CN115211465A

CN115211465A - Compound leavening agent for relieving blood pressure rise, brown lactobacillus beverage and preparation method thereof

Info

Publication number: CN115211465A
Application number: CN202210869020.7A
Authority: CN
Inventors: 方曙光; 王命荣; 郭晓娟; 汪欣; 朱建国
Original assignee: WeCare Probiotics Co Ltd
Current assignee: WeCare Probiotics Co Ltd
Priority date: 2022-07-22
Filing date: 2022-07-22
Publication date: 2022-10-21
Anticipated expiration: 2042-07-22
Also published as: CN115211465B

Abstract

The invention provides a compound leaven for relieving blood pressure rise, a brown lactobacillus beverage and a preparation method thereof. The compound leaven for relieving the rise of blood pressure comprises lactobacillus plantarum LP90, lactobacillus helveticus LH76, lactobacillus rhamnosus LRa05, lactobacillus casei LC89 and lactococcus lactis subsp. The invention decomposes milk protein by producing protease, peptidase and the like to supply basic carbon source for strain propagation, and simultaneously produces a large amount of small peptides and amino acids which also provide important nitrogen source for lactic acid bacteria coexisting with the small peptides and the amino acids, and the combined fermentation mode greatly enhances the ACE inhibitory activity of the lactic acid bacteria. The invention also provides a brown lactobacillus beverage, which is prepared from the raw materials of skim milk, skim milk powder, glucose, white granulated sugar, a seabuckthorn fruit extract, a stabilizer, a compound leavening agent and water. The brown lactic acid bacteria beverage has high ACE inhibitory activity, and thus has a function of anti-hypertension activity.

Description

Compound leavening agent for relieving blood pressure rise, brown lactobacillus beverage and preparation method thereof

Technical Field

The invention belongs to the technical field of fermented foods, and particularly relates to a compound leavening agent for relieving blood pressure rise, a brown lactobacillus beverage, and a preparation method and application thereof.

Background

Hypertension is a clinical syndrome characterized by increased systolic and diastolic blood pressure in blood vessels, and is also an important cause of coronary heart disease, myocardial infarction, brain death, heart and renal failure, etc., in which essential hypertension accounts for more than 95% of all hypertensive patients, and is an important disease affecting human physical and mental health. Angiotensin Converting Enzyme (ACE) produces Renin, angiotensin system (RAS) and kinins in vivo. In a kallikrein system (kallikemi. Kininstem, KKS), the kallikrein plays a key role in regulating and controlling blood pressure, and has a vasoconstriction effect on side effects of angiotensin I and bradykinin at a natural base, so that the blood pressure is increased, and the control of the ACE activity also has positive significance in lowering the blood pressure. Therefore, the research on foods having ACE controlling activity has been greatly regarded by human beings. Many scientific studies have demonstrated that foods containing large amounts of GABA have the efficacy of controlling ACE activity and lowering blood pressure in humans.

In recent years, brown lactic acid bacteria beverages have been favored by consumers because of their refreshing cheese flavor. The active lactobacillus beverage can improve the absorption rate of protein, saccharide, calcium, magnesium and other nutrients, and produce great amount of vitamin B and other beneficial matters. Is beneficial to keeping the flora balance in the digestive tract of the human body and establishing an antibacterial ecological barrier, thereby protecting the health of the human body. In addition, lactic acid bacteria generate lactic acid through metabolism in the digestive tract of a human body, and can inhibit the growth and reproduction of harmful bacteria and digest toxins formed by putrefying bacteria, so that the bowel is relaxed, and gastrointestinal waste is discharged.

The research direction of the brown lactobacillus beverage focuses on strain selection, good taste, mouthfeel and shelf life. In recent years, people pursue a more mellow fermentation taste by low-temperature long-time fermentation, add fruit juice and functional ingredients to enrich the variety of the food, optimize food fermentation inoculants, and achieve better mouthfeel and taste and obtain functional peptides.

The brown lactobacillus beverage rich in bioactive polypeptide has casein modified by browning to change enzyme cutting site, so as to reduce hydrophobic bitter polypeptide content in bioactive polypeptide obviously. Research shows that the lactobacillus helveticus is added into skim milk to carry out anaerobic fermentation, and then the bioactive peptides obtained are crudely extracted, analyzed, purified and collected. The fermented milk with the angiotensin converting enzyme inhibitory activity can be obtained by inoculating bacillus subtilis, fermenting at 37 ℃ in a shaking table for 48h in a shaking table, and then inoculating lactobacillus plantarum to ferment at 37 ℃ in a shaking table for 48 h.

After certain special strains such as lactobacillus helveticus (lactobacillus helveticus) and Saccharomyces Cerevisiae (Saccharomyces Cerevisiae) are propagated in a matrix which takes mammal milk as a base material, the protease hydrolysis formed by the special strains can be used, or after the special strains are digested and absorbed by extracellular protease, various polypeptide fragments are formed by utilizing mammal milk protein, and the formed polypeptide fragments further form polypeptide with lower molecular weight through the interaction of peptidase, so that the polypeptide fragments are released, and the blood pressure reducing effect is formed in a human body.

CN111296568A discloses a brown lactobacillus beverage, which comprises the following components in percentage by mass: 5-10% of milk powder; lactase 0.01-0.06%; 0.1 to 0.5 percent of stabilizer; 1.01 to 2.1 percent of functional sweetener; 0.001 to 0.1 percent of lactobacillus paracasei; the balance of water; the total mass percentage is 100 percent. The invention has the functions of low sugar content and low calorie of brown lactobacillus beverage, but has no antihypertensive activity.

