CN107987995A

CN107987995A - A kind of production method of Organic grape fruit wine

Info

Publication number: CN107987995A
Application number: CN201711486308.1A
Authority: CN
Inventors: 王依轩; 梁涵
Original assignee: Yantai Chi Chi Chemical Technology Co Ltd
Current assignee: Yantai Chi Chi Chemical Technology Co Ltd
Priority date: 2017-12-29
Filing date: 2017-12-29
Publication date: 2018-05-04

Abstract

The present invention relates to a kind of production method of Organic grape fruit wine, by the plants interplanting with expelling parasite insecticidal effect in orchard, manages orchard according to organic planting requirement, grape is made wine after harvesting ripe, up to Organic grape fruit wine.The advantages of present invention is made wine using Organic grape, has nutrition, health, safety, non agricultural chemical residuum.

Description

A kind of production method of Organic grape fruit wine

Technical field

The present invention relates to a kind of production method of Organic grape fruit wine.

Background technology

Bread is the staff of life, strengthens food security work, the health and life security of relation China more than 1,300,000,000, it is necessary to Grab tightly and tightly.These years, Party and government have descended very big strength to grab food security, and food security situation constantly improves, but deposits The problem of it is still many, common people still have many expectations, it is necessary to redouble one's efforts, work is run business into particular one reality, it is ensured that Ren Minqun Many " safety on the tip of the tongue ".

Grape wine is with a long history, it is acted on aid digestion, beautifying face and moistering lotion and extra-nutrition, is favored by people, at present The grape to make grape wine is more using common plantation, containing farm chemical ingredients, and poor taste.The taste of traditional wine cannot Meet Contemporary young Man's Demands, Wine Enterprises are sought to make the transition.

The content of the invention

The technical problem to be solved by the invention is to provide a kind of production method of Organic grape fruit wine, the present invention uses The advantages of Organic grape is made wine, and has nutrition, health, safety, non agricultural chemical residuum.

In order to solve the above-mentioned technical problem, the present invention uses following technical scheme：

A kind of production method of Organic grape fruit wine, it is characterised in that follow the steps below：

By the plants interplanting with expelling parasite insecticidal effect in orchard, orchard is managed according to organic planting requirement, picks fruit Mature fruit prepares fruit juice in garden, picks ripe grape in orchard and carries out vintage wine, by fruit juice and grape wine according to mass ratio 1 ~75:25~99 allotments, up to Organic grape fruit wine；

The plant with expelling parasite insecticidal effect includes garlic, lavender, fish pelargonium, common nepenthes, shichongcao and folium artemisiae argyi In one kind or arbitrary proportion it is two or more；

The fruit juice is prepared and followed the steps below：

1) harvest：Organic orchard harvests fresh, healthy, disease-free rotten fruit；

2) cool down：The fruit of harvesting is reduced into temperature to 0~5 DEG C；

3) squeeze the juice：60~120mg/kg of sulfur dioxide concentration is added in fruit, flexibility is broken, squeezes the juice；Juice container is contained to exist Nitrogen or carbon dioxide are filled with before pan feeding, air-discharging is anti-oxidation；

4) clarify：Enzyme is added in fruit juice, enzyme addition is 20~50mg/L, and 5~8 DEG C of temperature, adds fining agent, stands 24~48h is precipitated, during which inflated with nitrogen or carbon dioxide remove oxygen；

5) low tempertaure storage：Fruit juice in step 4) is stored at 0~5 DEG C, inflated with nitrogen or carbon dioxide bind anti-oxidation；

The preparation of the grape base liquor follows the steps below：

6) collecting period determines：After grape fruit annesl, the change of the pol, acidity index of grape fruit is measured by sampling, When acidity reaches 6.0~6.5g/L, start to harvest；

7) harvest：The grape fruit of fresh, healthy, the disease-free decayed fruit of harvesting and raw green fruit in organic orchard, pol 180~ 200g/L；

8) destemming crushes：The grape of harvesting is reduced into temperature to 5~8 DEG C, flexible removing carpopodium is carried out, crushes, before crushing 60~120mg/kg of sulfur dioxide concentration is added, 20~60mg/L of enzyme is added when crushing into the container sterilized, container is in pan feeding Before be filled with nitrogen or carbon dioxide, air-discharging is anti-oxidation, obtains grape mash；

9) cold soaking stain：Grape mash is reduced into temperature to 5~8 DEG C, impregnates 12~96h, daily Sprayer Circulation 1~2 time, every time Spray the grape mash of 1/2~1/3 times of vessel volume amount；

10) extracting juice is pressed：The grape mash of cold soaking stain is pressed into extracting juice, the turbid juice of grape is obtained, during which rushes nitrogen or carbon dioxide Except oxygen；

11) low temperature clarification：Temperature is controlled to add fining agent to 5~8 DEG C, staticly settle 24~48h in the turbid juice of grape, point From grape juice, 150~250NTU of turbidity, during which inflated with nitrogen or carbon dioxide remove oxygen；

12) the fragrant fermentation of production：Grape juice is risen again to 10~12 DEG C, fragrant Non-Saccharomyces are produced in inoculation, and yeast quantity is not low In 10⁶Cfu/ml, 10~12 DEG C of fermentation temperature, 24~72h of fermentation time；

13) alcoholic fermentation：The grape juice for producing fragrant fermentation is risen again to 15~25 DEG C, is inoculated with saccharomyces cerevisiae, yeast quantity is not Less than 10⁶Cfu/ml, 16~20 DEG C of fermentation temperature, 24~72h of fermentation time；

14) wine foot is separated：The proportion and temperature of grape mash, when proportion is down to 0.993, are cooled to when monitoring alcoholic fermentation 5~10 DEG C of separation wine feet, container are filled with nitrogen or carbon dioxide air-discharging before pan feeding；

15) malo-lactic fermentation：Be inoculated with the grape wine obtained to step i) Oneococcus onei and lactic acid bacteria 20~ 60mg/L, 20~25 DEG C, 24~480h of fermentation time of fermentation temperature, completes malo-lactic fermentation；

16) store：The grape wine of malo-lactic fermentation is stored for 10~15 DEG C in temperature, period of storage 3~6 months, It is 20~30ml/L to control free sulur dioxide concentration；

