CN104585762A - Dendrobium officinale fermented product and preparation method thereof - Google Patents
Dendrobium officinale fermented product and preparation method thereof Download PDFInfo
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Abstract
The invention provides a dendrobium officinale fermented product and a preparation method thereof. The method comprises the following steps: (1) crushing dendrobium officinale, adding water and extracting to prepare an extract liquid; (2) adding culture medium components to the extract liquid, inoculating leuconostoc mesenteroides and carrying out first fermentation to prepare a first fermentation liquor; (3) adding pectinase to a fruit and vegetable liquid with the pH value being 4.5-6, and carrying out enzymolysis treatment to prepare an enzymatic fruit and vegetable liquid; (4) adding the culture medium components to the enzymatic fruit and vegetable liquid, inoculating compound lactobacillus and carrying out second fermentation to prepare a second fermentation liquor; (5) adding the culture medium components to the second fermentation liquor, inoculating composite saccharomycetes and carrying out third fermentation to prepare a third fermentation liquor; and (6) mixing the first fermentation liquor with the third fermentation liquor, centrifuging, homogenizing and sterilizing centrifugal supernate to prepare the dendrobium officinale fermented product. The dendrobium officinale fermented product prepared by adopting the method is good in taste and unique in flavor, has good functions of immunoregulation and antioxidation, and is suitable for being taken by wide people.
Description
Technical field
The present invention relates to a kind of fermented product, particularly relate to a kind of dendrobium candidum fermented product and preparation method thereof.
Background technology
Dendrobium candidum, has another name called ribbed hedyotis herb, Yunnan iron sheet, belongs to micro-specific item, and the perennial herbaceous plant that grows nonparasitically upon another plant of the orchid family, is mainly distributed in the ground such as China Anhui, Zhejiang, Fujian.Dendrobium candidum has and promotes the production of body fluid and hypoglycemic activity, and it can promote glandular secretion and internal organs motion, reduces streptozotocin and brings out glycosuria blood glucose value; In addition, dendrobium candidum can also strengthen immunity of organisms, promote the phagocytic function of tumor animal macrophage, strengthen the activity of the lymphocytic propagation of T and differentiation and NK cell, Nonspecific immunity, specific cellular immunity and humoral immune function are all improved effect.
At present, a small amount of relevant report dendrobium candidum being processed into the products such as goods is had.Such as, publication number is that the Chinese patent of CN 103734824A discloses a kind of dendrobium officinale healthcare green juice, comprises the fresh bar of dendrobium candidum 200 ~ 250 parts, aloe 100 ~ 150 parts, carambola 50 ~ 150 parts, Chinese grooseberry 50 ~ 100 parts, green apple 50 ~ 150 parts, acacia honey 30 ~ 60 parts, stabilizing agent 5 ~ 10 parts, citric acid 1 ~ 2 part, surplus be water.This patent mixes after mainly being squeezed the juice by the raw materials such as the fresh bar of dendrobium candidum, after adding acacia honey, citric acid and stabilizing agent, blend water obtain, although it is rich in multiple fruit juice, nutrition is comparatively balanced, but it utilizes rate variance to dendrobium candidum raw material, product health care is limited.
Publication number is the preparation method that the Chinese patent of CN 102940038A discloses a kind of dendrobium officinale plant protein lactic acid beverage, extract after by iron-sheet dendrobe fresh product fragmentation, dendrobium candidum juice is formed by milling treatment of colloid through leaching filtrate, prepare soymilk simultaneously, and access Lactobacillus casei after being mixed with dendrobium candidum juice by soymilk and streptococcus thermophilus carries out fermented and cultured, until the pH value of soymilk is 4.3 ~ 4.5, finally the soymilk after fermented and cultured is refrigerated 24h at 1 DEG C ~ 4 DEG C, obtained product.The method is extracted dendrobium candidum, thus improves raw material availability, but its nutritional labeling is relatively single, and mouthfeel, local flavor and health care all have much room for improvement.
Summary of the invention
The invention provides a kind of dendrobium candidum fermented product and preparation method thereof, single for solving dendrobium candidum series products composition of the prior art, mouthfeel, local flavor, health care are limited and cannot carry out the technological deficiencies such as effectively utilization to dendrobium candidum.
The invention provides a kind of preparation method of dendrobium candidum fermented product, comprise the steps:
1) by extracting in water after dendrobium candidum fragmentation, obtained extract;
2), after adding medium component in described extract, access Leuconostoc mesenteroides carries out the first fermentation, when the pH value of zymotic fluid reduces by more than 0.5, and the content of reducing sugar of zymotic fluid lower than 1.5% time terminate described first fermentation, obtained first zymotic fluid;
3) to pH value be 4.5 ~ 6 fruits and vegetables liquid in add pectase and carry out enzymolysis processing, obtained enzymolysis fruits and vegetables liquid;
4) after adding medium component in described enzymolysis fruits and vegetables liquid, access compound lactobacillus carries out the second fermentation, terminates described second fermentation, obtained second zymotic fluid when the pH value of zymotic fluid reduces by more than 0.5; Wherein, described compound lactobacillus is selected from more than four kinds in streptococcus thermophilus, Lactobacillus delbrueckii, lactobacillus acidophilus, Lactobacillus casei, Lactobacillus plantarum, lactobacillus fermenti, Lactobacillus rhamnosus, Lactobacillus brevis, Lactobacillus helveticus, Pediococcus acidilactici, Pediococcus pentosaceus;
5), after adding medium component in described second zymotic fluid, access mulriple yeasts carries out the 3rd fermentation, when zymotic fluid total sugar content lower than 2% time terminate described 3rd fermentation, obtained 3rd zymotic fluid; Wherein, what described mulriple yeasts was selected from rice wine yeast, brewer's yeast, yellow wine yeast and fruit wine and wine yeast is two or more;
6) by centrifugal after described first zymotic fluid and described 3rd zymotic fluid mixing, after centrifuged supernatant homogeneous, sterilizing, obtained dendrobium candidum fermented product.