CN105613740A discloses a brown lactobacillus beverage of kudzuvine root and a preparation method thereof, wherein the raw materials comprise, by weight, 1000 parts: 200-380 parts of raw milk, 3-10 parts of kudzu root powder, 2-5 parts of a stabilizer, 10-42 parts of reducing sugar, 30-132 parts of a sweetening agent, 1-3 parts of an acidity regulator, 3000-4000U of amylase and the balance of water; the protein content in the raw milk is 3.5-4.6%; the leaven is Lactobacillus casei and Lactobacillus acidophilus. The brown lactobacillus beverage containing the kudzuvine root contains more than 10 percent ⁸ The cfu/mL active lactobacillus and the radix puerariae powder have the effects of relieving alcoholism and reducing blood pressure, and the effects of relieving alcoholism and reducing blood pressure are achieved by adding the radix puerariae which has the effects of reducing blood pressure and reducing myocardial oxygen consumption, but the radix puerariae is a traditional Chinese medicine, the taste of the radix puerariae is not suitable for being added into a beverage, the taste is excellent, in addition, the radix puerariae powder has higher density, the stable system of the brown lactobacillus beverage is complex, the radix puerariae powder is easy to separate out after being placed for a long time, and more importantly, the blood pressure of a person cannot be reduced from the aspect of controlling the ACE activity.

CN109517763A discloses Lactobacillus helveticus H11 and a method for preparing a brown active milk beverage, which comprises the steps of MRS culture medium preparation, strain activation, strain culture, preparation of a glucose-containing skim emulsion, preparation of a brown active lactic acid bacteria beverage and the like. The prepared brown active milk beverage produces ACE inhibitory activity and bioactive peptides Val-Pro-Pro and Ile-Pro-Pro with the function of lowering blood pressure during fermentation, but the ACE inhibitory rate is only 70% at the highest, and the ACE inhibitory rate is still to be improved.

Therefore, the development of a microbial agent capable of further improving the ACE inhibitory activity of fermented milk is a major research focus in the art.

Disclosure of Invention

Aiming at the defects of the prior art, the invention aims to provide a compound leaven for relieving the blood pressure rise, a brown lactobacillus beverage, and a preparation method and application thereof. The invention provides a brown lactobacillus beverage with angiotensin converting enzyme inhibitory activity prepared by a co-fermentation technology, which lays a foundation for developing functional brown lactobacillus beverages with antihypertensive activity in the future.

In order to achieve the purpose, the invention adopts the following technical scheme:

in a first aspect, the invention provides a compound leaven for relieving blood pressure rise, which comprises: lactobacillus plantarum LP90, lactobacillus helveticus LH76, lactobacillus rhamnosus LRa05, lactobacillus casei LC89, and lactococcus lactis subsp.

In the invention, a compound leaven is obtained by compounding lactobacillus plantarum LP90, lactobacillus helveticus LH76, lactobacillus rhamnosus LRa05, lactobacillus casei LC89 and lactococcus lactis subsp cremoris LLc46, all the components are matched with each other for synergy, the activity of angiotensin converting enzyme inhibitory peptide (ACE) can be more effectively maintained, and the angiotensin converting enzyme inhibitory peptide (ACE) is used for relieving the increase of blood pressure.

Preferably, the viable count ratio of the lactobacillus plantarum LP90, the lactobacillus helveticus LH76, the lactobacillus rhamnosus LRa05, the lactobacillus casei LC89 and the lactococcus lactis subsp butterfat LLc46 is (3-5): (16-20): (3-5): (8-12): (0.4-0.6);

wherein "3 to 5" may be, for example, 3, 3.5, 4, 4.5, 5, etc.;

wherein, "16-20" may be, for example, 16, 17, 18, 19, 20, etc.;

wherein "3 to 5" may be, for example, 3, 3.5, 4, 4.5, 5, etc.;

here, "8 to 12" may be, for example, 8, 9, 10, 11, 12, etc.

Here, "0.4 to 0.6" may be, for example, 0.4, 0.45, 0.5, 0.55, 0.6 or the like.

In a second aspect, the invention provides an application of the compound fermentation agent for relieving the blood pressure increase in the first aspect in preparing a beverage.

The content of Lactobacillus plantarum LP90 in the beverage is 5X 10 ⁷ -8×10 ⁷ CFU/mL, for example, may be 5X 10 ⁷ CFU/mL、6×10 ⁷ CFU/mL、7×10 ⁷ CFU/mL、8×10 ⁷ CFU/mL, etc.

The content of Lactobacillus helveticus LH76 in the beverage is 3 × 10 ⁸ -5×10 ⁸ CFU/mL, for example, may be 3X 10 ⁸ CFU/mL、4×10 ⁸ CFU/mL、5×10 ⁸ CFU/mL, etc.

In the beverage, the content of Lactobacillus rhamnosus LRa05 is 5 × 10 ⁷ -1×10 ⁸ CFU/mL, for example, may be 5X 10 ⁷ CFU/mL、6×10 ⁷ CFU/mL、7×10 ⁷ CFU/mL、8×10 ⁷ CFU/mL、 9×10 ⁷ CFU/mL、1×10 ⁸ CFU/mL, etc.

The content of Lactobacillus casei LC89 in the beverage is 1 × 10 ⁸ -3×10 ⁸ CFU/mL, for example, may be 1X 10 ⁸ CFU/mL、2×10 ⁸ CFU/mL、3×10 ⁸ CFU/mL, etc.

The beverage contains lactococcus lactis with milk fat subspecies LLc46 content of 6 × 10 ⁶ -9×10 ⁶ CFU/mL, for example, may be 6X 10 ⁶ CFU/mL、7×10 ⁶ CFU/mL、8×10 ⁶ CFU/mL、9×10 ⁶ CFU/mL, etc.

In a third aspect, the present invention provides a brown lactic acid bacteria beverage, which is prepared from the following raw materials: skim milk, skim milk powder, glucose, white granulated sugar, a seabuckthorn fruit extract, a stabilizer, the compound leaven for relieving the blood pressure rise in the first aspect and water.

