CN106722943A - A kind of apple ferment and preparation method thereof - Google Patents
A kind of apple ferment and preparation method thereof Download PDFInfo
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- CN106722943A CN106722943A CN201611109027.XA CN201611109027A CN106722943A CN 106722943 A CN106722943 A CN 106722943A CN 201611109027 A CN201611109027 A CN 201611109027A CN 106722943 A CN106722943 A CN 106722943A
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- zymotic fluid
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- 238000000855 fermentation Methods 0.000 title claims description 47
- 238000002360 preparation method Methods 0.000 title claims description 30
- 239000007788 liquid Substances 0.000 claims abstract description 45
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 42
- 239000012530 fluid Substances 0.000 claims abstract description 36
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 claims abstract description 30
- 241000235058 Komagataella pastoris Species 0.000 claims abstract description 29
- 239000000203 mixture Substances 0.000 claims abstract description 28
- 230000001954 sterilising effect Effects 0.000 claims abstract description 22
- 239000000047 product Substances 0.000 claims abstract description 18
- 235000011389 fruit/vegetable juice Nutrition 0.000 claims abstract description 15
- 238000004659 sterilization and disinfection Methods 0.000 claims abstract description 13
- 229930006000 Sucrose Natural products 0.000 claims abstract description 12
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims abstract description 12
- 239000012467 final product Substances 0.000 claims abstract description 12
- 239000005720 sucrose Substances 0.000 claims abstract description 12
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims abstract description 11
- 239000008103 glucose Substances 0.000 claims abstract description 11
- DPXJVFZANSGRMM-UHFFFAOYSA-N acetic acid;2,3,4,5,6-pentahydroxyhexanal;sodium Chemical compound [Na].CC(O)=O.OCC(O)C(O)C(O)C(O)C=O DPXJVFZANSGRMM-UHFFFAOYSA-N 0.000 claims abstract description 9
- 229920003123 carboxymethyl cellulose sodium Polymers 0.000 claims abstract description 9
- 229940063834 carboxymethylcellulose sodium Drugs 0.000 claims abstract description 9
- 230000004151 fermentation Effects 0.000 claims description 44
- 235000019987 cider Nutrition 0.000 claims description 21
- 235000015097 nutrients Nutrition 0.000 claims description 13
- 238000005119 centrifugation Methods 0.000 claims description 12
- 238000004108 freeze drying Methods 0.000 claims description 12
- 238000001556 precipitation Methods 0.000 claims description 12
- 239000003381 stabilizer Substances 0.000 claims description 11
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 claims description 9
- 239000000463 material Substances 0.000 claims description 9
- 239000011734 sodium Substances 0.000 claims description 9
- 229910052708 sodium Inorganic materials 0.000 claims description 9
- 239000001965 potato dextrose agar Substances 0.000 claims description 8
- 238000001035 drying Methods 0.000 claims description 7
- 238000002386 leaching Methods 0.000 claims description 7
- 235000013372 meat Nutrition 0.000 claims description 7
- 239000008363 phosphate buffer Substances 0.000 claims description 7
- 239000000843 powder Substances 0.000 claims description 7
- 235000002639 sodium chloride Nutrition 0.000 claims description 7
- 235000014347 soups Nutrition 0.000 claims description 7
- FRXSZNDVFUDTIR-UHFFFAOYSA-N 6-methoxy-1,2,3,4-tetrahydroquinoline Chemical compound N1CCCC2=CC(OC)=CC=C21 FRXSZNDVFUDTIR-UHFFFAOYSA-N 0.000 claims description 6
- 238000004140 cleaning Methods 0.000 claims description 6
- 239000002054 inoculum Substances 0.000 claims description 6
- 238000009630 liquid culture Methods 0.000 claims description 6
- 238000011218 seed culture Methods 0.000 claims description 6
- 238000004321 preservation Methods 0.000 claims description 5
- 235000005979 Citrus limon Nutrition 0.000 claims description 4
- 244000131522 Citrus pyriformis Species 0.000 claims description 4
- 239000002253 acid Substances 0.000 claims description 4
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 claims 8
- 229910000029 sodium carbonate Inorganic materials 0.000 claims 4
- 241000186427 Cutibacterium acnes Species 0.000 claims 1
- BVKZGUZCCUSVTD-UHFFFAOYSA-N carbonic acid Chemical compound OC(O)=O BVKZGUZCCUSVTD-UHFFFAOYSA-N 0.000 claims 1
