CN104585762B - A kind of Herba Dendrobii fermented product and preparation method thereof - Google Patents
A kind of Herba Dendrobii fermented product and preparation method thereof Download PDFInfo
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- CN104585762B CN104585762B CN201510025608.4A CN201510025608A CN104585762B CN 104585762 B CN104585762 B CN 104585762B CN 201510025608 A CN201510025608 A CN 201510025608A CN 104585762 B CN104585762 B CN 104585762B
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Abstract
The present invention provides a kind of Herba Dendrobii fermented product and preparation method thereof.The method comprises the steps: 1) by broken for Herba Dendrobii rear extracting in water, prepare extracting solution；2), after adding medium component in extracting solution, access Leuconostoc mesenteroides and carry out the first fermentation, prepare the first fermentation liquid；3) in the fruit and vegerable liquid that pH value is 4.5～6, add pectase and carry out enzymolysis processing, prepare enzymolysis fruit and vegerable liquid；4), after adding medium component in enzymolysis fruit and vegerable liquid, access compound lactobacillus and carry out the second fermentation, prepare the second fermentation liquid；5), after adding medium component in the second fermentation liquid, access mulriple yeasts and carry out the 3rd fermentation, prepare the 3rd fermentation liquid；6) it is centrifuged after the first fermentation liquid and the 3rd fermentation liquid being mixed, after centrifuged supernatant homogenizing, sterilizing, prepares Herba Dendrobii fermented product.Herba Dendrobii fermented product prepared by the present invention is in good taste, unique flavor, has the functions such as good immunomodulating and antioxidation, is suitable for crowd in extensive range.
Description
Technical field
The present invention relates to a kind of fermented product, particularly relate to a kind of Herba Dendrobii fermented product and preparation method thereof.
Background technology
Herba Dendrobii, has another name called Herba hedyotis costatae, Yunnan iron sheet, belongs to micro-specific item, the perennial herbaceous plant that grows nonparasitically upon another plant of the orchid family, is mainly distributed
In China Anhui, Zhejiang, Fujian and other places.Herba Dendrobii has and promotes the production of body fluid and hypoglycemic activity, and it can promote glandular secretion and internal organs
Motion, reduces streptozotocin induction glycosuria blood glucose value；Additionally, Herba Dendrobii can also enhancing human body immunity power, promote tumor animal
The phagocytic function of macrophage, the propagation of enhancing T lymphocyte and differentiation and the activity of NK cell, non-specific cell is exempted from
Epidemic disease, specific cellular immunity and humoral immune function all improve effect.
At present, there is a small amount of relevant report that Herba Dendrobii is processed into the products such as goods.Such as, Publication No. CN
The Chinese patent of 103734824A discloses a kind of dendrobium officinale healthcare green juice, comprises the fresh bar of Herba Dendrobii 200～250
Part, Aloe 100～150 parts, carambola 50～150 parts, kiwi fruit 50～100 parts, green apple 50～150 parts, acacia honey 30～60
Part, stabilizer 5～10 parts, citric acid 1～2 parts, surplus are water.After the raw materials such as the fresh bar of Herba Dendrobii are mainly squeezed the juice by this patent
Mixing, blends water after adding acacia honey, citric acid and stabilizer and prepares, although it more equalizes rich in multiple fruit juice, nutrition,
But it utilizes rate variance to Herba Dendrobii raw material, product health care is limited.
The Chinese patent of Publication No. CN 102940038A discloses the system of a kind of dendrobium officinale plant protein lactic acid beverage
Preparation Method, extracts after iron-sheet dendrobe fresh product is broken, forms Herba Dendrobii juice through leaching filtrate by milling treatment of colloid,
Prepare bean milk simultaneously, and access lactobacillus casei after being mixed with Herba Dendrobii juice by bean milk and streptococcus thermophilus carries out fermentation training
Support, until the pH value of bean milk is 4.3～4.5, finally by the cold preservation 24h at 1 DEG C～4 DEG C of the bean milk after fermentation culture, prepare and produce
Product.Herba Dendrobii is extracted by the method, thus improves raw material availability, but its nutritional labeling is the most single, mouth
Sense, local flavor and health care all have much room for improvement.
Summary of the invention
The present invention provides a kind of Herba Dendrobii fermented product and preparation method thereof, is used for solving iron sheet stone of the prior art
Dry measure used in former times series products composition is single, mouthfeel, local flavor, health care limited and Herba Dendrobii cannot be carried out the technology such as effective utilization lack
Fall into.
The present invention provides the preparation method of a kind of Herba Dendrobii fermented product, comprises the steps:
1) by broken for Herba Dendrobii rear extracting in water, extracting solution is prepared；
2), after adding medium component in described extracting solution, access Leuconostoc mesenteroides and carry out the first fermentation, work as fermentation
The pH value of liquid reduces by more than 0.5, and the content of reducing sugar of fermentation liquid less than 1.5% time terminate described first fermentation, prepare the
One fermentation liquid；
3) in the fruit and vegerable liquid that pH value is 4.5～6, add pectase and carry out enzymolysis processing, prepare enzymolysis fruit and vegerable liquid；
4) after adding medium component in described enzymolysis fruit and vegerable liquid, access compound lactobacillus and carry out the second fermentation, when sending out
The pH value of ferment liquid terminates described second fermentation when reducing by more than 0.5, prepare the second fermentation liquid；Wherein, described compound lactobacillus choosing
From streptococcus thermophilus, Deshi Lactobacillus, bacillus acidophilus, lactobacillus casei, Lactobacillus plantarum, Lactobacillus fermenti, rhamnose breast
In bacillus, Lactobacillus brevis, lactobacillus helveticus, pediococcus acidilactici, Pediococcus pentosaceus more than four kinds；
5) after adding medium component in described second fermentation liquid, access mulriple yeasts and carry out the 3rd fermentation, when sending out
Terminate described 3rd fermentation when the total sugar content of ferment liquid is less than 2%, prepare the 3rd fermentation liquid；Wherein, described mulriple yeasts choosing
From two or more with wine yeast of rice wine yeast, beer yeast, yellow wine yeast and fruit wine；
6) by centrifugal after described first fermentation liquid and described 3rd fermentation liquid mixing, after centrifuged supernatant homogenizing, sterilizing,
Prepare Herba Dendrobii fermented product.
