CN106616993A - Preparation method of high-activity functional ingredient enzyme - Google Patents
Preparation method of high-activity functional ingredient enzyme Download PDFInfo
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- CN106616993A CN106616993A CN201611207876.9A CN201611207876A CN106616993A CN 106616993 A CN106616993 A CN 106616993A CN 201611207876 A CN201611207876 A CN 201611207876A CN 106616993 A CN106616993 A CN 106616993A
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Abstract
The invention relates to a preparation method of high-activity functional ingredient enzyme. The preparation method comprises the following steps of using fruits as raw materials, and disinfecting by ultraviolet rays; under the clean condition, cutting; adding honey, fermenting for 30 days at normal temperature, and filtering, so as to obtain a fruit enzyme fermenting raw liquor; inoculating the enzyme fermenting raw liquor with bifidobacterium and lactobacillus acidophilus, and fermenting for 30 to 60 days, so as to obtain the high-activity functional ingredient enzyme; storing into a sealing jar. The preparation method has the advantages that the fermentation time is greatly shortened; the content of functional active ingredient in an enzyme product is increased, the fermenting enzyme production capability of a strain is furthest utilized, the function of the enzyme is stronger, and the health-care functions of the enzyme are effectively improved.
Description
Technical field
The present invention relates to ferment field, more particularly to a kind of preparation of high activity functional component ferment and measure.
Background technology
Ferment is also called " enzyme ", is a kind of a kind of protein less than cell-unit, is a kind of biocatalyst, and it holds
Various chemical reactions in metabolism in load human body, can increase the various biochemical reactions of tissue (oxidation, reduction, decompose, synthesize, turn
Change) speed, the metabolism of every cell in human body is newborn, decomposes, digestion, and synthesis etc. depends on ferment to complete, so ferment
Element is referred to as " spring of life " again.
Along with the growth at people's age, the ferment in people's body can be reduced increasingly, raw due to modern society's dog-eat-dog
Tempo increase living, environmental pollution is increasingly serious, and the activity of ferment is also declining, and metabolic function is also constantly failing.Ferment subtracts
Less and activity decline, the appearance of various symptoms can be caused, due to zymosthenic unicity, to prevent and treat these diseases
Disease, we must absorb more ferment from meals, but ferment is very fragile, and in high-temperature cooking, (more than 60 DEG C just
Lose activity completely), will lose activity in process, the ferment for causing people to absorb from food is very low, it is impossible to meet
The normal need of people.
The production technology of existing ferment is typically all to be formed with the mixing spontaneous fermentation such as various miscellaneous fruits, cereal or mushroom,
Fermentation required time long (fermentation adds the general most short needs of ageing 6 months, the even needs having a year), and fermented what is obtained
Beneficial viable bacteria content is than relatively low in ferment product, and health-care efficacy is not notable enough.
The content of the invention
It is an object of the invention to provide a kind of preparation method of the ferment of high activity functional component, the technique can be greatly shortened
Fermentation time, and the content of functional activity composition in ferment product can be improved, the enzymatic production of bacterial classification can be maximally utilized
Ability, makes ferment with better function, effectively improves the various health cares of ferment.
To solve above-mentioned technical problem, the technical solution used in the present invention is：
A kind of preparation method of high activity functional component ferment, comprises the steps：
(1) enzyme stoste is prepared：Water intaking fruit after carrying out ultraviolet disinfection process, under cleaning condition, is shredded as raw material,
Honey is subsequently adding, normal temperature bottom fermentation 30 days is filtered, and obtains fruit enzyme stoste；Herein in addition to selecting fruit, it is also possible to
Other food materials such as vegetables, edible mushroom, cereal, beans, Chinese medicine are selected as raw material.
(2) secondary fermentation：By enzyme stoste, inoculating bifidobacterium and lactobacillus acidophilus, after fermenting 30-60 days, this is obtained
The high activity functional component ferment of invention, in being stored in hermetically sealed can.Fermentation time according to sampling survey lipase, protease,
Superoxide dismutase activity changes to determine, usually 30-60 days.
