CN112680302B - Low-intoxication-causing refined beer containing hyaluronic acid oligosaccharide and process thereof - Google Patents
Low-intoxication-causing refined beer containing hyaluronic acid oligosaccharide and process thereof Download PDFInfo
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Abstract
The refined beer is prepared by selecting high-concentration concentrated wort and a high-grade alcohol low-yield condensed yeast strain, prolonging the hop boiling time and carrying out a micro-pressure and pressure fermentation process, and can reduce the content of high-grade alcohol in the finished product refined beer, improve the content of ester substances, reduce the alcohol ester ratio and further reduce the intoxication under the condition of not influencing the style and the flavor; in addition, a fermentation process of recombinant bacillus subtilis as a genetically engineered bacterium for producing hyaluronic acid oligosaccharide with high yield is added, the hyaluronic acid oligosaccharide can be produced by metabolism in the fermentation process, the yield is 30-70mg/L, and the average molecular weight is about 0.8 multiplied by 104Da, it is easy to be absorbed by human body when orally taken. The refined beer provided by the invention is rich in taste, is nutritional and healthy, has low intoxication, and has the effects of improving human immunity, maintaining beauty and keeping young, and protecting internal organs such as liver.
Description
Technical Field
The invention belongs to the technical field of alcoholic beverages, and particularly relates to low-intoxication brewed beer containing hyaluronic acid oligosaccharide and a process thereof.
Background
Beer is the beverage ranked third in global sales volume after water and tea are drunk, China is the biggest beer producing country in the world, the domestic industry mass beer market tends to be saturated, and the brewed beer is the new high-end beer industry, has richer flavor characteristics, mellow taste and longer aftertaste, caters to the change of pursuit of consumers on consumption concept, health concept and personalized features, and presents the trend of rapid development.
Consumers have worries about hurting, getting up and getting drunk while intoxicating in the strong aroma and the complex and changeable taste of the refined beer. The refined beer is brewed with pure malt, the concentration of the malt extract is higher, and higher alcohol is one of yeast metabolites in the fermentation process of the malt extract. The high alcohol in the refined beer has the function of highlighting the characteristic fragrance and full taste of the beer, and the drunkenness phenomenon of headache, dizziness, nausea and vomiting can be caused after the content of the high alcohol exceeds a certain limit. Because higher alcohol is slowly oxidized and metabolized, the physiological function of the central nervous system of the brain can be inhibited by too high content of the higher alcohol, and partial brain injury, liver injury and stomach injury are caused. The high content of ester substances can stretch nerves, mediate alcohol metabolism, and the two resist each other, so that the perception of the human body to the phenomenon of 'top' is adjusted. Therefore, the research on the low-intoxication brewed beer capable of reducing the content of higher alcohols and improving the content of esters has a far-reaching practical significance.
Disclosure of Invention
Therefore, the invention provides the low-intoxication home-brewed beer containing the hyaluronic acid oligosaccharide and the process thereof, which can reduce the content of higher alcohol in the home-brewed beer and improve the low-intoxication of ester content.
In order to achieve the above purpose, the invention provides the following technical scheme: the low-intoxication home-brewed beer containing the hyaluronic acid oligosaccharide comprises the following components in percentage by weight: 8-11% high concentration concentrated wort; 0.16-0.55% of concentrated fruit juice; 80-90% of water;
hops accounting for 0.03-5% of the weight of the high-concentration concentrated wort are added into the low-intoxication fine-brewed beer;
inoculating recombinant bacillus subtilis seed solution to the low-intoxication refined beer according to the volume ratio of 1: 10000;
the low-intoxication refined beer is added with the high alcohol low-yield condensed yeast seed liquid with the weight of 0.1-0.3 per mill of the weight of the high-concentration concentrated wort.
As a preferable scheme of the low-intoxication concentrated brewed beer containing the hyaluronic acid oligosaccharide, the concentrated fruit juice is at least one of strawberry concentrated juice, passion fruit concentrated juice, cranberry concentrated juice, blackcurrant concentrated juice and blueberry concentrated juice.