In the preparation process in accordance with the present invention, described Leuconostoc mesenteroides can select Leuconostoc mesenteroides dextran sub-
Plant (L.mesenteroides subsp.dextranicum) or leuconostoc mesenteroides sub species cremoris (L.mesenteroides
And each bacterium of the present invention can use in the way of using mycopowder or bacterium solution subsp.cremoris),.Using mycopowder
Time, the viable count of described each lactic acid bacteria mycopowder can be 109About CFU/g, the viable count of each yeast mycopowder can be
1010About CFU/g.Additionally, the inoculum concentration of described Leuconostoc mesenteroides (mycopowder) can be 0.05～0.2%, such as 0.1%,
The inoculum concentration of described compound lactobacillus (mycopowder) can be 0.05～0.2%, such as 0.1%, described mulriple yeasts (mycopowder)
Inoculum concentration can be 0.01～0.02%.When using bacterium solution, the viable count and the inoculum concentration that can refer to above-mentioned mycopowder connect
Kind.Each bacterial strain conventionally can activate according to needs before use.
Herba Dendrobii raw material is not made considered critical by the present invention, and it can be fresh Herba Dendrobii, it is also possible to be dry ferrum
Skin Herba Dendrobii.In concrete scheme of the present invention, dry Herba Dendrobii can be used as raw material, its moisture content is≤15.0%.Enter one
Step ground, step 1) in, Herba Dendrobii is crushed to 40～80 mesh, and controls the quality proportioning of the Herba Dendrobii after crushing and water
For 1:40～60, the temperature of described extraction is 90～120 DEG C, and extraction time is 5～30min.System is extracted by described Herba Dendrobii
The pH of the extracting solution obtained is 4.5～6.Extraction process can more efficiently make the active component in Herba Dendrobii raw material discharge,
Thus further increase raw material availability.
In the present invention, described medium component can include carbon source, nitrogen source, inorganic salt, trace element etc., this area skill
Art personnel can select suitable medium component according to the concrete bacterial strain cultivated or compound bacteria.At concrete scheme of the present invention
In, described medium component includes one or more in sugar, peptide, inorganic salt and surfactant, average in wherein said peptide
Gross mass content >=80% of the molecular weight component less than 1000Da.
That is, another aspect of the present invention additionally provides a kind of fluid medium for Leuconostoc mesenteroides fermentation, and it includes
Herba Dendrobii extracting solution, sugar, peptide, inorganic salt and surfactant, wherein sugar mass content in described extracting solution can be 5
～10%, peptide mass content in described extracting solution can be 0.3～0.8%, inorganic salt quality in described extracting solution
Content can be 0.1～0.3%, and surfactant mass content in described extracting solution can be 0.01～0.1%；This ferrum
Skin dendrobium extract can be by preparing broken for Herba Dendrobii rear extracting in water.
Another further aspect, present invention also offers a kind of liquid training fermented for compound lactobacillus and/or mulriple yeasts
Supporting base, it includes enzymolysis fruit and vegerable liquid, sugar, peptide, inorganic salt and surfactant, wherein sugar quality in described enzymolysis fruit and vegerable liquid
Content can be 5～10%, and peptide mass content in described enzymolysis fruit and vegerable liquid can be 0.3～0.8%, and inorganic salt is described
Mass content in enzymolysis fruit and vegerable liquid can be 0.1～0.3%, surfactant mass content in described enzymolysis fruit and vegerable liquid
Can be 0.01～0.1%；This enzymolysis fruit and vegerable liquid can be by adding pectinase enzymatic hydrolysis system in the fruit and vegerable liquid that pH value is 4.5～6
?.
Further, described sugar can be selected from one or more in glucose, sucrose and lactose, such as sucrose；Described
Inorganic salt can be selected from one or more in sodium salt, calcium salt, manganese salt, potassium salt and magnesium salt, such as calcium carbonate；Described surface activity
Agent can be selected from that fatty glyceride, fatty acid Pyrusussuriensis be smooth and one or more in Polysorbate, such as Tween 80 etc..Especially
It is that the component of heretofore described peptide is not particularly limited, if the wherein mean molecule quantity total matter of component less than 1000Da
Amount content >=80%, further >=90%, use described peptide as nitrogen source can remarkably promote Leuconostoc mesenteroides and
Compound lactobacillus and the growth of mulriple yeasts.
The step 2 of concrete scheme of the present invention) in, in described extracting solution, add sugar, peptide and inorganic salt, make described sugar in institute
Stating the mass content in extracting solution is 5～10%, and described peptide mass content in described extracting solution is 0.3～0.8%, described
Inorganic salt mass content in described extracting solution is 0.1～0.3%, and the temperature controlling described first fermentation is 22～28
DEG C, rotating speed is 80～120r/min.Under this condition Leuconostoc mesenteroides is fermented, it is possible to improve the mouthfeel of product and promote
Enter the performance of product health care.Additionally, the pH value controlling this fermentation liquid reduces by more than 0.5, (pH value of such as fermentation liquid is 4.5
～5), and the content of reducing sugar of fermentation liquid is less than 1.5%, not only contributes to avoid product tart flavour overweight and affect palatability,
And low content of reducing sugar can make product have more wide applicability, particularly can drink by diabetic.
