CN107307086A - A kind of application process of β glucuroides in soymilk - Google Patents
A kind of application process of β glucuroides in soymilk Download PDFInfo
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- CN107307086A CN107307086A CN201710425478.2A CN201710425478A CN107307086A CN 107307086 A CN107307086 A CN 107307086A CN 201710425478 A CN201710425478 A CN 201710425478A CN 107307086 A CN107307086 A CN 107307086A
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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
- A23C11/00—Milk substitutes, e.g. coffee whitener compositions
- A23C11/02—Milk substitutes, e.g. coffee whitener compositions containing at least one non-milk component as source of fats or proteins
- A23C11/10—Milk substitutes, e.g. coffee whitener compositions containing at least one non-milk component as source of fats or proteins containing or not lactose but no other milk components as source of fats, carbohydrates or proteins
- A23C11/103—Milk substitutes, e.g. coffee whitener compositions containing at least one non-milk component as source of fats or proteins containing or not lactose but no other milk components as source of fats, carbohydrates or proteins containing only proteins from pulses, oilseeds or nuts, e.g. nut milk
- A23C11/106—Addition of, or treatment with, microorganisms
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Abstract
The invention discloses a kind of application process of β glucuroides in soymilk, the Lactobacillus plantarum of culture is specifically accessed in soymilk, the β glucuroides produced using Lactobacillus plantarum in soymilk fermentation process are directly converted the isoflavone glucoside compound of soymilk and match somebody with somebody based compound for isoflavone glucoside, improve the biological effectiveness of soymilk.Using application process of the present invention, β glucosidase activities are up to 9.24U/mL, and pH is 4.12, lactic acid content is 92.45mM, and acetic acid content is 10.01mM, and the content of daidzein is 9.06mg/L, 1.9 times are improved, genistein content is 34.14mg/L, improves 4.0 times.The present invention effectively raises the biological effectiveness of soymilk, and more conducively human body is digested and assimilated, and lactic acid and acetic acid have adjusted the local flavor of soymilk, while the addition of probiotics Lactobacillus plantarum, promotes human body intestinal canal health.
Description
Technical field
The present invention relates to application of the beta-glucosidase in soymilk.
Background technology
Soymilk and bean powder are traditional bean products, there is long developing history.Start the seventies in last century it is prevailing in
The Asian countries such as China, Japan, South Korea.Why soymilk and bean powder are widely recognized as, because it is rich in soybean protein and unsaturation
Aliphatic acid, and without lactose and cholesterol.Soybean is the staple crops of China, and the processing industry of soybean, which is in, to be continued to develop
Cheng Zhong.The big-and-middle-sized processing enterprise of current domestic production soymilk and bean powder is about 90, and annual production is 850,000 tons, year soybean processing
650000 tons, annual output reconstitutes 350,000 tons of bean powder and soy milk powder, drinks 500,000 tons of soymilk, accounts for 18% of soy protein products or so.
Consumption of China town dweller in 2013 to dairy products reaches 25kg or so per capita, and growth rate reaches 18.3%.Due to China
Animal milk supply is limited, and Imported dairy products hold at high price, and the exploitation for plant protein source will play positive role,
Especially the development of soymilk and bean powder industry by be animal sources dairy produce important supplement.Foreign countries, such as U.S., Japan and Germany
More pay attention to the research and development of soymilk and bean powder.Soymilk and bean powder are processed further, it is assigned more beneficial to health
Feature by be application innovation important directions.
Recent study is found in soybean containing abundant isoflavones.There is isoflavones certain estrogen to live
Property, it is a kind of excellent natural, there is significant prevention to aging, atherosclerosis, cancer, osteoporosis etc.
Effect.As the soymilk and bean powder of main bean product because of the isoflavones containing special physiological activity by more and more
Concern.Isoflavones in soybean is mainly deposited with isoflavone glucoside compound and isoflavone glucoside with two kinds of forms of based compound
.Research shows that bioactivity of the isoflavone glucoside with based compound, apparently higher than isoflavone glucoside compound, is that human body is utilized
Effective form.And the content of the isoflavone glucoside compound in soybean matches somebody with somebody based compound far above isoflavone glucoside, so, day
Isoflavones biological effectiveness is relatively low in right soybean and its product.