In the preparation process in accordance with the present invention, described Leuconostoc mesenteroides can select Leuconostoc mesenteroides dextran subspecies (L.mesenteroides subsp.dextranicum) or leuconostoc mesenteroides sub species cremoris (L.mesenteroides subsp.cremoris), and each bacterium of the present invention can adopt the mode of bacterium powder or bacterium liquid to use.When adopting bacterium powder, the viable count of described each lactic acid bacteria bacterium powder can be 10
9about CFU/g, the viable count of each saccharomycete bacterium powder can be 10
10about CFU/g.In addition, the inoculum concentration of described Leuconostoc mesenteroides (bacterium powder) can be 0.05 ~ 0.2%, such as 0.1%, the inoculum concentration of described compound lactobacillus (bacterium powder) can be 0.05 ~ 0.2%, such as 0.1%, the inoculum concentration of described mulriple yeasts (bacterium powder) can be 0.01 ~ 0.02%.When adopting bacterium liquid, the viable count and the inoculum concentration that can refer to above-mentioned bacterium powder are inoculated.Each bacterial strain conventionally can activate according to needs before use.
The present invention does not make considered critical to dendrobium candidum raw material, and it can be fresh dendrobium candidum, also can be dry dendrobium candidum.In concrete scheme of the present invention, dry dendrobium candidum can be adopted as raw material, its moisture content is≤15.0%.Further, step 1) in, dendrobium candidum is crushed to 40 ~ 80 orders, and the quality proportioning of dendrobium candidum after controlled fragmentation and water is 1:40 ~ 60, the temperature of described extraction is 90 ~ 120 DEG C, and extraction time is 5 ~ 30min.The pH being extracted obtained extract by described dendrobium candidum is 4.5 ~ 6.Extraction process can make the active component in dendrobium candidum raw material discharge more effectively, thus further increases raw material availability.
In the present invention, described medium component can comprise carbon source, nitrogenous source, inorganic salts, trace element etc., and those skilled in the art can select according to the concrete bacterial strain cultivated or compound bacteria the medium component that is suitable for.In concrete scheme of the present invention, described medium component comprise in sugar, peptide, inorganic salts and surfactant one or more, in wherein said peptide, mean molecule quantity is less than gross mass content >=80% of the component of 1000Da.
Namely, the present invention additionally provides a kind of fluid nutrient medium for Leuconostoc mesenteroides fermentation on the other hand, it comprises dendrobium candidum extract, sugar, peptide, inorganic salts and surfactant, wherein the mass content of sugar in described extract can be 5 ~ 10%, the mass content of peptide in described extract can be 0.3 ~ 0.8%, the mass content of inorganic salts in described extract can be 0.1 ~ 0.3%, and the mass content of surfactant in described extract can be 0.01 ~ 0.1%; This dendrobium candidum extract is by obtaining extracting in water after dendrobium candidum fragmentation.
Again on the one hand, present invention also offers a kind of fluid nutrient medium fermented for compound lactobacillus and/or mulriple yeasts, it comprises enzymolysis fruits and vegetables liquid, sugar, peptide, inorganic salts and surfactant, wherein the mass content of sugar in described enzymolysis fruits and vegetables liquid can be 5 ~ 10%, the mass content of peptide in described enzymolysis fruits and vegetables liquid can be 0.3 ~ 0.8%, the mass content of inorganic salts in described enzymolysis fruits and vegetables liquid can be 0.1 ~ 0.3%, and the mass content of surfactant in described enzymolysis fruits and vegetables liquid can be 0.01 ~ 0.1%; This enzymolysis fruits and vegetables liquid by pH value be 4.5 ~ 6 fruits and vegetables liquid in add pectinase enzymatic hydrolysis obtain.
Further, described sugar can be selected from glucose, sucrose and lactose one or more, such as sucrose; Described inorganic salts can be selected from sodium salt, calcium salt, manganese salt, sylvite and magnesium salts one or more, such as calcium carbonate; Described surfactant can be selected from the smooth and polysorbate of fatty glyceride, aliphatic acid sorb one or more, such as Tween 80 etc.Particularly, the component of peptide described in the present invention is not particularly limited, as long as wherein mean molecule quantity is less than gross mass content >=80% of the component of 1000Da, further >=90%, adopts described peptide significantly can promote the growth of Leuconostoc mesenteroides and compound lactobacillus and mulriple yeasts as nitrogenous source.
The step 2 of concrete scheme of the present invention) in, sugar, peptide and inorganic salts are added in described extract, the mass content of described sugar in described extract is made to be 5 ~ 10%, the mass content of described peptide in described extract is 0.3 ~ 0.8%, the mass content of described inorganic salts in described extract is 0.1 ~ 0.3%, and the temperature controlling described first fermentation is 22 ~ 28 DEG C, and rotating speed is 80 ~ 120r/min.Under this condition Leuconostoc mesenteroides is fermented, the mouthfeel of product can be improved and promote the performance of product health care.In addition, the pH value controlling this zymotic fluid reduces by more than 0.5 (pH value of such as zymotic fluid is 4.5 ~ 5), and the content of reducing sugar of zymotic fluid is lower than 1.5%, not only be conducive to avoiding product tart flavour overweight and affecting palatability, and low content of reducing sugar can make product have applicability more widely, particularly can drink by diabetic.
