for 3 times, placing in an oven, and drying at 105 ℃ for 20min to obtain pretreated oat grains.
(2) Baking the obtained pretreated oat grains for 10min at 180 ℃ by infrared rays, and turning over for one time after 2min to ensure that the pretreated oat grains have rich baking fragrance; coarsely crushing the roasted seeds for 3 times by using a universal crusher, sieving by using a 40-mesh sieve, and grinding for 15min by using a ball mill (the ball milling frequency is 4000 Hz) to obtain the whole oat superfine powder.
(3) Adding 6 times of 55 ℃ deionized water into the whole oat superfine powder, stirring the oat milk for 20min at the rotating speed of 500rpm/min, shearing at the high speed of 6000rpm for 15min, and gelatinizing the oat milk completely sheared at 85 ℃ for 15min. And then carrying out enzymolysis, namely adding 0.5% of alpha-amylase, 0.6% of beta-amylase and 0.5% of cellulase into the gelatinized oat milk, wherein the enzymolysis temperature is 55 ℃, the time is 120min, the pH is regulated to 8 by sodium hydroxide, the temperature is regulated to 60 ℃, and 0.15% of alkaline protease is added for heating reaction for 30min to obtain an enzymolysis liquid.
(4) After enzyme deactivation, 7% sucrose was added to the oat milk. Homogenizing after uniformly stirring, wherein the homogenizing condition is 25 ℃, 35MPa, and homogenizing for 3 times; sterilizing at 95deg.C for 10min, and rapidly cooling to 45deg.C to obtain uniform stable system.
(5) Inoculating strain, fermenting, measuring pH and acid value at the whole point, and after fermentation, finishing fermentation to obtain oat plant-based fermented milk when pH value of fermentation liquor is less than or equal to 4.5 and acidity is more than or equal to 30 deg.T. The fermentation strain is lactobacillus casei and lactobacillus acidophilus, and the mixing ratio is 1:1, the concentration of the strains is 1 multiplied by 10 8 CFU/mL; the total inoculum size of the strain is 1% (v/v), the fermentation temperature is 37 ℃, the post-maturation temperature is 4 ℃, and the time is 18h. Experimental results of physicochemical indexes related to oat plant-based fermented milk are shown in the following table1。
Example 3:
the oat plant-based fermented milk enriched in polyphenol by the combined enzymolysis-fermentation technology comprises the following steps:
(1) Carefully selecting whole oat grains with good quality, soaking for half an hour, repeatedly cleaning for 3 times, placing in an oven, and drying at 105 ℃ for 30min to obtain pretreated oat grains.
(2) Baking the obtained pretreated oat grains for 10min at 180 ℃ by infrared rays, and turning over for one time after 2min to ensure that the pretreated oat grains have rich baking fragrance; coarsely crushing the roasted seeds for 3 times by using a universal crusher, sieving by using a 40-mesh sieve, and grinding for 15min by using a ball mill (the ball milling frequency is 4000 Hz) to obtain the whole oat superfine powder.
(3) Adding 6 times of 55 ℃ deionized water into the whole oat superfine powder, stirring the oat milk for 20min at the rotating speed of 500rpm/min, shearing at the high speed of 6000rpm for 15min, and gelatinizing the oat milk completely sheared at 85 ℃ for 15min. And then carrying out enzymolysis, namely adding 1.5% of alpha-amylase, 0.6% of beta-amylase and 0.5% of cellulase into the gelatinized oat milk, wherein the enzymolysis temperature is 55 ℃, the time is 120min, the pH is regulated to 8 by sodium hydroxide, the temperature is regulated to 60 ℃, and 0.15% of alkaline protease is added for heating reaction for 30min to obtain an enzymolysis liquid.
(4) After enzyme deactivation, 7% sucrose was added to the oat milk. Homogenizing after uniformly stirring, wherein the homogenizing condition is 25 ℃, 35MPa, and homogenizing for 3 times; sterilizing at 95deg.C for 10min, and rapidly cooling to 45deg.C to obtain uniform stable system.
(5) Inoculating strain, fermenting, measuring pH and acid value at the whole point, and after finishing fermentation when pH value of fermentation liquid is less than or equal to 4.5 and acidity is more than or equal to 30 DEG T, performing after-ripening to obtain oat plant-based fermented milk. The fermentation strain is lactobacillus casei and lactobacillus acidophilus, and the mixing ratio is 1:1, the concentration of the strains is 1 multiplied by 10 8 CFU/mL; the total inoculum size of the strain is 1% (v/v), the fermentation temperature is 37 ℃, the post-maturation temperature is 4 ℃, and the time is 18h. The experimental results of the physicochemical indexes related to the oat plant-based fermented milk are shown in table 1 below.
