CN104522817B - A kind of witloof fermented product and preparation method thereof - Google Patents

A kind of witloof fermented product and preparation method thereof Download PDF

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CN104522817B
CN104522817B CN201510025580.4A CN201510025580A CN104522817B CN 104522817 B CN104522817 B CN 104522817B CN 201510025580 A CN201510025580 A CN 201510025580A CN 104522817 B CN104522817 B CN 104522817B
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zymotic fluid
lactobacillus
fermentation
witloof
fruits
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CN104522817A (en
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蔡木易
谷瑞增
鲁军
陆路
潘兴昌
董哲
林峰
马勇
徐亚光
马永庆
金振涛
陈亮
刘文颖
魏颖
张海欣
刘艳
马涛
曹珂璐
姜思萌
王憬
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China National Research Institute of Food and Fermentation Industries
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China National Research Institute of Food and Fermentation Industries
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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
    • A23L2/38—Other non-alcoholic beverages
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
    • A23L2/70—Clarifying or fining of non-alcoholic beverages; Removing unwanted matter
    • A23L2/84—Clarifying or fining of non-alcoholic beverages; Removing unwanted matter using microorganisms or biological material, e.g. enzymes
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/11—Lactobacillus
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/11—Lactobacillus
    • A23V2400/161—Mesenteroides

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Nutrition Science (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Food Science & Technology (AREA)
  • Polymers & Plastics (AREA)
  • Microbiology (AREA)
  • Molecular Biology (AREA)
  • Zoology (AREA)
  • Coloring Foods And Improving Nutritive Qualities (AREA)
  • Non-Alcoholic Beverages (AREA)

Abstract

The invention provides a kind of witloof fermented product and preparation method thereof.The method comprises the steps: 1) witloof is soaked in water, crushing and beating, homogeneous, obtained slurries; 2) after adding medium component in slurries, access Leuconostoc mesenteroides carries out the 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 enzymolysis fruits and vegetables liquid, access compound lactobacillus carries out the second fermentation, obtained second zymotic fluid; 5) after adding medium component in the second zymotic fluid, access mulriple yeasts carries out the 3rd fermentation, obtained 3rd zymotic fluid; 6) by centrifugal after the first zymotic fluid and the mixing of the 3rd zymotic fluid, after centrifuged supernatant homogeneous, sterilizing, obtained witloof fermented product.This fermented product mouthfeel is good, unique flavor, has good immunological regulation and the function such as anti-oxidant, is suitable for crowd in extensive range.

