CN106172856A - A kind of functional fermented soybean milk and preparation method and application - Google Patents

A kind of functional fermented soybean milk and preparation method and application Download PDF

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Publication number
CN106172856A
CN106172856A CN201610575321.3A CN201610575321A CN106172856A CN 106172856 A CN106172856 A CN 106172856A CN 201610575321 A CN201610575321 A CN 201610575321A CN 106172856 A CN106172856 A CN 106172856A
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functional
soyabean milk
lactobacillus plantarum
milk
preparation
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CN106172856B (en
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谢静莉
吴惠玲
郎充
周汉康
周吉
郭之桢
沈张磊
王旭阳
魏东芝
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SHANGHAI HANKANG BEANS FOOD CO Ltd
East China University of Science and Technology
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SHANGHAI HANKANG BEANS FOOD CO Ltd
East China University of Science and Technology
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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
    • A23C20/00—Cheese substitutes
    • A23C20/005—Cheese substitutes mainly containing proteins from pulses or oilseeds
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
    • A23C20/00—Cheese substitutes
    • A23C20/02—Cheese substitutes containing neither milk components, nor caseinate, nor lactose, as sources of fats, proteins or carbohydrates
    • A23C20/025—Cheese substitutes containing neither milk components, nor caseinate, nor lactose, as sources of fats, proteins or carbohydrates mainly containing proteins from pulses or oilseeds
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/11—Lactobacillus

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  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Food Science & Technology (AREA)
  • Polymers & Plastics (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)

Abstract

The present invention provides a kind of functional soyabean milk, ferments through Lactobacillus plantarum, and described Lactobacillus plantarum is Lactobacillus plantarum X7021, and the preserving number of described Lactobacillus plantarum X7021 is: CCTCC NO:M2015039.The functional soyabean milk of the present invention has good uric acid resisting, blood pressure lowering effect and good curdled milk effect；Not only have common soyabean milk drinks function, also has the prebiotic health care of lactobacillus, simultaneously without any preservative, and has more preferable mouthfeel, has broad application prospects.