The brown lactobacillus beverage with the activity of inhibiting the angiotensin converting enzyme is mainly prepared by co-fermenting lactobacillus plantarum LP90, lactobacillus helveticus LH76, lactobacillus rhamnosus LRa05, lactobacillus casei LC89 and lactococcus lactis subsp.

Preferably, the raw materials for preparing the brown lactobacillus beverage comprise the following components in parts by mass per thousand: 1-2 per mill of skim milk, 0.1-0.5 per mill of skim milk powder, 0.1-0.5 per mill of glucose, 0.1-1.5 per mill of white granulated sugar, 0.05-0.5 per mill of seabuckthorn fruit extract, 0.01-0.1 per mill of stabilizer, 0.01-0.1 per mill of compound leaven for relieving blood pressure rise according to claim 1 or 2, and the balance of water.

The skim milk content is 1-2%, for example, 1%, 1.1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1.7%, 1.8%, 1.9%, 2% and the like, based on 1000% of the total mass of the raw materials for preparing the brown lactic acid bacteria beverage.

The content of the skim milk powder is 0.1 to 0.5%, for example, 0.1%, 0.15%, 0.2%, 0.25%, 0.3%, 0.35%, 0.4%, 0.45%, 0.5%, etc., based on 1000% by mass of the total raw materials for preparing the brown lactic acid bacteria beverage.

The total mass of the raw materials for preparing the brown lactobacillus beverage is 1000 per mill, and the content of the glucose is 0.1-0.5 per mill, such as 0.1 per mill, 0.15 per mill, 0.2 per mill, 0.25 per mill, 0.3 per mill, 0.35 per mill, 0.4 per mill, 0.45 per mill, 0.5 per mill and the like.

The content of the white granulated sugar is 0.1-1.5%, for example, 0.1%, 0.2%, 0.4%, 0.5%, 0.6%, 0.8%, 1.0%, 1.2%, 1.5%, etc., based on 1000% of the total mass of the raw materials for preparing the brown lactic acid bacteria beverage.

The content of fructus Hippophae extract is 0.05-0.5%, such as 0.05%, 0.1%, 0.15%, 0.2%, 0.25%, 0.3%, 0.35%, 0.4%, 0.45%, 0.5%, etc., based on 1000% of the total weight of raw materials for preparing the brown lactobacillus beverage.

The content of the stabilizer is 0.01-0.1%, for example, 0.01%, 0.02%, 0.04%, 0.06%, 0.08%, 0.1%, etc., based on 1000% of the total mass of the raw materials for preparing the brown lactic acid bacteria beverage.

The content of the compound leaven for relieving the blood pressure rise is 0.01-0.1 per mill, for example, 0.01 per mill, 0.02 per mill, 0.04 per mill, 0.06 per mill, 0.08 per mill, 0.1 per mill and the like based on the total mass of the preparation raw materials of the brown lactobacillus beverage.

Preferably, the raw materials for preparing the brown lactobacillus beverage comprise the following components in parts by mass per thousand: 14.5-15.7 per mill of skim milk, 1-3 per mill of skim milk powder, 0.5-2 per mill of glucose, 6-10 per mill of white granulated sugar, 1-1.7 per mill of seabuckthorn fruit extract, 0.3-0.45 per mill of stabilizer, 0.35-0.5 per mill of compound leaven for relieving blood pressure rise and the balance of water.

Preferably, the stabilizer comprises pectin and soy polysaccharide.

Preferably, the mass ratio of the pectin to the soybean polysaccharide is (1-1.5) to (2-3);

wherein "1 to 1.5" may be, for example, 1, 1.1, 1.2, 1.3, 1.4, 1.5, etc.;

here, "2-3" may be, for example, 2, 2.2, 2.4, 2.6, 2.8, 3, etc.

In the invention, the structure of the soybean polysaccharide is similar to that of pectin and contains more galacturonic acid and branched structures through the stabilizer with the specific proportion, and when the stabilizer is compounded with the pectin for use, the lactobacillus beverage of the invention has the advantages of more uniform brown color, better stability in a storage period, difficult occurrence of layering and bleeding phenomena in the storage period and the like.

Preferably, the fat content of the skim milk is 0.3wt% or less, for example, 0.3wt%, 0.25 wt%, 0.2wt%, 0.15wt%, 0.1wt%, 0.05wt%, etc., and the protein content of the skim milk is 3.3wt% or more, for example, 3.3wt%, 3.4wt%, 3.5wt%, 3.6wt%, 3.7wt%, 4wt%, 5wt%, etc.

In a fourth aspect, the present invention provides a method for preparing the brown lactic acid bacteria beverage according to the third aspect, the method comprising the steps of:

(1) Preparing a base material: mixing skim milk, skim milk powder, glucose and part of water, and standing for hydration to obtain a fermentation base material; mixing white granulated sugar, a stabilizer, a seabuckthorn fruit extract and the rest water to obtain a sugar water base material;

(2) Homogenizing and browning: homogenizing the fermentation base material, and carrying out browning reaction to obtain a browning fermentation base material;

(3) Inoculating and fermenting: inoculating the compound leaven for relieving the blood pressure rise to the browning fermentation base material, and fermenting to obtain fermentation liquor;

(4) Mixing and homogenizing: and mixing the fermentation liquor and the sugar water base material, and homogenizing to obtain the brown lactobacillus beverage.

The brown lactobacillus beverage containing angiotensin converting enzyme inhibitory peptide (ACE) is obtained by using skim milk and skim milk powder as main raw materials and through skim milk quality control, stabilizer preparation, base material blending, heat preservation browning, compound leavening agent preparation, mixed inoculation fermentation, sugar water blending, mixing, low-temperature refrigeration and storage. The beverage has the advantages of excellent taste, good stability in shelf life, capability of keeping the activity of angiotensin converting enzyme inhibitory peptide (ACE), integration of the advantages of the angiotensin converting enzyme inhibitory peptide (ACE), the sea buckthorn fruit extract, a brown lactic acid bacteria beverage and the like, and capability of adapting to the consumption requirements of the market.