Description

A kind of witloof fermented product and preparation method thereof
Technical field
The present invention relates to a kind of fermented product, particularly relate to a kind of witloof fermented product and preparation method thereof.
Background technology
Witloof is composite family herbaceos perennial, bitter, and another name hare's-lettuce, bitter dish etc., originate in Europe, and also there is plantation on the many ground of existing China.Witloof can clearing heat and detoxicating, inducing diuresis to reduce edema, raising immunity, nourishing the stomach to improve the production of body fluid etc., and instructions of taking comprises and boils, stewes, stir-fries and eats.
Though witloof is rich in various active composition, but be made up of cellulose, hemicellulose etc. due to the cell membrane of plant cell, its compact structure, above-mentioned eating method is generally difficult to be the nutritional labeling in absorption of human body witloof, and bioavilability is low.At present, a small amount of relevant report witloof being processed into beverage is had.Such as, notification number be CN102085014A patent discloses a kind of health drink containing witloof, it is soaked by witloof to obtain witloof liquid, add longan juice, honey, pure water again, this beverage can help human body to the absorption of calcium, but still do not discharge more nutritional labeling in witloof, effect is too single, and taste is common.
Notification number is the preparation method that patent discloses a kind of witloof sliced bread of CN103621592A, it is by high-strength flour 45 ~ 55 parts, 15 ~ 25 parts, water, Chicory syrup 4 ~ 12 parts, 3 ~ 8 parts, egg, 2 ~ 6 parts, butter, white granulated sugar 2 ~ 6 parts, 2 ~ 6 parts, witloof powder, inulin 1 ~ 3 part, milk powder 0.5 ~ 0.9 part, face powder 0.4 ~ 0.8 part, 0.3 ~ 0.6 part, yeast, salt 0.2 ~ 0.6 part, modifying agent 0.1 ~ 0.4 part, baking powder 0.1 ~ 0.4 part combination is obtained through toasting, the witloof powder that the method adopts and Chicory syrup, still all cannot discharge the active ingredient in witloof, and this product albumen fats is too much, too much eat and can bring burden for human body.
Summary of the invention
The invention provides a kind of witloof fermented product and preparation method thereof, for solving the technological deficiencies such as witloof series products composition of the prior art is single, taste is common, health care is limited.
The invention provides a kind of preparation method of witloof fermented product, comprise the steps:
1) witloof is soaked in water, crushing and beating, homogeneous, obtained slurries;
2), after adding medium component in described slurries, 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 be selected from streptococcus thermophilus, Lactobacillus delbrueckii, lactobacillus acidophilus, Lactobacillus casei, Lactobacillus plantarum, Lactobacillus rhamnosus, Lactobacillus brevis, Lactobacillus pentosus, Pediococcus acidilactici, more than four kinds of lactobacillus fermenti;
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 saccharomyces sake, rice wine yeast, yellow wine yeast, white 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 witloof fermented product.
In the preparation process in accordance with the present invention, described Leuconostoc mesenteroides can select Leuconostoc mesenteroides dextran subspecies (L.mesenteroidessubsp.dextranicum) or leuconostoc mesenteroides sub species cremoris (L.mesenteroidessubsp.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 described witloof, and it can be fresh witloof, also can be dry witloof.In concrete scheme of the present invention, dry witloof can be adopted as raw material, its moisture content is 10 ~ 15%.Further, the quality proportioning controlling described witloof and described water is 1:10 ~ 20, and described witloof is crushed to 40 ~ 80 orders.The pH of the slurries obtained by described witloof is 4.5 ~ 6.
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 witloof slurries, sugar, peptide, inorganic salts and surfactant, wherein the mass content of sugar in described slurries can be 5 ~ 10%, the mass content of peptide in described slurries can be 0.3 ~ 0.8%, the mass content of inorganic salts in described slurries can be 0.1 ~ 0.3%, and the mass content of surfactant in described slurries can be 0.01 ~ 0.1%; These witloof slurries are by obtaining witloof crushing and beating, homogeneous in water.
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 sodium sulphate; Described surfactant can be selected from the smooth and polysorbate of fatty glyceride, aliphatic acid sorb one or more, such as Arlacel-60 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 slurries, the mass content of described sugar in described slurries is made to be 5 ~ 10%, the mass content of described peptide in described slurries is 0.3 ~ 0.8%, the mass content of described inorganic salts in described slurries 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 banana, mango, sweet orange, mangosteen, dragon fruit, cherry, 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 carbon 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 Lactobacillus plantarum and Lactobacillus rhamnosus, and the weight proportion of Lactobacillus plantarum and Lactobacillus rhamnosus is (0.5 ~ 1.5): (0.5 ~ 1.5);
Three components: comprise Pediococcus acidilactici and Lactobacillus pentosus, and the weight proportion of Pediococcus acidilactici and Lactobacillus pentosus is (0.5 ~ 1): (0.5 ~ 1);
Four composition: comprise lactobacillus fermenti and Lactobacillus brevis, and the weight proportion between lactobacillus fermenti and Lactobacillus brevis is (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 saccharomyces sake and fruit wine and wine yeast, and saccharomyces sake, weight proportion between fruit wine and wine yeast are 1:(0.8 ~ 1.2);
Second component: comprise rice wine yeast and saccharomyces sake, and the weight proportion between rice wine yeast, saccharomyces sake is 1:(0.8 ~ 1.2);