- 229940055019 propionibacterium acne Drugs 0.000 claims 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 abstract description 20
- 230000000694 effects Effects 0.000 abstract description 12
- 239000002994 raw material Substances 0.000 abstract description 12
- 238000000034 method Methods 0.000 abstract description 11
- 230000003064 anti-oxidating effect Effects 0.000 abstract description 6
- 229910052799 carbon Inorganic materials 0.000 abstract description 6
- 238000001914 filtration Methods 0.000 abstract 1
- 239000000523 sample Substances 0.000 description 19
- 239000000243 solution Substances 0.000 description 18
- 235000019441 ethanol Nutrition 0.000 description 16
- 241000235648 Pichia Species 0.000 description 12
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 11
- 230000000052 comparative effect Effects 0.000 description 9
- 235000010724 Wisteria floribunda Nutrition 0.000 description 6
- HHEAADYXPMHMCT-UHFFFAOYSA-N dpph Chemical compound [O-][N+](=O)C1=CC([N+](=O)[O-])=CC([N+]([O-])=O)=C1[N]N(C=1C=CC=CC=1)C1=CC=CC=C1 HHEAADYXPMHMCT-UHFFFAOYSA-N 0.000 description 6
- 238000002156 mixing Methods 0.000 description 6
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 5
- 241000235342 Saccharomycetes Species 0.000 description 5
- 238000006243 chemical reaction Methods 0.000 description 5
- 230000002269 spontaneous effect Effects 0.000 description 5
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 4
- 239000002537 cosmetic Substances 0.000 description 4
- 244000005700 microbiome Species 0.000 description 4
- 241000894006 Bacteria Species 0.000 description 3
- 241000196324 Embryophyta Species 0.000 description 3
- 102000004190 Enzymes Human genes 0.000 description 3
- 108090000790 Enzymes Proteins 0.000 description 3
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 3
- 238000002835 absorbance Methods 0.000 description 3
- 235000013361 beverage Nutrition 0.000 description 3
- 235000013399 edible fruits Nutrition 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 239000000796 flavoring agent Substances 0.000 description 3
- 235000019634 flavors Nutrition 0.000 description 3
- 239000001963 growth medium Substances 0.000 description 3
- 210000004209 hair Anatomy 0.000 description 3
- 239000004615 ingredient Substances 0.000 description 3
- 238000004519 manufacturing process Methods 0.000 description 3
- 229920000609 methyl cellulose Polymers 0.000 description 3
- 239000001923 methylcellulose Substances 0.000 description 3
- 235000010981 methylcellulose Nutrition 0.000 description 3
- 235000016709 nutrition Nutrition 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- 244000170916 Paeonia officinalis Species 0.000 description 2
- 235000006484 Paeonia officinalis Nutrition 0.000 description 2
- 230000032683 aging Effects 0.000 description 2
- 230000001476 alcoholic effect Effects 0.000 description 2
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 2
- 239000003963 antioxidant agent Substances 0.000 description 2
- 230000003078 antioxidant effect Effects 0.000 description 2
- 235000006708 antioxidants Nutrition 0.000 description 2
- 239000007864 aqueous solution Substances 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 239000000872 buffer Substances 0.000 description 2
- 239000012153 distilled water Substances 0.000 description 2
- 230000006872 improvement Effects 0.000 description 2
- 239000002609 medium Substances 0.000 description 2
- 230000004060 metabolic process Effects 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 229940057059 monascus purpureus Drugs 0.000 description 2
- 238000009938 salting Methods 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- MARUHZGHZWCEQU-UHFFFAOYSA-N 5-phenyl-2h-tetrazole Chemical compound C1=CC=CC=C1C1=NNN=N1 MARUHZGHZWCEQU-UHFFFAOYSA-N 0.000 description 1
- 244000153158 Ammi visnaga Species 0.000 description 1
- 235000010585 Ammi visnaga Nutrition 0.000 description 1
- 239000007836 KH2PO4 Substances 0.000 description 1
- 240000003183 Manihot esculenta Species 0.000 description 1
- 235000016735 Manihot esculenta subsp esculenta Nutrition 0.000 description 1
- 230000002292 Radical scavenging effect Effects 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 239000011149 active material Substances 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 229930014669 anthocyanidin Natural products 0.000 description 1
- 150000001452 anthocyanidin derivatives Chemical class 0.000 description 1
- 235000008758 anthocyanidins Nutrition 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 230000000975 bioactive effect Effects 0.000 description 1
