CN108719976A - A kind of walnut grouts ferment production method - Google Patents
A kind of walnut grouts ferment production method Download PDFInfo
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- CN108719976A CN108719976A CN201810425861.2A CN201810425861A CN108719976A CN 108719976 A CN108719976 A CN 108719976A CN 201810425861 A CN201810425861 A CN 201810425861A CN 108719976 A CN108719976 A CN 108719976A
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- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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Abstract
A kind of walnut grouts ferment production method.This method is directed in walnut enterprise production process and generates a large amount of grouts as offal treatment,Not only cause the waste of resource,The problem of polluting the environment again simultaneously,It is proposed on the basis of analyzing and researching walnut grouts main chemical compositions and its content,Raw material is wanted based on walnut grouts,Reasonable compatibility has the Ways of Special Agricultural Products raw material Rhizoma Gastrodiae powder of health-care efficacy,Kudzu-vine root powder,Radix Notoginseng powder,Matrimony vine etc.,Include the enzymolysis of walnut grouts,Compound bacteria-fermented,Zymotic fluid hyperfiltration treatment,The high macro-molecular protein for being difficult to digest of content in walnut grouts is hydrolyzed into the small-molecule substance for being easy to human body and utilizing,Make full use of other nutritional ingredients therein and functional components again simultaneously,Its product has different functionalities and flavor,Protease,Superoxide dismutase,Lipase,Lactobacillus casei content is higher,Improve value-added content of product,The comprehensive utilization of walnut is expanded,Farthest realize the complete utilization of walnut.
Description
Technical field
The invention belongs to functional food processing technique field, more particularly to a kind of preparation method of walnut grouts ferment.
Background technology
Yunnan is the big province of walnut main product, by 2016, Walnut in Yunnan area up to 42,800,000 mu, 1,020,000 tons of yield,
30,500,000,000 yuan of the output value, the equal position of yield, area are the first in the nation.Walnut Industry has become great Yunnan Guanyuan characteristic, the whole nation or even
The world has an impact, competitive strong industry.Grouts residual rate is up to 70% after walnut oil expression, and zymogen is extracted from grouts
Liquid rate, with 1 ton of enzyme stoste ready market price, 50,000 yuan/ton of calculating, squeezes 1 ton of walnut fruit, can produce 280kg up to 40%
Enzyme stoste, for benefit up to 1.4 ten thousand yuan, i.e., walnut fruit extraction ferment per ton can add 1.4 ten thousand yuan of benefits, such as be produced to the whole province's walnut
Amount 5% extract oil after enzyme stoste extract, benefit is up to 5.95 hundred million yuan.
Making the basic principle of ferment is, micro enzyme is contained in the foods such as fresh fruit and vegetable, is placed on together with sugar
In air-tight bottle, it may occur that violent glycolysis reaction and enzymatic reaction make the enzymatic activity in food greatly improve, and the quantity of enzyme swashs
Increase, ultimately forms the ferment beneficial to human body.It is " extract of the enzyme of plant " or " essence of phytoenzyme extraction that ferment, which is translated into Chinese,
The meaning of China " is with plant materials such as fruit and vegetables, and with the made fermented food of lactic acid bacteria or yeast fermentation, why food has
Beneficial health, contains flavonoids, phytochrome and superoxide dismutase (SOD) isoreactivity ingredient mainly in the extract, can
Eliminate active oxygen harmful to health in human body.The whole intestines effect for additionally having benefited from lactic acid bacteria therein and yeast, can inhibit big
Various vitamins, amino acid and the mineral element etc. that the generation of the toxin such as corruption and amine in intestines and plant material are provided
Trophism.
Invention content
In order to solve to generate a large amount of walnut grouts in existing walnut enterprise production process as offal treatment, not only make
At the waste of resource, while the problem of pollute the environment again, the present invention studies walnut grouts, walnut grouts
Main chemical compositions and its weight percentage are:Moisture 5%, thick protein 70%, crude fat 8%, ash content 1.0%, thick fibre
It is mainly potassium, calcium, magnesium, iron, zinc, flavones about 0.2mg/g and polysaccharide 0.4mg/g in dimension 10%, ash content.Walnut grouts moisture removal and
Outside water-soluble actives, the high insoluble solid of content is mainly the macro-molecular protein for being difficult to digest, for this purpose, of the invention
It is proposed it is a kind of the macro-molecular protein for being difficult to digest in walnut grouts can be made to be hydrolyzed into be easy to the small-molecule substance that human body utilizes,
Other nutritional ingredients contained therein and functional components can be made full use of again, it is in good taste, improve the walnut grouts utilization of resources
A kind of walnut grouts liquid ferment that wanting raw material based on walnut grouts and solid ferment production method of rate.