The content of the invention
Based on above-mentioned technical problem, the present invention provides a kind of application process of beta-glucosidase in soymilk.
The adopted technical solution is that：
A kind of application process of beta-glucosidase in soymilk, is specifically the newborn bar of plant that culture is accessed in soymilk
Bacterium, the beta-glucosidase produced using Lactobacillus plantarum in soymilk fermentation process directly converts the isoflavone glucoside of soymilk
Compound is that isoflavone glucoside matches somebody with somebody based compound, improves the biological effectiveness of soymilk.
It is preferred that, the inoculum concentration of the Lactobacillus plantarum of the culture is 4%, with volume basis, and cultivation temperature is 37 DEG C, training
The time of supporting is 24h.
A kind of application process of the above-mentioned beta-glucosidase in soymilk, comprises the following steps：
(1) soymilk is prepared：Soybean is placed in soaked overnight in distilled water；Soybean skin is removed, and is cleaned with distilled water；Remove
Skin soybean 200g adds 500mL distilled water, is ground using soy bean milk making machine；Gained slurry is boiled after 15min in 80 DEG C, uses double gauze mistake
Filter, removes insoluble residue；The soya-bean milk that will filter out is added in conical flask, 121 DEG C of sterilizing 15min；Room temperature is cooled to, sugarcane is added
Sugar juice, obtains soymilk culture medium；Daidzin, genistin, daidzein and dyestuff in initial soymilk culture medium is measured by sampling
The content of lignin, remaining 4 DEG C of refrigerator are saved backup；
(2) Spawn incubation：Go bail for the μ L of Lactobacillus plantarum bacterium solution 300 being stored in glycerol tube, is inoculated in MRS culture mediums,
Anaerobism bottle cap is covered, in quiescent culture 20h at 37 DEG C；
(3) fermented and cultured：Lactobacillus plantarum seed liquor is taken to be inoculated in standby soymilk culture medium, inoculum concentration is 4%, with
Volume fraction, in cultivating 24h at 37 DEG C, take sample be used for determine activity of beta-glucosidase, pH, daidzin, genistin,
Daidzein, genistein, lactic acid, the content of acetic acid.
Above-mentioned activity of beta-glucosidase assay method is as follows：Lactobacillus plantarum fermentation produces the enzymatic of beta-glucosidase
Reaction system be 1.2mL 0.1M pH be 5.0 phosphoric acid-citrate buffer solution, 0.4mL 8mM p-NPG, 0.4mL enzyme liquid, 45
DEG C reaction 30min after add 2mL 0.5M sodium carbonate terminating reactions, 400nm determine p-nitrophenol light absorption value；With difference
The p-nitrophenol standard specimen of concentration draws standard curve, calculates content of p-nitrophenol；Enzyme-activity unit is defined as at 45 DEG C,
Enzyme amount needed for catalyzing hydrolysis p-NPG produces 1 μM of p-nitrophenol in 1min is 1 enzyme-activity unit.
Above-mentioned organic acid content testing method is as follows：Sample 1mL is taken to add in 1.5mL centrifuge tubes, 10000rpm centrifugations
1min, takes supernatant to be determined with 0.22 μm of membrane filtration for HPLC；Chromatographic condition：High performance liquid chromatograph, Aminex HPX-
87H organic acids and alcohol analytical column, mobile phase are 0.005M H2SO4The aqueous solution, flow velocity is 0.6mL/min, and Detection wavelength is
210nm, column temperature is 35 DEG C.
Above-mentioned isoflavone content assay method is as follows：5mL samples are taken to add the ethanol solution of 5mL mass percent concentrations 70%
After mixing, ultrasonic extraction 40min at 60 DEG C, 10000rpm centrifugation 5min take supernatant to be used for HPLC through 0.22 μm of membrane filtration
Determine；Chromatographic condition is：High performance liquid chromatograph, C18Chromatographic column, mobile phase is acetonitrile and 0.2% formic acid, gradient elution, flow velocity
For 0.8mL/min, Detection wavelength is 260nm, and column temperature is 35 DEG C.
The Lactobacillus plantarum that the present invention is inoculated with has the characteristic of exocytosis beta-glucosidase, so that will be different in soymilk
Flavone glycoside compound is converted into isoflavone glucoside with based compound.