- 235000010290 biphenyl Nutrition 0.000 description 1
- 239000004305 biphenyl Substances 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 239000003086 colorant Substances 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 238000000205 computational method Methods 0.000 description 1
- 230000003750 conditioning effect Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 229910000397 disodium phosphate Inorganic materials 0.000 description 1
- 238000004821 distillation Methods 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 125000001495 ethyl group Chemical group [H]C([H])([H])C([H])([H])* 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 229930003935 flavonoid Natural products 0.000 description 1
- 235000017173 flavonoids Nutrition 0.000 description 1
- 150000002215 flavonoids Chemical class 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 230000007760 free radical scavenging Effects 0.000 description 1
- 235000012055 fruits and vegetables Nutrition 0.000 description 1
- 238000004817 gas chromatography Methods 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 238000003780 insertion Methods 0.000 description 1
- 230000037431 insertion Effects 0.000 description 1
- 239000003550 marker Substances 0.000 description 1
- 230000005499 meniscus Effects 0.000 description 1
- 239000002207 metabolite Substances 0.000 description 1
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 150000007524 organic acids Chemical class 0.000 description 1
- 230000008520 organization Effects 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 239000011049 pearl Substances 0.000 description 1
- ZUOUZKKEUPVFJK-UHFFFAOYSA-N phenylbenzene Natural products C1=CC=CC=C1C1=CC=CC=C1 ZUOUZKKEUPVFJK-UHFFFAOYSA-N 0.000 description 1
- 150000008442 polyphenolic compounds Chemical class 0.000 description 1
- 235000013824 polyphenols Nutrition 0.000 description 1
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 description 1
- 239000006041 probiotic Substances 0.000 description 1
- 235000018291 probiotics Nutrition 0.000 description 1
- 150000003254 radicals Chemical class 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000029058 respiratory gaseous exchange Effects 0.000 description 1
- 239000012488 sample solution Substances 0.000 description 1
- 230000002000 scavenging effect Effects 0.000 description 1
- 150000003384 small molecules Chemical class 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 230000003068 static effect Effects 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
-
- 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
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
- A61K2800/85—Products or compounds obtained by fermentation, e.g. yoghurt, beer, wine
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- General Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Microbiology (AREA)
- Birds (AREA)
- Epidemiology (AREA)
- Mycology (AREA)
- Botany (AREA)
- Biotechnology (AREA)
- Engineering & Computer Science (AREA)
- Dermatology (AREA)
- Non-Alcoholic Beverages (AREA)
- Preparation Of Fruits And Vegetables (AREA)
Abstract
A kind of apple ferment, with apple as primary raw material, mixes after squeezing the juice, filtering with pure water, it is the glucose of raw mixture gross weight 3% to be added in gained raw mixture, then it is 6.0 7.0 to adjust pH value, and sterilizing is inoculated with Pichia pastoris subspecies in the proferment feed liquid for obtainingPichia kudriavzeviiPKD3001, then it is 120r/min to control rotating speed, temperature carries out 24 72h that ferment for 25 35 DEG C, in the zymotic fluid for obtaining, is calculated by every liter of zymotic fluid, add the 80g of sucrose 40 successively wherein, the 10g of citric acid 5, the 4g of carboxymethyl cellulose sodium 2, mix after 121 DEG C of sterilization 20min, then it is canned, obtain final product apple ferment finished product.Compared with the apple ferment obtained by natural fermentating method, its antioxidation activity index improves 75% to gained apple ferment finished product, and alcohol output is less than 1%.
Description
Technical field
The present invention relates to a kind of apple ferment and preparation method thereof, belong to technical field of bioengineering.
Background technology
According to the definition of Chinese biological fermentation industry association, ferment refers to, as raw material, to adopt with fruits and vegetables or other animal and plant etc.