In the present invention, described fruit and vegerable liquid is serosity, juice of making through fruit and/or fruits and vegetables etc., the most permissible
Use commercial fruit and vegetable juice etc., the most preferably fruit and vegetable juice without additives such as any preservative.Further, it is also possible to
It is crushed to 40～80 mesh, such as 60 mesh after choosing fruit and vegetable materials, prepares described fruit and vegerable liquid；This particle size range both can improve fermentation
Speed, also help ensure fermented product mouthfeel.Additionally, the kind of fruit and vegetable materials is the most strictly limited by the present invention, special
Be not can choose locality abound with and/or be easy to processing fruits and vegetables as raw material, such as Fructus Ananadis comosi, Citrullus vulgaris, Fructus Fragariae Ananssae, Herba Marsileae Quadrifoliae
Really, Fructus Musae, Fructus Mangifera Indicae, Fructus Chaenomelis, Radix Raphani, Fructus Cucumidis sativi, Fructus Lycopersici esculenti etc.；Further, when choosing fruit and vegetable materials, can be for raw material itself
Characteristic (such as pH value) carries out reasonably combined, and make the fruit and vegerable liquid pH value made in the range of 4.5～6, thus without add
Other pH adjusting agent regulates the pH value of fruit and vegerable liquid, in order to ensure the genuineness of product.
Pectase of the present invention can by ordinary city available from.In described fruit and vegerable liquid, add pectase mainly use
In decomposing pectin etc., so that the nutritional labeling of fruit and vegetable materials discharges more thorough, to improve raw material availability.Further
Ground, the consumption of described pectase is every gram of fruit and vegerable liquid 2～3 unit, and the temperature controlling described enzymolysis processing is 40～50 DEG C,
Time is 2～3h.
Step 4 of the present invention) in, in described enzymolysis fruit and vegerable liquid, add sugar and peptide, make described sugar at described enzymolysis fruit and vegerable liquid
In mass content be 3～5%, described peptide mass content in described enzymolysis fruit and vegerable liquid is 0.3～0.8%；Described compound
Lactic acid bacteria at least includes streptococcus thermophilus and Deshi Lactobacillus, described streptococcus thermophilus, Deshi Lactobacillus and remaining lactic acid bacteria
Weight proportion is 3:2:(2～5), and control described second fermentation temperature be 18～23 DEG C, rotating speed is 80～120r/min.
Further, one or more during remaining lactic acid bacteria described can be following first to fourth component:
First component: include bacillus acidophilus and lactobacillus casei, and the weight of bacillus acidophilus and lactobacillus casei
Proportioning is (0.5～1.5): (0.5～1.5)；
Second component: include pediococcus acidilactici and Pediococcus pentosaceus, and the weight of pediococcus acidilactici and Pediococcus pentosaceus
Proportioning is (0.5～1.5): (0.5～1.5)；
3rd component: include Lactobacillus plantarum and lactobacillus rhamnosus, and Lactobacillus plantarum and lactobacillus rhamnosus
Weight proportion is (0.5～1): (0.5～1)；
4th component: include Lactobacillus fermenti, Lactobacillus brevis and lactobacillus helveticus, and Lactobacillus fermenti, Lactobacillus brevis
With the weight proportion between lactobacillus helveticus is (0.5～1): (0.5～1): (0.5～1).
Step 5 of the present invention) in, in described second fermentation liquid, add sugar and peptide, make described sugar at described second fermentation liquid
In mass content be 4～8%, described peptide mass content in described second fermentation liquid is 0.3～0.8%；And control
The weight proportion between two primary yeasts in described mulriple yeasts is 1:(0.8～1.2), the temperature of described 3rd fermentation is 16
～20 DEG C, rotating speed is 40～60r/min.In the case of particularly each yeast in mulriple yeasts all uses mycopowder, multiple
When in conjunction yeast, each yeast weight is identical, the local flavor of fermented product is splendid.
Further, any one during described mulriple yeasts is selected from following first to fourth component:
First component: include rice wine yeast and fruit wine and wine yeast, and rice wine yeast, fruit wine and wine yeast
Between weight proportion be 1:(0.8～1.2)；
Second component: include beer yeast and fruit wine and wine yeast, and rice wine yeast, fruit wine and wine yeast
Between weight proportion be 1:(0.8～1.2)；
3rd component: include rice wine yeast, beer yeast and yellow wine yeast, and rice wine yeast, beer yeast and yellow wine
Weight proportion between yeast is 1:(0.8～1.2): (0.8～1.2)；
4th component: include rice wine yeast, yellow wine yeast and fruit wine and wine yeast, and rice wine yeast, yellow wine ferment
Weight proportion between mother and fruit wine and wine yeast is 1:(0.8～1.2): (0.8～1.2).
Step 6 of the present invention) in, the weight proportion of described first fermentation liquid and described 3rd fermentation liquid is (4～6): (4～
6).Further, before described homogenizing, optionally centrifuged supernatant can be allocated, such as, can add appropriate sugar, peptide etc. and come
Improving the mouthfeel of product and improve the health care etc. of product, wherein said sugar mass content in centrifuged supernatant can be
8～16%, described peptide mass content in centrifuged supernatant can be 0.5～2%.
Further, described sugar can include white sugar and brown sugar, and described white sugar and brown sugar quality in described sugar
Proportioning can be 1:0.5～2.Additionally, described peptide can be that in collagen peptide, and the former peptide of this peptide, mean molecule quantity is less than 1000Da
Gross mass content >=80% of component, further >=90%.
The present invention also provides for a kind of Herba Dendrobii fermented product, prepares according to any of the above-described described preparation method.