Example 4:
the oat plant-based fermented milk enriched in polyphenol by the combined enzymolysis-fermentation technology comprises the following steps:
(1) Carefully selecting whole oat grains with good quality, soaking for half an hour, repeatedly cleaning for 3 times, placing in an oven, and drying at 105 ℃ for 40min to obtain pretreated oat grains.
(2) Baking the obtained pretreated oat grains for 10min at 180 ℃ by infrared rays, and turning over for one time after 2min to ensure that the pretreated oat grains have rich baking fragrance; coarsely crushing the roasted seeds for 3 times by using a universal crusher, sieving by using a 40-mesh sieve, and grinding for 15min by using a ball mill (the ball milling frequency is 4000 Hz) to obtain the whole oat superfine powder.
(3) Adding 6 times of 55 ℃ deionized water into the whole oat superfine powder, stirring the oat milk for 20min at the rotating speed of 500rpm/min, shearing at the high speed of 6000rpm for 15min, and gelatinizing the oat milk completely sheared at 85 ℃ for 15min. And then carrying out enzymolysis, namely adding 2% of alpha-amylase, 0.6% of beta-amylase and 0.5% of cellulase into the gelatinized oat milk, wherein the enzymolysis temperature is 55 ℃, the time is 120min, the pH is regulated to 8 by sodium hydroxide, the temperature is regulated to 60 ℃, and 0.15% of alkaline protease is added for heating reaction for 30min to obtain an enzymolysis liquid.
(4) After enzyme deactivation, 7% sucrose was added to the oat milk. Homogenizing the mixture after uniform stirring, wherein the homogenizing condition is 25 ℃, 35MPa, and homogenizing for 3 times; sterilizing at 95deg.C for 10min, and rapidly cooling to 45deg.C to obtain uniform stable system.
(5) Inoculating strain, fermenting, measuring pH and acid value at the whole point, and after finishing fermentation when pH value of fermentation liquid is less than or equal to 4.5 and acidity is more than or equal to 30 DEG T, performing after-ripening to obtain oat plant-based fermented milk. The fermentation strain is lactobacillus casei and lactobacillus acidophilus, and the mixing ratio is 1:1, the concentration of the strains is 1 multiplied by 10 8 CFU/mL; the total inoculum size of the strain is 1% (v/v), the fermentation temperature is 37 ℃, the time is 16h, the post-maturation temperature is 4 ℃, and the time is 18h. The experimental results of the physicochemical indexes related to the oat plant-based fermented milk are shown in table 1 below.
Comparative example 1:
the procedure was as in example 1, except that the enzymatic hydrolysis treatment, fermentation time, etc. (see Table 1) were not performed, and the other treatment conditions were the same. The experimental results of the physicochemical indexes related to the obtained oat plant-based fermented milk are shown in the following table 1.
Comparative example 2:
the same as in example 1, except that the enzymes used in example 1 were changed to 1% alpha-amylase and 0.6% beta-amylase, the fermentation time was as shown in Table 1, and the other treatment conditions were the same. The experimental results of the physicochemical indexes related to the obtained oat plant-based fermented milk are shown in the following table 1.
Comparative example 3:
the same as in example 1, except that the enzymes used in example 1 were changed to 1% alpha-amylase, 0.2% pullulanase and 0.15% alkaline protease, and the fermentation time was as shown in Table 1, and the other treatment conditions were the same. The experimental results of the physicochemical indexes related to the obtained oat plant-based fermented milk are shown in the following table 1.
Comparative example 4:
the enzymes used in example 1 were changed to 1% alpha-amylase, 0.6% beta-amylase and 0.5% cellulase, the fermentation time is shown in Table 1, and the other treatment conditions were the same. The experimental results of the physicochemical indexes related to the obtained oat plant-based fermented milk are shown in the following table 1.
Comparative example 5:
the strain used in example 1 was changed to Lactobacillus plantarum (i.e., lactobacillus plantarum subspecies, accession number: CICC 22158), and the fermentation time was as shown in Table 1, except that the treatment conditions were the same. The experimental results of the physicochemical indexes related to the obtained oat plant-based fermented milk are shown in the following table 1.
Comparative example 6:
the strain used in example 1 was changed to lactobacillus acidophilus, the fermentation time was shown in table 1, and other treatment conditions were the same. The experimental results of the physicochemical indexes related to the obtained oat plant-based fermented milk are shown in the following table 1.
Comparative example 7:
the strain used in example 1 was changed to lactobacillus casei, the fermentation time was shown in table 1, and other treatment conditions were the same. The experimental results of the physicochemical indexes related to the obtained oat plant-based fermented milk are shown in the following table 1.