With nature or artificial infection microorganism fermentation process, by the fermented product with some specific functions obtained by extraction.Ferment
Element not only saves original nutriment in fermentation raw material, such as Polyphenols, flavonoids, anthocyanidin, and by beneficial bacterium
Fermentating metabolism generate some new bioactive ingredients, such as organic acid, amino acid etc..Compared with before fermentation, microorganism
Conversion causes that these small-molecule substances are easier to absorption of human body.
Last decade, enzyme microb functional article industry starts to start in the whole world, and industry size constantly expands.
The areas such as Japan, Taiwan start to be produced using pure-blood ferment.Pure-blood ferment is in the single fermentation substrate of composition, only to connect
Enter one or more microorganisms, by the culture to the microorganism, obtain purity unicity product higher.Different from sending out naturally
Ferment, its fermentation time is short, easy to control, is very suitable for the industrialization production of ferment.But to be smoothed out fermentation, it is desirable to artificial
The strain of expansion must not be contaminated.It is thus higher to Zymolysis Equipment and environmental requirement, there are strict sterilizing measure and air net
Change system.Fermentation strain also must be the strain for being best suitable for production filtered out after strict research.
Apple ferment not only maintains apple originally nutrition and local flavor, will also assign the anti-oxidant work produced by its probiotics
Property, for consumer provides new functional product.But current fruit ferment makes and is primarily present：Fermentation time is long, influences water
Fruit raw material stability；Fermented bacterium performance and stability are poor, in influence ferment the problems such as natural active material；Fermented bacterium wine
Smart yield is higher, is not suitable for being applied to beverage and cosmetic field.
Pertinent literature
[1] bright Yue is hooked, Liu Liang, Zhang Chunzhi determine the 26 kinds of antioxidation activity of plant [J] food with fermentation work using DPPH methods
Industry, 2010（03）：148-150.
The such as [2] Sun Haiyan, Liu Enshi, Zhao Pingjuan, Li Juanhua gas chromatographies and distillation-alcohol meter method determine cassava zymotic fluid
Comparing [J] brewing science and technologies of middle alcohol content, 2012（11）：105-107.
The content of the invention
An object of the present invention is to solve the long influence ferment of fermentation time in existing fruit ferment production process
Activity, alcohol output is high, provides a kind of preparation method of apple ferment the problems such as strain unstable properties, the apple ferment
Preparation method has fermentation time short, and the apple ferment antioxidation activity of final gained is high, and alcohol content is low, is suitably applied drink
Material and cosmetic field.
The second object of the present invention is to provide the apple ferment obtained by above-mentioned preparation method.
Technical scheme
A kind of apple ferment, i.e., with the good apple of fresh mature degree as main material, use Pichia pastorisPichia kudriavzevii
PKD3001 carries out fermentation and is prepared from, and its preparation process specifically includes following steps：
（1）, strain selection
Strain used, is September in 2016 29 in China typical culture collection center preservation, and its deposit number is
CCTCC NO:The Pichia pastoris subspecies of M 2016537Pichia kudriavzeviiPKD3001 freeze-drying pipes, preservation machine
Structure address：Wuchang, wuhan Luo Jia Shan China typical culture collection center (Wuhan University), postcode：430072；
（2）, proferment feed liquid preparation
Through drying up, going after the elements in Fuji apple in Shaanxi province running water that fresh, maturity is good, natural ferment content is high is cleaned up
Core, squeezed the juice with juice extractor, obtained pure cider with 100 mesh filtered through gauze, then being mixed with pure water, being obtained raw material mixing
Thing, wherein the ratio of pure cider and pure water, is calculated, pure cider by percent by volume：Pure water is 10%:90%；
It is the glucose of raw mixture gross weight 3% to be added in the raw mixture of above-mentioned gained, and carbon is eaten through 1mol/L
It is 6.0-7.0 that acid sodium aqueous solution adjusts pH value, and the 10min that then sterilized in 105 DEG C obtains proferment feed liquid；
（3）, actication of culture