Preferably, the mass of the portion of water is 20-30% of the total mass of the water, for example, it may be 20%, 22%, 24%, 26%, 28%, 30%, etc.

Preferably, in step (1), the temperature of the static hydration is 40-50 ℃, such as 40 ℃, 42 ℃, 44 ℃, 46 ℃, 48 ℃, 50 ℃ and the like, and the time of the static hydration is 30-50min, such as 30min, 35min, 40min, 45min, 50min and the like.

In the present invention, the skim milk and the skim milk powder are mixed by sieving with a sieve having a mesh number of 80 or more (for example, 80 mesh, 100 mesh, 120 mesh, 150 mesh, 200 mesh, etc.), then subjecting the mixture to a pre-sterilization treatment at 82 to 88 ℃ (for example, 82 ℃, 83 ℃, 84 ℃, 85 ℃, 86 ℃, 87 ℃, 88 ℃, etc.) for 10 or more (for example, 10s, 12s, 14s, 16s, 18s, 20s, etc.), and then finally cooling the mixture to 2 to 6 ℃ (for example, 2 ℃, 3 ℃, 4 ℃,5 ℃, 6 ℃, etc.) for storage.

In the present invention, the stabilizer is: the pectin and the soybean polysaccharide are fully and dryly mixed according to the mass proportion of the total formula.

In the invention, the compound leaven for relieving the blood pressure rise is obtained by mixing lactobacillus plantarum LP90, lactobacillus helveticus LH76, lactobacillus rhamnosus LRa05, lactobacillus casei LC89 and lactococcus lactis subsp.

In the invention, the prepared syrup base materials are mixed after being sterilized: heating to 90-98 deg.C (such as 90 deg.C, 92 deg.C, 94 deg.C, 96 deg.C, 98 deg.C, etc.), stirring, maintaining temperature, and sterilizing for 20-30min (such as 20min, 22min, 24min, 26min, 28min, 30min, etc.).

Preferably, in the step (2), the temperature for homogenization is 55 to 60 ℃, for example, 55 ℃, 56 ℃, 57 ℃, 58 ℃, 59 ℃, 60 ℃ and the like, and the pressure for homogenization is 18 to 20MPa, for example, 18MPa, 18.5MPa, 19MPa, 19.5MPa, 20MPa and the like.

Preferably, in the step (2), the temperature of the browning reaction is 95-99 ℃, for example, 95 ℃, 96 ℃, 97 ℃, 98 ℃, 99 ℃ and the like, and the time of the browning reaction is 2-3h, for example, 2h, 2.2h, 2.4h, 2.6h, 2.8h, 3h and the like.

Preferably, in the step (3), the browning fermentation base material is cooled to 35-39 ℃ (for example, 35 ℃, 36 ℃, 37 ℃, 38 ℃, 39 ℃ and the like) and then inoculated.

Preferably, in the step (3), after the compound leaven for relieving the blood pressure rise is inoculated, stirring is carried out for 8-10min, such as 8min, 8.5min, 9min, 9.5min, 10min and the like, at a rotating speed of 50-150rpm (such as 50rpm, 60rpm, 80rpm, 100rpm, 120rpm, 150rpm and the like).

Preferably, in step (3), the fermentation temperature is 35-39 ℃, for example 35 ℃, 36 ℃, 37 ℃, 38 ℃, 39 ℃ and the like, the fermentation time is 70-120h, for example 70h, 80h, 90h, 100h, 110h, 120h and the like, and the fermentation broth pH at the end of the fermentation is 3.6-3.8, for example 3.6, 3.7, 3.8 and the like.

Preferably, in step (4), the temperature of the mixing is 10 to 20 ℃, and may be, for example, 10 ℃, 12 ℃, 14 ℃, 16 ℃, 18 ℃, 20 ℃ or the like.

Preferably, in step (4), the temperature for homogenization is 20 to 25 ℃, for example, 20 ℃, 21 ℃, 22 ℃, 23 ℃, 24 ℃, 25 ℃ and the like, and the pressure for homogenization is 18 to 20MPa, for example, 18MPa, 18.5MPa, 19MPa, 19.5MPa, 20MPa and the like.

Compared with the prior art, the invention has the following beneficial effects:

(1) The invention utilizes the lactobacillus plantarum and the lactobacillus helveticus to decompose milk protein by producing protease, peptidase and the like so as to supply basic carbon sources for bacterial reproduction, and simultaneously the produced small peptides and amino acids also provide important nitrogen sources for lactic acid bacteria coexisting with the small peptides and amino acids;

(2) Compared with the single strain fermentation effect, the combined fermentation technology of five lactic acid bacteria is utilized, and the co-fermentation of lactobacillus plantarum LP90, lactobacillus helveticus LH76, lactobacillus rhamnosus LRa05, lactobacillus casei LC89 and lactococcus lactis subsp lactis LLc46 improves the ACE inhibitory activity of the fermented milk, so that the invention lays a foundation for developing a novel functional brown lactic acid bacteria beverage with antihypertensive activity in the future;

(3) The brown lactobacillus beverage is prepared by adding lactobacillus helveticus and lactococcus lactis milk fat subspecies for co-fermentation, and adding the low-temperature cold green sea buckthorn fruit extract, so that the brown lactobacillus beverage has richer mouthfeel compared with the commercially available lactobacillus beverage.

Detailed Description

The technical solution of the present invention is further explained by the following embodiments. It should be understood by those skilled in the art that the examples are only for the understanding of the present invention and should not be construed as the specific limitation of the present invention.