As a preferable embodiment of the low-intoxicating home-brewed beer containing the hyaluronic acid oligosaccharide, the hop is at least one of pearl and cassett.
As a preferable scheme of the low-intoxication home-brewed beer containing the hyaluronic acid oligosaccharide, the recombinant bacillus subtilis is a high-hyaluronic acid oligosaccharide-producing genetically engineered bacterium, namely recombinant bacillus subtilis, and the recombinant bacillus subtilis generates the hyaluronic acid oligosaccharide through self metabolism.
As a preferable embodiment of the low-intoxicating home-brewed beer containing the hyaluronan oligosaccharide, the higher alcohol low-production condensing yeast strain is QC1118 yeast strain.
The invention also provides a process for brewing the low-intoxication home-brewed beer containing the hyaluronic acid oligosaccharide, which comprises the following steps:
diluting the high-concentration concentrated wort with water until the sugar content is 7-12 DEG P, adding hops accounting for 0.03-5% of the weight of the high-concentration concentrated wort, prolonging the boiling time, and cooling to 37-39 ℃;
inoculating the recombinant bacillus subtilis seed solution according to the volume of 1:10000 for fermentation, and after the fermentation is finished, carrying out high-temperature sterilization and cooling to 16-18 ℃;
inoculating high-concentration concentrated wort 0.1-0.3 wt% of high-alcohol low-yield condensed yeast seed liquid, and the number of yeast in the full tank is 3-5 × 107and/mL, carrying out pressurized fermentation at the temperature of 18-20 ℃, and obtaining the low-intoxication brewed beer after the sugar degree is reduced to 4-6P degrees.
As a preferred embodiment of the process for brewing a low-intoxicating concentrated beer containing hyaluronan oligosaccharide, the high-concentration concentrated wort is prepared by:
crushing malt to the particle size of 400-800 um, preserving heat at 45-50 ℃ for 20min, performing low-temperature extraction, saccharifying at 60-65 ℃ for 80-120 min, filtering to obtain beer wort with over 20wt% of dry matter, performing vacuum concentration at 50-80 ℃ until the dry matter is over 75wt%, filling, refrigerating and storing to obtain the beer wort.
As a preferred scheme of the process for brewing the low-intoxication fine beer containing the hyaluronic acid oligosaccharide, after hops accounting for 0.03-5% of the weight of the high-concentration concentrated wort are added, the boiling time is prolonged from 60min to 90-120 min;
after fermentation, high-temperature sterilization is carried out for 5-10 min under the conditions of 121-125 ℃.
As a preferred scheme of the process for brewing the low-intoxication home-brewed beer containing the hyaluronic acid oligosaccharide, the preparation process of the recombinant bacillus subtilis seed solution comprises the following steps:
and (3) picking the bacterial colony cultured on the fresh solid culture medium into a seed shake flask, and culturing at 37 ℃ and 170-180 rpm for 16-18 h to serve as seed liquid.
As a preferable scheme of the process for brewing the low-intoxication home-brewed beer containing the hyaluronic acid oligosaccharide, the fermentation conditions of the recombinant bacillus subtilis are as follows: the pressure of the tank is 0.02-0.05 MPa, the temperature is 37 ℃, and the culture time is 24-36 h.
In a preferable embodiment of the process for brewing the low-intoxication fine beer containing the hyaluronic acid oligosaccharide, the fermentation condition of the high alcohol low-production condensed yeast under pressure is 0.04 to 0.05 MPa.