A kind of walnut grouts liquid ferment production method proposed by the invention, includes the following steps:
(1) enzymolysis of walnut grouts
Press walnut grouts:The weight ratio of pure water is 1:2 ratio mixes walnut grouts with pure water, adjusts pH and is
5.0~6.0, walnut grouts solution is obtained, on the basis of walnut grouts weight, pepsin is added in walnut grouts solution
1.5%, trypsase 1.0% and pectase 0.5%, heating water bath digest 15~16 hours controlled at 50 DEG C~55 DEG C
Obtain walnut grouts enzymolysis liquid;
(2) compound bacteria-fermented
The pH for adjusting walnut grouts enzymolysis liquid is 6, is 0.5% by lactobacillus acidophilus inoculum concentration, Lactobacillus casei inoculum concentration
It is 0.5%, lactobacillus acidophilus and Lactobacillus casei is added in walnut grouts enzymolysis liquid, then, brown sugar or ice brown sugar is added, makes
Pol is 30%, adds Rhizoma Gastrodiae powder, kudzu-vine root powder, Radix Notoginseng powder or matrimony vine, is uniformly mixed to be placed at aeration-drying and ferment, fermentation
30 DEG C, 9~11d of fermentation time of temperature makes zymotic fluid production is fragrant to obtain zymotic fluid, Rhizoma Gastrodiae powder, kudzu-vine root powder, Radix Notoginseng powder or the matrimony vine of addition
Amount be 5% of walnut grouts enzymolysis liquid weight described in step (1);
(3) zymotic fluid hyperfiltration treatment
After step (2) gained zymotic fluid natural sedimentation 24 hours, hyperfiltration treatment is carried out, gained filtrate is walnut grouts liquid
Body ferment.
Further, the hyperfiltration treatment described in step (3) carries out hyperfiltration treatment, plate and frame using plate and frame ultrafiltration apparatus
The filter plate ultrafiltration specification of ultrafiltration apparatus:Ceramic membrane membrane aperture is 0.5 μm, filter pressure:0.5MPa.
A kind of solid walnut grouts ferment production method provided by the invention first uses a kind of above-mentioned walnut grouts liquid ferment
After plain production method obtains walnut grouts liquid ferment, walnut grouts liquid ferment is subjected to rotary evaporation concentration, rotary evaporation
Bath temperature control is 30 DEG C~40 DEG C, the walnut grouts liquid ferment that walnut grouts liquid ferment volume concentration 50% must be concentrated
Then the walnut grouts liquid ferment of concentration is packed into pallet, the pallet of the walnut grouts liquid ferment equipped with concentration is put by element
The progress vacuum freeze drying processing in vacuum freeze drier is set, keeps the moisture of the walnut grouts liquid ferment of concentration small
It is solid walnut grouts ferment to obtain walnut grouts ferment powder in 5%w/w.
Further, the vacuum freeze drying condition of the vacuum freeze drying processing is:Cryogenic temperature is -26 DEG C, vacuum
Degree is 10~20Pa, and condenser temperature is -40 DEG C.
Further, it is the core that each pallet fills 500mL concentrations that the walnut grouts liquid ferment by concentration, which is packed into pallet,
6 pallets of the walnut grouts liquid ferment equipped with concentration are placed on vacuum freeze drier by peach grouts liquid ferment every time
Middle progress vacuum freeze drying processing.
With the method for the present invention, four kinds of walnut grouts liquid ferment products containing Rhizoma Gastrodiae, pueraria lobata, Radix Notoginseng or matrimony vine are can get
Or four kinds of solid walnut grouts ferment containing Rhizoma Gastrodiae, pueraria lobata, Radix Notoginseng or matrimony vine.