The enforcement of the present invention program, at least has the advantage that
1, the present invention uses Leuconostoc mesenteroides to ferment Herba Dendrobii extracting solution, the most more efficient releases ferrum
The activity of skin Herba Dendrobii and nutritional labeling, improve the utilization rate of Herba Dendrobii, additionally enhance the health care of fermentation liquid；With
Time, the present invention uses compound lactobacillus and mulriple yeasts to ferment fruit and vegerable liquid successively, not only ensure that in fermented product
Nutritional labeling comprehensively and equilibrium, additionally improve the mouthfeel of fermented product and improve the local flavor of fermented product.
2, the condition of fermentation technology is optimized by the present invention, not only increases fermenting speed, shortens fermentation time,
In addition the mouthfeel of fermented product, local flavor and the function such as immunomodulating and antioxidation are significantly improved.
3, the Herba Dendrobii fermented product that prepared by the present invention is in good taste, unique flavor, nutritional labeling general equilibrium, edible side
Just, this fermented product does not contains any additive, green health；Additionally, it also has the merits such as good immunomodulating and antioxidation
Can, it is suitable for crowd in extensive range.
Accompanying drawing explanation
Fig. 1 is the mouse T lymphocyte multiplication capacity that ConA is induced by the fermented product of the embodiment of the present invention and reference examples
Evaluation result；
Fig. 2 is the mouse T lymphocyte multiplication capacity that LPS is induced by the fermented product of the embodiment of the present invention and reference examples
Evaluation result；
Fig. 3 is the anti-oxidation function evaluation result of the fermented product of the embodiment of the present invention and reference examples.
Detailed description of the invention
For making the object, technical solutions and advantages of the present invention clearer, below in conjunction with accompanying drawing and the enforcement of the present invention
Example, is clearly and completely described the technical scheme in the embodiment of the present invention, it is clear that described embodiment is the present invention
A part of embodiment rather than whole embodiments.Based on the embodiment in the present invention, those of ordinary skill in the art are not having
Make the every other embodiment obtained under creative work premise, broadly fall into the scope of protection of the invention.
Each lactic acid bacteria that various embodiments of the present invention are used is all from Research for Industrial Microbial Germ preservation administrative center (CICC),
Each lactic acid bacteria is respectively as follows: leuconostoc mesenteroides sub species cremoris (CICC 22181), streptococcus thermophilus (CICC 6222), De Shi breast
Bacillus lactic acid subspecies (CICC 6077), bacillus acidophilus (CICC 6081), lactobacillus casei (CICC 6117), plant breast bar
Bacterium (CICC 22132), Lactobacillus fermenti (CICC 22538), lactobacillus rhamnosus (CICC 6147), Lactobacillus brevis (CICC
23474), lactobacillus helveticus (CICC 22172), pediococcus acidilactici (CICC 10346), Pediococcus pentosaceus (CICC 20536),
The viable count of each lactic acid bacteria is each about 109CFU/g；
Each yeast is all from Angel Yeast Co., Ltd, and each saccharomycetic viable count is each about 1010CFU/g；
Gallus Domesticus peptide and collagen peptide, purchased from Beijing Zhongshi Haishi Biotechnology Co., Ltd., in two kinds of peptides, mean molecule quantity is little
In 1000Da component gross mass content all >=80%；
Pectase: purchased from Pangbo Bioengineering Co Ltd, Nanning；
Embodiment 1
1, the first fermentation
The Herba Dendrobii of moisture content about 13% is crushed to 60 mesh, adds water (i.e. solid-liquid ratio according to weight proportion 1:50
For 1:50) after, at a temperature of about 105 DEG C, extracting about 5min, prepared pH value is about the extracting solution of 5.5；
In above-mentioned extracting solution, add sucrose, Gallus Domesticus peptide and calcium carbonate, make sucrose mass content in extracting solution be about
7%, Gallus Domesticus peptide mass content in extracting solution is about 0.6%, and calcium carbonate mass content in extracting solution is about 0.2%,
After stirring and evenly mixing, the inoculum concentration according to about 0.1% accesses Leuconostoc mesenteroides in extracting solution and carries out the first fermentation, controls the
The temperature of one fermentation is about 24 DEG C, and speed of agitator is about 110r/min, when the pH value reduction by more than 0.5 of fermentation liquid, and
Terminate the first fermentation when the content of reducing sugar of fermentation liquid is less than 1.5%, prepare the first fermentation liquid (pH value is 4.5～5).
Use the cell concentration of spectrophotometry the first fermentation liquid, the results are shown in Table 1.
2, enzymolysis processing
After being 2:1 mixing by fresh Fructus Mangifera Indicae and Radix Dauci Sativae according to weight proportion, being crushed to 60 mesh, prepared pH value is about 5.5
Fruit and vegerable liquid；
Adding pectase in above-mentioned fruit and vegerable liquid and carry out enzymolysis processing, wherein the consumption of pectase is every gram of fruit and vegerable liquid 2.5
About unit, and the temperature that controlled enzymatic hydrolysis processes is about 45 DEG C, and the time is about 3h, prepares enzymolysis fruit and vegerable liquid.
3, the second fermentation
In described enzymolysis fruit and vegerable liquid, add sucrose and Gallus Domesticus peptide, make sucrose mass content in enzymolysis fruit and vegerable liquid be about
4%, Gallus Domesticus peptide mass content in enzymolysis fruit and vegerable liquid is about 0.5%, after stirring and evenly mixing, according to the inoculum concentration of about 0.1%
In enzymolysis fruit and vegerable liquid access compound lactobacillus carry out the second fermentation, compound lactobacillus by streptococcus thermophilus, Deshi Lactobacillus, addicted to
Lactobacillus lactis and lactobacillus casei composition, wherein between streptococcus thermophilus, Deshi Lactobacillus, bacillus acidophilus and lactobacillus casei
Weight proportion be 3:2:1.5:1.5, and the temperature controlling the second fermentation is about 20 DEG C, and speed of agitator is that 100r/min is left
The right side, terminates the second fermentation when the pH value of fermentation liquid reduces by more than 0.5, prepares the second fermentation liquid (pH value is 4.5～5).