Description

A kind of functional fermented soybean milk and preparation method and application
Technical field
The invention belongs to functional food technical field, specifically, be about a kind of functional fermented soybean milk and system thereof Preparation Method and application.
Background technology
Along with raising and the living-pattern preservation of living standards of the people, hypertension, the prevalence of hyperuricemia significantly increase Add.In addition to using traditional medical treatment regime, it is also possible to preferably prevent by healthy dietary habit and reduce generation The incidence rate of thanking property disease.Therefore, increasing people starts edible health food such as: Organic food, functional food etc..Merit Energy property food refers to the food with specific function, is suitable for specific crowd and eats, and the function of scalable body, the most not with treatment For the purpose of and human body do not produced any acute, the subacute or food of chronic hazard.
Soyabean milk is welcome by consumers in general deeply because its protein content is high and has abundant nutritional labeling, usually through Lactic acid bacteria fermentation is prepared as functional soyabean milk, and the functional soyabean milk using different lactic acid bacteria fermentations to obtain has different mouths Sense, nutritional labeling and function.Substantial amounts of researcher has filtered out various plants lactobacillus, and has prepared several functions bean Breast, such as: Chinese patent CN104694409A discloses a kind of Lactobacillus plantarum, preservation entitled Lactobacillus plantarum Y-9, and preservation is compiled Number be CCTCC NO:M2013391, and disclose this Lactobacillus plantarum in preparing medicine or food for reduce cholesterol, Blood pressure lowering, raising immunity, fat-reducing or the purposes of regulating intestinal canal flora.CN1927005A disclose one have increase bone close Degree, the fermentation soybean milk and preparation method thereof of Angiotensin-converting enzyme inhibition function, wherein use lactobacillus helveticus TUST005 The composite ferment being main with diacetyl streptococcus ferments, and the fermentation soybean milk tool of preparation has efficacy in lowering high blood pressure.
Prior art does not the most find the fermented soybean milk with xanthine oxidase (XO) suppression function.XO is that human body is fast Purine metabolism generates the key enzyme of uric acid, when XO is suppressed, is effectively reduced the formation of internal uric acid.Soyabean milk is a kind of purine content Higher food, has patient's general recommendations fasting of hyperuricemia and gout.If soyabean milk is after lactic acid bacteria fermentation, produce The rejection ability of XO, can effectively reduce soyabean milk eater and the risk of hyperuricemia occurs.Additionally, two or more metabolism Syndrome (hyperlipemia, hypertension, hyperglycemia and hyperuricemia are commonly called as " four is high ") simultaneous situation is in obese people Much.Therefore, research and develop a kind of functional fermented soybean milk with higher XO suppression ratio, there is blood pressure reduction effect simultaneously, and Good mouthfeel just tool is kept to be of great significance.
Summary of the invention
First purpose of the present invention is to provide a kind of functional soyabean milk, has higher XO and angiotensin-converter Enzyme (ACE) suppression ratio, and keep good mouthfeel.
Second object of the present invention is to provide the preparation method of described functional soyabean milk.
For achieving the above object, the present invention adopts the following technical scheme that
A kind of functional soyabean milk, obtains through Lactobacillus plantarum fermentation, and described Lactobacillus plantarum is Lactobacillus plantarum X7021, its preserving number is: CCTCC NO:M2015039.
The preparation method of described functional soyabean milk, comprises the technical steps that:
1) Lactobacillus plantarum X7021 lyophilizing mycopowder is taken out defrosting, be seeded to the MRS training of sterilizing by the volume ratio of 2-6% Support in base, be placed in 37 DEG C, cultivate under the conditions of 200rpm and take out to during exponential phase, obtain activating bacterium solution；
2) by step 1) activation bacterium solution be seeded in the soyabean milk of sterilizing according to the volume ratio of 2-6%, in 33-42 DEG C of standing Fermentation 6-12h, after-ripening 2-48h under the conditions of 0-10 DEG C, obtain functional soyabean milk.
Preferably, described step 2) in volume ratio be 5%.
Preferably, described step 2) in fermentation temperature be 40 DEG C.
Preferably, described step 2) in fermentation time be 9h.
Third object of the present invention is to provide the application in reducing uric acid of described functional soyabean milk.
Fourth object of the present invention is to provide the application in blood pressure lowering of described functional soyabean milk.
Compared with prior art, there is advantages that
1) the functional soyabean milk of the present invention has good uric acid resisting and blood pressure lowering effect.
2) the Lactobacillus plantarum acid producing ability that the functional soyabean milk of the present invention uses is strong, the curdled milk of the functional soyabean milk of preparation Respond well.
3) what the functional soyabean milk of the present invention not only had a common soyabean milk drinks function, also has the prebiotic health care of lactobacillus Function, and without any preservative, meet the trend of beverage industry " green, nature, health ", there is wide application Prospect.
Preservation explanation
Lactobacillus plantarum X7021 (the Lactobacillus plantarum X7021) bacterial strain of the present invention, has another name called plant breast Bacillus S1L12 (Lactobacillus plantarum S1L12), is saved in Chinese Typical Representative on January 16th, 2015 and cultivates Thing preservation center (China Center for Type Culture Collection is called for short CCTCC), depositary institution address: China. Wuhan. Wuhan University, deposit number is CCTCC NO:M2015039.
Detailed description of the invention
Below in conjunction with specific embodiment, the present invention will be further described.Should be understood that following example are merely to illustrate this Invent not for limiting the scope of the present invention.