The walnut grouts liquid ferment appearance transparent of gained, rufous, sweet mouthfeel is agreeable to the taste, has the distinctive faint scent of walnut
Taste, protease content > 1500U/g, superoxide dismutase (SOD) content > 200U/g, lipase content > 100U/g are done
Lactobacillus paracasei quantity > 2 × 108CFU/g。
The solid walnut grouts ferment appearance dark-brown of gained, band sweet tea in mouthfeel acid, has fermenting aroma, protease content
> 4500U/g, superoxide dismutase (SOD) content > 600U/g, lipase content >
200U/g, Lactobacillus casei quantity > 4 × 108CFU/g。
Containing Rhizoma Gastrodiae, pueraria lobata, Radix Notoginseng or the walnut of matrimony vine grouts liquid ferment product or containing Rhizoma Gastrodiae, pueraria lobata, Radix Notoginseng or
The solid walnut grouts ferment of matrimony vine also contains corresponding Gastrodin, pueraria isoflavone, notoginsenoside respectively in addition to These characteristics
Or the multiple efficacies ingredient such as polysaccharides, there is plurality of health care functions and flavor.
Compared with prior art, the invention has the advantages that:
The present invention be using walnut extract oil by-product grouts as primary raw material, will be in walnut grouts using suitable complex enzyme hydrolysis
The high-leveled and difficult macro-molecular protein with digestion of content be hydrolyzed into and be easy to the small-molecule substance that human body utilizes, while taking full advantage of again
Wherein other nutritional ingredients and functional components contained in walnut grouts, other characteristic agricultures with health-care efficacy of reasonable selection
Products material carries out compatibility (Rhizoma Gastrodiae powder, kudzu-vine root powder, Radix Notoginseng powder, matrimony vine), produces ferment using suitable probiotics fermention, passes through
The difference of compatibility raw material develops the walnut grouts ferment product of different functionalities and flavor type, makes walnut oil expression by-product
A large amount of protein in grouts are utilized effectively, protease, superoxide dismutase, lipase, Lactobacillus casei in product
Content is higher, is conducive to health, improves the added value of food, has expanded the comprehensive utilization of walnut, farthest
Realize the complete utilization of walnut.
Specific implementation mode
Below by specific implementation mode, the present invention is further illustrated, pepsin, tryptose in each embodiment
The various substances such as enzyme, pectase (i.e. alkaline endo polygalacturonase), lactobacillus acidophilus, Lactobacillus casei are commercially available
(including equipment).Without specified otherwise it is conventional method in each embodiment.
Embodiment 1 makes the walnut grouts liquid ferment containing Rhizoma Gastrodiae
(1) enzymolysis of walnut grouts
It is mixed with 2000g pure water with 1000g walnuts grouts, with the citric acid of a concentration of 0.1%w/w or a concentration of
It is 5.5 that the sodium hydroxide solution of 0.15%w/w, which adjusts pH, obtains walnut grouts solution, adds 15g pepsin (i.e. stomach cardias
The addition of enzyme is the 1.5% of walnut grouts weight), (i.e. the addition of trypsase is walnut grouts weight to 10g trypsase
1.0%), 5g pectases (i.e. the addition of pectase be walnut grouts weight 0.5%) mixing after heating water bath in temperature
Under the conditions of 50 DEG C~55 DEG C, enzymolysis obtains walnut grouts enzymolysis liquid in 16 hours;
(2) compound bacteria-fermented
Walnut grouts enzyme is adjusted with the citric acid of a concentration of 0.1%w/w or the sodium hydroxide solution of a concentration of 0.15%w/w
The pH for solving liquid is 6, is that 0.5% (amount for the lactobacillus acidophilus being added is described in step (1) by lactobacillus acidophilus inoculum concentration
The 0.5% of walnut grouts enzymolysis liquid weight), Lactobacillus casei inoculum concentration is 0.5% (amount for the Lactobacillus casei being added is step
Suddenly 0.5% of the walnut grouts enzymolysis liquid weight described in (1)), lactobacillus acidophilus is added in walnut grouts enzymolysis liquid and does
Then brown sugar dissolving is added, it is 30% to make pol, adds Rhizoma Gastrodiae powder in Lactobacillus paracasei (Rhizoma Gastrodiae powder was the Rhizoma Gastrodiae powder of 80 mesh)
It is uniformly mixed to be placed at aeration-drying and ferment, 30 DEG C, fermentation time 10d of fermentation temperature, zymotic fluid production is fragrant to obtain zymotic fluid, is added
Rhizoma Gastrodiae powder amount be step (1) described in walnut grouts enzymolysis liquid weight 5%;
(3) zymotic fluid hyperfiltration treatment
After zymotic fluid natural sedimentation obtained by step (2) 24 hours, using plate and frame ultrafiltration apparatus to gained zymotic fluid into
Row hyperfiltration treatment is flowed through zymotic fluid using oil pump among a kind of special ultrafiltration membrane, to remove the macromolecular in zymotic fluid
Substance, floating material and other impurity, gained filtrate are walnut grouts liquid ferment.Filter plate ultrafiltration specification:Ceramic membrane membrane aperture
It is 0.5 μm, filter area 0.64m2, hold-down mechanism:Mechanical compaction, feeding manner:Middle upper feeding, filter pressure:0.5MPa.