4, the 3rd fermentation
In above-mentioned second fermentation liquid, add sucrose and Gallus Domesticus peptide, make sucrose mass content in the second fermentation liquid be about
6%, Gallus Domesticus peptide mass content in the second fermentation liquid is about 0.5%, after stirring and evenly mixing, according to the inoculum concentration of about 0.01%
Carrying out the 3rd fermentation to accessing mulriple yeasts, described mulriple yeasts is made up of with wine yeast rice wine yeast and fruit wine,
Wherein the weight proportion of rice wine yeast and fruit wine and wine yeast is 1:0.8；And the temperature controlling the 3rd fermentation is 18 DEG C
Left and right, speed of agitator is about 50r/min, terminates the 3rd fermentation when the total sugar content of fermentation liquid is less than 2%, prepares the 3rd
Ferment liquid.
5, preparation Herba Dendrobii fermented product
After above-mentioned first fermentation liquid and the 3rd fermentation liquid are mixed according to weight proportion 1:1, under 4000r/min from
The heart about 15min, carries out homogenizing, sterilizing to centrifuged supernatant, prepares Herba Dendrobii fermented product (pH value is 3.8～4.2).
50 experimenters making age level be distributed in 20 years old～80 years old eat the Herba Dendrobii fermented product of above-mentioned preparation,
And respectively this Herba Dendrobii fermented product is carried out taste and flavor marking according to following standard:
Mouthfeel scoring criterion: full marks 10 points, wherein sweet taste and persistence length total score meter 3.5 points, tart flavour and persistence length are total
Divide meter 3.5 points, smooth degree total score meter 1.5 points, lingering taste total score meter 1.5 points in mouth；
Local flavor scoring criterion: full marks 10 points, wherein raw material peculiar fragrance total score meter 3 points, ferment peculiar fragrance total score meter 3
Point, fragrance mixability total score meter 3 points, other miscellaneous taste total score meters 1 point；
The average score of Herba Dendrobii fermented product be the results are shown in Table 1.
Embodiment 2
1, the first fermentation
The Herba Dendrobii of moisture content about 10% is crushed to 40 mesh, adds water (i.e. solid-liquid ratio according to weight proportion 1:40
For 1:40) after, at a temperature of about 120 DEG C, extracting about 30min, prepared pH value is about the extracting solution of 5；
In above-mentioned extracting solution, add glucose, Gallus Domesticus peptide and manganese sulfate, make glucose mass content in extracting solution
Being about 10%, Gallus Domesticus peptide mass content in extracting solution is about 0.8%, and manganese sulfate mass content in extracting solution is about
0.3%, after stirring and evenly mixing, the inoculum concentration according to about 0.15% accesses Leuconostoc mesenteroides in extracting solution and carries out first
Ferment, control first fermentation temperature be about 28 DEG C, speed of agitator is about 80r/min, when fermentation liquid pH value reduce by 0.5 with
On, and the content of reducing sugar of fermentation liquid less than 1.5% time terminate the first fermentation, prepare the first fermentation liquid (pH value be 4.2～
4.5)；The cell concentration of the first fermentation liquid is shown in Table 1.
2, enzymolysis processing
After fresh Fructus Musae peeling, it is that 1:1:1 mixes with Fructus Chaenomelis, Fructus Lycopersici esculenti according to weight proportion, subsequently breaks up to 40
Mesh, prepared pH value is about the fruit and vegerable liquid of 4.5；
Adding pectase in above-mentioned fruit and vegerable liquid and carry out enzymolysis processing, wherein the consumption of pectase is that every gram of fruit and vegerable liquid 3 is single
About Wei, and the temperature that controlled enzymatic hydrolysis processes is about 50 DEG C, and the time is about 2h, prepares enzymolysis fruit and vegerable liquid.
3, the second fermentation
In described enzymolysis fruit and vegerable liquid, add glucose and Gallus Domesticus peptide, make glucose mass content in enzymolysis fruit and vegerable liquid
Being about 5%, Gallus Domesticus peptide mass content in enzymolysis fruit and vegerable liquid is about 0.8%, after stirring and evenly mixing, according to about 0.1% connect
Amount of planting accesses compound lactobacillus in enzymolysis fruit and vegerable liquid and carries out the second fermentation, and compound lactobacillus is by streptococcus thermophilus, De Shi breast bar
Bacterium, Lactobacillus plantarum and lactobacillus rhamnosus composition, wherein streptococcus thermophilus, Deshi Lactobacillus, Lactobacillus plantarum and rhamnose
Weight proportion between lactobacillus is 3:2:1:1, and the temperature controlling the second fermentation is 22 DEG C, and speed of agitator is 80r/min,
Terminate the second fermentation when the pH value of fermentation liquid reduces by more than 0.5, prepare the second fermentation liquid (pH value is 3.8～4).
4, the 3rd fermentation
In above-mentioned second fermentation liquid, add glucose and Gallus Domesticus peptide, make glucose mass content in the second fermentation liquid
Being about 8%, oligopeptide mass content in the second fermentation liquid is about 0.7%, after stirring and evenly mixing, according to about 0.02% connect
Amount of planting carries out the 3rd fermentation to accessing mulriple yeasts, and described mulriple yeasts is by rice wine yeast, beer yeast and yellow wine yeast
Composition, wherein the weight proportion between rice wine yeast, beer yeast and yellow wine yeast is 1:1.2:0.8, and controls the 3rd
The temperature of ferment is 20 DEG C, and speed of agitator is 40r/min, terminates the 3rd fermentation when the total sugar content of fermentation liquid is less than 2%, prepares
3rd fermentation liquid.