Three components: comprise rice wine yeast, yellow wine yeast and white wine yeast, and rice wine yeast, weight proportion between yellow wine yeast and white wine yeast are 1:(0.8 ~ 1.2): (0.8 ~ 1.2);
Four composition: comprise yellow wine yeast and fruit wine and wine yeast, and yellow wine yeast and the weight proportion between fruit wine and wine yeast are 1:(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 witloof 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 witloof slurries, not only more effectively releases activity and the nutritional labeling of witloof, improves the utilization rate of witloof, add 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 witloof 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 embodiments of the 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 (CICC22181), streptococcus thermophilus (CICC6218), Lactobacillus delbrueckii subsp. lactis (CICC22170), lactobacillus acidophilus (CICC6082), Lactobacillus casei (CICC6114), Lactobacillus plantarum (CICC22138), Lactobacillus rhamnosus (CICC6154), Lactobacillus brevis (CICC23474), Pediococcus acidilactici (CICC10344), Lactobacillus pentosus (CICC21811), lactobacillus fermenti (CICC22551), 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
By moisture content be 11% witloof be soaked in (namely solid-liquid ratio is 1:1) in water according to weight proportion 1:1, witloof crushing and beating to 60 order, homogeneous, obtained pH value is about the slurries of 5.5;
Sucrose is added in above-mentioned slurries, black-bone chicken peptide and sodium sulphate, the mass content of sucrose in slurries is made to be about 8%, the mass content of black-bone chicken peptide in slurries is about 0.5%, the mass content of sodium sulphate in slurries is about 0.18%, after stirring and evenly mixing, inoculum concentration according to about 0.1% accesses Leuconostoc mesenteroides and carries out the first fermentation in slurries, the temperature controlling the first fermentation is about 25 DEG C, speed of agitator is about 100r/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 4.7 ~ 5).
Adopt the cell concentration of spectrophotometry first zymotic fluid, the results are shown in Table 1.
2, enzymolysis processing
After fresh peeling banana and dragon fruit are 1:1 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
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 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 4.7 ~ 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 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 comprises saccharomyces sake and fruit wine and wine yeast, wherein saccharomyces sake, weight proportion between fruit wine and wine yeast is 1:0.8, and the temperature controlling the 3rd fermentation is about 18 DEG C, 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, witloof 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 witloof fermented product (pH value is 4.5 ~ 5).
50 experimenters making age level be distributed in 20 years old ~ 80 years old eat the witloof fermented product of above-mentioned preparation, and carry out mouthfeel and local flavor marking to this witloof 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 witloof fermented product.
Embodiment 2
1, the first fermentation
Witloof is soaked in (namely solid-liquid ratio is 1:0.5) in water according to weight proportion 1:0.5, witloof crushing and beating to 50 order, homogeneous, obtained pH value is about the slurries of 5.3;
Sucrose is added in above-mentioned slurries, black-bone chicken peptide and potassium phosphate, the mass content of sucrose in slurries is made to be about 6.5%, the mass content of black-bone chicken peptide in slurries is about 0.3%, the mass content of potassium phosphate in slurries is about 0.3%, after stirring and evenly mixing, inoculum concentration according to about 0.05% accesses Leuconostoc mesenteroides and carries out the first fermentation in slurries, the temperature controlling the first fermentation is about 22 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 4.5 ~ 4.8), the cell concentration of the first zymotic fluid is in table 1.
2, enzymolysis processing
After fresh sweet orange and dragon fruit are 1:3 mixing according to weight proportion, be crushed to 70 orders, obtained pH value is about the fruits and vegetables liquid of 4.7;
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 42 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 3.5%, the mass content of black-bone chicken peptide in enzymolysis fruits and vegetables liquid is about 0.6%, 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 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 about 22 DEG C, speed of agitator is about 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 3.9 ~ 4.2).
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 0.7%, after stirring and evenly mixing, inoculum concentration according to about 0.015% carries out the 3rd fermentation to access mulriple yeasts, described mulriple yeasts bag is made up of rice wine yeast and saccharomyces sake, wherein rice wine yeast, weight proportion between saccharomyces sake is 1:0.8, and the temperature controlling the 3rd fermentation is about 16 DEG C, 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, witloof fermented product is prepared
Above-mentioned first zymotic fluid and the 3rd zymotic fluid are mixed centrifugal 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 witloof fermented product (pH value is 4.2 ~ 4.6); The mouthfeel of this witloof fermented product and the average score of local flavor the results are shown in Table 1.
Embodiment 3
1, the first fermentation
Witloof is soaked in (namely solid-liquid ratio is 1:0.5) in water according to weight proportion 1:1.5, witloof crushing and beating to 70 order, homogeneous, obtained pH value is about the slurries of 5;