(1) The Lactobacillus plantarum LP90 related to the following is a Lactobacillus plantarum LP90 strain, the preservation unit is China general microbiological culture Collection center (CGMCC) with the preservation number of CGMCC No.10453, the preservation date of 2015 is 01-27 months, and the preservation address is No. 3 of Xilu No.1 Chen of the rising area of Beijing.

(2) The related Lactobacillus helveticus LH76 is Lactobacillus helveticus LH76 strain, the preservation unit is China general microbiological culture Collection center, the preservation number is CGMCC No.18796, the preservation date is 11/4/2019, and the preservation address is No. 3 of Xilu 1 of the sunward area of Beijing.

(3) The Lactobacillus rhamnosus LRa05 is a Lactobacillus rhamnosus LRa05 strain, the preservation unit is China general microbiological culture Collection center, the preservation number is CGMCC No.1.12734, the preservation date is 07-20 days in 2020, and the preservation address is NO. 3 of Xilu No.1 Beijing North Cheng Yang-oriented district in Beijing.

(4) The Lactobacillus casei LC89 is Lactobacillus casei LC89 strain, the preservation unit is China general microbiological culture Collection center, the preservation number is CGMCC No.15409, the preservation date is 03 and 05 days in 2018, and the preservation address is No. 3 of Xilu No.1 Beijing-oriented northward time district in Beijing city.

(5) Lactococcus lactis subsp. Cremoris LLc46 strain is Lactococcus lactis subsp. Cremoris LLc46 strain, the preservation unit is China general microbiological culture Collection center, the preservation number is CGMCC No.20549, the preservation date is 2020, 08 and 25 days, and the preservation address is No. 3 of No.1 Xilu-Beijing & Inward-Yang district of Beijing.

And other bacterial species are from microbiology and technology companies.

The sources of the components in the following examples are as follows:

raw materials	Manufacturer of the product	Specification of
			Skimmed milk	Guangming Dairy Co Ltd	Fat is less than or equal to 0.3 percent, and protein is more than or equal to 3.3 percent
Defatted milk powder	Fonterra Group	Fat is less than or equal to 1.0%, and protein is greater than or equal to 32%.9％
			Glucose	LIAO Biotechnology Ltd in Anhui	Superior product
White granulated sugar	GUANGXI XIANGGUI SUGAR INDUSTRY GROUP Co.,Ltd.	First level
			Hippophae rhamnoides fruit extract	Shanxi Guben Biotechnology Co., ltd	The content of fructus Hippophae is greater than or equal to 99%
Pectin	ANHUI YUNING PECTIN Co.,Ltd.	The main component of alpha-D-galacturonic acid/HM
			Soybean polysaccharide	Shandong Jiyuan Biotech Co Ltd	Type B

Example 1

The present example provides a brown lactic acid bacteria beverage, which is composed of the following components:

the brown lactobacillus beverage of the embodiment is prepared by the following preparation method:

(1) Preparing a base material:

fermentation base material: screening skim milk qualified by inspection, performing pre-sterilization treatment at 85 ℃ for 15s according to the filter mesh number of 80 meshes, and cooling to 4 ℃ for storage; heating purified water to 45 deg.C, dissolving skimmed milk, skimmed milk powder and glucose in purified water under stirring, diluting with purified water to 250g, standing the raw material liquid at 45 deg.C, and hydrating for 40min to obtain fermented base material;

syrup base material: fully and dryly mixing pectin and soybean polysaccharide according to the proportion to obtain a compound stabilizer; heating purified water to 75 ℃, adding white granulated sugar and a compound stabilizer into the purified water, stirring for 9min, then adding the seabuckthorn fruit extract, stirring for 6min, fixing the volume to 750g, heating the prepared feed liquid to 95 ℃, stirring, preserving heat and sterilizing for 25min to obtain a sugar water base material;

(2) Homogenizing and browning:

heating the fermentation base material to 60 ℃, homogenizing under 19Mpa, keeping the temperature of the homogenized feed liquid at 96 ℃ for 2.5h for browning, and thus, sterilizing the fermentation base material in the browning treatment process to obtain a browning fermentation base material;

(3) Inoculating and fermenting:

cooling the browning fermentation base material to a temperature of 36 ℃ for inoculation, adding the compound leavening agent under the aseptic protection condition, stirring for 9min, fermenting at 36 ℃ for about 90h until the pH value is 3.7, and stopping fermentation to obtain fermentation liquor;

(4) Mixing and homogenizing: respectively cooling the fermentation liquor and the base material of the sugar water to 20 ℃, stirring, homogenizing the uniformly mixed feed liquid under an aseptic environment by 19Mpa, simultaneously carrying out aseptic filling on the homogenized feed liquid, and storing the aseptically filled feed liquid in an environment of 4 ℃ to obtain the brown lactobacillus beverage.

Example 2

(1) Preparing a base material:

fermentation base material: screening the qualified skim milk, performing pre-sterilization treatment at 85 ℃ for 15s according to the filter mesh number of 100 meshes, and cooling to 2 ℃ for storage; heating purified water to 40 ℃, stirring and dissolving the skimmed milk, the skimmed milk powder and the glucose into the purified water, fixing the volume to 250g by using the purified water, and standing and hydrating the raw material liquid at 40 ℃ for 50min to obtain a fermentation base material;

base material of syrup: fully and dryly mixing pectin and soybean polysaccharide according to the proportion to obtain a compound stabilizer; heating purified water to 80 ℃, adding white granulated sugar and compound stabilizer into the purified water, stirring for 8min, then adding the seabuckthorn fruit extract, stirring for 5min, fixing the volume to 750g, heating the prepared feed liquid to 90 ℃, stirring, preserving the temperature and sterilizing for 30min to obtain a sugar water base material;