17) clarify：The grape wine that step 16) is obtained is clarified using fining agent, obtains grape base liquor；

The preparation of aerating fruity Organic grape fruit wine follows the steps below：

18) allocate：Both grape base liquors of the fruit juice of step 5) and step 17) are subjected to allotment fusion, obtain fruity Portugal Grape alcoholic drink mixed with fruit juice, 2~8%vol of alcoholic strength, 50~80g/L of pol, acidity 5~6g/L, more than sugar-free extract 17g/L；

19) cold stabilization：Grape alcoholic drink mixed with fruit juice is freezed in temperature for -5~0 DEG C, cooling time 5~15 days, filtering clarification, during which It is filled with nitrogen or carbon dioxide removes oxygen；

20) it is degerming：It is degerming that 0.45 and 0.20 micron of film device is respectively adopted in the grape alcoholic drink mixed with fruit juice of cold stabilization；

21) it is filled with carbon dioxide：Degerming grape alcoholic drink mixed with fruit juice is cooled down, food-grade carbon-dioxide is filled with, it is dissolved in dioxy Pressure produced by changing carbon reaches 0.3~0.5MPa；

22) it is filling：By above-mentioned grape alcoholic drink mixed with fruit juice after the assay was approved, it is filling；

The grape is Cabernet Sauvignon and Mei Le, Cabernet Gernischt, Cabernet franc, hila, PINOT NOIR, Chardonnay, Italian Riesling and Lei Si One kind or arbitrary proportion in order it is two or more；

The enzyme is two of one kind or arbitrary proportion in pectase, cellulase, papain, alginase and amylase More than kind；

The fining agent is the two or more of one kind in gelatin, bentonite, PVPP and egg white or arbitrary proportion；

Fertilizer used in organic plantation is one kind or arbitrary proportion in organic fertilizer, organic and inorganic fertilizer and biological organic fertilizer It is two or more；

Ecological seaweed biostimulant or seaweed residue are added in the organic fertilizer, organic and inorganic fertilizer and biological organic fertilizer；

The ecology seaweed biostimulant is prepared according to following steps：

A) seaweed fragment, is prepared：Seaweed after cleaning is crushed, obtains seaweed fragment；

B) enzymolysis seaweed, is prepared：Seaweed fragment and water are fitted into agitator tank, under the conditions of 60~120 DEG C sterilize 5~ 40min, is cooled to 27~45 DEG C of access enzymes, digests 30~60min, the mass ratio 5~30 of seaweed fragment, water and enzyme:70~ 95:0.02~1, seaweed must be digested；

C) seaweed filtrate and seaweed slag, are prepared：Enzymolysis seaweed is separated, gained filtrate is seaweed filtrate, residue obtained to be Seaweed slag；

D) fermentation of seaweed thing, is prepared：By seaweed slag, urea, magnesium sulfate, sodium chloride, potassium dihydrogen phosphate and glucose according to Mass ratio 95.5:1:0.5:1:1:1 is added in fermentation tank and mixes, and water content is adjusted to water as 50~70%, at 70~150 DEG C Under the conditions of sterilize 5~40min, be cooled to 27~36 DEG C, obtain mixed culture medium, access rhodotorula mucilaginosa, rhodotorula mucilaginosa and mixing are trained The mass ratio for supporting base is 0.1~2:98~99.9, it is 27~36 DEG C in temperature, oxygen concentration is 8~21% in fermentation tank, fermentation 6~144h, sterilize 20~40min under the conditions of being 70~150 DEG C in temperature, is cooled to 20~40 DEG C, obtains fermentation of seaweed thing；

E) ecological seaweed liquid biostimulant and seaweed residue, are prepared：The seaweed filtrate that step 3) obtains is added to Stirred and evenly mixed in the fermentation of seaweed thing that step 4) obtains, obtain mixture, corynebacterium ammoniagenes prepared by access thawing method, mixture and The mass ratio of corynebacterium ammoniagenes prepared by thawing method is 95~99.8:0.2~5, be 25~35 DEG C in temperature, oxygen content for 1~ Under the conditions of 8mg/L, ferment 1~6 day, gained filtrate is ecological seaweed liquid biostimulant after press filtration；Filter residue is seaweed Residue；

F) ecological seaweed biostimulant, is prepared：Ecological seaweed liquid biostimulant it is concentrated and spray drying after, i.e., Obtain ecological seaweed biostimulant；

The seaweed is that one kind in Enteromorpha, kelp, sargassum, opotism and kelp or arbitrary proportion are two or more.

Pesticide used in the orchard is lime sulfur, Bordeaux mixture, mineral oil, trichoderma harzianum, quinolinone suspending agent, second Alcohol, soda, China tree alkali, rotenone, Dalmatian chrysanthemum missible oil and one kind in matrine or arbitrary proportion are two or more.

The preparation of thawing method corynebacterium ammoniagenes follows the steps below：

I) expansion of strain is further cultured for：Using the thalline in exponential phase as seed, it is inoculated with 10% inoculum concentration In fluid nutrient medium, 48h is cultivated in 30 DEG C of shaking table 160r/min, after 8000r/min centrifuges 10min, expansion is obtained and is further cultured for Strain；

II) thawing method improves the permeability of cell：Thawing method is at least repeated once, its step is：Step 1) is obtained Expansion is further cultured for strain and is put into -20 DEG C of refrigerator, freezes 4h, takes out, dissolves at room temperature；

PH7.2~7.4 of the fluid nutrient medium, and be made into according to following mass ratio：Beef infusion broth 1.0L, peptone 10.0g, NaCl 5.0g, 20.0~25.0g of agar；

The preparation method of wherein beef infusion broth follows the steps below：Lean beef is cleaned, and chopping, weighs 550 grams of immersions In 1375ml water, soak a night, filtering, 0.6kg/cm after filtrate packing²Sterilize 40min.