In step (2), the inoculum concentration of Bifidobacterium and lactobacillus acidophilus is 1-10%.It is preferred that Bifidobacterium and acidophilus breast
The inoculum concentration of bacillus is 5-8%.
In step (1), the fruit selected from apple, pears, Kiwi berry, pomegranate, dragon fruit, shaddock, banana, lemon, mango,
The mixture of one or more arbitrary proportions in pawpaw, grape, orange, hawthorn, strawberry.
In step (1), the raw fruit is 1 with the weight ratio of honey:0.5-2, preferably 1:1.
In step (2), the Bifidobacterium and lactobacillus acidophilus are commercially available commercially available, it is also possible to by following method systems
：In taking the conical flask that Birid Triple Viable live triple capsule 3 is added to after sterilizing, 50ml physiological saline is added to mix, from taper
1ml is taken in test tube, add the mixing of 9ml SPSSs, so dilution 3 times in bottle；
Under aseptic condition, 0.5ml bacterium solutions are taken out from the bacterium solution after third time dilution, are coated on BBL culture medium plates,
Anaerobism is inverted culture 48h in 37 DEG C of constant incubators after half an hour, and bacterium colony in plate is carried out into gram stain microscopy, takes feminine gender
On BBL culture medium flat plates, 37 DEG C of constant-temperatureanaerobic anaerobic culture 18h are obtained Bifidobacterium to colony inoculation；
Under aseptic condition, 0.5ml bacterium solutions are taken out from the bacterium solution after third time dilution, coat acidifying MRS culture mediums and put down
On ware, anaerobism is inverted culture 48h in 37 DEG C of constant incubators after half an hour, and bacterium colony in plate is carried out into gram stain microscopy,
Take positive bacterium colony to be inoculated on the MRS culture medium flat plates of acidifying, 37 DEG C of constant-temperatureanaerobic anaerobic culture 24h, lactobacillus acidophilus is obtained.
Beneficial effect：The preparation method of the present invention effectively shorten fermentation time (fermentation time is usually 3-4 month,
Totally shorten more than half the time compared with prior art), production efficiency is substantially increased, by activity during compound bacteria-fermented
The monitoring at any time of composition, finally determines the Best Times of the whole fermentation process termination of ferment, so that it is guaranteed that active component reaches
Peak, significantly improves its health-care efficacy.
Specific embodiment
With reference to embodiment, the invention will be further described.
Remarks：1. in following embodiments, the method for testing of lipase, protease and superoxide dismutase content is as follows：
(1) lipase content assay method refers to GB/T 23535-2009；
Enzyme activity is defined：1g solid enzyme powders (or 1mL liquid enzymes), under the conditions of 40 DEG C of temperature and pH7.5,1min hydrolysis substrates
The titratable aliphatic acid of 1umol is produced, as 1 enzyme activity unit is represented with u/g (u/ml).
Lipase under certain condition, can make triglyceride be hydrolyzed into aliphatic acid, diglyceride, monoglycerides and glycerine,
The aliphatic acid available standards aqueous slkali for being discharged is neutralized titration, is counted with PH or phenol peptide indicator solution Indicator Reaction terminal, according to
The iodine number of consumption, calculates its enzyme activity.Reaction equation is：RCOOH+NaOH→RCOONa+H2O。
(2) protease content assay method refers to GB/T23527-2009；
Enzyme activity is defined：1g solid enzyme powders (or 1ml liquid enzymes), in 40 DEG C of (acid pH=3.0, neutral pH=7.5, alkalescence
PH=10.5 under the conditions of), it is an enzyme activity unit that 1min hydrolyzed caseins produce 1 μ g tyrosine.
With the ferment solution to be measured that distilled water compound concentration is 2%, regulation 1g solid enzyme powders (or 1mL liquid enzymes),
Under the conditions of constant temperature degree and pH, it is an enzyme activity unit that 1min hydrolyzed caseins produce 1 μ g tyrosine, is represented with μ g (U/mL).Egg
White enzyme activity is represented with " ferment 1min water casein generates 1 μ g amino acid as an enzyme activity unit ".