(2) Homogenizing and browning:

heating the fermentation base material to 60 ℃, homogenizing under the pressure of 20Mpa, keeping the temperature of the homogenized material liquid at 99 ℃ for 2h for browning, and thus, sterilizing the fermentation base material in the browning treatment process to obtain a browning fermentation base material;

(3) Inoculating and fermenting:

cooling the browning fermentation base material to 37 ℃, inoculating, adding the compound leavening agent under the aseptic protection condition, stirring for 8min, fermenting at 37 ℃ for about 100h until the pH value is 3.6, and stopping fermentation to obtain fermentation liquor;

(4) Mixing and homogenizing: respectively cooling the fermentation liquor and the base material of the sugar water to 20 ℃, stirring, homogenizing the uniformly mixed feed liquid under an aseptic environment by 20Mpa, simultaneously carrying out aseptic filling on the homogenized feed liquid, and storing the aseptically filled feed liquid in an environment of 2 ℃ to obtain the brown lactobacillus beverage.

Example 3

This example provides a brown lactic acid bacteria beverage, which is composed of the following components:

(1) Preparing a base material:

fermentation base material: screening qualified skim milk, performing 85 ℃ and 15s pre-sterilization treatment according to the filter mesh number of 120 meshes, and cooling to 3 ℃ for storage; heating purified water to 50 deg.C, dissolving skimmed milk, skimmed milk powder and glucose in purified water under stirring, diluting with purified water to 250g, standing at 50 deg.C, and hydrating for 30min to obtain fermented base material;

syrup base material: fully dry-mixing pectin and soybean polysaccharide according to the proportion to obtain a compound stabilizer; heating purified water to 70 deg.C, adding white sugar and compound stabilizer into purified water, stirring for 10min, adding fructus Hippophae extract, stirring for 7min, adding water to a constant volume of 750g, heating the mixed liquid to 98 deg.C, stirring, maintaining the temperature, and sterilizing for 20min to obtain syrup base material;

(2) Homogenizing and browning:

heating the fermentation base material to 55 ℃, homogenizing under the pressure of 18Mpa, keeping the temperature of the homogenized material liquid at 95 ℃ for 3h for browning, and thus, sterilizing the fermentation base material during the browning treatment process to obtain a browning fermentation base material;

(3) Inoculating and fermenting:

cooling the browning fermentation base material to a temperature of 35 ℃ for inoculation, adding the compound leavening agent under the aseptic protection condition, stirring for 8min, fermenting at 35 ℃ for about 75h until the pH value is 3.8, and stopping fermentation to obtain fermentation liquor;

(4) Mixing and homogenizing: respectively cooling the fermentation liquor and the base material of syrup to 10 ℃, stirring, homogenizing the uniformly mixed feed liquid under 18Mpa in an aseptic environment, carrying out aseptic filling on the homogenized feed liquid, and storing the aseptically filled feed liquid in an environment of 6 ℃ to obtain the brown lactobacillus beverage.

Example 4

This example provides a brown lactic acid bacteria beverage differing from example 1 only in that the pectin content was 0.005g and the soybean polysaccharide content was 0.04g.

Example 5

This example provides a brown lactic acid bacteria beverage differing from example 1 only in that the pectin content was 0.04g and the soybean polysaccharide content was 0.005g.

Example 6

This example provides a brown lactic acid bacteria beverage differing from example 1 only in that the content of lactobacillus plantarum LP90 was reduced to 0.01g, the content of lactobacillus helveticus LH76 was increased to 0.19U, the content of lactobacillus rhamnosus LRa05 was increased to 0.05U, the content of lactobacillus casei LC89 was increased to 0.11U, and the content of lactococcus lactis subsp.

Example 7

This example provides a brown lactic acid bacteria beverage differing from example 1 only in that the content of lactobacillus plantarum LP90 was reduced to 0.05g, the content of lactobacillus helveticus LH76 was increased to 0.15U, the content of lactobacillus rhamnosus LRa05 was increased to 0.02U, the content of lactobacillus casei LC89 was increased to 0.07U, and the content of lactococcus lactis subsp.

Example 8

This example provides a brown lactic acid bacteria beverage which differs from example 1 only in that the content of lactobacillus plantarum LP90 was reduced to 0.03g, the content of lactobacillus helveticus LH76 was increased to 0.14U, the content of lactobacillus rhamnosus LRa05 was increased to 0.06U, the content of lactobacillus casei LC89 was increased to 0.20U, and the content of lactococcus lactis subsp.

Example 9

This example provides a brown lactic acid bacteria beverage, which is different from example 1 only in that the skim milk content is increased to 2g without adding skim milk powder.

Example 10

This example provides a brown lactic acid bacteria beverage differing from example 1 only in that the skim milk content was increased to 2g without adding skim milk.

Example 11

This example provides a brown lactic acid bacteria beverage, differing from example 1 only in that the content of white sugar was increased to 1g without adding glucose.

Example 12

This example provides a brown lactic acid bacteria beverage, differing from example 1 only in that the glucose content was increased to 0.3g without adding the seabuckthorn fruit extract.