In the present invention, described fruits and vegetables liquid is the slurries, juice etc. made through fruit and/or fruits and vegetables, such as, can adopt commercial Juice etc., particularly preferably not containing the fruit and vegetable juice of the additives such as any anticorrisive agent.Further, after fruit and vegetable materials can also be chosen, be crushed to 40 ~ 80 orders, such as 60 orders, obtained described fruits and vegetables liquid; This particle size range both can improve the speed of fermentation, also helped the mouthfeel ensureing fermented product.In addition, the present invention does not do strict restriction to the kind of fruit and vegetable materials, particularly can choose locality and abound with and/or be convenient to the fruits and vegetables of processing as raw material, such as pineapple, watermelon, strawberry, apple, banana, mango, pawpaw, radish, cucumber, tomato etc.; And, when choosing fruit and vegetable materials, can carry out reasonably combined for the characteristic of raw material itself (such as pH value), and make the fruits and vegetables liquid pH value made in the scope of 4.5 ~ 6, thus without the need to add other pH adjusting agent to regulate the pH value of fruits and vegetables liquid, to ensure the genuineness of product.
Pectase of the present invention can by common commercial acquisition.In described fruits and vegetables liquid, add pectase be mainly used in decompose pectin etc., thus make the nutritional labeling of fruit and vegetable materials discharge more thorough, to improve raw material availability.Further, the consumption of described pectase is every gram of fruits and vegetables liquid 2 ~ 3 unit, and the temperature controlling described enzymolysis processing is 40 ~ 50 DEG C, and the time is 2 ~ 3h.
Step 4 of the present invention) in, in described enzymolysis fruits and vegetables liquid, add sugar and peptide, make the mass content of described sugar in described enzymolysis fruits and vegetables liquid be 3 ~ 5%, the mass content of described peptide in described enzymolysis fruits and vegetables liquid is 0.3 ~ 0.8%; Described compound lactobacillus at least comprises streptococcus thermophilus and Lactobacillus delbrueckii, the weight proportion of described streptococcus thermophilus, Lactobacillus delbrueckii and all the other lactic acid bacterias is 3:2:(2 ~ 5), and the temperature controlling described second fermentation is 18 ~ 23 DEG C, and rotating speed is 80 ~ 120r/min.
Further, all the other lactic acid bacterias described can be one or more in following first to fourth component:
First component: comprise lactobacillus acidophilus and Lactobacillus casei, and the weight proportion of lactobacillus acidophilus and Lactobacillus casei is (0.5 ~ 1.5): (0.5 ~ 1.5);
Second component: comprise Pediococcus acidilactici and Pediococcus pentosaceus, and the weight proportion of Pediococcus acidilactici and Pediococcus pentosaceus is (0.5 ~ 1.5): (0.5 ~ 1.5);
Three components: comprise Lactobacillus plantarum and Lactobacillus rhamnosus, and the weight proportion of Lactobacillus plantarum and Lactobacillus rhamnosus is (0.5 ~ 1): (0.5 ~ 1);
Four composition: comprise lactobacillus fermenti, Lactobacillus brevis and Lactobacillus helveticus, and lactobacillus fermenti, weight proportion between Lactobacillus brevis and Lactobacillus helveticus are (0.5 ~ 1): (0.5 ~ 1): (0.5 ~ 1).
Step 5 of the present invention) in, in described second zymotic fluid, add sugar and peptide, make the mass content of described sugar in described second zymotic fluid be 4 ~ 8%, the mass content of described peptide in described second zymotic fluid is 0.3 ~ 0.8%; And the weight proportion controlled 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.Each saccharomycete particularly in mulriple yeasts all adopts bacterium powder, when in mulriple yeasts, each yeast weight is identical, the local flavor of fermented product is splendid.
Further, described mulriple yeasts can be selected from any one in following first to fourth component:
First component: comprise rice wine yeast and fruit wine and wine yeast, and rice wine yeast, weight proportion between fruit wine and wine yeast are 1:(0.8 ~ 1.2);
Second component: comprise brewer's yeast and fruit wine and wine yeast, and rice wine yeast, weight proportion between fruit wine and wine yeast are 1:(0.8 ~ 1.2);
Three components: comprise rice wine yeast, brewer's yeast and yellow wine yeast, and rice wine yeast, weight proportion between brewer's yeast and yellow wine yeast are 1:(0.8 ~ 1.2): (0.8 ~ 1.2);
Four composition: comprise rice wine yeast, yellow wine yeast and fruit wine and wine yeast, and rice wine yeast, yellow wine yeast and the weight proportion between fruit wine and wine yeast are 1:(0.8 ~ 1.2): (0.8 ~ 1.2).
Step 6 of the present invention) in, the weight proportion of described first zymotic fluid and described 3rd zymotic fluid is (4 ~ 6): (4 ~ 6).And, before described homogeneous, can optionally allocate centrifuged supernatant, such as can add appropriate sugar, peptide etc. improve the mouthfeel of product and improve the health care etc. of product, the mass content of wherein said sugar in centrifuged supernatant can be 8 ~ 16%, and the mass content of described peptide in centrifuged supernatant can be 0.5 ~ 2%.