Compared with the prior art, the invention has the beneficial effects that:
firstly, a high-concentration concentrated wort and a high-alcohol low-yield condensed yeast strain are selected, the boiling time is prolonged, and a micro-pressure pressurized fermentation process is adopted, so that the content of high-alcohol in finished product refined beer can be reduced, the content of ester substances can be improved, the alcohol ester ratio can be reduced under the condition of not influencing the style and the flavor, the content of the high-alcohol in the refined beer obtained by the traditional fermentation process is 40-50%, the alcohol ester ratio can be effectively monitored, and the intoxication can be reduced;
secondly, selecting high-concentration concentrated wort: compared with the traditional direct malt fermentation, the method has the advantages that two-step extraction is added, so that the protein content and volatile flavor impurities in the wort are reduced, the impurities available for yeast fermentation are reduced, and the content of higher alcohol generated in the yeast metabolic process is reduced;
thirdly, selecting a high-alcohol low-yield condensed yeast strain: compared with common beer fermentation yeast, the method regulates and controls the metabolic pathway of higher alcohol, and obviously reduces the generation amount of the higher alcohol;
fourthly, prolonging the boiling time: the hops contain 4-8% of polyphenol substances, and when the hops are boiled, proteins in the wort can be separated out in a complexing mode to form a thermal coagulum, the boiling time is prolonged, the proteins in the wort can be separated out more completely, and the content of higher alcohols generated in an amino acid degradation way is effectively reduced;
fifthly, a micro-pressure belt pressing process: compared with conventional fermentation, the method can reduce the content of higher alcohol, effectively promote the generation of ester substances in the refined beer, help to reduce the alcohol ester ratio of the refined beer and weaken the intoxication;
sixthly, the fermentation process of the recombinant bacillus subtilis which is the genetically engineered bacterium for increasing the high yield of the hyaluronic acid oligosaccharide can be used for producing the hyaluronic acid oligosaccharide by metabolism in the fermentation process, the yield is 30-70mg/L, and the average molecular weight is about 0.8 multiplied by 104Da, it is easy to be absorbed by human body when orally taken.
Detailed Description
The present invention is described in terms of particular embodiments, other advantages and features of the invention will become apparent to those skilled in the art from the following disclosure, and it is to be understood that the described embodiments are merely exemplary of the invention and that it is not intended to limit the invention to the particular embodiments disclosed. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Example 1
The embodiment 1 of the invention provides a low-intoxication home-brewed beer containing hyaluronic acid oligosaccharide, and the formula of the low-intoxication home-brewed beer comprises the following components in percentage by weight: 8% high-concentration concentrated wort; 0.55% concentrated juice; 90% water;
the low-intoxication fine-brewed beer contains hops 0.03 wt% of the high-concentration concentrated wort;
inoculating recombinant bacillus subtilis seed solution to the low-intoxication refined beer according to the volume ratio of 1: 10000;
the low-intoxication refined beer is added with a high alcohol low-yield coagulation type yeast seed solution with the weight of 0.1 per mill of the weight of the high-concentration concentrated wort.
In this embodiment, the concentrated juice is passion fruit concentrated juice. The hops are pearls and cassettes, and the content ratio of the pearls to the cassettes is 1: 2.
In the embodiment, the recombinant bacillus subtilis is a high-yield hyaluronic acid oligosaccharide gene engineering bacterium, a Hyaluronidase (HAS) gene in streptococcus zooepidemicus is cloned, and bacillus subtilis 168 is used as a host to construct a new metabolic pathway to form the recombinant bacillus subtilis, so that hyaluronic acid oligosaccharide can be generated through self metabolism of the recombinant bacillus subtilis and purchased in Jilin university.
In this example, the higher alcohol low-yield condensed yeast strain QC1118 is an EC1118 strain whose higher alcohol metabolic pathway is regulated by genome rearrangement and homologous recombination techniques to obtain a higher alcohol low-yield condensed yeast strain purchased from tianjin university.
The process for brewing low-intoxicating beer containing hyaluronan oligosaccharide of example 1, comprising the steps of:
diluting the high concentration concentrated wort with water until the sugar degree is 7 ° P, adding hops 0.03 wt% of the high concentration concentrated wort, boiling for a prolonged period, and cooling to 37 deg.C;
inoculating the recombinant bacillus subtilis seed solution according to the volume of 1:10000 for fermentation, and after the fermentation is finished, carrying out high-temperature sterilization and cooling to 16 ℃;
inoculating high-concentration concentrated wort 0.1 ‰ of high alcohol low-yield coagulated yeast seed liquid with full-tank yeast number of 3 × 107and/mL, fermenting under pressure at 18 ℃, and obtaining the low-intoxication brewed beer after the sugar degree is reduced to 4 ℃ P and stabilized.