In the present invention, the Lactobacillus plantarum X7021 of application derives from bean curd with odor saltwater brine.
MRS culture medium prescription: 10.0g peptone, 5.0g beef powder, 4.0g yeast powder, 20.0g glucose, 1.0mL tween 80,2.0g dipotassium hydrogen phosphate, 5.0g sodium acetate, 2.0g Triammonium citrate, 0.2g magnesium sulfate, 0.05g manganese sulfate, 15.0g agar Powder and 1000mL distilled water.
Other raw materials of the present invention and equipment are commercially available.
Product skyr standard: the pH value of functional soyabean milk when measuring fermentation termination, it is the most that pH value the lowest explanation system produces acid, The curdled milk effect of functional soyabean milk is the best.
Embodiment 1: the preparation of Lactobacillus plantarum lyophilizing mycopowder
1) in aseptic operating platform, a small amount of bacterium solution of picking from the glycerol pipe of Lactobacillus plantarum X7021, put down at MRS solid Lining out, cultivates 36-48h in 37 DEG C, and from MRS solid plate, picking list colony inoculation is in MRS culture medium, detests in 37 DEG C Oxygen cultivates 18h, is then inoculated in the fermentation tank containing MRS culture medium, anaerobic fermentation 24h, is then centrifuged 10 under the conditions of 6000rpm Minute, collect thalline, and wash 2 times with the sterile phosphate buffer that pH value is 6.4, recentrifuge, obtain bacterium mud.
2) by step 1) the bacterium mud prepared is sufficiently mixed emulsifying with sterilized freeze drying protectant by weight 1:2, obtains Emulsion mixture.
3) by step 2) emulsion mixture pre-freeze more than 4h below 45 DEG C of preparing, place into lyophilizing in freeze dryer, To Lactobacillus plantarum X7021 lyophilizing mycopowder.
4) after measured, step 3) viable count in the Lactobacillus plantarum lyophilizing mycopowder prepared is 1.0 × 1010-1.5× 1011CFU/g。
Wherein, above-mentioned steps 2) in freeze drying protectant contain 20% defatted milk powder, 6% trehalose, 2.5% glutamic acid Sodium, 2% glycerol and 0.5% cysteine hydrochloride (being mass percent above).
Embodiment 2: the preparation of soyabean milk raw material
Soyabean milk is made in Non-transgenic soybean making beating, boiling sterilization 30 minutes in 100 DEG C of boiling water baths, it is prepared as sterilizing bean Breast, standby.
The preparation process of soyabean milk is as follows: selected Non-transgenic soybean → cleaning → according to bean: water=1:3 (volume ratio) adds steaming The hot water of 85 DEG C of distilled water soaked overnight → addition certain volume carries out defibrator process → filtered through gauze → boil 30min → cooling.
Embodiment 3-5: Lactobacillus plantarum is used for preparing functional fermented soybean milk
The preparation method of functional soyabean milk is as follows:
1) Lactobacillus plantarum X7021 lyophilizing mycopowder embodiment 1 prepared takes out and thaws, and is seeded to by the volume ratio of 2% In the MRS culture medium of sterilizing, it is placed in 37 DEG C, cultivates under the conditions of 200rpm and take out to when entering exponential phase, obtain activating bacterium Liquid.
2) by step 1) activation bacterium solution according to the volume ratio of 2-6% be seeded to embodiment 2 preparation sterilizing soyabean milk in, Standing for fermentation 6-12h in 33-42 DEG C of constant incubator, after-ripening 2-48h under the conditions of 0-10 DEG C, obtain functional soyabean milk.
3) pH value of functional soyabean milk during detection fermentation termination.
It is observed that the functional soyabean milk curdled milk of preparation is in order.
The concrete fermentation condition of embodiment 3-5 is as shown in table 1:
PH value after the fermentation condition of table 1 embodiment 3-11 and fermentation
Embodiment 6: functional soyabean milk uric acid resisting ability detects
Xanthine oxidase (XO) is can be catalyzed hypoxanthine in human body to be oxidized to xanthine, and xanthine oxidase turns to uric acid Key enzyme, being therefore that treatment is gentle solves gout or the drug target of hyperuricemia disease.The activity of suppression XO can reduce gout Or the uric acid content that hyperuricemia disease is in the patient, thus slow down gout symptom.The present embodiment is fermented bean by detection functionality The external XO suppression ratio of breast measures the uric acid resisting vigor of the functional soyabean milk of the present invention.The concrete assay method of XO suppression ratio is such as Under:
The hydrochloric acid adjustment pH value of functional soyabean milk 2mol/L embodiment 3-5 prepared, to 4.0, is subsequently placed in centrifugal Guan Zhong, 8000rpm are centrifuged 10 minutes, obtain supernatant, take part supernatant and with 2mol/L NaOH adjustment pH value to 7.5, then Secondary 8000rpm is centrifuged 10 minutes, and the supernatant obtained is as testing sample.
Test group 1: draw testing sample 25 μ L, the xanthine solution adding 0.5mmol/L (is dissolved in 20m mol/L In the sodium phosphate buffer of pH7.5) 25 μ L, and temperature bath 5 minutes under the conditions of 25 DEG C, being subsequently added concentration is the molten of 100mU/mL Solution xanthine oxidase 50 μ L in same sodium phosphate buffer, starts reaction, reacts 30 minutes under the conditions of 25 DEG C, Rear addition 25 μ L concentration is that the HCl of 1mol/L terminates reaction, the light absorption value A of product uric acid under the conditions of mensuration 290nm1。
Test group 2: draw testing sample 25 μ L, adds 25 μ L xanthine solution, and in 25 DEG C of temperature bath 5min, is subsequently added The 50 above-mentioned sodium phosphate buffers of μ L, react 30 minutes under the conditions of 25 DEG C, are eventually adding the HCl that 25 μ L concentration are 1mol/L and terminate Reaction, measures the light absorption value A under the conditions of 290nm2。
Matched group 1: draw above-mentioned sodium phosphate buffer 25 μ L, adds 25 μ L xanthine solution, and in 25 DEG C of temperature baths 5 points Clock, is subsequently added xanthine oxidase (XO) 50 μ L and starts reaction, react 30 minutes, be eventually adding 25 μ L dense under the conditions of 25 DEG C The HCl that degree is 1mol/L terminates reaction, the light absorption value A of product uric acid under the conditions of mensuration 290nm3。