The enzyme is lysozyme and pectase according to mass ratio 5:1 composition, wherein lysozyme according to following steps into OK：

(A) add the water of 5 times of weight to egg white or whole egg liquid, after stirring evenly add homogenizer in, at least it is homogeneous once, it is even Matter temperature is 30~60 DEG C, and homogeneous pressure is 2~40MPa, and the homogeneous time is 10~40min, is filtered after homogeneous, gained filtrate is Dilution；

(B) the dilution molecular cut off for obtaining step (A) is the ultrafiltration membrane ultrafiltration of 15000~30000Da, is obtained Thin liquid and dope；

(C) add resin in the thin liquid that step (B) obtains to be adsorbed, the part by weight of the thin liquid and resin is 1: 3~8, the resin after being adsorbed；

(D) resin after the absorption for obtaining step (C) is eluted with the NaCl aqueous solutions of 0.001~0.5mol/L, Obtain eluent；

(E) by the eluent that step (D) obtains ultrafiltration membrane ultrafiltration desalination, ultrafiltrate, the retention of the ultrafiltration membrane are obtained Molecular weight is 2000~10000Da；

(F) after the ultrafiltrate drying obtained step (E), that is, lysozyme is obtained；

The resin is that the resin is cation exchange resin or macroporous absorbent resin；The model of cation exchange resin For 001*7,732, AmberliteIR-120, Dowex-50, Lewatit-100, Diaion SK-1, AllassionCS, One or more in Duolite C-20, SDB-3, macroporous absorbent resin AB-8, D101, D3520, X-5, NKA-II, One or more in NKA-9, S-8, XDA-1, H-20, H-30, H-40.

Step c) the separation is included more than one or both of press filtration, filtering and centrifugation.

Step d) the mixed culture mediums to water content is 60%, and the mass ratio of rhodotorula mucilaginosa and mixed culture medium is 0.5: 99.5, it is 30 DEG C in temperature, oxygen concentration 12%, ferment 120h, and sterilize 20min under the conditions of being 120 DEG C in temperature.

The mass ratio of corynebacterium ammoniagenes prepared by step e) mixtures and thawing method is 98:2, it is 28 DEG C in temperature, it is oxygen-containing Measure under the conditions of 3mg/L, to ferment 5 days.

The rhodotorula mucilaginosa and corynebacterium ammoniagenes are purchased from Chinese industrial Microbiological Culture Collection administrative center, numbering difference For CICC31192 and CICC 10168.

Step A) described in it is homogeneous be homogeneous 2 times, homogeneous temperature be 40 DEG C, the homogeneous time be 15min, wherein first time it is even Matter pressure is 25MPa, and second of homogeneous pressure is 4MPa.

The mode that ecological seaweed biostimulant is added in fertilizer is：With the interior addition manner of fertilizer material mixing granulation With the outer addition manner for spraying to fertiliser granulates surface.

Invention has following advantageous effects：

1st, the present invention uses organic fruits and Organic grape as raw material, and the wine set that it is prepared is nutritious, health, peace Entirely, the advantages of non agricultural chemical residuum.

2nd, grape base liquor technique of the present invention is unique, grape wine is had strong, pure and fresh fragrance, and mouthfeel is more compliant, and Nutriment retains abundant；First using color and benefit materials in low temperature cold impregnation technology controlling extraction pericarp and fruit seed； Then squeezing goes pericarp to carry out alcoholic fermentation using clarification grape juice, in the process order inoculation Non-Saccharomyces and wine brewing ferment Mother fermentation generation aromatic substance and alcohol etc., increase the fragrance complexity of grape base liquor and strong degree, and pass through cold fermentation, storage Hiding technique prevents the volatilization loss of aroma substance, increases the sophistication in wine；It is anti-by whole inflated with nitrogen or carbon dioxide etc. Oxidation technology, increases the freshness and nutriment activity of grape base liquor to greatest extent；The present invention need not add preservative substance.

3rd, the present invention can reach expelling parasite and reduce pesticide by interplanting the plant with expelling parasite insecticidal effect in orchard The purpose of usage amount.

4th, ecological seaweed biostimulant uses new fresh seaweed in the present invention, and life can largely be stimulated by containing in new fresh seaweed Long stimulin and resistance material, activity is good, can achieve the purpose that to reduce Fertilizer application amount.

5th, ecological seaweed biostimulant is fermented under anaerobic using aerobic and anaerobism rhodotorula mucilaginosa in the present invention, It can prevent the loss of seaweed nutritive component, and the alcohol that anaerobic fermentation produces is a supporting role seaweed extraction.

6th, ecological seaweed biostimulant is sent out under anoxic conditions using aerobic and anaerobism corynebacterium ammoniagenes in the present invention Ferment, produces ATP, and ATP is a kind of high energy phosphate compound, and in cell, it can realize energy storage and put with mutually converting for ADP Can, so as to ensure that the energy supply of cell items vital movement, promote plant growth.It is unreasonable due to long-term fertilization, make Into soil acidification, corynebacterium ammoniagenes can reduce N0₃ ^-And urea, producing ammonia makes tunning be in alkalescent, can improve acidifying soil Earth problem.

7th, ecological seaweed biostimulant has heavy metal certain fixation in the present invention.Contain in fermentate of the present invention There are polypeptide, alginic acid and other protides, heavy metal can make albumen, alginic acid and polypeptide inactivation, while heavy metal also can Fixed by albumen, alginic acid and polypeptide, so as to prevent crop from absorbing.

8th, the present invention is using aerobic and oxygen bacterium, and soil hardening is serious at present, and the microorganism applied in ground is often in anoxic shape State, present soil environment is suitable for using aerobic and oxygen bacterium.

9th, ecological seaweed biostimulant improves the permeability of corynebacterium ammoniagenes cell by thawing method in the present invention, can be with The ATP that corynebacterium ammoniagenes produce is set to be easier to penetrate into from cell membrane extracellularly, its product effects is significantly better than with without place The product of the corynebacterium ammoniagenes of reason.

Embodiment

The present invention is further illustrated with reference to instantiation.