Specifically, the preparation process of the high-concentration concentrated wort is as follows:
crushing malt to the particle size of 400-500 um, preserving heat at 45 ℃ for 20min, performing low-temperature extraction, saccharifying at 60 ℃ for 120min, filtering to obtain beer wort with over 20wt% of dry matter, performing vacuum concentration at 50 ℃ until the dry matter is over 75wt%, filling, refrigerating and storing to obtain the beer wort.
In this example, the boiling time was extended from 60min to 90min after the addition of hops in an amount of 0.03% by weight of the high concentrated wort; after fermentation, high temperature sterilization is carried out at 121 ℃ for 10 min.
In this embodiment, the preparation process of the recombinant bacillus subtilis seed solution is as follows:
the colony cultured from the fresh solid culture medium is selected into a seed shake flask and cultured at 37 ℃ and 170rpm for 18h to serve as seed liquid.
In this embodiment, the fermentation conditions of the recombinant bacillus subtilis are as follows: the pressure of the tank is 0.02MPa, the temperature is 37 ℃, and the culture time is 24 h.
In the embodiment, the high alcohol low-yield condensed yeast seed solution is obtained by inoculating an activated strain on a flat plate into a primary seed culture medium, performing static culture at 30 ℃ for 24 hours, inoculating into a secondary seed culture medium according to 10% of the inoculation amount, and performing static culture at 30 ℃ for 16 hours. The pressure fermentation condition of the high alcohol low-yield condensed yeast is 0.04 MPa.
Example 2
The embodiment 2 of the invention provides a low-intoxication home-brewed beer containing hyaluronic acid oligosaccharide, and the formula of the low-intoxication home-brewed beer comprises the following components in percentage by weight: 10% high-concentration concentrated wort; 0.35% concentrated juice; 85% of water;
the low-intoxication fine-brewed beer contains hops 2.8% by weight of the high-concentration concentrated wort;
inoculating recombinant bacillus subtilis seed solution to the low-intoxication refined beer according to the volume ratio of 1: 10000;
the low-intoxication refined beer is added with a high alcohol low-yield coagulation type yeast seed solution with the weight of 0.2 per mill of the weight of the high-concentration concentrated wort.
In this embodiment, the concentrated juice is strawberry concentrated juice. The hops are pearls and cassettes, and the content ratio of the pearls to the cassettes is 1: 2.
In the embodiment, the recombinant bacillus subtilis is a high-yield hyaluronic acid oligosaccharide gene engineering bacterium, a Hyaluronidase (HAS) gene in streptococcus zooepidemicus is cloned, and bacillus subtilis 168 is used as a host to construct a new metabolic pathway to form the recombinant bacillus subtilis, so that hyaluronic acid oligosaccharide can be generated through self metabolism of the recombinant bacillus subtilis and purchased in Jilin university.
In this example, the higher alcohol low-yield condensed yeast strain QC1118 is an EC1118 strain whose higher alcohol metabolic pathway is regulated by genome rearrangement and homologous recombination techniques to obtain a higher alcohol low-yield condensed yeast strain purchased from tianjin university.
The process for brewing low-intoxicating beer containing hyaluronan oligosaccharide of example 2, comprising the steps of:
diluting the high concentration concentrated wort with water until the sugar degree is 10 ° P, adding hops 2.8% of the high concentration concentrated wort, boiling for a prolonged period, and cooling to 38 deg.C;
inoculating the recombinant bacillus subtilis seed solution according to the volume of 1:10000 for fermentation, and after the fermentation is finished, carrying out high-temperature sterilization and cooling to 17 ℃;
inoculating high-concentration concentrated wort 0.2 ‰ of high alcohol low-yield coagulated yeast seed liquid with full yeast number of 4 × 107and/mL, fermenting under pressure at 19 ℃, and obtaining the low-intoxication refined beer after the sugar degree is reduced to 5 DEG P.