Further, described sugar can comprise white sugar and brown sugar, and described white sugar and the brown sugar quality proportioning in described sugar can be 1:0.5 ~ 2.In addition, described peptide can be collagen peptide, and in the former peptide of this peptide, mean molecule quantity is less than gross mass content >=80% of the component of 1000Da, further >=90%.
The present invention also provides a kind of dendrobium candidum fermented product, obtains according to above-mentioned arbitrary described preparation method.
The enforcement of the present invention program, at least has following advantage:
1, the present invention adopts Leuconostoc mesenteroides to ferment to dendrobium candidum extract, not only more effectively releases activity and the nutritional labeling of dendrobium candidum, improves the utilization rate of dendrobium candidum, enhance the health care of zymotic fluid in addition; Meanwhile, the present invention adopts compound lactobacillus and mulriple yeasts to ferment to fruits and vegetables liquid successively, not only ensure that the comprehensive and balanced of the nutritional labeling in fermented product, improves the mouthfeel of fermented product in addition and improves the local flavor of fermented product.
2, the condition of the present invention to zymotechnique is optimized, and not only increases fermenting speed, shortens fermentation time, and the mouthfeel of fermented product, local flavor and immunological regulation and the function such as anti-oxidant are significantly improved in addition.
3, the dendrobium candidum fermented product mouthfeel prepared of the present invention good, unique flavor, nutritional labeling general equilibrium, instant, this fermented product not containing any additive, green health; In addition, it also has good immunological regulation and the function such as anti-oxidant, is suitable for crowd in extensive range.
Accompanying drawing explanation
Fig. 1 is that the fermented product of the embodiment of the present invention and reference examples is to the evaluation result of the mouse T lymphocyte multiplication capacity that ConA induces;
Fig. 2 is that the fermented product of the embodiment of the present invention and reference examples is to the evaluation result of the mouse T lymphocyte multiplication capacity that LPS induces;
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 clearly, below in conjunction with drawings and Examples of the present invention, the technical scheme in the embodiment of the present invention is clearly and completely described, obviously, described embodiment is the present invention's part embodiment, instead of whole embodiments.Based on the embodiment in the present invention, those of ordinary skill in the art, not making the every other embodiment obtained under creative work prerequisite, belong to the scope of protection of the invention.
Each lactic acid bacteria that various embodiments of the present invention adopt is all from Research for Industrial Microbial Germ preservation administrative center (CICC), each lactic acid bacteria is respectively: leuconostoc mesenteroides sub species cremoris (CICC 22181), streptococcus thermophilus (CICC 6222), Lactobacillus delbrueckii subsp. lactis (CICC 6077), lactobacillus acidophilus (CICC 6081), Lactobacillus casei (CICC 6117), Lactobacillus plantarum (CICC 22132), lactobacillus fermenti (CICC 22538), Lactobacillus rhamnosus (CICC 6147), Lactobacillus brevis (CICC23474), Lactobacillus helveticus (CICC 22172), Pediococcus acidilactici (CICC 10346), Pediococcus pentosaceus (CICC 20536), the viable count of each lactic acid bacteria is all about 10
9cFU/g,
Each saccharomycete is all from Angel Yeast Co., Ltd, and each saccharomycetic viable count is all about 10
10cFU/g;
Black-bone chicken peptide and collagen peptide, purchased from Beijing Zhongshi Haishi Biotechnology Co., Ltd., in two kinds of peptides, mean molecule quantity is less than the gross mass content of the component of 1000Da all >=80%;
Pectase: purchased from Pangbo Bioengineering Co Ltd, Nanning;
Embodiment 1
1, the first fermentation
Dendrobium candidum moisture content being about 13% is crushed to 60 orders, and after adding water (namely solid-liquid ratio is 1:50) according to weight proportion 1:50, at the temperature of about 105 DEG C, extract about 5min, obtained pH value is about the extract of 5.5;
Sucrose is added in above-mentioned extract, black-bone chicken peptide and calcium carbonate, the mass content of sucrose in extract is made to be about 7%, the mass content of black-bone chicken peptide in extract is about 0.6%, the mass content of calcium carbonate in extract is about 0.2%, after stirring and evenly mixing, inoculum concentration according to about 0.1% accesses Leuconostoc mesenteroides and carries out the first fermentation in extract, the temperature controlling the first fermentation is about 24 DEG C, speed of agitator is about 110r/min, when the pH value of zymotic fluid reduces by more than 0.5, and the content of reducing sugar of zymotic fluid lower than 1.5% time terminate the first fermentation, obtained first zymotic fluid (pH value is 4.5 ~ 5).
Adopt the cell concentration of spectrophotometry first zymotic fluid, the results are shown in Table 1.
2, enzymolysis processing
After fresh mango and carrot are 2:1 mixing according to weight proportion, be crushed to 60 orders, obtained pH value is about the fruits and vegetables liquid of 5.5;
In above-mentioned fruits and vegetables liquid, add pectase carry out enzymolysis processing, wherein the consumption of pectase is about every gram of fruits and vegetables liquid 2.5 unit, and the temperature of controlled enzymatic hydrolysis process is about 45 DEG C, and the time is about 3h, obtained enzymolysis fruits and vegetables liquid.