Embodiment 1

By fish pelargonium interplanting in orchard, orchard is managed according to organic planting requirement, mature fruit in orchard is picked and prepares Fruit juice, picks ripe grape in orchard and carries out vintage wine, by fruit juice and grape wine according to mass ratio 5:95 allotments, up to organic Grape fruit wine；

The fruit juice is prepared and followed the steps below：

2) cool down：The fruit of harvesting is reduced into temperature to 3 DEG C；

3) squeeze the juice：Sulfur dioxide concentration 45mg/kg is added in fruit, flexibility is broken, squeezes the juice；Juice container is contained before pan feeding Nitrogen is filled with, air-discharging is anti-oxidation；

4) clarify：Enzyme is added in fruit juice, enzyme addition is 40mg/L, and 6 DEG C of temperature, adds fining agent, staticly settle 24h, during which inflated with nitrogen removal oxygen；

5) low tempertaure storage：Fruit juice is stored at 2 DEG C, inflated with nitrogen binds anti-oxidation；

The preparation of the grape base liquor follows the steps below：

8) destemming crushes：The grape of harvesting is reduced into temperature to 5~8 DEG C, flexible removing carpopodium is carried out, crushes, before crushing Sulfur dioxide concentration 80mg/kg is added, enzyme 40mg/L is added when crushing into the container sterilized, container is filled with nitrogen before pan feeding Gas, air-discharging is anti-oxidation, obtains grape mash；

9) cold soaking stain：Grape mash is reduced into temperature to 5~8 DEG C, impregnates 48h, daily Sprayer Circulation 2 times, sprays 1/2 every time The grape mash of times vessel volume amount；

10) extracting juice is pressed：The grape mash of cold soaking stain is pressed into extracting juice, the turbid juice of grape is obtained, during which rushes nitrogen and remove oxygen；

11) low temperature clarification：Control temperature to add fining agent to 5~8 DEG C in the turbid juice of grape, staticly settle 24h, separate Portugal Grape juice, turbidity 200NTU, during which inflated with nitrogen removal oxygen；

12) the fragrant fermentation of production：Grape juice is risen again to 10~12 DEG C, the fragrant Non-Saccharomyces of inoculation production, yeast quantity is 2* 10⁶Cfu/ml, 10~12 DEG C of fermentation temperature, fermentation time 36h；

13) alcoholic fermentation：The grape juice for producing fragrant fermentation is risen again to 16~20 DEG C, is inoculated with saccharomyces cerevisiae, yeast quantity is 3*10⁶Cfu/ml, 16~20 DEG C of fermentation temperature, fermentation time 30h；

14) wine foot is separated：The proportion and temperature of grape mash, when proportion is down to 0.993, are cooled to when monitoring alcoholic fermentation 5~10 DEG C of separation wine feet, container are filled with nitrogen purge gas before pan feeding；

15) malo-lactic fermentation：Oneococcus onei and lactic acid bacteria 20mg/ are inoculated with the grape wine obtained to step i) L, 20~25 DEG C, fermentation time 300h of fermentation temperature, completes malo-lactic fermentation；

16) store：By the grape wine of malo-lactic fermentation in 10~15 DEG C of storages of temperature, period of storage 5 months, control Free sulur dioxide concentration processed is 20ml/L；

18) allocate：Both grape base liquors of the fruit juice of step 5) and step 17) are subjected to allotment fusion, obtain fruity Portugal Grape alcoholic drink mixed with fruit juice, alcoholic strength 4%vol, pol 60g/L, 5~6g/L of acidity, sugar-free extract 18g/L；

19) cold stabilization：Grape alcoholic drink mixed with fruit juice is freezed in temperature for -5~0 DEG C, cooling time 10 days, filtering clarification, during which fills Enter nitrogen and remove oxygen；

21) it is filled with carbon dioxide：Degerming grape alcoholic drink mixed with fruit juice is cooled down, food-grade carbon-dioxide is filled with, it is dissolved in dioxy Pressure produced by changing carbon reaches 0.4MPa；

The fruit is strawberry；

The grape is PINOT NOIR；

The enzyme is pectase and cellulase according to mass ratio 1:2 composition；

The fining agent is gelatin.

The orchard uses the organic fertilizer for adding ecological seaweed biostimulant；

The ecology seaweed biostimulant is prepared according to following steps：

A) seaweed fragment, is prepared：Fresh Laminaria Japonica after cleaning is crushed, obtains seaweed fragment；

B) enzymolysis seaweed, is prepared：Seaweed fragment and water are fitted into agitator tank, sterilize 15min under the conditions of 100 DEG C, 30 DEG C of access enzymes are cooled to, digest 40min, the mass ratio 20 of seaweed fragment, water and enzyme:79.94:0.06, seaweed must be digested, The enzyme is lysozyme and pectase according to mass ratio 5:1 composition；

C) seaweed filtrate and seaweed slag, are prepared：The press filtration of enzymolysis seaweed is separated, gained filtrate is seaweed filtrate, and gained is residual Slag is seaweed slag；

D) fermentation of seaweed thing, is prepared：By seaweed slag, urea, magnesium sulfate, sodium chloride, potassium dihydrogen phosphate and glucose according to Mass ratio 95.5:1:0.5:1:1:1 is added in fermentation tank and mixes, and is adjusted to water content as 60% with water, goes out under the conditions of 120 DEG C Bacterium 20min, is cooled to 30 DEG C, obtains mixed culture medium, accesses rhodotorula mucilaginosa, and the mass ratio of rhodotorula mucilaginosa and mixed culture medium is 0.5:99.5, it is 30 DEG C in temperature, oxygen concentration is 12% in fermentation tank, and ferment 120h, is sterilized under the conditions of being 120 DEG C in temperature 20min, is cooled to 35 DEG C, obtains fermentation of seaweed thing；

E) ecological seaweed liquid biostimulant and seaweed residue, are prepared：The seaweed filtrate that step 3) obtains is added to Stirred and evenly mixed in the fermentation of seaweed thing that step 4) obtains, obtain mixture, corynebacterium ammoniagenes prepared by access thawing method, mixture and The mass ratio of corynebacterium ammoniagenes prepared by thawing method is 98:2, it is 28 DEG C in temperature, under the conditions of oxygen content is 3mg/L, fermentation 5 My god, press filtration, gained filtrate is ecological seaweed liquid biostimulant；Filter residue is seaweed residue；

The preparation of wherein thawing method corynebacterium ammoniagenes follows the steps below：

II) thawing method improves the permeability of cell：In triplicate, its step is thawing method：The expansion that step 1) is obtained It is further cultured for strain to be put into -20 DEG C of refrigerator, freezes 4h, takes out, dissolve, be placed again into -20 DEG C of refrigerator at room temperature, it is cold Freeze 4h, take out, dissolve, be placed again into -20 DEG C of refrigerator at room temperature, freeze 4h, take out, dissolve at room temperature；