Comparative examples 1 to 5

Comparative examples 1-5 provide five different brown lactic acid bacteria beverages, differing from example 1 only in that the formulation of the starter was fermented with a single microbial inoculum, as shown below:

content (wt.)	Comparative example 1	Comparative example 2	Comparative example 3	Comparative example 4	Comparative example 5
						Lactobacillus plantarum LP90	0.05g	0	0	0	0
Lactobacillus helveticus LH76	0	0.3U	0	0	0
						Lactobacillus rhamnosus LRa05	0	0	0.3U	0	0
Lactobacillus casei LC89	0	0	0	0.3U	0
						Lactococcus lactis milk fat subspecies LLc46	0	0	0	0	0.3U

Comparative examples 6 to 10

Comparative examples 6-10 provide five different brown lactic acid bacteria beverages, differing from example 1 only in that the formulation of the starter was fermented with a single microbial inoculum, as shown below:

test example 1

Test samples: examples 1 to 12 and comparative examples 1 to 10 provided brown lactic acid bacteria beverages;

the test method comprises the following steps:

(1) Respectively putting 10mL of each group of brown lactobacillus beverage into a sterilized centrifugal tube, and centrifuging to obtain supernate; adjusting pH to 8.3 with 1moL/L NaOH, centrifuging again to obtain supernatant, filtering with 0.45 μm, and measuring ACE inhibitory activity, wherein each sample is repeated three times;

(2) ACE inhibition was determined as follows:

an aqueous solution of 5mmoL/L of marurethanylleucine (HHL) was prepared: HHL solubility in 300mmoL/L NaCl in 50mmoL/L HEPES buffer pH 8.3 to form HHL aqueous solution;

dissolving ACE solution in HEPES buffer solution with pH of 8.3 and containing NaCL 300mmoL/L at a final content of about 25mU/mL to obtain ACE solution;

taking 100. Mu.L of HHL solution, placing in 1.5mL centrifuge tube, adding 40. Mu.L of filtered brown lactobacillus beverage, shaking, reacting at 37 deg.C for 5min, adding 40. Mu.L of ACE solution, stirring, reacting at 37 deg.C for 30min, decocting in boiling water for 10min to inactivate enzyme, and stopping reaction.

Detecting hippuric acid content in reactants by using an RPHPLC method, wherein the chromatographic column comprises the following steps: kromasil C18 (150X 4.6mm,5 μm, E102427); mobile phase: 25% acetonitrile +75% water (0.1% TFA); column temperature: 30 ℃; flow rate: 1mL/min; sample introduction amount: 20 mu L of the solution; elution was monitored at 228 nm; the completion time is about 10min; preparing 5mg/mL hippuric acid solution, and respectively diluting to 0.1, 0.2, 0.4, 0.6, 0.8 and 1.0 in mg/mL;

the ACE inhibition ratio formula is calculated as:

wherein, B is hippuric acid concentration (ACE + HHL + buffer solution) measured when no sample is added, A sample group (ACE + HHL + detection sample) is hippuric acid concentration measured after the sample to be detected is added, and C is hippuric acid concentration (HHL + buffer solution) measured by using the buffer solution instead of enzyme.

The specific test results are shown in table 1, wherein the blank control is a beverage prepared without adding a microbial inoculum in the formulation of example 1):

TABLE 1

The test results in table 1 show that the ACE inhibition rate of the brown lactobacillus beverage reaches more than 80%, and the results show that the ACE inhibition activity of the fermented milk is improved by utilizing the five lactobacillus composite fermentation technologies and the co-fermentation of lactobacillus plantarum LP90, lactobacillus helveticus LH76, lactobacillus rhamnosus LRa05, lactobacillus casei LC89 and lactococcus lactis milk fat subspecies LLc46 compared with the fermentation effect of single strains.

Test example 2

test items: the invention carries out taste and flavor evaluation tests. The sensory examination items include tissue state, color, refreshing degree, and flavor. About 40 persons participating in the test are selected, the score of the score item in the sensory examination is averaged, the score is higher, the best character of the commodity is indicated to be closer, and the preference degree of the tested person to the commodity is counted;

and (4) testing standard: each item is full of 10 points;

the specific test results are shown in table 2:

TABLE 2

Evaluation item	8 to 10 minutes	6 to 8 minutes	3 to 5 points
				Tissue state	Uniformity	Is still uniform	Is not uniform enough
Colour(s)	Dark brown color	Light brown color	Slightly white in color
				Degree of refreshing	Refreshing	Yet fresh and cool	Not cool enough
Flavor (I) and flavor (II)	Is obviously outstanding	It is also clearly prominent	Is not outstanding

The specific test results are shown in table 3 below, where the blank control is a beverage prepared without adding a microbial inoculum in the formulation of example 1:

TABLE 3

Item	Tissue state	Colour(s)	Degree of refreshing	Flavor (I) and flavor (II)	Comprehensive evaluation
						Example 1	8.7	8.6	8.8	9.5	8.90
Example 2	8.8	8.5	9.0	9.1	8.85
						Example 3	8.6	8.7	8.9	9.2	8.85
Example 4	7.8	8.5	8.2	8.2	8.17
						Example 5	7.5	8.2	7.8	8.0	7.87
Example 6	8.5	8.8	8.0	8.5	8.45
						Example 7	8.3	8.6	8.2	8.4	8.37
Example 8	8.2	8.9	7.8	8.5	8.35
						Example 9	8.6	8.4	8.6	9.3	8.72
Example 10	8.7	8.3	8.5	9.2	8.67
						Example 11	8.5	6	8.0	8.0	7.62
Example 12	8.5	7.8	8.7	8.0	8.25
						Comparative example 1	8.4	8.6	6.2	5.5	7.17
Comparative example 2	8.3	8.3	5.8	6.0	7.10
						Comparative example 3	8.6	8.5	5.4	5.2	6.92
Comparative example 4	8.5	8.4	6.3	5.4	7.15
						Comparative example 5	8.3	8.6	6.0	4.5	7.13
Comparative example 6	7.8	8.4	6.5	6.4	7.27
						Comparative example 7	8.1	8.4	6.2	6.0	7.17
Comparative example 8	8.5	8.7	5.5	5.0	6.92
						Comparative example 9	8.3	8.5	5.3	5.2	6.82
Comparative example 10	8.2	8.4	6.0	6.3	7.22
						Blank control	8.0	8.5	3.0	4.2	5.92

As can be seen from the test results in Table 3, the products of the examples of the present invention all had the highest texture, color, refreshing, flavor, and overall score. The single strain is used in comparative examples 1-5, and the complex microbial inoculum which is lack of any necessary strain is used in comparative examples 6-10, and the color, the taste and the fineness are obviously lower than those of the products provided in examples 1-10.