The walnut grouts liquid ferment of 1 gained of embodiment is the walnut grouts liquid ferment containing Rhizoma Gastrodiae.
Embodiment 2 makes the walnut grouts liquid ferment containing pueraria lobata
For embodiment 2 in addition to following steps are different, remaining step is same as Example 1:
The time of step (1) enzymolysis is 15 hours, obtains walnut grouts enzymolysis liquid;The day that will be added in 1 step of embodiment (2)
Numb powder replaces with kudzu-vine root powder (kudzu-vine root powder was the kudzu-vine root powder of 80 mesh, and the addition of kudzu-vine root powder is identical as the addition of Rhizoma Gastrodiae powder),
The brown sugar being added in 1 step of embodiment (2) replaces with ice brown sugar (pol is constant), fermentation time 11d, and zymotic fluid production perfume (or spice) obtains
Zymotic fluid;Filtrate obtained by step (3) zymotic fluid hyperfiltration treatment is the walnut grouts liquid ferment containing pueraria lobata, has pueraria lobata light
The flavor of light time sweet tea and walnut fragrance.
Embodiment 3 makes the walnut grouts liquid ferment containing Radix Notoginseng
For embodiment 3 in addition to following steps are different, remaining step is same as Example 1:
By the Rhizoma Gastrodiae powder being added in 1 step of embodiment (2) replace with Radix Notoginseng powder (Radix Notoginseng powder was the Radix Notoginseng powder of 80 mesh, three
The addition of seven powder is identical as the addition of Rhizoma Gastrodiae powder), step (3) zymotic fluid hyperfiltration treatment gained filtrate is to contain Radix Notoginseng
Walnut grouts liquid ferment, with the slightly bitter flavor of Radix Notoginseng.
Embodiment 4 makes the walnut grouts liquid ferment containing matrimony vine
For embodiment 4 in addition to following steps are different, remaining step is same as Example 1:
The time of step (1) enzymolysis is 15 hours, obtains walnut grouts enzymolysis liquid;The Rhizoma Gastrodiae being added in 1 step of embodiment (2)
Powder replaces with matrimony vine (addition of matrimony vine is identical as the addition of Rhizoma Gastrodiae powder), fermentation time 9d, and zymotic fluid production perfume (or spice) is fermented
Liquid;Filtrate obtained by step (3) zymotic fluid hyperfiltration treatment is the walnut grouts liquid ferment containing matrimony vine, is mainly walnut egg
White oil Flavor.
Embodiment 5 makes the solid walnut grouts ferment containing Rhizoma Gastrodiae
For embodiment 5 in addition to following steps are different, remaining step is same as Example 1.
The walnut grouts liquid ferment containing Rhizoma Gastrodiae of gained in embodiment 1 is subjected to rotary evaporation concentration, rotary evaporation
Bath temperature control is 30 DEG C~40 DEG C, and the walnut grouts liquid ferment volume concentration 50% containing Rhizoma Gastrodiae (is contained Rhizoma Gastrodiae
The volume of walnut grouts liquid ferment subtracted half) the walnut grouts liquid ferment containing Rhizoma Gastrodiae that must concentrate, then will be dense
The walnut grouts liquid ferment containing Rhizoma Gastrodiae of contracting is packed into pallet, the walnut cakes containing Rhizoma Gastrodiae of each pallet dress 500mL concentrations
6 pallets of the walnut grouts liquid ferment containing Rhizoma Gastrodiae equipped with concentration are placed on vacuum refrigeration by dregs of rice liquid ferment every time
Vacuum freeze drying processing is carried out in drying machine, vacuum freeze drying condition is:Sample (the walnut grouts containing Rhizoma Gastrodiae of concentration
Liquid ferment) cryogenic temperature be -26 DEG C, vacuum degree 20Pa, condenser temperature be -40 DEG C, make the walnut grouts liquid ferment of concentration
It is the solid walnut grouts ferment containing Rhizoma Gastrodiae that the moisture of element obtains walnut grouts ferment powder less than 5%w/w.