Sucrose is added in above-mentioned slurries, black-bone chicken peptide, potassium phosphate and Arlacel-60, the mass content of sucrose in slurries is made to be about 8.5%, the mass content of black-bone chicken peptide in slurries is about 0.7%, the mass content of potassium phosphate in slurries is about 0.2%, the mass content of Arlacel-60 in slurries is about 0.05%, after stirring and evenly mixing, inoculum concentration according to about 0.2% accesses Leuconostoc mesenteroides and carries out the first fermentation in slurries, the temperature controlling the first fermentation is about 26 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 4.1 ~ 4.5), the cell concentration of the first zymotic fluid is in table 1.
2, enzymolysis processing
By fresh mangosteen, cherry and cucumber according to weight proportion be 1:1:2 mixing after, be crushed to 80 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 2.7 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 0.45%, after stirring and evenly mixing, inoculum concentration according to about 0.15% accesses compound lactobacillus and carries out the second fermentation in enzymolysis fruits and vegetables liquid, compound lactobacillus is by streptococcus thermophilus, Lactobacillus delbrueckii, Lactobacillus pentosus and Pediococcus acidilactici composition, wherein streptococcus thermophilus, Lactobacillus delbrueckii, weight proportion between Lactobacillus pentosus and Pediococcus acidilactici is 3:2:1:1, and the temperature controlling the second fermentation is about 21 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 4.7 ~ 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 6.5%, the mass content of black-bone chicken peptide in the second zymotic fluid is about 0.75%, 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, yellow wine yeast and white wine yeast composition, wherein rice wine yeast, yellow wine yeast, weight proportion between white wine yeast is 1:1.2:1.2, and the temperature controlling the 3rd fermentation is about 20 DEG C, speed of agitator is about 40r/min, when zymotic fluid total sugar content lower than 2% time terminate the 3rd fermentation, obtained 3rd zymotic fluid.
5, witloof 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 witloof fermented product (pH value is 4.4 ~ 4.8); The mouthfeel of this witloof fermented product and local flavor marking 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 plantarum, Lactobacillus rhamnosus, Lactobacillus brevis and lactobacillus fermenti, and wherein streptococcus thermophilus, Lactobacillus delbrueckii, Lactobacillus plantarum, Lactobacillus rhamnosus, weight proportion between Lactobacillus brevis and lactobacillus fermenti are 3:2:1:1:1:1; In 3rd fermentation step, mulriple yeasts is that outside the yellow wine 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 casei, lactobacillus acidophilus, Pediococcus acidilactici and Lactobacillus pentosus, wherein streptococcus thermophilus, Lactobacillus delbrueckii, Lactobacillus casei, lactobacillus acidophilus, weight proportion between Pediococcus acidilactici and Lactobacillus pentosus are outside 3:2:1:1:0.5:0.5, 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 slurries 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 slurries of compound lactobacillus to embodiment 1 that the Bifidobacterium of 1:1 and lactobacillus bulgaricus form ferment, 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.
Table 1 Fermentation Process of Parameter measures and fermented product marking result
Remarks: "-" expression does not detect.
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 10mLPBS 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 streptomysin and 10mMHepes), 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, 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), 10 μ LLPS (5 μ g/mL), 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, witloof 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 test example 2 anti-oxidation function
The fermented product of embodiment 1, 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 '-dichlorofluorescindiacetate, 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.
Adopt ROS content in flow cytomery cell, the oxidation resistance of each sample selects EC50 value to represent, EC50 is the amount of the antioxidant required for the generation of 50%ROS in T suppression cell, wherein, EC50 is less, then represent that the non-oxidizability of this antioxidant is better.Result as shown in Figure 3.As shown in Figure 3: relative to the fermented product of reference examples 1 and reference examples 2, witloof 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 witloof fermented product, is characterized in that, comprises the steps:
1) witloof is soaked in water, crushing and beating, homogeneous, obtained slurries;
2), after adding medium component in described slurries, 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 be selected from streptococcus thermophilus, Lactobacillus delbrueckii, lactobacillus acidophilus, Lactobacillus casei, Lactobacillus plantarum, Lactobacillus rhamnosus, Lactobacillus brevis, Lactobacillus pentosus, Pediococcus acidilactici, more than four kinds of lactobacillus fermenti;
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 saccharomyces sake, rice wine yeast, yellow wine yeast, white 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 witloof fermented product.
2. preparation method according to claim 1, is characterized in that, the quality proportioning controlling described witloof and described water is 1:0.5 ~ 1.5, and described witloof is crushed to 40 ~ 80 orders.
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 slurries, make the mass content of described sugar in described slurries be 5 ~ 10%, the mass content of described peptide in described slurries is 0.3 ~ 0.8%, and the mass content of described inorganic salts in described slurries 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, be crushed to 40 ~ 80 orders by after the two or more fruit and vegetable materials mixing be selected from banana, mango, sweet orange, mangosteen, dragon fruit, cherry, cucumber and tomato, 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 witloof fermented product, is characterized in that, according to claim 1 to 9, arbitrary described preparation method obtains.
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