PH7.2~7.4 of the fluid nutrient medium, and be made into according to following mass ratio：Beef infusion broth 1.0L, peptone 10.0g, NaCl 5.0g, agar 25.0g；

Wherein lysozyme follows the steps below：

(A) add the water of 5 times of weight to egg white or whole egg liquid, added after stirring evenly in homogenizer, homogeneous 2 times, homogeneous temperature Degree is 40 DEG C, and the homogeneous time is 15min, and wherein first time homogeneous pressure is 25MPa, and second of homogeneous pressure is 4MPa, Filtering after homogeneous, gained filtrate is dilution；

(B) the ultrafiltration membrane ultrafiltration for being 28000Da with molecular cut off by the dilution that step (A) obtains, obtain thin liquid and Dope；

(C) add resin in the thin liquid that step (B) obtains to be adsorbed, the part by weight of the thin liquid and resin is 1: 5, the resin after being adsorbed；

(D) resin after the absorption for obtaining step (C) is eluted with the NaCl aqueous solutions of 0.003mol/L, is washed De- liquid；

(E) by the eluent that step (D) obtains ultrafiltration membrane ultrafiltration desalination, ultrafiltrate, the retention of the ultrafiltration membrane are obtained Molecular weight is 8000Da；

(F) after the ultrafiltrate drying obtained step (E), that is, lysozyme is obtained.

Embodiment 2

By fish pelargonium interplanting in orchard, orchard is managed according to organic planting requirement, mature fruit in orchard is picked and prepares Fruit juice, picks ripe grape in orchard and carries out vintage wine, by fruit juice and grape wine according to mass ratio 10:90 allotments, have to obtain the final product Machine grape fruit wine；

The fruit juice is prepared and followed the steps below：

3) squeeze the juice：Sulfur dioxide concentration 80mg/kg is added in fruit, flexibility is broken, squeezes the juice；Juice container is contained before pan feeding Nitrogen is filled with, air-discharging is anti-oxidation；

The preparation of the grape base liquor follows the steps below：

The fruit is strawberry；

The grape is PINOT NOIR；

The enzyme is pectase；

The fining agent is gelatin.

The ecology seaweed biostimulant is prepared according to following steps：

Wherein lysozyme follows the steps below：

(A) add the water of 5 times of weight to egg white or whole egg liquid, added after stirring evenly in homogenizer, homogeneous 2 times, homogeneous temperature Degree is 35 DEG C, and the homogeneous time is 15min, and wherein first time homogeneous pressure is 25MPa, and second of homogeneous pressure is 4MPa, Filtering after homogeneous, gained filtrate is dilution；

Embodiment 3

By fish pelargonium interplanting in orchard, orchard is managed according to organic planting requirement, mature fruit in orchard is picked and prepares Fruit juice, picks ripe grape in orchard and carries out vintage wine, by fruit juice and grape wine according to mass ratio 15:85 allotments, have to obtain the final product Machine grape fruit wine；

The fruit juice is prepared and followed the steps below：

The preparation of the grape base liquor follows the steps below：

11) low temperature clarification：Control temperature to add fining agent to 5~8 DEG C in the turbid juice of grape, staticly settle 24h, separate Portugal Grape juice, turbidity 190NTU, during which inflated with nitrogen removal oxygen；

18) allocate：Both grape base liquors of the fruit juice of step 5) and step 17) are subjected to allotment fusion, obtain fruity Portugal Grape alcoholic drink mixed with fruit juice, alcoholic strength 6%vol, pol 70g/L, acidity 5g/L, sugar-free extract 20g/L；

The fruit is cherry；

The grape is PINOT NOIR and Riesling according to mass ratio 2:1 composition；

The enzyme is pectase and cellulase according to mass ratio 3:1 composition；

The fining agent is gelatin and PVPP according to mass ratio 3:1 composition.

The organic fertilizer that the orchard is prepared using seaweed residue；

The seaweed residue is prepared according to following steps：

Wherein lysozyme follows the steps below：

(A) add the water of 5 times of weight to egg white or whole egg liquid, added after stirring evenly in homogenizer, homogeneous 2 times, homogeneous temperature Degree is 45 DEG C, and the homogeneous time is 15min, and wherein first time homogeneous pressure is 25MPa, and second of homogeneous pressure is 4MPa, Filtering after homogeneous, gained filtrate is dilution；

Embodiment 4

By fish pelargonium interplanting in orchard, orchard is managed according to organic planting requirement, mature fruit in orchard is picked and prepares Fruit juice, picks ripe grape in orchard and carries out vintage wine, by fruit juice and grape wine according to mass ratio 20:80 allotments, have to obtain the final product Machine grape fruit wine；

The fruit juice is prepared and followed the steps below：

The preparation of the grape base liquor follows the steps below：

The fruit is cherry；

The grape is PINOT NOIR and Riesling according to mass ratio 2:1 composition；

The enzyme is pectase and cellulase according to mass ratio 3:1 composition；

The fining agent is gelatin and PVPP according to mass ratio 1:1 composition.

The organic fertilizer that the orchard is prepared using seaweed residue；

The seaweed residue is prepared according to following steps：

Wherein lysozyme follows the steps below：

(A) add the water of 5 times of weight to egg white or whole egg liquid, added after stirring evenly in homogenizer, homogeneous 1 time, homogeneous temperature Degree is 40 DEG C, and the homogeneous time is 30min, and homogeneous pressure 25MPa, is filtered, gained filtrate is dilution after homogeneous；

Embodiment 5

By fish pelargonium interplanting in orchard, orchard is managed according to organic planting requirement, mature fruit in orchard is picked and prepares Fruit juice, picks ripe grape in orchard and carries out vintage wine, by fruit juice and grape wine according to mass ratio 25:75 allotments, have to obtain the final product Machine grape fruit wine；

The fruit juice is prepared and followed the steps below：

The preparation of the grape base liquor follows the steps below：

The fruit is strawberry；

The grape is PINOT NOIR；

The enzyme is pectase and cellulase according to mass ratio 1:2 composition；

The fining agent is gelatin.