Comparative example 8:
the strain used in example 1 was changed to Lactobacillus bulgaricus (i.e., lactobacillus delbrueckii subspecies bulgaricus, purchased from Bio-technology Co., ltd., bio-52481, beijing) and the fermentation time was as shown in Table 1, and the other treatment conditions were the same. The experimental results of the physicochemical indexes related to the obtained oat plant-based fermented milk are shown in the following table 1.
Comparative example 9:
the strains used in example 1 were changed to Streptococcus thermophilus (from Bio-72807, biotechnology Co., european Bowei, beijing) and the fermentation times were as shown in Table 1, with the other treatment conditions being the same. The experimental results of the physicochemical indexes related to the obtained oat plant-based fermented milk are shown in the following table 1.
Comparative example 10:
the strain used in example 1 was changed to lactobacillus acidophilus and lactobacillus bulgaricus mix 1:1, fermentation time is shown in table 1, and other treatment conditions are the same. The experimental results of the physicochemical indexes related to the obtained oat plant-based fermented milk are shown in the following table 1.
Comparative example 11:
the strain used in example 1 was changed to lactobacillus acidophilus and streptococcus thermophilus mix 1:1, fermentation time is shown in table 1, and other treatment conditions are the same. The experimental results of the physicochemical indexes related to the obtained oat plant-based fermented milk are shown in the following table 1.
Comparative example 12:
the strain used in example 1 was changed to lactobacillus casei and lactobacillus bulgaricus mix 1:1, fermentation time is shown in table 1, and other treatment conditions are the same. The experimental results of the physicochemical indexes related to the obtained oat plant-based fermented milk are shown in the following table 2.
Comparative example 13:
the strain used in example 1 was changed to lactobacillus casei and streptococcus thermophilus mix 1:1, fermentation time is shown in table 1, and other treatment conditions are the same. The experimental results of the physicochemical indexes related to the obtained oat plant-based fermented milk are shown in the following table 1.
Comparative example 14:
the strain used in example 1 was changed to lactobacillus bulgaricus and streptococcus thermophilus blend 1:1, fermentation time is shown in table 1, and other treatment conditions are the same. The experimental results of the physicochemical indexes related to the obtained oat plant-based fermented milk are shown in the following table 1.
Test example:
the lactic acid bacteria prepared in examples 1 to 4 and comparative examples 1 to 14 were tested. The specific test method is as follows:
extraction of phenolic substances: samples (4 mL) of examples and comparative examples were collected, mixed with 20mL80% ethanol, and placed in an ultrasonic cleaner at 25℃for 20 minutes. The obtained liquid was centrifuged at 4000rpm at 4℃for 10 minutes, and the above steps were repeated three times. The supernatant was collected and concentrated at 40 ℃ using a vacuum rotary evaporator, and the liquid containing free phenolics was kept with methanol at a constant volume of 10mL and stored under dark conditions. The remaining precipitate was added to hexane to remove lipids. Then, the precipitate was hydrolyzed by adding 20mL of 4M NaOH, shaking for 1 hour, and adjusting the pH to 2.0-3.0 with 6M HCl. The mixture was extracted with 20mL ethyl acetate, then sonicated for 20 minutes, and centrifuged at 4000rpm for 10 minutes. The procedure was repeated three times to collect the supernatant and evaporated in vacuo at 40 ℃. The liquid containing bound phenolics was kept at a constant volume of 10mL with methanol and stored under dark conditions.
And (3) phenol content determination: 0.25mL of the extract was mixed with 1mL of distilled water, and 0.25mL of Folin-Ciocalteu phenol reagent was added to conduct a reaction for 6 minutes. Then, 2.5mL of 7% Na 2 CO 3 And methanol was added to a total volume of 10 mL. The obtained liquid was incubated in the dark at room temperature for 90 minutes. Subsequently, the absorbance was measured at 760nm on a spectrophotometer. Phenolic compounds were quantified using a gallic acid standard calibration curve. Results are expressed in milligrams Gallic Acid Equivalent (GAE)/1L sample.
Determination of the pH value: the pH value was measured with a PB-10 type high-precision acidometer.
Determination of acidity: the acidity of the fermented milk was measured by the first method using the phenolphthalein indicator method, which was performed with reference to GB 5009.239-2016, determination of acidity in food safety national Standard food.
Measurement of viable cell count: the viable count of lactic acid bacteria is determined by referring to the method in GB 4789.35-2016 "food safety national Standard food microorganism test lactic acid bacteria test".
The test results are shown in Table 1.