Specifically, the preparation process of the high-concentration concentrated wort is as follows:
crushing malt to the particle size of 500-600 um, preserving heat at 48 ℃ for 20min for low-temperature extraction, saccharifying at 62 ℃ for 90min, filtering to obtain beer wort with over 20wt% of dry matter, vacuum concentrating at 65 ℃ until over 75wt% of dry matter, filling, refrigerating and storing to obtain the beer wort.
In this example, after addition of hops in an amount of 2.8% by weight of the high concentrated wort, the boiling time was increased from 60min to 100 min; after fermentation, high temperature sterilization is carried out at 123 ℃ for 8 min.
In this embodiment, the preparation process of the recombinant bacillus subtilis seed solution is as follows:
the colony cultured from the fresh solid culture medium is selected into a seed shake flask and cultured at 37 ℃ and 175rpm for 17 hours to serve as seed liquid.
In this embodiment, the fermentation conditions of the recombinant bacillus subtilis are as follows: the pressure of the tank is 0.04MPa, the temperature is 37 ℃, and the culture time is 28 h.
In the embodiment, the high alcohol low-yield condensed yeast seed solution is obtained by inoculating an activated strain on a flat plate into a primary seed culture medium, performing static culture at 30 ℃ for 24 hours, inoculating into a secondary seed culture medium according to 10% of inoculation amount, and performing static culture at 30 ℃ for 16 hours. The pressure fermentation condition of the high alcohol low-yield condensed yeast is 0.05 MPa.
Example 3
The embodiment 3 of the invention provides a low-intoxication home-brewed beer containing hyaluronic acid oligosaccharide, and the formula of the low-intoxication home-brewed beer comprises the following components in percentage by weight: 11% high-strength concentrated wort; 0.16% concentrated juice; 80% water;
the low-intoxication fine-brewed beer is added with hops with a concentration of 5% by weight of concentrated wort;
inoculating recombinant bacillus subtilis seed solution to the low-intoxication refined beer according to the volume ratio of 1: 10000;
the low-intoxication refined beer is added with a high alcohol low-yield coagulation type yeast seed solution with the weight of 0.3 per mill of the weight of the high-concentration concentrated wort.
In this embodiment, the concentrated juice is cranberry concentrated juice, blackcurrant concentrated juice, or blueberry concentrated juice.
The hops are pearls and cassettes, and the content ratio of the pearls to the cassettes is 1: 2.
In the embodiment, the recombinant bacillus subtilis is a high-yield hyaluronic acid oligosaccharide gene engineering bacterium, a Hyaluronidase (HAS) gene in streptococcus zooepidemicus is cloned, and bacillus subtilis 168 is used as a host to construct a new metabolic pathway to form the recombinant bacillus subtilis, so that hyaluronic acid oligosaccharide can be generated through self metabolism of the recombinant bacillus subtilis and purchased in Jilin university.
In this example, the higher alcohol low-yield condensed yeast strain QC1118 is an EC1118 strain whose higher alcohol metabolic pathway is regulated by genome rearrangement and homologous recombination techniques to obtain a higher alcohol low-yield condensed yeast strain purchased from tianjin university.
The process for brewing low-intoxicating beer containing hyaluronan oligosaccharide of example 3, comprising the steps of:
diluting the high concentration concentrated wort with water until the sugar degree is 12 ° P, adding hops 5% of the high concentration concentrated wort weight, prolonging boiling time, and cooling to 39 deg.C;
inoculating the recombinant bacillus subtilis seed solution according to the volume of 1:10000 for fermentation, and after the fermentation is finished, carrying out high-temperature sterilization and cooling to 18 ℃;
inoculating high-concentration concentrated malt juice with 0.1 ‰ weight of higher alcohol, and fermenting at 5 × 107and/mL, fermenting under pressure at the temperature of 20 ℃, and obtaining the low-intoxication refined beer after the sugar degree is reduced to 6 DEG P.
In this example, the process for preparing the high-strength concentrated wort was:
crushing malt to the particle size of 600-800 um, keeping the temperature at 50 ℃ for 20min for low-temperature extraction, saccharifying at 60-65 ℃ for 80min, filtering to obtain beer wort with over 20wt% of dry matter, vacuum concentrating at 80 ℃ until over 75wt% of dry matter, filling, refrigerating and storing to obtain the beer wort.