Matched group 2: draw 25 μ L xanthine solution, and in 37 DEG C of temperature baths 5 minutes, be subsequently added the 75 above-mentioned sodium phosphates of μ L and delay Rush liquid, react 30 minutes under the conditions of 37 DEG C, be eventually adding the HCl that 25 μ L concentration are 1mol/L and terminate reaction, measure 290nm bar Light absorption value A under part4。
Calculated the suppression ratio of XO by the change of light absorption value, computing formula is as follows:
XO suppression ratio (%)=[(A1-A2)-(A3-A4)]/(A3-A4) × 100%
Wherein: A1、A2、A3And A4Product under the conditions of expression test group 1, test group 2, matched group 1 and matched group 2 respectively Light absorption value.
Each testing sample is made three Duplicate Samples respectively, and the meansigma methods of the XO suppression ratio taking each Duplicate Samples is sample XO suppression ratio, result is as shown in table 2.
As can be seen from Table 2, without the soyabean milk of Lactobacillus plantarum fermentation without XO inhibitory activity, and through Lactobacillus plantarum The soyabean milk of X7021 fermentation has stronger XO inhibitory activity, illustrates that the functional soyabean milk of the present invention has good uric acid resisting merit Effect；Compared with the soyabean milk fermented without Lactobacillus plantarum.
Embodiment 7: functional soyabean milk blood pressure lowering ability detects
Angiotensin converting enzyme (ACE) is a kind of dipeptides carboxylic acid played an important role during human blood-pressure regulates, main The regulation of renin angiotensin aldosterone system to be participated in.ACE catalyzing hydrolysis angiotensin is transformed into and has strong boosting The Angiotensin II of effect, resolves into inactive fragments by the bradykinin playing vasorelaxation action simultaneously, causes elevation of the blood pressure. The activity of suppression ACE can reduce the generation of Angiotensin II and the inactivation of bradykinin, thus reduces the mesh of blood pressure 's.The present embodiment measures the blood pressure lowering energy of Lactobacillus plantarum X7021 Fermentation Function soyabean milk by the outer ACE suppression ratio of detection bodies Power.The concrete detection method of ACE suppression ratio is as follows:
The hydrochloric acid adjustment pH value of functional soyabean milk 2mol/L embodiment 3-5 prepared, to 4.0, is subsequently placed in centrifuge tube In, 8000rpm is centrifuged 10min, obtains supernatant, then the NaOH of the pH value 2mol/L of part supernatant is adjusted pH value extremely 8.3,8000rpm is centrifuged 10min again, obtains supernatant, be stored in-20 DEG C stand-by as testing sample.
Test group: add testing sample 25 μ L in 1.5mLEP pipe, add the ACE that concentration is 310mU/mL and (be dissolved in 0.1mol/L NaCl Han 0.5mol/L, pH value is in the sodium borate buffer solution of 8.3) 10 μ L, and temperature bath 5min at 37 DEG C, (HHL is dissolved in 0.1mol/L containing 0.5mol/L to be subsequently adding the hippuroyl histidyl leucine that 50 μ L concentration are 5.0m mol/L NaCl, pH value is in the sodium borate buffer solution of 8.3) starting reaction, isothermal reaction 1h at 37 DEG C, then 100 DEG C of conditions Lower heating in water bath 10min makes enzyme inactivation terminate reaction.
Matched group: the testing sample in test group is replaced with the above-mentioned sodium borate buffer liquid of same volume.
Use after high-efficient liquid phase chromatogram technique analysis reaction the content of hippuric acid in test group and matched group system, with high-efficient liquid The content of the peak area A correspondence hippuric acid in phase chromatograph collection of illustrative plates, and according to below equation calculating ACE suppression ratio:
ACE suppression ratio (%)=(AComparison-ATest)/AComparison× 100%
In formula: AComparisonPeak area for matched group；ATestPeak area for test group.
The condition determination of high performance liquid chromatography is: chromatographic column: WondaSil C18 chromatographic column (250 × 4.6mm, 5 μm)；Stream Dynamic phase: A be ultra-pure water (containing 0.1%TFA), B be acetonitrile；Elution requirement: 0min, 20%B；10min, 100%B；16min, 20%B；20min, 20%B；Flow velocity: 0.8mL/min；Column temperature: 30 DEG C；Detection wavelength: 228nm；Sample size: 10 μ L.
Result of the test is as shown in table 2.
Functional soyabean milk effect prepared by table 2 embodiment 3-5 compares
As can be seen from Table 2, soyabean milk is after Lactobacillus plantarum X7021 ferments, and ACE suppression ratio all raises, and the present invention is described Functional fermented soybean milk there is obvious blood pressure reduction effect.
Embodiment 8, functional soyabean milk mouthfeel are tested
Randomly choose 30 citizen and foretaste the functional soyabean milk (test prepared according to the preparation method of the present invention simultaneously Group), commercially available common soyabean milk (contrast groups 1) and commercially available usual functionality soyabean milk (contrast groups 2), add up its satisfaction, such as table 3 Shown in:
Table 3
By the result of table 3 it can be seen that the functional satisfaction than commercially available soyabean milk and functional soyabean milk of the present invention all Height, the functional soyabean milk of the present invention has good mouthfeel.
In sum, the functional soyabean milk that the present invention uses Lactobacillus plantarum fermentation to prepare has obvious uric acid resisting And blood pressure, it is suitable for metabolic arthritis and edible for patients with elevated blood pressure；The Lactobacillus plantarum acid producing ability used is strong, and curdled milk is respond well；No Only have common soyabean milk drinks function, also has the prebiotic health care of lactobacillus, simultaneously without any preservative, and tool There is more preferable mouthfeel, therefore there is prospects for commercial application widely.
Being described in detail the specific embodiment of the present invention above, but it is only used as example, the present invention is not limiting as In particular embodiments described above.To those skilled in the art, any equivalent modifications that this invention is carried out and replacing In generation, is the most all among scope of the invention.Therefore, the equalization made without departing from the spirit and scope of the invention converts and repaiies Change, all should contain within the scope of the invention.