Test example 3

the test method comprises the following steps: storing at 2-6 deg.C under refrigeration, visually observing tissue state, and testing taste to check water-milk decomposition degree and protein stability of test product;

the specific test results are shown in table 4 below, where the blank control is a beverage prepared without adding a microbial inoculum in the formulation of example 1:

TABLE 4

As can be seen from the results in Table 4, the brown lactic acid bacteria beverage prepared by the method has good shelf stability, and the taste change test of the product in the shelf life is mainly carried out according to the change of the flavor and the taste in the shelf life.

Test example 4

the test method comprises the following steps: 230 mild and moderate obese family women aged 30-45 years are selected and divided into 23 groups, 10 persons in each group are subjected to the hypertension reducing experiment, and the test time is 90 days. The test method comprises the following steps: recording various data of each person before taking the beverage, and then taking each person for 2 times a day, 100g each time;

the specific test results are shown in table 5 below, where the blank control is a beverage prepared without adding a microbial inoculum in the formulation of example 1:

TABLE 5

The test results in Table 5 show that the compound leaven and the brown lactobacillus beverage for relieving the blood pressure rise can effectively relax blood vessels, reduce blood pressure and condition the blood pressure within a normal range.

The applicant states that the compound leaven for relieving the blood pressure rise, the brown lactic acid bacteria beverage and the preparation method and application thereof are illustrated by the above examples, but the invention is not limited by the above examples, i.e. the invention is not limited by the above examples. It should be understood by those skilled in the art that any modifications of the present invention, equivalent substitutions of the raw materials of the product of the present invention, and the addition of auxiliary components, selection of specific modes, etc., are within the scope and disclosure of the present invention.

Claims

1. The compound leaven for relieving the blood pressure rise is characterized by comprising the following components in parts by weight: lactobacillus plantarum LP90, lactobacillus helveticus LH76, lactobacillus rhamnosus LRa05, lactobacillus casei LC89 and lactococcus lactis subsp.

2. The compound leaven for relieving the blood pressure rise according to claim 1, wherein the viable count ratio of the lactobacillus plantarum LP90, the lactobacillus helveticus LH76, the lactobacillus rhamnosus LRa05, the lactobacillus casei LC89 and the lactococcus lactis subsp.

3. Use of a built starter culture according to claim 1 or 2 for relieving an increase in blood pressure in the preparation of a beverage.

4. The brown lactobacillus beverage is characterized in that the brown lactobacillus beverage is prepared from the following raw materials: skim milk, skim milk powder, glucose, white granulated sugar, seabuckthorn fruit extract, a stabilizer, the compound leaven for relieving the blood pressure rise according to claim 1 or 2, and water.

5. The brown lactic acid bacteria beverage according to claim 4, characterized in that the raw material for preparing the brown lactic acid bacteria beverage comprises the following components by mass per thousand: 1-2 per mill of skim milk, 0.1-0.5 per mill of skim milk powder, 0.1-0.5 per mill of glucose, 0.1-1.5 per mill of white granulated sugar, 0.05-0.5 per mill of seabuckthorn fruit extract, 0.01-0.1 per mill of stabilizer, 0.01-0.1 per mill of compound leaven for relieving blood pressure rise according to claim 1 or 2, and the balance of water;

preferably, the raw materials for preparing the brown lactobacillus beverage comprise the following components in parts by mass per thousand: skim milk 1.45-1.57%, skim milk powder 0.1-0.3%, glucose 0.05-0.2%, white granulated sugar 0.6-1.0%, fructus Hippophae extract 0.1-0.17%, stabilizer 0.03-0.045%, compound leaven for relieving blood pressure rise 0.035-0.05%, and water in balance.

6. The brown lactic acid bacteria beverage according to claim 4 or 5, characterized in that the stabilizer comprises pectin and soy polysaccharide;

preferably, the fat content of the skim milk is 0.3wt% or less, and the protein content of the skim milk is 3.3wt% or more.

7. A method of preparing a brown lactic acid bacteria beverage according to any of claims 4-6, characterized in that the method comprises the steps of:

(3) Inoculating and fermenting: inoculating the browning fermentation base material into the compound leavening agent for relieving the blood pressure rise, and fermenting to obtain fermentation liquor;

8. The method for preparing a brown lactic acid bacteria beverage according to claim 7, wherein the mass of the portion of water is 20-30% of the total mass of the water;

preferably, in the step (1), the temperature of the standing hydration is 40-50 ℃, and the time of the standing hydration is 30-50min.

9. The brown lactic acid bacteria beverage according to claim 7 or 8, wherein in the step (2), the temperature of the homogenization is 55-60 ℃, and the pressure of the homogenization is 18-20MPa;

preferably, in the step (2), the temperature of the browning reaction is 95-99 ℃, and the time of the browning reaction is 2-3h.

10. The brown lactic acid bacteria beverage preparation method according to any one of claims 7 to 9, characterized in that, in the step (3), the browning fermentation base material is inoculated after being cooled to 35 to 39 ℃;

preferably, in the step (3), after the compound leaven for relieving the rise of blood pressure is inoculated, stirring for 8-10min at the rotating speed of 50-150 rpm;

preferably, in the step (3), the fermentation temperature is 35-39 ℃, the fermentation time is 70-120h, and the pH value of the fermentation liquid at the end of fermentation is 3.6-3.8;

preferably, in the step (4), the temperature of the mixing is 10-20 ℃;

preferably, in the step (4), the temperature for homogenization is 20-25 ℃, and the pressure for homogenization is 18-20MPa.