3, the second fermentation
Sucrose and black-bone chicken peptide is added in described enzymolysis fruits and vegetables liquid, the mass content of sucrose in enzymolysis fruits and vegetables liquid is made to be about 4%, the mass content of black-bone chicken peptide in enzymolysis fruits and vegetables liquid is about 0.5%, after stirring and evenly mixing, inoculum concentration according to about 0.1% accesses compound lactobacillus and carries out the second fermentation in enzymolysis fruits and vegetables liquid, compound lactobacillus is by streptococcus thermophilus, Lactobacillus delbrueckii, lactobacillus acidophilus and Lactobacillus casei composition, wherein streptococcus thermophilus, Lactobacillus delbrueckii, weight proportion between lactobacillus acidophilus and Lactobacillus casei is 3:2:1.5:1.5, and the temperature controlling the second fermentation is about 20 DEG C, speed of agitator is about 100r/min, the second fermentation is terminated when the pH value of zymotic fluid reduces by more than 0.5, obtained second zymotic fluid (pH value is 4.5 ~ 5).
4, the 3rd fermentation
Sucrose and black-bone chicken peptide is added in above-mentioned second zymotic fluid, the mass content of sucrose in the second zymotic fluid is made to be about 6%, the mass content of black-bone chicken peptide in the second zymotic fluid is about 0.5%, after stirring and evenly mixing, inoculum concentration according to about 0.01% carries out the 3rd fermentation to access mulriple yeasts, described mulriple yeasts is made up of rice wine yeast and fruit wine and wine yeast, and 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 about 18 DEG C, and speed of agitator is about 50r/min, when zymotic fluid total sugar content lower than 2% time terminate the 3rd fermentation, obtained 3rd zymotic fluid.
5, dendrobium candidum fermented product is prepared
After above-mentioned first zymotic fluid and the 3rd zymotic fluid being mixed according to weight proportion 1:1, centrifugal about 15min under 4000r/min, carries out homogeneous, sterilizing to centrifuged supernatant, obtained dendrobium candidum 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 dendrobium candidum fermented product of above-mentioned preparation, and carry out mouthfeel and local flavor marking to this dendrobium candidum fermented product respectively 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 total score 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, its Raw peculiar fragrance total score meter 3 points, ferment peculiar fragrance total score meter 3 points, fragrance mixability total score meter 3 points, other assorted taste total score meters 1 point;
1 be the results are shown in Table to the average score of dendrobium candidum fermented product.
Embodiment 2
1, the first fermentation
Dendrobium candidum moisture content being about 10% is crushed to 40 orders, and after adding water (namely solid-liquid ratio is 1:40) according to weight proportion 1:40, at the temperature of about 120 DEG C, extract about 30min, obtained pH value is about the extract of 5;
Glucose is added in above-mentioned extract, black-bone chicken peptide and manganese sulfate, the mass content of glucose in extract is made to be about 10%, the mass content of black-bone chicken peptide in extract is about 0.8%, the mass content of manganese sulfate in extract is about 0.3%, after stirring and evenly mixing, inoculum concentration according to about 0.15% accesses Leuconostoc mesenteroides and carries out the first fermentation in extract, the temperature controlling the first fermentation is about 28 DEG C, speed of agitator is about 80r/min, when the pH value of zymotic fluid reduces by more than 0.5, and the content of reducing sugar of zymotic fluid lower than 1.5% time terminate the first fermentation, obtained first zymotic fluid (pH value is 4.2 ~ 4.5), the cell concentration of the first zymotic fluid is in table 1.
2, enzymolysis processing
After being removed the peel by fresh banana, be that 1:1:1 mixes with pawpaw, tomato according to weight proportion, be crushed to 40 orders subsequently, obtained pH value is about the fruits and vegetables liquid of 4.5;
In above-mentioned fruits and vegetables liquid, add pectase carry out enzymolysis processing, wherein the consumption of pectase is about every gram of fruits and vegetables liquid 3 unit, and the temperature of controlled enzymatic hydrolysis process is about 50 DEG C, and the time is about 2h, obtained enzymolysis fruits and vegetables liquid.
3, the second fermentation
Glucose and black-bone chicken peptide is added in described enzymolysis fruits and vegetables liquid, the mass content of glucose in enzymolysis fruits and vegetables liquid is made to be about 5%, the mass content of black-bone chicken peptide in enzymolysis fruits and vegetables liquid is about 0.8%, after stirring and evenly mixing, inoculum concentration according to about 0.1% accesses compound lactobacillus and carries out the second fermentation in enzymolysis fruits and vegetables liquid, compound lactobacillus is by streptococcus thermophilus, Lactobacillus delbrueckii, Lactobacillus plantarum and Lactobacillus rhamnosus composition, wherein streptococcus thermophilus, Lactobacillus delbrueckii, weight proportion between Lactobacillus plantarum and Lactobacillus rhamnosus is 3:2:1:1, and the temperature controlling the second fermentation is 22 DEG C, speed of agitator is 80r/min, the second fermentation is terminated when the pH value of zymotic fluid reduces by more than 0.5, obtained second zymotic fluid (pH value is 3.8 ~ 4).
4, the 3rd fermentation
Glucose and black-bone chicken peptide is added in above-mentioned second zymotic fluid, the mass content of glucose in the second zymotic fluid is made to be about 8%, the mass content of oligopeptide in the second zymotic fluid is about 0.7%, after stirring and evenly mixing, inoculum concentration according to about 0.02% carries out the 3rd fermentation to access mulriple yeasts, described mulriple yeasts is by rice wine yeast, brewer's yeast and yellow wine yeast composition, wherein rice wine yeast, weight proportion between brewer's yeast and yellow wine yeast is 1:1.2:0.8, and the temperature controlling the 3rd fermentation is 20 DEG C, speed of agitator is 40r/min, when zymotic fluid total sugar content lower than 2% time terminate the 3rd fermentation, obtained 3rd zymotic fluid.