5, preparation Herba Dendrobii fermented product
It is centrifuged, in centrifuged supernatant after above-mentioned first fermentation liquid and the 3rd fermentation liquid are mixed according to weight proportion 2:3
Add white sugar, brown sugar and collagen peptide, make white sugar and the brown sugar mass content in centrifuged supernatant be about 6%, collagen peptide
Mass content in centrifuged supernatant is about 1%, carries out homogenizing, sterilizing subsequently, prepares Herba Dendrobii fermented product (pH value
It is 3.8～4.2)；The average score of the taste and flavor of this Herba Dendrobii fermented product the results are shown in Table 1.
Embodiment 3
1, the first fermentation
The Herba Dendrobii of moisture content about 15% is crushed to 80 mesh, adds water (i.e. solid-liquid ratio according to weight proportion 1:60
For 1:60) after, at a temperature of about 90 DEG C, extracting about 15min, prepared pH value is about the extracting solution of 6；
In above-mentioned extracting solution, add lactose, Gallus Domesticus peptide, dipotassium hydrogen phosphate and Tween 80, make lactose matter in soak
Amount content is about 5%, and Gallus Domesticus peptide mass content in soak is about 0.3%, dipotassium hydrogen phosphate quality in soak
Content is about 0.15%, and Tween 80 mass content in soak is about 0.05%, after stirring and evenly mixing, according to about 0.05%
Inoculum concentration in extracting solution, access Leuconostoc mesenteroides carry out the first fermentation, the temperature controlling the first fermentation is about 22 DEG C,
Speed of agitator is about 120r/min, and when the pH value of fermentation liquid reduces by more than 0.5, and the content of reducing sugar of fermentation liquid is less than
Terminate the first fermentation when 1.5%, prepare the first fermentation liquid (pH value is 5～5.5)；The cell concentration of the first fermentation liquid is shown in Table 1.
2, enzymolysis processing
By Fructus Fragariae Ananssae, Fructus Mali pumilae, Fructus Cucumidis sativi according to weight proportion be 2:1:2 mixing, subsequently break up to 80 mesh, prepared pH value is about 6
Fruit and vegerable liquid；
Adding pectase in above-mentioned fruit and vegerable liquid and carry out enzymolysis processing, wherein the consumption of pectase is that every gram of fruit and vegerable liquid 2 is single
About Wei, and the temperature that controlled enzymatic hydrolysis processes is about 40 DEG C, and the time is about 3h, prepares enzymolysis fruit and vegerable liquid.
3, the second fermentation
In described enzymolysis fruit and vegerable liquid, add glucose and Gallus Domesticus peptide, make glucose mass content in enzymolysis fruit and vegerable liquid
Being about 3%, Gallus Domesticus peptide mass content in enzymolysis fruit and vegerable liquid is about 0.3%, after stirring and evenly mixing, according to about 0.05% connect
Amount of planting accesses compound lactobacillus in enzymolysis fruit and vegerable liquid and carries out the second fermentation, and compound lactobacillus is by streptococcus thermophilus, De Shi breast bar
Bacterium, Lactobacillus fermenti, Lactobacillus brevis and lactobacillus helveticus composition, wherein streptococcus thermophilus, Deshi Lactobacillus, Lactobacillus fermenti,
Weight proportion between Lactobacillus brevis and lactobacillus helveticus is 3:2:1.5:1.5:1.5, and the temperature controlling the second fermentation is
18 DEG C, speed of agitator is 100r/min, terminates the second fermentation when the pH value of fermentation liquid reduces by more than 0.5, prepares the second fermentation
Liquid (pH value is 5～5.5).
4, the 3rd fermentation
In above-mentioned second fermentation liquid, add glucose and Gallus Domesticus peptide, make glucose mass content in the second fermentation liquid
Being about 4%, Gallus Domesticus peptide mass content in the second fermentation liquid is about 0.4%, after stirring and evenly mixing, according to about 0.01% connect
Amount of planting carries out the 3rd fermentation to accessing mulriple yeasts, and described mulriple yeasts is by rice wine yeast, yellow wine yeast and fruit wine and Portugal
Grape brewer yeast forms, and wherein the weight proportion between rice wine yeast, yellow wine yeast, fruit wine and wine yeast is 1:1:1, and
The temperature controlling the 3rd fermentation is 16 DEG C, and speed of agitator is 60r/min, terminates the when the total sugar content of fermentation liquid is less than 2%
Three fermentations, prepare the 3rd fermentation liquid.
5, preparation Herba Dendrobii fermented product
By centrifugal according to weight proportion 3:2 mixing to above-mentioned first fermentation liquid and the 3rd fermentation liquid, centrifuged supernatant is carried out
Homogenizing, sterilizing, prepare Herba Dendrobii fermented product (pH value is 3.8～4.2)；The taste and flavor of this Herba Dendrobii fermented product
Average score the results are shown in Table 1.
Embodiment 4
Except, in the second fermentation step, compound lactobacillus is by streptococcus thermophilus, Deshi Lactobacillus, bacillus acidophilus, cheese milk
Bacillus, pediococcus acidilactici and Pediococcus pentosaceus composition, wherein streptococcus thermophilus, Deshi Lactobacillus, bacillus acidophilus, cheese milk bar
Weight proportion between bacterium, pediococcus acidilactici and Pediococcus pentosaceus is 3:2:1:1:1:1；In 3rd fermentation step, composite yeast
Outside bacterium is made up of with wine yeast the beer yeast that weight proportion is 1:1 and fruit wine, remaining is same as in Example 1, respectively surveys
Surely the results are shown in Table 1.