Embodiment 6 makes the solid walnut grouts ferment containing pueraria lobata
For embodiment 6 in addition to following steps are different, remaining step is same as Example 2.
The walnut grouts liquid ferment containing pueraria lobata of gained in embodiment 2 is subjected to rotary evaporation concentration, rotary evaporation
Bath temperature control is 30 DEG C~40 DEG C, and the walnut grouts liquid ferment volume concentration 50% containing pueraria lobata (is contained pueraria lobata
The volume of walnut grouts liquid ferment subtracted half) the walnut grouts liquid ferment containing pueraria lobata that must concentrate, then will be dense
The walnut grouts liquid ferment containing pueraria lobata of contracting is packed into pallet, the walnut cakes containing pueraria lobata of each pallet dress 500mL concentrations
6 pallets of the walnut grouts liquid ferment containing pueraria lobata equipped with concentration are placed on vacuum refrigeration by dregs of rice liquid ferment every time
Vacuum freeze drying processing is carried out in drying machine, vacuum freeze drying condition is:Sample (the walnut grouts containing pueraria lobata of concentration
Liquid ferment) cryogenic temperature be -26 DEG C, vacuum degree 15Pa, condenser temperature be -40 DEG C, make the walnut containing pueraria lobata of concentration
It is the solid walnut grouts containing pueraria lobata that the moisture of grouts liquid ferment obtains walnut grouts ferment powder less than 5%w/w
Ferment.
Embodiment 7 makes the solid walnut grouts ferment containing Radix Notoginseng
For embodiment 7 in addition to following steps are different, remaining step is same as Example 3.
The walnut grouts liquid ferment containing Radix Notoginseng of gained in embodiment 3 is subjected to rotary evaporation concentration, rotary evaporation
Bath temperature control is 30 DEG C~40 DEG C, and the walnut grouts liquid ferment volume concentration 50% containing Radix Notoginseng (is contained Radix Notoginseng
The volume of walnut grouts liquid ferment subtracted half) the walnut grouts liquid ferment containing Radix Notoginseng that must concentrate, then will be dense
The walnut grouts liquid ferment containing Radix Notoginseng of contracting is packed into pallet, the walnut cakes containing Radix Notoginseng of each pallet dress 500mL concentrations
6 pallets of the walnut grouts liquid ferment containing Radix Notoginseng equipped with concentration are placed on vacuum refrigeration by dregs of rice liquid ferment every time
Vacuum freeze drying processing is carried out in drying machine, vacuum freeze drying condition is:Sample (the walnut grouts containing Radix Notoginseng of concentration
Liquid ferment) cryogenic temperature be -26 DEG C, vacuum degree 10Pa, condenser temperature be -40 DEG C, make the walnut containing Radix Notoginseng of concentration
It is the solid walnut grouts containing Radix Notoginseng that the moisture of grouts liquid ferment obtains walnut grouts ferment powder less than 5%w/w
Ferment.
Embodiment 8 makes the solid walnut grouts ferment containing matrimony vine
For embodiment 8 in addition to following steps are different, remaining step is same as Example 4.