Table 1 test data for examples and comparative examples
Examples 1-4 process optimization of alpha-amylase addition in enzymatic treatments it was necessary to study alpha-amylase addition because oat flour contained a large amount of starch. As shown in Table 1, the system of oat milk may be unstable due to light enzymolysis or excessive enzymolysis, which is unfavorable for strain fermentation, and results in reduced fermentation acidity and prolonged fermentation time. The added amount of the alpha-amylase has the greatest influence on the stability of the oat milk system, and meanwhile, the enzymolysis effect is relatively more obvious by using 1% of the added amount of the alpha-amylase.
As is clear from the comparative examples, the polyphenol content was significantly reduced when no enzymatic treatment was performed. When the enzyme type is changed, the fermentation time is longer than that of the examples, the acidity is lower, and the polyphenol content is lower. When the species of bacteria are changed or single bacteria are used for fermentation, the total polyphenol content is low, the acid production speed is low, and particularly in comparative example 9, when streptococcus thermophilus is singly used, the fermentation time is up to 20 hours, the acidity is low, and the polyphenol content is low. Meanwhile, as can be seen from examples and comparative examples, the presence of lactobacillus casei increases the polyphenol content, particularly the combination of lactobacillus casei and lactobacillus acidophilus, more advantageously increases the polyphenol content, and shortens the fermentation time, compared with other strains.
The preparation method of oat plant-based fermented milk based on the composite enzymolysis-fermentation combined technology for enriching polyphenol provided by the invention effectively improves the total polyphenol content and the lactic acid bacteria acid production rate, can improve the acidity of oat plant-based fermented milk in a short time, and shortens the fermentation time.
The above is only a preferred embodiment of the present invention, and the present invention is not limited to the above examples. It is to be understood that other modifications and variations which may be directly derived or contemplated by those skilled in the art without departing from the spirit and concepts of the present invention are deemed to be included within the scope of the present invention.

Claims (10)

1. The preparation method of the oat plant-based fermented milk for enriching polyphenol based on the combined enzymolysis-fermentation technology is characterized by comprising the following steps of:
(1) Soaking, cleaning and drying oat to obtain pretreated oat;
(2) Baking, crushing and ball milling the pretreated oat to obtain whole oat superfine powder;
(3) Adding water into whole wheat superfine powder to obtain oat milk, stirring, shearing at high speed, gelatinizing the completely dissolved oat milk, and adding enzyme for enzymolysis to obtain enzymolysis solution;
(4) After enzyme deactivation, preparing and homogenizing to obtain a homogeneous liquid, sterilizing and cooling to obtain a uniform and stable system;
(5) Adding strain for inoculation, fermentation and after-ripening to obtain the oat plant-based fermented milk.
2. The method of claim 1, wherein in step (1), the oat is whole oat kernel.
3. The method according to claim 1, wherein in the step (2), the baking temperature is 170-190 ℃ for 10-20min, and the baking is performed once every 2-4 min.
4. The method according to claim 1, wherein in the step (3), the volume ratio of the Quan Maichao micro powder to the water is 1:5-8; the temperature of the water is 50-55 ℃.
5. The method according to claim 1, wherein in the step (3), the stirring speed is 300 to 500r/min for 10 to 20min; the rotating speed of the high-speed shearing is 4000-7000r/min, and the shearing time is 10-20min; preferably, the rotating speed of the high-speed shearing is 6000rpm, and the shearing time is 10-15min; the gelatinization temperature is 80-90deg.C, and the time is 10-20min.
6. The method according to claim 1, wherein in the step (3), the enzyme comprises an α -amylase, a β -amylase, a cellulase, an alkaline protease; the addition amount of each enzyme is 0.1% -2%; the specific process of enzymolysis by adding enzyme is as follows: firstly adding alpha-amylase, beta-amylase and cellulase to react for 90-120min at 55-65 ℃, then changing enzymolysis conditions to carry out proteolysis, namely adjusting the pH value to 8 by sodium hydroxide, adjusting the temperature to 60 ℃, and adding alkaline protease to react for 30-60min.
7. The method of claim 1, wherein in step (4), the formulation is 1% -7% sucrose added.
8. The method according to claim 1, wherein in the step (4), the homogenizing pressure is 30 to 40MPa and the temperature is 20 to 30 ℃; the cooling refers to cooling to 43-48 ℃.
9. The method according to claim 1, wherein in the step (5), the strains are lactobacillus acidophilus and lactobacillus casei, and the ratio of the added two strains is 1:1 by volume.
10. An oat plant-based fermented milk prepared by the method of any one of claims 1-9.
CN202310708269.4A 2023-06-15 2023-06-15 Preparation method of oat plant-based fermented milk for enriching polyphenol based on composite enzymolysis-fermentation combined technology Pending CN117770317A (en)

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