The method have the benefit that：
Using application process of the present invention, activity of beta-glucosidase is up to 9.24U/mL, and pH is 4.12, and lactic acid content is
92.45mM, acetic acid content is 10.01mM, and the content of daidzein is 9.06mg/L, improves 1.9 times, genistein content is
34.14mg/L, improves 4.0 times.This method effectively raises the biological effectiveness of soymilk, and the more conducively digestion of human body is inhaled
Receipts, lactic acid and acetic acid have adjusted the local flavor of soymilk, while the addition of probiotics Lactobacillus plantarum, promotes human body intestinal canal health.
Embodiment
The isoflavone glucoside compound that beta-glucosidase can be catalyzed in soybean is converted into isoflavone glucoside aglucon chemical combination
Thing, more conducively absorption of human body are utilized, and can improve the biological effectiveness of isoflavones in bean product.Have no in the market using β-
The soymilk and bean powder product of glucuroide production occur.So, beta-glucosidase is applied in soymilk and bean powder to have
Larger market potential.
Probiotics plays an important role during absorption of human body is using isoflavones, a considerable amount of probiotics energy
It is that isoflavone glucoside matches somebody with somebody based compound, and some probiotics that enough secretion beta-glucosidase, which directly converts isoflavone glucoside compound,
Soybean isoflavone glucoside can also be matched somebody with somebody the daidzein in based compound and further be metabolized to equol, bioactivity is higher.But
The species of probiotics in human body intestinal canal is different because of the influence of the reasons such as age, disease, causes everyone to daily intake
The amount of isoflavones have very big difference.Probiotics is directly appended in soybean fermentive food, it is prebiotic during the fermentation
Bacterium secretion beta-glucosidase soybean transformation isoflavone glucoside compound is aglycone forms, beneficial to absorbing for enteron aisle, simultaneously
The growth of beneficial flora in enteron aisle can be promoted.
For the ease of understanding the present invention, further illustrated below in conjunction with specific embodiment, but these embodiments are only
It is to play a part of explanation, the invention is not limited in following embodiments.
Embodiment 1
Make soymilk by oneself：Soybean is placed in soaked overnight in distilled water；Soybean skin is removed, and is cleaned with distilled water；Remove skin
Soybean 200g adds 500mL distilled water, is ground using soy bean milk making machine；All slurries are boiled after 15min in 80 DEG C, use double gauze mistake
Filter, removes insoluble residue；The soya-bean milk that will filter out is added in conical flask, 121 DEG C of sterilizing 15min；Room temperature is cooled to, was added
The sucrose solution of bacterium is filtered out, the content of daidzin in initial soymilk, genistin, daidzein and genistein is measured by sampling,
Remaining 4 DEG C of refrigerator is saved backup.
Spawn incubation：Go bail for the μ L of Lactobacillus plantarum bacterium solution 300 being stored in glycerol tube, is inoculated in MRS culture mediums, covers
Anaerobism bottle cap, in quiescent culture 20h at 37 DEG C.Fermented and cultured：Lactobacillus plantarum seed liquor is taken to be inoculated in ready soymilk
In culture medium, inoculum concentration be 4% (v/v), in cultivating 12h at 37 DEG C, take sample be used for determine activity of beta-glucosidase, pH,
Daidzin, genistin, daidzein, genistein, lactic acid, the content of acetic acid.
Activity of beta-glucosidase is determined：Lactobacillus plantarum fermentation produce beta-glucosidase enzymatic reaction system be
Phosphoric acid-citrate buffer solution, 0.4mL 8mM p-NPG, 0.4mL enzyme liquid that 1.2mL0.1M pH are 5.0,45 DEG C of reaction 30min
2mL 0.5M sodium carbonate terminating reactions are added afterwards, and the light absorption value of p-nitrophenol is determined in 400nm.With various concentrations to nitro
Phenol standard specimen draws standard curve, calculates content of p-nitrophenol.Enzyme-activity unit (U) is defined as at 45 DEG C, is catalyzed in 1min
The enzyme amount hydrolyzed needed for p-NPG produces 1 μM of p-nitrophenol is 1 enzyme-activity unit.