The ecology seaweed biostimulant is prepared according to following steps：

Wherein lysozyme follows the steps below：

(A) add the water of 5 times of weight to egg white or whole egg liquid, added after stirring evenly in homogenizer, homogeneous 3 times, homogeneous temperature Degree is 40 DEG C, and the homogeneous time is 15min, and wherein first time homogeneous pressure is 25MPa, and second of homogeneous pressure is 4MPa, Third time homogeneous pressure is 4MPa, is filtered after homogeneous, gained filtrate is dilution；

Embodiment 6

The fruit juice is prepared and followed the steps below：

The preparation of the grape base liquor follows the steps below：

The fruit is strawberry；

The grape is PINOT NOIR；

The enzyme is pectase and cellulase according to mass ratio 1:2 composition；

The fining agent is gelatin.

The ecology seaweed biostimulant is prepared according to following steps：

A) seaweed fragment, is prepared：Fresh Laminaria Japonica after cleaning is dried, dry kelp is crushed using pulverizer, is obtained Seaweed fragment；

Wherein lysozyme follows the steps below：

Beneficial effects of the present invention are further illustrated with reference to experimental data：

Experiment one

1st, material to be tested is tested

1 materials and methods：

1.1 test sites and experimental subjects：Yantai university chemistry analysis inspection center.

1.2 experiment detections：Survey the activity and the rate of recovery of lysozyme.

1.3 material to be tested：It is (even without homogenizer after the preparation method of lysozyme is except stirring evenly for common process Matter, other production methods are consistent with embodiment 1), embodiment 1, embodiment 2, embodiment 3, prepare in embodiment 4 and embodiment 5 Lysozyme do effect and compare.

1.4 experimental design：Using high performance liquid chromatography to sample treatment.

This experiment is consistent with other operations in addition to processing difference except testing.

2 results and analysis

Common process (homogeneous without homogenizer after stirring evenly, other production methods are consistent with embodiment 1), implement Bacteriolyze enzymatic treatment prepared by example 1, embodiment 2, embodiment 3, embodiment 4 and embodiment 5 is compared, and data are shown in Table 1

Table 1

Compared by 1 embodiment 1 of table with common process, the present invention is either in terms of the lysozyme rate of recovery, or enzymatic activity All it is significantly better than common process；Compared by embodiment 1 to embodiment 3 it can be seen that the rate of recovery and work of the homogeneous temperature to lysozyme Property still have certain influence, wherein embodiment 1 is best using 40 DEG C of general effects；Compared by embodiment 1, embodiment 4 and embodiment 5 Relatively it can be seen that homogeneous pressure and number also have certain influence to the rate of recovery and activity of lysozyme, and non-pressure is bigger, the time It is more long better, 1 best results of Integrated comparative embodiment.

Experiment two

1st, material to be tested is tested

1 materials and methods：

1.1 test site：Laboratory.

1.2 material to be tested：The water in fruit and grape and the organic orchard of the present invention in common orchard (non-organic orchard) Fruit and grape, compare.

This experiment is consistent with other management in addition to processing difference except testing.

1.3 experimental design：The fruit in common orchard (non-organic orchard) and grape and the organic fruit of the present invention are picked respectively Fruit and grape in garden make grape fruit wine according to the method for embodiment 2, detect the NO in grape fruit wine₂ ^-And pesticide residue.

2 results and analysis

	NO in grape fruit wine₂ ^-(mg/kg)	Pesticide residue (mg/kg)
			Common orchard grape fruit wine	0.7	0.8
The organic orchard grape fruit wine of the present invention	0	0

As can be seen from the above data, the grape fruit wine made of the present invention and common glucose fruit wine in terms of food health more Tool advantage.

Experiment three

1st, material to be tested is tested

1 materials and methods：

1.1 test site：Yantai Qixia City Su Jiadian towns vineyard.

1.2 material to be tested：Common orchard (in addition to not there is no the plant of expelling parasite insecticidal effect, other production methods and this hair It is bright consistent), contrast 1 is (when preparing ecological seaweed liquid biostimulant, except corynebacterium ammoniagenes are without thawing method processing directly use Outside, other production methods are consistent with the present invention) and embodiment 1, compare.

1.3 experimental design：The consistent vineyard of 6 mu of age of trees is selected, 2 is parallel, and 1 mu is 1 processing unit.For common orchard (in addition to the plant with expelling parasite insecticidal effect is not interplanted, other production methods are consistent with the present invention), contrast 1 (prepare ecological sea During algae solution body biostimulant, except corynebacterium ammoniagenes are without the processing of thawing method directly in addition to use, other production methods with this hair It is bright consistent) and embodiment 1 contrast, observe orchard insect pest situation, statistics orchard vintage.

2 results and analysis

	Grape average product (kg)	Orchard insect pest situation
			Common orchard	2028.6	Insect pest is serious
Contrast 1	2371.4	Substantially without worm
			The present invention	2611.5	Substantially without worm

Compared by common orchard with the present invention, it can be seen that the present invention has obvious anthelminthic effect, using the teaching of the invention it is possible to provide grape Yield；Being compared by embodiment 1 and contrast 1 can show that corynebacterium ammoniagenes through thawing method high treating effect, can improve vintage.

" managing orchard according to organic planting requirement " of the present invention refers to according to GB/T19630《Organic products》Mark Quasi- management orchard.