5, dendrobium candidum fermented product is prepared
It is centrifugal after above-mentioned first zymotic fluid and the 3rd zymotic fluid are mixed according to weight proportion 2:3, white sugar, brown sugar and collagen peptide is added in centrifuged supernatant, white sugar and the brown sugar mass content in centrifuged supernatant is made to be about 6%, the mass content of collagen peptide in centrifuged supernatant is about 1%, carry out homogeneous, sterilizing subsequently, obtained dendrobium candidum fermented product (pH value is 3.8 ~ 4.2); The mouthfeel of this dendrobium candidum fermented product and the average score of local flavor the results are shown in Table 1.
Embodiment 3
1, the first fermentation
Dendrobium candidum moisture content being about 15% is crushed to 80 orders, and after adding water (namely solid-liquid ratio is 1:60) according to weight proportion 1:60, at the temperature of about 90 DEG C, extract about 15min, obtained pH value is about the extract of 6;
Lactose is added in above-mentioned extract, black-bone chicken peptide, dipotassium hydrogen phosphate and Tween 80, the mass content of lactose in soak is made to be about 5%, the mass content of black-bone chicken peptide in soak is about 0.3%, the mass content of dipotassium hydrogen phosphate in soak is about 0.15%, the mass content of Tween 80 in soak is about 0.05%, after stirring and evenly mixing, inoculum concentration according to about 0.05% accesses Leuconostoc mesenteroides and carries out the first fermentation in extract, the temperature controlling the first fermentation is about 22 DEG C, speed of agitator is about 120r/min, when the pH value of zymotic fluid reduces by more than 0.5, and the content of reducing sugar of zymotic fluid lower than 1.5% time terminate the first fermentation, obtained first zymotic fluid (pH value is 5 ~ 5.5), the cell concentration of the first zymotic fluid is in table 1.
2, enzymolysis processing
By strawberry, apple, cucumber according to weight proportion be 2:1:2 mixing, be crushed to 80 orders subsequently, obtained pH value is about the fruits and vegetables liquid of 6;
In above-mentioned fruits and vegetables liquid, add pectase carry out enzymolysis processing, wherein the consumption of pectase is about every gram of fruits and vegetables liquid 2 unit, and the temperature of controlled enzymatic hydrolysis process is about 40 DEG C, and the time is about 3h, obtained enzymolysis fruits and vegetables liquid.
3, the second fermentation
Glucose and black-bone chicken peptide is added in described enzymolysis fruits and vegetables liquid, the mass content of glucose in enzymolysis fruits and vegetables liquid is made to be about 3%, the mass content of black-bone chicken peptide in enzymolysis fruits and vegetables liquid is about 0.3%, after stirring and evenly mixing, inoculum concentration according to about 0.05% accesses compound lactobacillus and carries out the second fermentation in enzymolysis fruits and vegetables liquid, compound lactobacillus is by streptococcus thermophilus, Lactobacillus delbrueckii, lactobacillus fermenti, Lactobacillus brevis and Lactobacillus helveticus composition, wherein streptococcus thermophilus, Lactobacillus delbrueckii, 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, the second fermentation is terminated when the pH value of zymotic fluid reduces by more than 0.5, obtained second zymotic fluid (pH value is 5 ~ 5.5).
4, the 3rd fermentation
Glucose and black-bone chicken peptide is added in above-mentioned second zymotic fluid, the mass content of glucose in the second zymotic fluid is made to be about 4%, the mass content of black-bone chicken peptide in the second zymotic fluid is about 0.4%, after stirring and evenly mixing, inoculum concentration according to about 0.01% carries out the 3rd fermentation to access mulriple yeasts, described mulriple yeasts is by rice wine yeast, yellow wine yeast and fruit wine and wine yeast form, wherein rice wine yeast, yellow wine yeast, weight proportion between fruit wine and wine yeast is 1:1:1, and the temperature controlling the 3rd fermentation is 16 DEG C, speed of agitator is 60r/min, when zymotic fluid total sugar content lower than 2% time terminate the 3rd fermentation, obtained 3rd zymotic fluid.
5, dendrobium candidum fermented product is prepared
Above-mentioned first zymotic fluid and the 3rd zymotic fluid are mixed centrifugal according to weight proportion 3:2, homogeneous, sterilizing are carried out to centrifuged supernatant, obtained dendrobium candidum fermented product (pH value is 3.8 ~ 4.2); The mouthfeel of this dendrobium candidum fermented product and the average score of local flavor the results are shown in Table 1.
Embodiment 4
Except in the second fermentation step, compound lactobacillus is made up of streptococcus thermophilus, Lactobacillus delbrueckii, lactobacillus acidophilus, Lactobacillus casei, Pediococcus acidilactici and Pediococcus pentosaceus, and wherein streptococcus thermophilus, Lactobacillus delbrueckii, lactobacillus acidophilus, Lactobacillus casei, weight proportion between Pediococcus acidilactici and Pediococcus pentosaceus are 3:2:1:1:1:1; In 3rd fermentation step, mulriple yeasts is that outside the brewer's yeast of 1:1 and fruit wine and wine yeast form, all the other are identical with embodiment 1, and each measurement result is in table 1 by weight proportion.