In this example, after the addition of 5% by weight of hops based on the high concentrated wort, the boiling time was extended from 60min to 120 min; after fermentation, high temperature sterilization is carried out at 125 ℃ for 5 min.
In this embodiment, the preparation process of the recombinant bacillus subtilis seed solution is as follows:
the colony cultured from the fresh solid culture medium is selected into a seed shake flask and cultured for 16h at 37 ℃ and 180rpm as seed liquid.
In this embodiment, the fermentation conditions of the recombinant bacillus subtilis are as follows: the pressure of the tank is 0.05MPa, the temperature is 37 ℃, and the culture time is 36 h.
In the embodiment, the high alcohol low-yield condensed yeast seed solution is obtained by inoculating an activated strain on a flat plate into a primary seed culture medium, performing static culture at 30 ℃ for 24 hours, inoculating into a secondary seed culture medium according to 10% of inoculation amount, and performing static culture at 30 ℃ for 16 hours. The pressure fermentation condition of the high alcohol low-yield condensed yeast is 0.05 MPa.
Comparative example 1
This comparative example 1 is different from example 1 in that a high concentrated wort is not selected and conventional malt fermentation is used. Crushing the malt to the grain size of 400-500 um, preserving heat at 45 ℃ for 20min, saccharifying at 60 ℃ for 120min, filtering, adding hops and boiling, wherein the steps of the method are the same.
Comparative example 2
This comparative example 2 is different from example 1 in that a higher alcohol low-yield coacervation type yeast QC1118 was not selected, but a common Saccharomyces cerevisiae EC1118 was used, except that the process steps were the same.
Comparative example 3
Comparative example 3 is compared with example 1 with the difference that the boiling time is 60min, except that the process steps are identical.
Comparative example 4
Comparative example 4 is different from example 1 in that the micropressure and pressure process is not used, but the fermentation is not carried out under pressure, except that the other steps are the same.
Comparative example 5
Compared with the example 1, the comparative example 5 is different from the example 1 in that the fermentation process of the recombinant bacillus subtilis which is a genetically engineered bacterium for increasing the high-yield hyaluronic acid oligosaccharide is eliminated, except that the steps of the method are the same.
Control group
The method comprises the steps of adopting a conventional fermentation process of refined beer, crushing malt to 400-500 um in particle size, preserving heat at 45 ℃ for 20min, adding water, saccharifying at 60 ℃ for 120min, filtering, adding hops accounting for 0.03% of the weight of concentrated wort, boiling for 60min, cooling to 16 ℃, inoculating conventional saccharomyces cerevisiae EC1118 strain seed liquid accounting for 0.1% of the weight of the concentrated wort, fermenting at 18 ℃ without pressure until the sugar degree is reduced to 4-degree P stably, and obtaining the beer.
Examples of the experiments
9 kinds of refined beers provided by example 1, example 2, example 3, comparative example 1, comparative example 2, comparative example 3, comparative example 4, comparative example 5 and a control group are respectively sampled, the contents of higher alcohols and total esters are determined by gas chromatography, and the content of the total esters is determined by an ultraviolet spectrophotometer, wherein the content of the higher alcohols is the sum of the contents of tertiary butanol, normal propanol, isobutanol, normal butanol, isoamyl alcohol, normal pentanol, benzyl alcohol and beta-phenylethyl alcohol, and the content of the total esters is the sum of the contents of ethyl acetate and isoamyl acetate. The detection results are shown in table 1:
TABLE 1 nine kinds of refined beer sampling and testing results
According to the above table, the detection results of the embodiment 1, the embodiment 2, the embodiment 3 and the control group show that the low-intoxication brewed beer containing hyaluronic acid oligosaccharide and the process provided by the invention can effectively reduce the content of higher alcohol by more than 35.3%, improve the total ester content by more than 22.2%, reduce the alcohol ester ratio by more than 46.2% and increase the content of hyaluronic acid oligosaccharide by more than 54.41 mg/L.