Organic acid content testing：Take sample (Lactobacillus plantarum fermented in soymilk production beta-glucosidase directly convert it is different
Flavone glycoside chemical combination thing liquid) 1mL add 1.5mL centrifuge tubes in, 10000rpm centrifugation 1min, take supernatant to pass through 0.22 μm of filter membrane
Filter and determined for HPLC.Chromatographic condition：Shimadzu LC-20A high performance liquid chromatographs, Aminex HPX-87H organic acids and alcohol point
Post is analysed, mobile phase is 0.005M H2SO4The aqueous solution, flow velocity is 0.6mL/min, and Detection wavelength is 210nm, and column temperature is 35 DEG C.
Isoflavone content is determined：5mL samples are taken to add after the mixing of the ethanol of 5mL 70%, ultrasonic extraction 40min at 60 DEG C,
10000rpm centrifuges 5min, takes supernatant to be determined through 0.22 μm of membrane filtration for HPLC.Chromatographic condition is：Shimadzu LC-20A is high
Effect liquid phase chromatogram instrument, C18Chromatographic column (150mm × 4.6mm, 5.0 μm), mobile phase is acetonitrile and 0.2% formic acid (A：Acetonitrile, B：
0.2% formic acid), gradient elution, flow velocity is 0.8mL/min, and Detection wavelength is 260nm, and column temperature is 35 DEG C.
After measured, original soy glycosides is 5.91mg/mL in soymilk, and genistin is 28.30mg/mL, and daidzein is
3.09mg/mL, after Lactobacillus plantarum fermentation 12h, the daidzin in soymilk is 3.71mg/mL, and genistin is
11.62mg/mL, daidzein is 5.36mg/mL, and genistein is 23.53mg/mL, and beta-glucosidase is 8.64U/mL, breast
Acid is 44.58mM, and acetic acid is 8.79mM, and pH is 5.11.
Embodiment 2
Make soymilk by oneself：Soybean is placed in soaked overnight in distilled water；Soybean skin is removed, and is cleaned with distilled water；Remove skin
Soybean 200g adds 500mL distilled water, is ground using soy bean milk making machine；All slurries are boiled after 15min in 80 DEG C, use double gauze mistake
Filter, removes insoluble residue；The soya-bean milk that will filter out is added in conical flask, 121 DEG C of autoclaving 15min；Room temperature is cooled to, plus
The sucrose solution of the filtration sterilization entered makes concentration be 2% (w/v), and daidzin, genistin, soybean in initial soymilk is measured by sampling
The content of aglycon and genistein, remaining 4 DEG C of refrigerator are saved backup.
Spawn incubation：Go bail for the μ L of Lactobacillus plantarum LYT-3 bacterium solutions 300 being stored in glycerol tube, is inoculated in MRS culture mediums
In, anaerobism bottle cap is covered, in quiescent culture 20h at 37 DEG C.
Fermented and cultured：Lactobacillus plantarum LYT-3 seed liquors are taken to be inoculated in ready soymilk culture medium, inoculum concentration is
4% (v/v), in cultivating 12h at 37 DEG C, takes sample to be used for enzymatic activity, pH, daidzin, genistin, daidzein, dyewood
Element, lactic acid, the measure of acetic acid.
Activity of beta-glucosidase is determined：Lactobacillus plantarum LYT-3 fermentations produce the enzymatic reaction body of beta-glucosidase
System be 1.2mL 0.1M pH be 5.0 phosphoric acid-citrate buffer solution, 0.4mL 8mM p-NPG, 0.4mL enzyme liquid, 45 DEG C reaction
2mL 0.5M sodium carbonate terminating reactions are added after 30min, the light absorption value of p-nitrophenol is determined in 400nm.With various concentrations
P-nitrophenol standard specimen draws standard curve, calculates content of p-nitrophenol.Enzyme-activity unit (U) is defined as at 45 DEG C, in 1min
Enzyme amount needed for interior catalyzing hydrolysis p-NPG produces 1 μM of p-nitrophenol is 1 enzyme-activity unit.
Organic acid content testing：Take sample (Lactobacillus plantarum LYT-3 fermented in soymilk production beta-glucosidase directly turn
Changing isoflavone glucoside chemical combination thing liquid) 1mL added in 1.5mL centrifuge tubes, and 10000rpm centrifugation 1min take supernatant in 0.22 μm of filter
Membrane filtration is determined for HPLC.Chromatographic condition：Shimadzu LC-20A high performance liquid chromatographs, Aminex HPX-87H organic acids and alcohol
Analytical column, mobile phase is 0.005M H2SO4The aqueous solution, flow velocity is 0.6mL/min, and Detection wavelength is 210nm, and column temperature is 35 DEG C.