Embodiment 5
Except, in the second fermentation step, compound lactobacillus is by streptococcus thermophilus, Deshi Lactobacillus, Lactobacillus plantarum, rhamnose
Lactobacillus, Lactobacillus fermenti, Lactobacillus brevis and lactobacillus helveticus composition, wherein streptococcus thermophilus, Deshi Lactobacillus, plant breast bar
Weight proportion between bacterium, lactobacillus rhamnosus, Lactobacillus fermenti, Lactobacillus brevis and lactobacillus helveticus is 3:2:1:1:0.5:
Outside 0.5:0.5, remaining is same as in Example 2, and each measurement result is shown in Table 1.
Reference examples 1
Except, in the first fermentation step, accessing Leuconostoc mesenteroides after adding MRS medium component in above-mentioned extracting solution and enter
Outside row first ferments, remaining is same as in Example 1, and each measurement result is shown in Table 1.
Reference examples 2
This reference examples only implements the first fermentation of embodiment 1, and use by the streptococcus thermophilus that weight proportion is 1:1 and
The extracting solution of embodiment 1 is fermented by the compound lactobacillus of lactobacillus casei composition, the of fermentation parameter and embodiment 1
One fermentation is identical, and the fermentation liquid after fermentation after centrifugal about 15min, carries out homogenizing, goes out under 4000r/min centrifuged supernatant
Bacterium, prepares fermented product, and each measurement result is shown in Table 1.
Table 1 Fermentation Process of Parameter measures and the average score result of fermented product
As seen from the results in Table 1:
Relative to conventional MRS culture medium, use Gallus Domesticus peptide can remarkably promote the life of Leuconostoc mesenteroides as nitrogen source
Long；Additionally, only use the compound lactic acid including streptococcus thermophilus, Deshi Lactobacillus and other two or more lactic acid bacterias simultaneously
Bacterium and simultaneously include that fruit and vegetable materials is fermented by the mulriple yeasts of two or more yeast successively, can make fermented product
There is preferably taste and flavor simultaneously.
Test example 1 immunoloregulation function evaluation
1, the preparation of mouse boosting cell suspension
Draw neck to put to death Balb/c mice, in 75% ethanol, soak 5min disinfection, the complete spleen of aseptic separation
Dirty, wash away floating blood with PBS, divest connective tissue and fat constituent.Use syringe to draw about 5mLPBS, blow in inserting spleen gently
Going out cell, repetitive operation is transparent to spleen adventitia for several times, and residue spleen tissue is cut into size 1mm3Fritter, shreds and is placed on small beaker
200 mesh stainless steel filtering nets on, grind with syringe nook closing member, PBS liquid washs, and collects flushing liquor to centrifuge tube, and 1200rpm is centrifuged
5min, abandons supernatant, adds 3mL erythrocyte cracked liquid, stands 2min, adds 10mL PBS and terminates reaction, and 1200rpm is centrifuged
5min, abandons supernatant, washes twice with PBS liquid, and 1200rpm is centrifuged 5min, washes once by the incomplete culture medium of RPMI-1640,
1200rpm is centrifuged 5min, abandons supernatant, adds appropriate RPMI-1640 complete medium (containing 10% hyclone, 100U/mL penicillium sp
Element, 100 μ g/mL streptomycin and 10mM Hepes), adjust cell concentration to 2 × 106Individual cell/mL, Trypan Blue detection is thin
Born of the same parents' survival rate is more than 95%.
2, each fermented product is on lymphopoietic impact
Fermented product pure water embodiment 1～5, reference examples 1 and reference examples 2 prepared carries out gradient dilution respectively,
Prepare respective gradient dilution liquid.
Above-mentioned splenocyte suspension is joined in 96 well culture plates by 100 μ L/ holes, then adds 10 μ L ConA (5 μ g/ respectively
ML) or 10 μ L LPS (5 μ g/mL) and the gradient dilution liquid of the above-mentioned each fermented product of 10 μ L, each gradient is done 6 repetitions, is incited somebody to action
Culture plate is placed in 37 DEG C, 5%CO2Incubator in cultivate 72h；Blank (i.e. adding 10 μ L pure water) is set simultaneously.
Use each fermented product of CCK-8 kit measurement that (impact of spleen lymphocyte proliferation ability is used stimulation index
(SI) represent), result is as depicted in figs. 1 and 2.From Fig. 1 and Fig. 2: relative to reference examples 1 and the fermented product of reference examples 2,
Herba Dendrobii fermented product prepared by the embodiment of the present invention can dramatically increase mouse T lymphocyte activity, has more excellent
Immunoloregulation function.
Reference examples 2 anti-oxidation function evaluation
The fermented product of embodiment 1～5, reference examples 1 and reference examples 2 is carried out ladder by the DMEM culture medium of serum-free respectively
Degree dilution, prepares gradient dilution liquid；
By HepG2 cell with cell density 2 × 105It is seeded to Tissue Culture Plate, every hole 2ml, is placed in 37 DEG C, 5%CO2Carefully
In born of the same parents' incubator cultivate 24h, the most carefully siphon away the cell supernatant in hole, with HBSS clean once, be separately added into above-mentioned respectively
The gradient dilution liquid 2ml of fermented product, in 37 DEG C, 5%CO2Cell culture incubator is cultivated 3.5h, is subsequently adding fluorescent dye 2 ',
7'-dihydro dichlorofluorescein sodium diacetate (2 ', 7 '-dichlorofluorescin diacetate, DCFH-DA, final concentration of
10 μMs), carefully clean with HBSS three times, be eventually adding antioxygen damage 600 μMs of AAPH of agent；Arrange blank (i.e. to add simultaneously
The DMEM culture medium of 2ml serum-free).
Flow cytomery intracellular ROS content is used (to use EC50Value represents), result is as shown in Figure 3.As shown in Figure 3:
Relative to reference examples 1 and the fermented product of reference examples 2, Herba Dendrobii fermented product prepared by the present invention has more excellent resisting
Oxidative function.