(3) method of testing of superoxide dismutase (SOD) refers to GB/T5009.171-2003.
SOD amounts required when suppressing mouse thymus cells speed 50% when 25 DEG C are a unit of activity.
Using pyrogallol in the basic conditions can autoxidation rapidly, discharge O2-, generate colored intermediate product.Instead
First become yellow green after should starting, yellow is switched to after a few minutes, linear session maintains 3～4min.Enzyme liquid is added then to suppress it
Autoxidation speed, determines the absorbance of solution at 325nm.Unit of enzyme activity suppresses adjacent benzene using per minute in 1mL reactant liquors
Triphenol autoxidation speed is up to the quantification of unit of activity of enzyme when 50%.
2. in following embodiments, the Bifidobacterium and lactobacillus acidophilus of employing are obtained by the following method：
In taking the conical flask that Birid Triple Viable live triple capsule 3 is added to after sterilizing, 50ml physiological saline is added to mix, from
1ml is taken in conical flask in test tube, the mixing of 9ml SPSSs, so dilution 3 times is added, with this be labeled as 10-1,
10-2、10-3；
Under aseptic condition, 0.5ml bacterium solutions are taken out from 10-3 test tubes, are coated on BBL culture mediums (market purchase) plate,
Anaerobism is inverted culture 48h in 37 DEG C of constant incubators after half an hour, and bacterium colony in plate is carried out into gram stain microscopy, takes feminine gender
On BBL culture medium flat plates, 37 DEG C of constant-temperatureanaerobic anaerobic culture 18h are obtained Bifidobacterium to colony inoculation；
Under aseptic condition, 0.5ml bacterium solutions are taken out from 10-3 test tubes, coat acidifying MRS culture mediums (market purchase) and put down
On ware, anaerobism is inverted culture 48h in 37 DEG C of constant incubators after half an hour, and bacterium colony in plate is carried out into gram stain microscopy,
Take positive bacterium colony to be inoculated on the MRS culture medium flat plates of acidifying, 37 DEG C of constant-temperatureanaerobic anaerobic culture 24h, lactobacillus acidophilus is obtained.
Embodiment 1
(1) enzyme stoste is prepared：Take apple, pears, Kiwi berry, pomegranate, dragon fruit, shaddock, banana, lemon, mango, pawpaw
Each 200g after carrying out ultraviolet disinfection process, under cleaning condition, is cut into sheet as raw material, is well mixed, and adds honey, honeybee
Honey is 1 with fruit weight ratio:1, normal temperature bottom fermentation 30 days is filtered, and obtains enzyme stoste；Determine lipase, protease, super oxygen
The enzyme activity of thing mutase, lipase activity reaches 11.5u/ml, and proteinase activity reaches 1056.6u/ml, superoxide dismutase
Enzyme activity reaches 526.5u/ml；
(2) secondary fermentation：Inoculating bifidobacterium and lactobacillus acidophilus, inoculum concentration is respectively 8%, and secondary fermentation 30 days is obtained final product,
Enzyme liquid is stored in hermetically sealed can.
Measure：Lipase activity reaches 52.8u/ml, and proteinase activity reaches 2859.3u/ml, superoxide dismutase enzyme
Work reaches 1347.4u/ml.
Comparative example
(1) enzyme stoste is prepared：Take apple, pears, Kiwi berry, pomegranate, dragon fruit, shaddock, banana, lemon, mango, pawpaw
Each 200g after carrying out ultraviolet disinfection process, under cleaning condition, is cut into sheet as raw material, is well mixed, and adds honey, honeybee
Honey is 1 with fruit weight ratio:1, normal temperature bottom fermentation 30 days is filtered, and obtains fruit enzyme stoste；
(2) the fruit enzyme stoste natural secondary for obtaining step (1) ferments 30 days, obtains final product, and enzyme liquid is stored in close
In sealed cans.
Measure：Lipase activity reaches 20.7u/ml, and proteinase activity reaches 1502.3u/ml, superoxide dismutase enzyme
Work reaches 701.8u/ml.