Isoflavone content is determined：5mL samples are taken to add after the mixing of the ethanol of 5mL 70%, ultrasonic extraction 40min at 60 DEG C,
10000rpm centrifuges 5min, takes supernatant to be determined through 0.22 μm of membrane filtration for HPLC.Chromatographic condition is：Shimadzu LC-20A is high
Effect liquid phase chromatogram instrument, C18Chromatographic column (150mm × 4.6mm, 5.0 μm), mobile phase is acetonitrile and 0.2% formic acid (A：Acetonitrile, B：
0.2% formic acid), gradient elution program is 0-10min, B 5%-80%；10-16min, B 80%；16-22min, B 80%-
5%, flow velocity is 0.8mL/min, and Detection wavelength is 260nm, and column temperature is 35 DEG C.
After measured, original soy glycosides is 5.91mg/mL in soymilk, and genistin is 28.30mg/mL, and daidzein is
3.09mg/mL, after Lactobacillus plantarum LYT-3 fermentations 18h, the daidzin in soymilk is 1.23mg/mL, and genistin is
2.97mg/mL, daidzein is 7.63mg/mL, and genistein is 32.45mg/mL, and beta-glucosidase is 8.49U/mL, breast
Acid is 74.24mM, and acetic acid is 9.05mM, and pH is 4.71.
Embodiment 3
Make soymilk by oneself：Soybean is placed in soaked overnight in distilled water；Soybean skin is removed, and is cleaned with distilled water；Remove skin
Soybean 200g adds 500mL distilled water, is ground using soy bean milk making machine；All slurries are boiled after 15min in 80 DEG C, use double gauze mistake
Filter, removes insoluble residue；The soya-bean milk that will filter out is added in conical flask, 121 DEG C of autoclaving 15min；Room temperature is cooled to, plus
The sucrose solution of the filtration sterilization entered makes concentration be 2% (w/v), and daidzin, genistin, soybean in initial soymilk is measured by sampling
The content of aglycon and genistein, remaining 4 DEG C of refrigerator are saved backup.
Spawn incubation：Go bail for the μ L of Lactobacillus plantarum LYT-3 bacterium solutions 300 being stored in glycerol tube, is inoculated in MRS culture mediums
In, anaerobism bottle cap is covered, in quiescent culture 20h at 37 DEG C.
Fermented and cultured：Lactobacillus plantarum LYT-3 seed liquors are taken to be inoculated in ready soymilk culture medium, inoculum concentration is
4% (v/v), in cultivating 24h at 37 DEG C, takes sample to be used for enzymatic activity, pH, daidzin, genistin, daidzein, dyewood
Element, lactic acid, the measure of acetic acid.
Activity of beta-glucosidase is determined：Lactobacillus plantarum LYT-3 fermentations produce the enzymatic reaction body of beta-glucosidase
System be 1.2mL 0.1M pH be 5.0 phosphoric acid-citrate buffer solution, 0.4mL 8mM p-NPG, 0.4mL enzyme liquid, 45 DEG C reaction
2mL 0.5M sodium carbonate terminating reactions are added after 30min, the light absorption value of p-nitrophenol is determined in 400nm.With various concentrations
P-nitrophenol standard specimen draws standard curve, calculates content of p-nitrophenol.Enzyme-activity unit (U) is defined as at 45 DEG C, in 1min
Enzyme amount needed for interior catalyzing hydrolysis p-NPG produces 1 μM of p-nitrophenol is 1 enzyme-activity unit.
Organic acid content testing：Take Lactobacillus plantarum LYT-3 fermented in soymilk production beta-glucosidase directly convert it is different
Flavone glycoside chemical combination thing liquid 1mL is added in 1.5mL centrifuge tubes, 10000rpm centrifugation 1min, takes supernatant in 0.22 μm of filter membrane mistake
Filter and determined for HPLC.Chromatographic condition：Shimadzu LC-20A high performance liquid chromatographs, Aminex HPX-87H organic acids and alcohol analysis
Post, mobile phase is 0.005M H2SO4The aqueous solution, flow velocity is 0.6mL/min, and Detection wavelength is 210nm, and column temperature is 35 DEG C.