Embodiment 5
Except in the second fermentation step, compound lactobacillus is made up of streptococcus thermophilus, Lactobacillus delbrueckii, Lactobacillus plantarum, Lactobacillus rhamnosus, lactobacillus fermenti, Lactobacillus brevis and Lactobacillus helveticus, wherein streptococcus thermophilus, Lactobacillus delbrueckii, Lactobacillus plantarum, Lactobacillus rhamnosus, lactobacillus fermenti, weight proportion between Lactobacillus brevis and Lactobacillus helveticus are outside 3:2:1:1:0.5:0.5:0.5, all the other are identical with embodiment 2, and each measurement result is in table 1.
Reference examples 1
Except in the first fermentation step, access Leuconostoc mesenteroides add MRS medium component in above-mentioned extract after and carry out outside the first fermentation, all the other are identical with embodiment 1, and each measurement result is in table 1.
Reference examples 2
This reference examples only implements the first fermentation of embodiment 1, and adopting by weight proportion is that the extract of compound lactobacillus to embodiment 1 that the streptococcus thermophilus of 1:1 and Lactobacillus casei form ferments, fermentation parameter ferments identical with first of embodiment 1, zymotic fluid after fermentation is under 4000r/min after centrifugal about 15min, homogeneous, sterilizing are carried out to centrifuged supernatant, obtained fermented product, each measurement result is in table 1.
The average score result of table 1 Fermentation Process of Parameter mensuration and fermented product
As seen from the results in Table 1:
Relative to the MRS culture medium of routine, adopt black-bone chicken peptide significantly can promote the growth of Leuconostoc mesenteroides as nitrogenous source; In addition, only have employing to comprise streptococcus thermophilus, the compound lactobacillus of Lactobacillus delbrueckii and other two or more lactic acid bacteria and the mulriple yeasts that simultaneously comprises two or more yeast to ferment to fruit and vegetable materials successively simultaneously, fermented product can be made simultaneously to have preferably mouthfeel and local flavor.
The evaluation of test example 1 immunoloregulation function
1, the preparation of mouse boosting cell suspension
Draw neck to put to death Balb/c mouse, soak 5min disinfection in 75% ethanol, the complete spleen of aseptic separation, washes away floating blood with PBS, divests connective tissue and fat constituent.Adopt syringe to draw about 5mLPBS, insert blowout cell in spleen gently, repeat transparent to spleen adventitia for several times, residue spleen tissue is cut into size 1mm
3fritter, shred on the 200 order stainless steel filtering nets that are placed on small beaker, grind with syringe nook closing member, PBS liquid washs, collect flushing liquor to centrifuge tube, the centrifugal 5min of 1200rpm, abandon supernatant, add 3mL erythrocyte cracked liquid, leave standstill 2min, add 10mL PBS cessation reaction, the centrifugal 5min of 1200rpm, abandon supernatant, twice is washed with PBS liquid, the centrifugal 5min of 1200rpm, wash once with the incomplete culture medium of RPMI-1640, the centrifugal 5min of 1200rpm, abandon supernatant, add appropriate RPMI-1640 complete medium (containing 10% hyclone, 100U/mL penicillin, 100 μ g/mL streptomysins and 10mM Hepes), adjustment cell concentration to 2 × 10
6individual cell/mL, Trypan Blue detects cell survival rate and is greater than 95%.
2, each fermented product is on lymphopoietic impact
Fermented product pure water prepared by embodiment 1 ~ 5, reference examples 1 and reference examples 2 is carried out gradient dilution respectively, obtained respective gradient dilution liquid.
Above-mentioned splenocyte suspension is joined in 96 well culture plates by 100 μ L/ holes, add the gradient dilution liquid of 10 μ LConA (5 μ g/mL) or 10 μ L LPS (5 μ g/mL) and the above-mentioned each fermented product of 10 μ L more respectively, each gradient does 6 repetitions, culture plate is placed in 37 DEG C, 5%CO
2incubator in cultivate 72h; Blank (namely adding 10 μ L pure water) is set simultaneously.
Adopt each fermented product of CCK-8 kit measurement on the impact (representing with SI (SI)) of spleen lymphocyte proliferation ability, result as depicted in figs. 1 and 2.From Fig. 1 and Fig. 2: relative to the fermented product of reference examples 1 and reference examples 2, dendrobium candidum fermented product prepared by the embodiment of the present invention can significantly increase mouse T lymphocyte activity, has more excellent immunoloregulation function.
The evaluation of reference examples 2 anti-oxidation function
The fermented product of embodiment 1 ~ 5, reference examples 1 and reference examples 2 is carried out gradient dilution with the DMEM culture medium of serum-free respectively, obtained gradient dilution liquid;
By HepG2 cell with cell density 2 × 10
5be seeded to Tissue Culture Plate, every hole 2ml, be placed in 37 DEG C, 5%CO
2cultivate 24h in cell culture incubator, carefully siphon away the cell conditioned medium liquid in hole subsequently, clean once with HBSS, add the gradient dilution liquid 2ml of above-mentioned each fermented product respectively, in 37 DEG C, 5%CO
23.5h is cultivated in cell culture incubator, then fluorescent dye 2 ' is added, 7'-dihydro dichlorofluorescein sodium Diacetate (2 ', 7 '-dichlorofluorescin diacetate, DCFH-DA, final concentration is 10 μMs), carefully clean three times with HBSS, finally add antioxygen damage agent 600 μMs of AAPH; Blank (namely adding the DMEM culture medium of 2ml serum-free) is set simultaneously.