As can be seen from example 1 and comparative example 1, the higher alcohol content of the selected high-concentration concentrated wort is reduced by 20.7% compared with that of the conventional malt fermentation;
as can be seen from the example 1 and the comparative example 2, compared with the selection of the common saccharomyces cerevisiae EC1118, the selection of the higher alcohol low-yield condensed yeast QC1118 has the advantage that the higher alcohol content is reduced by 22.2 percent;
as can be seen from example 1 and comparative example 3, the higher alcohol content is reduced by 10.9% by prolonging the boiling time to 90min as compared with the conventional boiling time of 60 min;
as can be seen from the example 1 and the comparative example 4, compared with the non-pressure fermentation, the total ester content is reduced by 16.4% by adopting the micro-pressure and pressure process;
as can be seen from the example 1 and the comparative example 5, the fermentation process of the recombinant bacillus subtilis which is a high-yield hyaluronic acid oligosaccharide genetic engineering bacterium is increased, and the content of hyaluronic acid oligosaccharide is increased by 54.41 mg/L;
hyaluronic Acid (HA), also known as hyaluronic acid, is now classified as an important medical application material and cosmetic additive raw material as a new food resource. The high molecular weight HA (molecular weight over 100 ten thousand Da) HAs good elasticity and moisture retention, and is not easy to be absorbed by human body when orally taken. HA oligosaccharide (molecular weight less than 1 ten thousand Da) HAs important physiological activity and special physiological function, and can promote skin to absorb nutrition, increase skin elasticity, delay aging, promote angiogenesis and wound healing, regulate immunocompetence and resist tumor, and is easier to absorb. Therefore, the invention can greatly meet the consumption requirements of people on the nutrition and health care of the brewed beer.
In conclusion, the low-intoxication refined beer containing hyaluronic acid oligosaccharide and the process thereof select the high-concentration concentrated wort, the high-grade alcohol low-yield condensed yeast strain, prolong the boiling time and carry out the fermentation process under the micro pressure and the pressure, can reduce the content of the high-grade alcohol in the finished product refined beer, improve the content of ester substances and reduce the alcohol ester ratio under the condition of not influencing the style and the flavor, are 40-50 percent of the content of the high-grade alcohol in the refined beer obtained by the traditional fermentation process, can effectively monitor the alcohol ester ratio and reduce the intoxication; selecting high-concentration concentrated wort: compared with the traditional direct malt fermentation, the method has the advantages that two-step extraction is added, so that the protein content and volatile flavor impurities in the wort are reduced, the impurities available for yeast fermentation are reduced, and the content of higher alcohol generated in the yeast metabolic process is reduced; selecting a high-alcohol low-yield condensed yeast strain: compared with common beer fermentation yeast, the method regulates and controls the metabolic pathway of higher alcohol, and obviously reduces the generation amount of the higher alcohol; and (3) prolonging the boiling time: the hops contain 4-8% of polyphenol substances, and when the hops are boiled, proteins in the wort can be separated out in a complexing mode to form a thermal coagulum, the boiling time is prolonged, the proteins in the wort can be separated out more completely, and the content of higher alcohols generated in an amino acid degradation way is effectively reduced; a micro-pressure belt pressing process: compared with conventional fermentation, the method can reduce the content of higher alcohol, effectively promote the generation of ester substances in the refined beer, help to reduce the alcohol ester ratio of the refined beer and weaken the intoxication; the fermentation process of recombinant Bacillus subtilis as high yield hyaluronic acid oligosaccharide gene engineering bacteria can produce hyaluronic acid oligosaccharide in 30-70mg/L amount and average molecular weight of 0.8X 104Da, it is easy to be absorbed by human body when orally taken. The invention can powerfully promote the development of the refined beer to the nutritional and healthy direction and is worthy of popularization and application.
Although the invention has been described in detail above with reference to a general description and specific examples, it will be apparent to one skilled in the art that modifications or improvements may be made thereto based on the invention. Accordingly, such modifications and improvements are intended to be within the scope of the invention as claimed.