Isoflavone content is determined：5mL samples are taken to add after the mixing of the ethanol of 5mL 70%, ultrasonic extraction 40min at 60 DEG C,
10000rpm centrifuges 5min, takes supernatant to be determined through 0.22 μm of membrane filtration for HPLC.Chromatographic condition is：Shimadzu LC-20A is high
Effect liquid phase chromatogram instrument, C18Chromatographic column (150mm × 4.6mm, 5.0 μm), mobile phase is acetonitrile and 0.2% formic acid (A：Acetonitrile, B：
0.2% formic acid), gradient elution program is 0-10min, B 5%-80%；10-16min, B 80%；16-22min, B 80%-
5%, flow velocity is 0.8mL/min, and Detection wavelength is 260nm, and column temperature is 35 DEG C.
After measured, original soy glycosides is 5.91mg/mL in soymilk, and genistin is 28.30mg/mL, and daidzein is
3.09mg/mL, after Lactobacillus plantarum LYT-3 fermentations 24h, the daidzin in soymilk is 0mg/mL, and genistin is 0mg/
ML, daidzein is 9.05mg/mL, and genistein is 34.14mg/mL, and beta-glucosidase is 7.52U/mL, and lactic acid is
92.45mM, acetic acid is 10.01mM, and pH is 4.12.
Claims (6)
1. a kind of application process of beta-glucosidase in soymilk, it is characterised in that：The plant breast of culture is accessed in soymilk
Bacillus, the beta-glucosidase produced using Lactobacillus plantarum in soymilk fermentation process directly converts the isoflavone glucoside of soymilk
Compound is that isoflavone glucoside matches somebody with somebody based compound, improves the biological effectiveness of soymilk.
2. a kind of application process of the beta-glucosidase according to claim 1 in soymilk, it is characterised in that：The training
The inoculum concentration of foster Lactobacillus plantarum is 4%, with volume basis, and cultivation temperature is 37 DEG C, and incubation time is 24h.
3. a kind of application process of the beta-glucosidase according to claim 1 in soymilk, it is characterised in that including with
Lower step：
(1) soymilk is prepared：Soybean is placed in soaked overnight in distilled water；Soybean skin is removed, and is cleaned with distilled water；Remove skin big
Beans 200g adds 500mL distilled water, is ground using soy bean milk making machine；Gained slurry is boiled after 15min in 80 DEG C, is filtered with double gauze,
Remove insoluble residue；The soya-bean milk that will filter out is added in conical flask, 121 DEG C of sterilizing 15min；Room temperature is cooled to, sucrose is added
Solution, obtains soymilk culture medium；Daidzin, genistin, daidzein and dyewood in initial soymilk culture medium is measured by sampling
The content of element, remaining 4 DEG C of refrigerator are saved backup；
(2) Spawn incubation：Go bail for the μ L of Lactobacillus plantarum bacterium solution 300 being stored in glycerol tube, is inoculated in MRS culture mediums, covers
Anaerobism bottle cap, in quiescent culture 20h at 37 DEG C；
(3) fermented and cultured：Lactobacillus plantarum seed liquor is taken to be inoculated in standby soymilk culture medium, inoculum concentration is 4%, with volume
Fraction meter, in cultivating 24h at 37 DEG C, takes sample to be used to determine activity of beta-glucosidase, pH, daidzin, genistin, soybean
Aglycon, genistein, lactic acid, the content of acetic acid.
4. a kind of application process of the beta-glucosidase according to claim 3 in soymilk, it is characterised in that β-grape
Glycosidase activity assay method is as follows：The enzymatic reaction system that Lactobacillus plantarum fermentation produces beta-glucosidase is 1.2mL
Add after phosphoric acid-citrate buffer solution, 0.4mL 8mM p-NPG, 0.4mL enzyme liquid that 0.1M pH are 5.0,45 DEG C of reaction 30min
Enter 2mL 0.5M sodium carbonate terminating reactions, the light absorption value of p-nitrophenol is determined in 400nm；With the p-nitrophenol of various concentrations
Standard specimen draws standard curve, calculates content of p-nitrophenol；Enzyme-activity unit is defined as at 45 DEG C, the catalyzing hydrolysis p- in 1min
Enzyme amount needed for NPG produces 1 μM of p-nitrophenol is 1 enzyme-activity unit.