Last it is noted that various embodiments above is only in order to illustrate technical scheme, it is not intended to limit；To the greatest extent
The present invention has been described in detail by pipe with reference to foregoing embodiments, it will be understood by those within the art that: it depends on
So the technical scheme described in foregoing embodiments can be modified, or the most some or all of technical characteristic is entered
Row equivalent；And these amendments or replacement, do not make the essence of appropriate technical solution depart from various embodiments of the present invention technology
The scope of scheme.
Claims (8)
1. the preparation method of a Herba Dendrobii fermented product, it is characterised in that comprise the steps:
1) by broken for Herba Dendrobii rear extracting in water, extracting solution is prepared；
2), after adding medium component in described extracting solution, access Leuconostoc mesenteroides and carry out the first fermentation, when fermentation liquid
PH value reduces by more than 0.5, and terminates described first fermentation when the content of reducing sugar of fermentation liquid is less than 1.5%, prepares first
Ferment liquid；
3) in the fruit and vegerable liquid that pH value is 4.5～6, add pectase and carry out enzymolysis processing, prepare enzymolysis fruit and vegerable liquid；
4), after adding medium component in described enzymolysis fruit and vegerable liquid, access compound lactobacillus and carry out the second fermentation, work as fermentation liquid
PH value reduce by more than 0.5 time terminate described second fermentation, prepare the second fermentation liquid；Wherein, described compound lactobacillus selected from addicted to
Hot streptococcus, Deshi Lactobacillus, bacillus acidophilus, lactobacillus casei, Lactobacillus plantarum, Lactobacillus fermenti, lactobacillus rhamnosus,
In Lactobacillus brevis, lactobacillus helveticus, pediococcus acidilactici, Pediococcus pentosaceus more than four kinds；
5), after adding medium component in described second fermentation liquid, access mulriple yeasts and carry out the 3rd fermentation, work as fermentation liquid
Total sugar content less than 2% time terminate described 3rd fermentation, prepare the 3rd fermentation liquid；Wherein, described mulriple yeasts is selected from rice
Two or more with wine yeast of brewer yeast, beer yeast, yellow wine yeast and fruit wine；
6) by centrifugal after described first fermentation liquid and described 3rd fermentation liquid mixing, after centrifuged supernatant homogenizing, sterilizing, prepare
Herba Dendrobii fermented product；
Described medium component includes in one or more in sugar, peptide, inorganic salt and surfactant, and described peptide average
Gross mass content >=80% of the molecular weight component less than 1000Da；
Step 4) in, in described enzymolysis fruit and vegerable liquid, add sugar and peptide, make described sugar quality in described enzymolysis fruit and vegerable liquid contain
Amount is 3～5%, and described peptide mass content in described enzymolysis fruit and vegerable liquid is 0.3～0.8%；Described compound lactobacillus is at least
Including streptococcus thermophilus and Deshi Lactobacillus, described streptococcus thermophilus, Deshi Lactobacillus with the weight proportion of remaining lactic acid bacteria are
3:2:(2～5), and control described second fermentation temperature be 18～23 DEG C, rotating speed is 80～120r/min.
Preparation method the most according to claim 1, it is characterised in that step 1) in, Herba Dendrobii is crushed to 40～80
Mesh, and control broken after the quality proportioning of Herba Dendrobii and water be 1:40～60, the temperature of described extraction is 90～120
DEG C, extraction time is 5～30min.
Preparation method the most according to claim 1, it is characterised in that step 2) in, in described extracting solution, add sugar, peptide
And inorganic salt, making described sugar mass content in described extracting solution is 5～10%, described peptide quality in described extracting solution
Content is 0.3～0.8%, and described inorganic salt mass content in described extracting solution is 0.1～0.3%, and controls described
The temperature of the first fermentation is 22～28 DEG C, and rotating speed is 80～120r/min.
Preparation method the most according to claim 1, it is characterised in that will selected from Fructus Ananadis comosi, Citrullus vulgaris, Fructus Fragariae Ananssae, Fructus Mali pumilae, Fructus Musae,
Two or more fruit and vegetable materials in Fructus Mangifera Indicae, Fructus Chaenomelis, Radix Raphani, Fructus Cucumidis sativi, Fructus Lycopersici esculenti is crushed to 40～80 mesh, prepared pH value be 4.5～
The described fruit and vegerable liquid of 6.
Preparation method the most according to claim 1, it is characterised in that the consumption of described pectase is every gram of fruit and vegerable liquid 2～3
Unit, and the temperature controlling described enzymolysis processing is 40～50 DEG C, and the time is 2～3h.
Preparation method the most according to claim 1, it is characterised in that step 5) in, add in described second fermentation liquid
Sugar and peptide, making described sugar mass content in described second fermentation liquid is 4～8%, and described peptide is in described second fermentation liquid
Mass content be 0.3～0.8%；And controlling the weight proportion between two primary yeasts in described mulriple yeasts is 1:
(0.8～1.2), the temperature of described 3rd fermentation is 16～20 DEG C, and rotating speed is 40～60r/min.
Preparation method the most according to claim 1, it is characterised in that step 6) in, described first fermentation liquid and described the
The weight proportion of three fermentation liquids is (4～6): (4～6).
8. a Herba Dendrobii fermented product, it is characterised in that prepare according to the arbitrary described preparation method of claim 1 to 7.
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| CN110623180A (en) * | 2019-09-12 | 2019-12-31 | 湖南农业大学 | Dendrobium officinale fermented probiotic beverage and preparation method thereof |
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| CN102940038B (en) * | 2012-11-06 | 2013-10-09 | 福建省农业科学院农业生物资源研究所 | Method for preparing dendrobium officinale plant protein lactic acid beverage |
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