Can be seen that by the testing result of embodiment 1 and comparative example：The ferment that secondary fermentation is obtained using spontaneous fermentation
In, the enzyme activity of three kinds of enzymes improves compared to one time fermentation ferment, but changes little；Add Bifidobacterium and acidophilus breast
In the ferment that the secondary fermentation of bacillus is obtained, the enzyme activity of three kinds of enzymes is significantly improved.
Embodiment 2
(1) enzyme stoste is prepared：Take Kiwi berry 300g, pomegranate 400g, orange 200g, hawthorn 100g, strawberry 300g, lemon
400g after carrying out ultraviolet disinfection process, under cleaning condition, is cut into sheet as raw material, is well mixed, and adds honey, honey
It is 1.5 with fruit weight ratio:1, normal temperature bottom fermentation 30 days is filtered, and obtains fruit enzyme stoste；Determine lipase, protease, surpass
The enzyme activity of superoxide dismutase, lipase activity reaches 13.5u/ml, and proteinase activity reaches 1508.7u/ml, superoxides discrimination
Change enzyme enzyme activity and reach 554.1u/ml；
(2) secondary fermentation：Inoculating bifidobacterium and lactobacillus acidophilus, inoculum concentration is respectively 5%, and secondary fermentation 35 days is obtained final product,
Enzyme liquid is stored in hermetically sealed can.
Measure：Lipase activity reaches 62.2u/ml, and proteinase activity reaches 2922.1u/ml, superoxide dismutase enzyme
Work reaches 1415.5u/ml.
Embodiment 3
(1) enzyme stoste is prepared：Take asparagus 200g, Hericium erinaceus 300g, agaric 200g, white fungus 100g, mushroom 300g, tea
Tree mushroom 100g after carrying out ultraviolet disinfection process, under cleaning condition, is cut into sheet as raw material, is well mixed, and adds honey,
Honey is 0.5 with edible mushroom weight ratio:1, normal temperature bottom fermentation 30 days is filtered, and obtains enzyme stoste；Determine lipase, protease,
The enzyme activity of superoxide dismutase, lipase activity reaches 5.5u/ml, and proteinase activity reaches 912.5u/ml, superoxides discrimination
Change enzyme enzyme activity and reach 658.2u/ml；
(2) secondary fermentation：Inoculating bifidobacterium and lactobacillus acidophilus, inoculum concentration is respectively 6%, and secondary fermentation 45 days is obtained final product,
Enzyme liquid is stored in hermetically sealed can.
Measure：Lipase activity reaches 8.2u/ml, and proteinase activity reaches 1204.6u/ml, superoxide dismutase enzyme
Work reaches 815.5u/ml.
Embodiment 4
(1) enzyme stoste is prepared：Take brown rice 400g, soybean 100g, maize germ 200g, seed of Job's tears 200g, barley 150g
As raw material, after carrying out ultraviolet disinfection process, under cleaning condition, it is well mixed, addition honey, honey is with cereal weight ratio
0.6:1, normal temperature bottom fermentation 30 days is filtered, and obtains enzyme stoste；Determine lipase, protease, the enzyme of superoxide dismutase
Living, lipase activity reaches 0.5u/ml, and proteinase activity reaches 655.5u/ml, and superoxide dismutase enzyme activity reaches
911.4u/ml；
(2) secondary fermentation：Inoculating bifidobacterium and lactobacillus acidophilus, inoculum concentration is respectively 8%, and secondary fermentation 50 days is obtained final product,
Enzyme liquid is stored in hermetically sealed can.
Measure：Lipase activity reaches 0.8u/ml, and proteinase activity reaches 774.1u/ml, superoxide dismutase enzyme activity
Reach 1155.8u/ml.
The above is not limited thereto for the more preferred specific embodiment of the present invention, but the scope of the present invention, root
All should being included within protection scope of the present invention of corresponding change is carried out according to technical scheme and inventive concept.