Claims (7)

1. a functional soyabean milk, obtains through Lactobacillus plantarum fermentation, it is characterised in that described Lactobacillus plantarum is plant breast Bacillus X7021, its preserving number is: CCTCC NO:M2015039.
The preparation method of functional soyabean milk the most as claimed in claim 1, comprises the technical steps that:
1) Lactobacillus plantarum X7021 lyophilizing mycopowder is taken out defrosting, be seeded to the MRS culture medium of sterilizing by the volume ratio of 2-6% In, it is placed in 37 DEG C, cultivates under the conditions of 200rpm and take out to during exponential phase, obtain activating bacterium solution；
2) by step 1) activation bacterium solution be seeded in sterilizing soyabean milk according to the volume ratio of 2-6%, in 33-42 DEG C of standing for fermentation 6- After-ripening 2-48 hour under the conditions of 12h, 0-10 DEG C, obtains functional soyabean milk.
3. preparation method as claimed in claim 2, it is characterised in that described step 2) in volume ratio be 5%.
4. preparation method as claimed in claim 2, it is characterised in that described step 2) in fermentation temperature be 40 DEG C.
5. preparation method as claimed in claim 2, it is characterised in that described step 2) in fermentation time be 9h.
The functional soyabean milk the most as claimed in claim 1 application in reducing uric acid.
The functional soyabean milk the most as claimed in claim 1 application in blood pressure lowering.
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Publication number Priority date Publication date Assignee Title
CN109504636A (en) * 2018-12-27 2019-03-22 内蒙古农业大学 A kind of lactobacillus plantarum P12 and application thereof
CN109517767A (en) * 2019-01-21 2019-03-26 北京农学院 A kind of lactobacillus plantarum strain, soymilk and its production method based on strain fermentation production
CN109517767B (en) * 2019-01-21 2020-07-10 北京农学院 Lactobacillus plantarum strain, soymilk produced based on fermentation of strain and production method of soymilk
CN110638037A (en) * 2019-08-14 2020-01-03 浙江黑金生物科技有限公司 Application of lactobacillus plantarum X7022, black garlic enzyme and preparation method and application of black garlic enzyme
CN111635870A (en) * 2020-05-26 2020-09-08 华东理工大学 Leavening agent, direct-feeding fermentation method of stinky tofu marinating liquid and stinky tofu marinating liquid
CN111635870B (en) * 2020-05-26 2022-07-01 华东理工大学 Leavening agent, direct-feeding fermentation method of stinky tofu marinating liquid and stinky tofu marinating liquid
CN114376152A (en) * 2022-01-12 2022-04-22 东北农业大学 Preparation method of fermented soybean milk and application of fermented soybean milk in enhancing immunocompetence

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