Claims

1. a kind of production method of Organic grape fruit wine, it is characterised in that follow the steps below：

By the plants interplanting with expelling parasite insecticidal effect in orchard, orchard is managed according to organic planting requirement, is picked in orchard Mature fruit prepares fruit juice, picks ripe grape in orchard and carries out vintage wine, by fruit juice and grape wine according to mass ratio 1~ 75:25~99 allotments, up to Organic grape fruit wine；

The plant with expelling parasite insecticidal effect is included in garlic, lavender, fish pelargonium, common nepenthes, shichongcao and folium artemisiae argyi A kind of or arbitrary proportion is two or more；

The fruit juice is prepared and followed the steps below：

1）Harvesting：Organic orchard harvests fresh, healthy, disease-free rotten fruit；

2）Cooling：The fruit of harvesting is reduced into temperature to 0~5 DEG C；

3）Squeeze the juice：60~120mg/kg of sulfur dioxide concentration is added in fruit, flexibility is broken, squeezes the juice；Juice container is contained in pan feeding Before be filled with nitrogen or carbon dioxide, air-discharging is anti-oxidation；

4）Clarification：Enzyme is added in fruit juice, enzyme addition is 20~50mg/L, and 5~8 DEG C of temperature, adds fining agent, staticly settle 24~48h, during which inflated with nitrogen or carbon dioxide remove oxygen；

5）Low tempertaure storage：By step 4）In fruit juice stored at 0~5 DEG C, inflated with nitrogen or carbon dioxide bind anti-oxidation；

The preparation of the grape base liquor follows the steps below：

6）Collecting period determines：After grape fruit annesl, the change of the pol, acidity index of grape fruit is measured by sampling, works as acid Degree reaches 6.0~6.5g/L, starts to harvest；

7）Harvesting：The grape fruit of fresh, healthy, disease-free decayed fruit and raw green fruit, 180~200g/ of pol are harvested in organic orchard L；

8）Destemming crushes：The grape of harvesting is reduced into temperature to 5~8 DEG C, carries out flexible removing carpopodium, broken, addition before crushing 60~120mg/kg of sulfur dioxide concentration, adds 20~60mg/L of enzyme when crushing into the container sterilized, container is filled before pan feeding Enter nitrogen or carbon dioxide, air-discharging is anti-oxidation, obtains grape mash；

9）Cold soaking stain：Grape mash is reduced into temperature to 5~8 DEG C, impregnates 12~96h, daily Sprayer Circulation 1~2 time, spray every time The grape mash of 1/2~1/3 times of vessel volume amount；

10）Press extracting juice：The grape mash of cold soaking stain is pressed into extracting juice, the turbid juice of grape is obtained, during which rushes nitrogen or carbon dioxide deoxygenation Gas；

11）Low temperature clarification：Control temperature to add fining agent to 5~8 DEG C in the turbid juice of grape, staticly settle 24~48h, separate Portugal Grape juice, 150~250NTU of turbidity, during which inflated with nitrogen or carbon dioxide remove oxygen；

12）The fragrant fermentation of production：Grape juice is risen again to 10~12 DEG C, the fragrant Non-Saccharomyces of inoculation production, yeast quantity is not less than 10⁶Cfu/ml, 10~12 DEG C of fermentation temperature, 24~72h of fermentation time；

13）Alcoholic fermentation：The grape juice for producing fragrant fermentation is risen again to 15~25 DEG C, is inoculated with saccharomyces cerevisiae, yeast quantity is not less than 10⁶Cfu/ml, 16~20 DEG C of fermentation temperature, 24~72h of fermentation time；

14）Separate wine foot：The proportion and temperature of grape mash when monitoring alcoholic fermentation, when proportion is down to 0.993, it is cooled to 5~ 10 DEG C of separation wine feet, container are filled with nitrogen or carbon dioxide air-discharging before pan feeding；

15）Malo-lactic fermentation：To step i）20~60mg/ of Oneococcus onei and lactic acid bacteria is inoculated with obtained grape wine L, 20~25 DEG C, 24~480h of fermentation time of fermentation temperature, completes malo-lactic fermentation；

16）Storage：By the grape wine of malo-lactic fermentation in 10~15 DEG C of storages of temperature, period of storage 3~6 months, control Free sulur dioxide concentration is 20~30ml/L；

17）Clarification：By step 16）Obtained grape wine is clarified using fining agent, obtains grape base liquor；

18）Allotment：By step 5）Fruit juice and step 17）Both grape base liquors carry out allotment fusion, obtain fruity grape syrup Wine, 2~8%vol of alcoholic strength, 50~80g/L of pol, acidity 5~6g/L, more than sugar-free extract 17g/L；

19）Cold stabilization：Grape alcoholic drink mixed with fruit juice is freezed in temperature for -5~0 DEG C, cooling time 5~15 days, filtering clarification, is during which filled with Nitrogen or carbon dioxide remove oxygen；

20）It is degerming：It is degerming that 0.45 and 0.20 micron of film device is respectively adopted in the grape alcoholic drink mixed with fruit juice of cold stabilization；

21）It is filled with carbon dioxide：Degerming grape alcoholic drink mixed with fruit juice is cooled down, food-grade carbon-dioxide is filled with, it is dissolved in carbon dioxide Produced pressure reaches 0.3~0.5MPa；

22）It is filling：By above-mentioned grape alcoholic drink mixed with fruit juice after the assay was approved, it is filling；

The grape is in Cabernet Sauvignon and Mei Le, Cabernet Gernischt, Cabernet franc, hila, PINOT NOIR, Chardonnay, Italian Riesling and Riesling One kind or arbitrary proportion it is two or more；

The enzyme be two kinds of one kind or arbitrary proportion in pectase, cellulase, papain, alginase and amylase with On；

Fertilizer used in organic plantation is two kinds of one kind or arbitrary proportion in organic fertilizer, organic and inorganic fertilizer and biological organic fertilizer More than;

Ecological seaweed biostimulant or seaweed residue are added in the organic fertilizer, organic and inorganic fertilizer and biological organic fertilizer;

The ecology seaweed biostimulant is prepared according to following steps：

a）, prepare seaweed fragment：Seaweed after cleaning is crushed, obtains seaweed fragment；

b）, prepare enzymolysis seaweed：Seaweed fragment and water are fitted into agitator tank, under the conditions of 60~120 DEG C sterilize 5~ 40min, is cooled to 27~45 DEG C of access enzymes, digests 30~60min, the mass ratio 5~30 of seaweed fragment, water and enzyme:70~ 95:0.02~1, seaweed must be digested；

c）, prepare seaweed filtrate and seaweed slag：Enzymolysis seaweed is separated, gained filtrate is seaweed filtrate, and residue obtained is seaweed Slag；