5. a kind of application process of the beta-glucosidase according to claim 3 in soymilk, it is characterised in that organic acid
Content assaying method is as follows：Sample 1mL is taken to add in 1.5mL centrifuge tubes, 10000rpm centrifugation 1min take supernatant with 0.22 μm
Membrane filtration is determined for HPLC；Chromatographic condition：High performance liquid chromatograph, Aminex HPX-87H organic acids and alcohol analytical column,
Mobile phase is 0.005M H2SO4The aqueous solution, flow velocity is 0.6mL/min, and Detection wavelength is 210nm, and column temperature is 35 DEG C.
6. a kind of application process of the beta-glucosidase according to claim 3 in soymilk, it is characterised in that isoflavones
Content assaying method is as follows：5mL samples are taken to add after the ethanol solution mixing of 5mL mass percent concentrations 70%, it is ultrasonic at 60 DEG C
40min, 10000rpm centrifugation 5min are extracted, takes supernatant to be determined through 0.22 μm of membrane filtration for HPLC；Chromatographic condition is：It is high
Effect liquid phase chromatogram instrument, C18Chromatographic column, mobile phase is acetonitrile and 0.2% formic acid, and gradient elution, flow velocity is 0.8mL/min, detects ripple
A length of 260nm, column temperature is 35 DEG C.
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| CN110699271A (en) * | 2018-07-10 | 2020-01-17 | 重庆第二师范学院 | Lactobacillus plantarum CQPC02 and application thereof in preparation of food for improving constipation |
| CN110692726A (en) * | 2018-07-10 | 2020-01-17 | 重庆第二师范学院 | Lactobacillus plantarum CQPC01 and application thereof in preparation of food for improving constipation |
| CN111616227A (en) * | 2020-03-30 | 2020-09-04 | 华南农业大学 | Sour pulp bean curd rich in aglycone type isoflavone and preparation method thereof |
| CN112772928A (en) * | 2021-01-29 | 2021-05-11 | 华中农业大学 | Preparation method of fermented bean curd juice and application of fermented bean curd juice in promoting intestinal health |
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| CN113558193A (en) * | 2021-06-24 | 2021-10-29 | 苏州大学 | Preparation method of fermented beverage rich in soybean isoflavone aglycone |
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| CN110699271A (en) * | 2018-07-10 | 2020-01-17 | 重庆第二师范学院 | Lactobacillus plantarum CQPC02 and application thereof in preparation of food for improving constipation |
| CN110692726A (en) * | 2018-07-10 | 2020-01-17 | 重庆第二师范学院 | Lactobacillus plantarum CQPC01 and application thereof in preparation of food for improving constipation |
| CN110699271B (en) * | 2018-07-10 | 2022-11-01 | 善恩康生物科技（苏州）有限公司 | Lactobacillus plantarum CQPC02 and application thereof in preparation of food for improving constipation |
| CN109402000A (en) * | 2018-10-31 | 2019-03-01 | 南京师范大学 | One plant of production beta-glucosidase lactic acid bacteria and its screening technique and the preparation method rich in active flavone aglycone Yoghourt |
| CN111616227A (en) * | 2020-03-30 | 2020-09-04 | 华南农业大学 | Sour pulp bean curd rich in aglycone type isoflavone and preparation method thereof |
| CN112772928A (en) * | 2021-01-29 | 2021-05-11 | 华中农业大学 | Preparation method of fermented bean curd juice and application of fermented bean curd juice in promoting intestinal health |
| CN112772928B (en) * | 2021-01-29 | 2023-04-07 | 华中农业大学 | Preparation method of fermented succus Bambusae semen Phaseoli clear liquid and its application in promoting intestinal health |
| CN113208058A (en) * | 2021-06-16 | 2021-08-06 | 齐鲁工业大学 | Preparation method of natto with easy storage and high content of active isoflavone |
| CN113558193A (en) * | 2021-06-24 | 2021-10-29 | 苏州大学 | Preparation method of fermented beverage rich in soybean isoflavone aglycone |
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