ROS content in flow cytomery cell is adopted (to use EC
50value represents), result is as shown in Figure 3.As shown in Figure 3: relative to the fermented product of reference examples 1 and reference examples 2, dendrobium candidum fermented product prepared by the present invention has more excellent anti-oxidation function.
Last it is noted that above each embodiment is only in order to illustrate technical scheme of the present invention, be not intended to limit; Although with reference to foregoing embodiments to invention has been detailed description, those of ordinary skill in the art is to be understood that: it still can be modified to the technical scheme described in foregoing embodiments, or carries out equivalent replacement to wherein some or all of technical characteristic; And these amendments or replacement, do not make the essence of appropriate technical solution depart from the scope of various embodiments of the present invention technical scheme.
Claims (10)
1. a preparation method for dendrobium candidum fermented product, is characterized in that, comprises the steps:
1) by extracting in water after dendrobium candidum fragmentation, obtained extract;
2), after adding medium component in described extract, access Leuconostoc mesenteroides carries out the first fermentation, when the pH value of zymotic fluid reduces by more than 0.5, and the content of reducing sugar of zymotic fluid lower than 1.5% time terminate described first fermentation, obtained first zymotic fluid;
3) to pH value be 4.5 ~ 6 fruits and vegetables liquid in add pectase and carry out enzymolysis processing, obtained enzymolysis fruits and vegetables liquid;
4) after adding medium component in described enzymolysis fruits and vegetables liquid, access compound lactobacillus carries out the second fermentation, terminates described second fermentation, obtained second zymotic fluid when the pH value of zymotic fluid reduces by more than 0.5; Wherein, described compound lactobacillus is selected from more than four kinds in streptococcus thermophilus, Lactobacillus delbrueckii, lactobacillus acidophilus, Lactobacillus casei, Lactobacillus plantarum, lactobacillus fermenti, Lactobacillus rhamnosus, Lactobacillus brevis, Lactobacillus helveticus, Pediococcus acidilactici, Pediococcus pentosaceus;
5), after adding medium component in described second zymotic fluid, access mulriple yeasts carries out the 3rd fermentation, when zymotic fluid total sugar content lower than 2% time terminate described 3rd fermentation, obtained 3rd zymotic fluid; Wherein, what described mulriple yeasts was selected from rice wine yeast, brewer's yeast, yellow wine yeast and fruit wine and wine yeast is two or more;
6) by centrifugal after described first zymotic fluid and described 3rd zymotic fluid mixing, after centrifuged supernatant homogeneous, sterilizing, obtained dendrobium candidum fermented product.
2. preparation method according to claim 1, is characterized in that, step 1) in, dendrobium candidum is crushed to 40 ~ 80 orders, and the quality proportioning of the dendrobium candidum after controlled fragmentation and water is 1:40 ~ 60, the temperature of described extraction is 90 ~ 120 DEG C, and extraction time is 5 ~ 30min.
3. preparation method according to claim 1, is characterized in that, described medium component comprise in sugar, peptide, inorganic salts and surfactant one or more, and in described peptide, mean molecule quantity is less than gross mass content >=80% of the component of 1000Da.
4. preparation method according to claim 3, it is characterized in that, step 2) in, sugar, peptide and inorganic salts are added in described extract, make the mass content of described sugar in described extract be 5 ~ 10%, the mass content of described peptide in described extract is 0.3 ~ 0.8%, and the mass content of described inorganic salts in described extract is 0.1 ~ 0.3%, and the temperature controlling described first fermentation is 22 ~ 28 DEG C, and rotating speed is 80 ~ 120r/min.
5. preparation method according to claim 1, it is characterized in that, the two or more fruit and vegetable materials be selected from pineapple, watermelon, strawberry, apple, banana, mango, pawpaw, radish, cucumber, tomato is crushed to 40 ~ 80 orders, and obtained pH value is the described fruits and vegetables liquid of 4.5 ~ 6.
6. preparation method according to claim 1, is characterized in that, the consumption of described pectase is every gram of fruits and vegetables liquid 2 ~ 3 unit, and the temperature controlling described enzymolysis processing is 40 ~ 50 DEG C, and the time is 2 ~ 3h.
7. preparation method according to claim 3, it is characterized in that, step 4) in, in described enzymolysis fruits and vegetables liquid, add sugar and peptide, make the mass content of described sugar in described enzymolysis fruits and vegetables liquid be 3 ~ 5%, the mass content of described peptide in described enzymolysis fruits and vegetables liquid is 0.3 ~ 0.8%; Described compound lactobacillus at least comprises streptococcus thermophilus and Lactobacillus delbrueckii, the weight proportion of described streptococcus thermophilus, Lactobacillus delbrueckii and all the other lactic acid bacterias is 3:2:(2 ~ 5), and the temperature controlling described second fermentation is 18 ~ 23 DEG C, and rotating speed is 80 ~ 120r/min.
8. preparation method according to claim 3, it is characterized in that, step 5) in, in described second zymotic fluid, add sugar and peptide, make the mass content of described sugar in described second zymotic fluid be 4 ~ 8%, the mass content of described peptide in described second zymotic fluid is 0.3 ~ 0.8%; And the weight proportion controlled 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.
9. preparation method according to claim 1, is characterized in that, step 6) in, the weight proportion of described first zymotic fluid and described 3rd zymotic fluid is (4 ~ 6): (4 ~ 6).
10. a dendrobium candidum fermented product, is characterized in that, according to claim 1 to 9, arbitrary described preparation method obtains.
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