The walnut grouts liquid ferment containing matrimony vine of gained in embodiment 4 is subjected to rotary evaporation concentration, rotary evaporation
Bath temperature control is 30 DEG C~40 DEG C, and the walnut grouts liquid ferment volume concentration 50% containing matrimony vine (is contained matrimony vine
The volume of walnut grouts liquid ferment subtracted half) the walnut grouts liquid ferment containing matrimony vine that must concentrate, then will be dense
The walnut grouts liquid ferment containing matrimony vine of contracting is packed into pallet, the walnut cakes containing matrimony vine of each pallet dress 500mL concentrations
6 pallets of the walnut grouts liquid ferment containing matrimony vine equipped with concentration are placed on vacuum refrigeration by dregs of rice liquid ferment every time
Vacuum freeze drying processing is carried out in drying machine, vacuum freeze drying condition is:Sample (the walnut grouts containing matrimony vine of concentration
Liquid ferment) cryogenic temperature be -26 DEG C, vacuum degree 15Pa, condenser temperature be -40 DEG C, make the walnut containing matrimony vine of concentration
It is the solid walnut grouts containing matrimony vine that the moisture of grouts liquid ferment obtains walnut grouts ferment powder less than 5%w/w
Ferment.
Claims (5)
1. a kind of walnut grouts liquid ferment production method, which is characterized in that include the following steps:
(1) enzymolysis of walnut grouts
Press walnut grouts:The weight ratio of pure water is 1:2 ratio mixes walnut grouts with pure water, adjust pH be 5.0~
6.0, walnut grouts solution is obtained, on the basis of walnut grouts weight, pepsin 1.5%, pancreas are added in walnut grouts solution
Protease 1.0% and pectase 0.5%, heating water bath controlled at 50 DEG C~55 DEG C, enzymolysis 15~16 hours walnut
Grouts enzymolysis liquid;
(2) compound bacteria-fermented
The pH for adjusting walnut grouts enzymolysis liquid is 6, is 0.5% by lactobacillus acidophilus inoculum concentration, and Lactobacillus casei inoculum concentration is
0.5%, lactobacillus acidophilus and Lactobacillus casei are added in walnut grouts enzymolysis liquid, then, brown sugar or ice brown sugar is added, makes sugar
Degree is 30%, adds Rhizoma Gastrodiae powder, kudzu-vine root powder, Radix Notoginseng powder or matrimony vine, is uniformly mixed to be placed at aeration-drying and ferment, fermentation temperature
30 DEG C, 9~11d of fermentation time of degree, makes fragrant the zymotic fluid of zymotic fluid production, the Rhizoma Gastrodiae powder of addition, kudzu-vine root powder, Radix Notoginseng powder or matrimony vine
Amount is 5% of walnut grouts enzymolysis liquid weight described in step (1);
(3) zymotic fluid hyperfiltration treatment
After step (2) gained zymotic fluid natural sedimentation 24 hours, hyperfiltration treatment is carried out, gained filtrate is walnut grouts liquid ferment
Element.
2. walnut grouts liquid ferment production method according to claim 1, which is characterized in that described in step (3)
Hyperfiltration treatment carries out hyperfiltration treatment, the filter plate ultrafiltration specification of plate and frame ultrafiltration apparatus using plate and frame ultrafiltration apparatus:Ceramic membrane
Membrane aperture is 0.5 μm, filter pressure:0.5MPa.
3. a kind of solid walnut grouts ferment production method, which is characterized in that with walnut grouts liquid as claimed in claim 1 or 2
After body ferment production method obtains walnut grouts liquid ferment, walnut grouts liquid ferment is subjected to rotary evaporation concentration, rotation
It is 30 DEG C~40 DEG C to evaporate bath temperature control, the walnut grouts liquid that walnut grouts liquid ferment volume concentration 50% must be concentrated
Then the walnut grouts liquid ferment of concentration is packed into pallet, by the support of the walnut grouts liquid ferment equipped with concentration by body ferment
Disk is placed on progress vacuum freeze drying processing in vacuum freeze drier, and the moisture of the walnut grouts liquid ferment of concentration is made to contain
It is solid walnut grouts ferment that amount obtains walnut grouts ferment powder less than 5%w/w.
4. solid walnut grouts ferment production method according to claim 3, which is characterized in that the vacuum freeze drying
The vacuum freeze drying condition of processing is:Cryogenic temperature is -26 DEG C, and vacuum degree is 10~20Pa, and condenser temperature is -40 DEG C.
5. solid walnut grouts ferment production method according to claim 3 or 4, which is characterized in that described by concentration
It is the walnut grouts liquid ferment that each pallet fills 500mL concentrations that walnut grouts liquid ferment, which is packed into pallet, will be equipped with every time dense
6 pallets of the walnut grouts liquid ferment of contracting are placed on progress vacuum freeze drying processing in vacuum freeze drier.
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