Claims (5)
1. a kind of preparation method of high activity functional component ferment, it is characterised in that comprise the steps：
(1) enzyme stoste is prepared：Water intaking fruit after carrying out ultraviolet disinfection process, under cleaning condition, is shredded, then as raw material
Add honey, normal temperature bottom fermentation 30 days to filter, obtain enzyme stoste；
(2) secondary fermentation：By enzyme stoste inoculating bifidobacterium and lactobacillus acidophilus, after fermenting 30-60 days, obtain the present invention's
High activity functional component ferment, in being stored in hermetically sealed can；
In step (2), the inoculum concentration of Bifidobacterium and lactobacillus acidophilus is 1-10%.
2. the preparation method of high activity functional component ferment as claimed in claim 1, it is characterised in that described in step (2)
The inoculum concentration of Bifidobacterium and lactobacillus acidophilus is 5-8%.
3. the preparation method of high activity functional component ferment as claimed in claim 1, it is characterised in that described in step (1)
Fruit is selected from apple, pears, Kiwi berry, pomegranate, dragon fruit, shaddock, banana, lemon, mango, pawpaw, grape, orange, hawthorn, grass
The mixture of one or more arbitrary proportions in the certain kind of berries.
4. the preparation method of the high activity functional component ferment as described in claim 1 or 2 or 3, it is characterised in that step (1)
In, the raw fruit is 1 with the weight ratio of honey:0.5-2.
5. the preparation method of high activity functional component ferment as claimed in claim 4, it is characterised in that the raw fruit with
The weight ratio of honey is 1:1.
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Cited By (8)
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|---|---|---|---|---|
| CN106983139A (en) * | 2017-05-16 | 2017-07-28 | 西昌学院 | Potato ferment made of potato and preparation method thereof |
| CN107173799A (en) * | 2017-06-29 | 2017-09-19 | 张萌 | Extend the preparation method of the shelf life of ferment |
| CN107307407A (en) * | 2017-08-01 | 2017-11-03 | 福建新之源生物制品有限公司 | A kind of fruit ferment drink |
| CN108713669A (en) * | 2018-05-24 | 2018-10-30 | 浙江竹孝宝生物科技有限公司 | A kind of preparation method of leaf of bamboo enzyme beverage |
| CN109222080A (en) * | 2018-10-10 | 2019-01-18 | 曹飞英 | Mixed fruit nutrition enzyme |
| CN109527553A (en) * | 2018-12-25 | 2019-03-29 | 南京林业大学 | A kind of food medicine woods fruit function ferment and preparation method thereof |
| CN109938338A (en) * | 2019-05-05 | 2019-06-28 | 河西学院 | A kind of high-performance "Hami" melon ferment and preparation method thereof |
| CN111955725A (en) * | 2020-08-27 | 2020-11-20 | 江润金 | Preparation method and application of lemon enzyme |
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106983139A (en) * | 2017-05-16 | 2017-07-28 | 西昌学院 | Potato ferment made of potato and preparation method thereof |
| CN107173799A (en) * | 2017-06-29 | 2017-09-19 | 张萌 | Extend the preparation method of the shelf life of ferment |
| CN107307407A (en) * | 2017-08-01 | 2017-11-03 | 福建新之源生物制品有限公司 | A kind of fruit ferment drink |
| CN108713669A (en) * | 2018-05-24 | 2018-10-30 | 浙江竹孝宝生物科技有限公司 | A kind of preparation method of leaf of bamboo enzyme beverage |
| CN109222080A (en) * | 2018-10-10 | 2019-01-18 | 曹飞英 | Mixed fruit nutrition enzyme |
| CN109527553A (en) * | 2018-12-25 | 2019-03-29 | 南京林业大学 | A kind of food medicine woods fruit function ferment and preparation method thereof |
| CN109938338A (en) * | 2019-05-05 | 2019-06-28 | 河西学院 | A kind of high-performance "Hami" melon ferment and preparation method thereof |
| CN111955725A (en) * | 2020-08-27 | 2020-11-20 | 江润金 | Preparation method and application of lemon enzyme |
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