d）, prepare fermentation of seaweed thing：By seaweed slag, urea, magnesium sulfate, sodium chloride, potassium dihydrogen phosphate and glucose according to quality Than 95.5:1:0.5:1:1:1 is added in fermentation tank and mixes, and water content is adjusted to water as 50~70%, in 70~150 DEG C of conditions 5~40min of lower sterilizing, is cooled to 27~36 DEG C, obtains mixed culture medium, accesses rhodotorula mucilaginosa, rhodotorula mucilaginosa and mixed culture medium Mass ratio be 0.1~2:98~99.9, it is 27~36 DEG C in temperature, oxygen concentration is 8~21% in fermentation tank, fermentation 6~ 144h, sterilize 20~40min under the conditions of being 70~150 DEG C in temperature, is cooled to 20~40 DEG C, obtains fermentation of seaweed thing；

e）, prepare ecological seaweed liquid biostimulant and seaweed residue：By step 3）Obtained seaweed filtrate is added to step 4）Stirred and evenly mixed in obtained fermentation of seaweed thing, obtain mixture, corynebacterium ammoniagenes prepared by access thawing method, mixture and thawing The mass ratio of corynebacterium ammoniagenes prepared by method is 95~99.8:0.2~5, it is 25~35 DEG C in temperature, oxygen content is 1~8mg/L Under the conditions of, ferment 1~6 day, gained filtrate is ecological seaweed liquid biostimulant after press filtration；Filter residue is seaweed residue；

f）, prepare ecological seaweed biostimulant：Ecological seaweed liquid biostimulant is concentrated with after spray drying, gives birth to obtain the final product State seaweed bio stimulin；

2. the production method of Organic grape fruit wine as claimed in claim 1, it is characterised in that：Pesticide used in the orchard is stone sulphur Mixture, Bordeaux mixture, mineral oil, trichoderma harzianum, quinolinone suspending agent, ethanol, soda, China tree alkali, rotenone, Dalmatian chrysanthemum breast One kind or arbitrary proportion in oil and matrine is two or more.

3. the production method of Organic grape fruit wine as claimed in claim 1, it is characterised in that：The system of thawing method corynebacterium ammoniagenes It is standby to follow the steps below：

Ⅰ）The expansion of strain is further cultured for：Using the thalline in exponential phase as seed, liquid is inoculated in 10% inoculum concentration In culture medium, 48h is cultivated in 30 DEG C of shaking table 160r/min, after 8000r/min centrifuges 10min, expansion is obtained and is further cultured for strain；

Ⅱ）Thawing method improves the permeability of cell：Thawing method is at least repeated once, its step is：By step 1）Obtained expansion It is further cultured for strain to be put into -20 DEG C of refrigerator, freezes 4h, takes out, dissolve at room temperature；

PH7.2~7.4 of the fluid nutrient medium, and be made into according to following mass ratio：Beef infusion broth 1.0L, peptone 10.0g, NaCl 5.0g, 20.0~25.0g of agar;

The preparation method of wherein beef infusion broth follows the steps below：Lean beef is cleaned, and chopping, weighs 550 grams and be soaked in In 1375ml water, soak a night, filtering, 0.6kg/cm after filtrate packing²Sterilize 40min.

4. the production method of Organic grape fruit wine as claimed in claim 1, it is characterised in that：The enzyme is lysozyme and pectin Enzyme is according to mass ratio 5:1 composition, wherein lysozyme follow the steps below：

(A) add the water of 5 times of weight to egg white or whole egg liquid, after stirring evenly add homogenizer in, at least it is homogeneous once, homogeneous temperature Spend for 30~60 DEG C, homogeneous pressure is 2~40MPa, and the homogeneous time is 10~40min, is filtered after homogeneous, and gained filtrate is dilution Liquid；

(B) the dilution molecular cut off for obtaining step (A) is the ultrafiltration membrane ultrafiltration of 15000~30000Da, obtains thin liquid And dope；

(C) add resin in the thin liquid that step (B) obtains to be adsorbed, the part by weight of the thin liquid and resin is 1:3~ 8, the resin after being adsorbed；

(D) resin after the absorption for obtaining step (C) is eluted with the NaCl aqueous solutions of 0.001~0.5mol/L, is obtained Wash

De- liquid；

(E) by the eluent that step (D) obtains ultrafiltration membrane ultrafiltration desalination, ultrafiltrate, the retention molecule of the ultrafiltration membrane are obtained Measure as 2000~10000Da；

The resin is that the resin is cation exchange resin or macroporous absorbent resin；The model of cation exchange resin 001*7、732、AmberliteIR-120、Dowex-50、Lewatit-100、Diaion SK-1、AllassionCS、 One or more in Duolite C-20, SDB-3, macroporous absorbent resin AB-8, D101, D3520, X-5, NKA-II, One or more in NKA-9, S-8, XDA-1, H-20, H-30, H-40.

5. the production method of Organic grape fruit wine as claimed in claim 1, it is characterised in that：Step c）The separation includes pressure It is more than one or both of filter, filtering and centrifugation.

6. the production method of Organic grape fruit wine as claimed in claim 1, it is characterised in that：Step d）The mixed culture medium It is 60% to water content, the mass ratio of rhodotorula mucilaginosa and mixed culture medium is 0.5:99.5, it is 30 DEG C in temperature, oxygen concentration is 12%, ferment 120h, and sterilize 20min under the conditions of being 120 DEG C in temperature.

7. the production method of Organic grape fruit wine as claimed in claim 1, it is characterised in that：Step e）Mixture and thawing method The mass ratio of the corynebacterium ammoniagenes of preparation is 98:2, it is 28 DEG C in temperature, under the conditions of oxygen content is 3mg/L, ferments 5 days.

8. the production method of Organic grape fruit wine as claimed in claim 1, it is characterised in that：The rhodotorula mucilaginosa and production ammonia rod Bacillus is purchased from Chinese industrial Microbiological Culture Collection administrative center, and numbering is respectively CICC 31192 and CICC 10168.

9. the production method of Organic grape fruit wine as claimed in claim 4, it is characterised in that：Step A）Described in it is homogeneous to be even Matter 2 times, homogeneous temperature are 40 DEG C, and the homogeneous time is 15min, and wherein first time homogeneous pressure is 25MPa, second of homogeneous pressure For 4MPa.

10. the production method of Organic grape fruit wine as claimed in claim 1, it is characterised in that：Ecological seaweed biostimulant The mode being added in fertilizer is：With the interior addition manner of fertilizer material mixing granulation and spraying to the outer of fertiliser granulates surface and adding Add mode.