Claims (8)
1. The low-intoxication home-brewed beer containing the hyaluronic acid oligosaccharide is characterized by comprising the following components in percentage by weight: 8-11% high concentration concentrated wort; 0.16-0.55% of concentrated fruit juice; 80-90% of water;
hops accounting for 0.03-5% of the weight of the high-concentration concentrated wort are added into the low-intoxication fine-brewed beer;
inoculating a recombinant bacillus subtilis seed solution into the low-intoxication refined beer according to the volume ratio of 1:10000, wherein the recombinant bacillus subtilis is a high-yield hyaluronic acid oligosaccharide genetic engineering bacterium-recombinant bacillus subtilis, and the recombinant bacillus subtilis generates hyaluronic acid oligosaccharide through self metabolism;
the low-intoxication refined beer is added with a high-alcohol low-yield condensed yeast seed solution with the weight of 0.1-0.3 per mill of the weight of high-concentration concentrated wort, the high-alcohol low-yield condensed yeast is fermented under pressure, and the fermentation condition under pressure is 0.04-0.05 MPa micro-pressure.
2. The low-intoxication brewed beer containing hyaluronan oligosaccharides according to claim 1, wherein the fruit juice concentrate is at least one of strawberry juice concentrate, passion fruit juice concentrate, cranberry juice concentrate, blackcurrant juice concentrate, and blueberry juice concentrate.
3. The low intoxication brewed beer containing hyaluronan oligosaccharide according to claim 1, wherein the hop is at least one of pearl and cassett.
4. The low-intoxication home-brewed beer containing hyaluronan oligosaccharides according to claim 1, wherein the higher alcohol low-production condensing yeast strain is QC1118 yeast strain.
5. The process for brewing the low-intoxication home-brewed beer containing the hyaluronic acid oligosaccharide is characterized by comprising the following steps of:
diluting the high-concentration concentrated wort with water until the sugar degree is 7-12 DEG P, adding hops accounting for 0.03-5% of the weight of the high-concentration concentrated wort, prolonging the boiling time from 60min to 90-120 min, and then cooling to 37-39 ℃;
inoculating a recombinant bacillus subtilis seed solution according to the volume of 1:10000 for fermentation, performing high-temperature sterilization after fermentation is completed, and cooling to 16-18 ℃, wherein the recombinant bacillus subtilis is a high-yield hyaluronic acid oligosaccharide genetically engineered bacterium, namely recombinant bacillus subtilis, and the recombinant bacillus subtilis generates hyaluronic acid oligosaccharide through self metabolism;
inoculating high-concentration concentrated wort 0.1-0.3 wt% of high-alcohol low-yield condensed yeast seed liquid, and the number of yeast in the full tank is 3-5 × 107and/mL, carrying out pressure fermentation at 18-20 ℃, wherein the pressure fermentation condition is 0.04-0.05 MPa of micro pressure, and obtaining the low-intoxication refined beer after the sugar degree is reduced to 4-6P degrees.
6. The process of claim 5, wherein the concentrated high gravity wort is prepared by:
crushing malt to the particle size of 400-800 um, preserving heat at 45-50 ℃ for 20min, performing low-temperature extraction, saccharifying at 60-65 ℃ for 80-120 min, filtering to obtain beer wort with over 20wt% of dry matter, performing vacuum concentration at 50-80 ℃ until the dry matter is over 75wt%, filling, refrigerating and storing to obtain the beer wort.
7. The process for brewing the low-intoxication concentrated beer containing the hyaluronic acid oligosaccharide according to claim 5, wherein the high-temperature sterilization is carried out at 121-125 ℃ for 5-10 min after the fermentation is finished.
8. The process of claim 5, wherein the recombinant Bacillus subtilis seed solution is prepared by the following steps:
picking bacterial colonies cultured on a fresh solid culture medium into a seed shake flask, and culturing at 37 ℃ and 170-180 rpm for 16-18 h to serve as seed liquid;
the fermentation conditions of the recombinant bacillus subtilis are as follows: the tank pressure is 0.02-0.05 MPa, the temperature is 37 ℃, and the fermentation time is 24-36 h.
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