The Pichia pastoris subspecies that the freeze-drying pipe for taking aseptic water dissolves with oese is preservedPichia kudriavzevii
In the flat lining out of potato dextrose agar, 30 DEG C of culture 24h, obtain the ring of PKD3001 strains one in incubator
Flat board activated spawn；
（4）, seed culture
Step is taken with oese（3）The ring of flat board activated spawn one of gained accesses and 50mL fructus hordei germinatus leaching powder meat soup Liquid Cultures is housed
In the triangular flask of the 250mL specifications of base, prepare several pieces, be placed in the incubator of 30 DEG C of temperature, then control the rotating speed to be
120rpm, incubated 36h, obtains nutrient solution；
The rotating speed is controlled to carry out centrifugation 20min for 5000r/min the nutrient solution of above-mentioned gained, the precipitation obtained by centrifugation is used
0.01mol/L sterile phosphate buffer salines solution is cleaned 3 times, then adds 50mL 0.01mol/L aseptic in precipitation after cleaning
PBS shakes resuspended thalline through whirlpool under 500rpm, obtains seed liquor, in the seed liquor of gained, every milliliter of seed
Subspecies containing Pichia pastoris in liquidPichia kudriavzevii PKD3001 is 1 × 108-1×1010cfu；
（5）, fermented and cultured
In step（2）In the proferment feed liquid of gained, by 1 × 105Cfu/mL inoculum concentrations are by step（4）The seed liquor for obtaining connects
Enter step（2）In the proferment feed liquid of gained, then control rotating speed for 120r/min, temperature carries out fermentation 24- for 25-35 DEG C
72h, obtains zymotic fluid；
（6）, comprehensive allotment
In step（5）In the zymotic fluid of gained, calculated by every liter of zymotic fluid, add sucrose 40-80g, lemon successively in zymotic fluid
Lemon acid 5-10g, stabilizer carboxymethyl cellulose sodium 2-4g, mix after 121 DEG C of sterilization 20min, then canned, obtain final product apple ferment
Finished product.
Beneficial effects of the present invention
A kind of apple ferment of the invention, because the raw material apple for using is pure natural, using cold sterilization, strict control hair
Ferment temperature, effectively reduces the loss of ferment nutritional ingredient, and enzyme activity is remained to greatest extent.
Further, a kind of apple ferment of the invention, because the fermentation time used by preparation process is controlled within 80h,
Avoid the loss of flavor substance and ferment material that apple contains in itself.
Further, a kind of apple ferment of the invention, is to use Pichia pastorisPichia kudriavzevii PKD3001
Carry out fermentation to form, compared with the apple ferment obtained by spontaneous fermentation, its antioxidation activity index is improved gained apple ferment
75%, alcohol output is less than 1%, therefore can be widely applied to beverage and cosmetic field.
Brief description of the drawings
Fig. 1, embodiment 1,2,3,4 are by adding Pichia pastoris subspeciesPichia kudriavzeviiPKD3001 is carried out
Alcohol in the apple ferment sample of apple ferment sample and comparative examples 1 obtained by fermentation as obtained by the method for spontaneous fermentation
Degree block diagram；
Fig. 2, embodiment 1,2,3,4 are by adding Pichia pastoris subspeciesPichia kudriavzeviiPKD 3001 is fermented
The clearance rate column of apple ferment sample of the apple ferment sample and comparative examples 1 of gained as obtained by the method for spontaneous fermentation
Figure.
Specific embodiment
The present invention is expanded on further below by specific embodiment, but is not intended to limit the present invention.
Raw material apple used in following embodiment is purchased from Shanghai Lian Hua supermarkets.
Potato dextrose agar used and fructus hordei germinatus leaching powder meat soup fluid nutrient medium are in following embodiment
Known to those skilled in the art, it is purchased from Beijing overpass technical concern Co., Ltd.
0.01mol/L sterile phosphate buffer saline solution used in following embodiment is those skilled in the art
Well known, specific compound method is：Weigh 8g NaCl, 0.2g KCl, 1.44g Na2HPO4With 0.24g KH2PO4, it is dissolved in
In 800mL distilled water, with the pH value of hydrochloric acid conditioning solution to 7.4, finally 1L, 121 DEG C of sterilizing 20min are settled to distilled water.
Pichia pastoris subspecies used in various embodiments of the present inventionPichia kudriavzeviiPKD3001 freeze-dryings
Pipe, deposit number CCTCC NO:M 2016537, is that the present inventor was stored in Chinese Typical Representative culture on 29th in September in 2016
Collection preservation, preservation organization address：Wuchang, wuhan Luo Jia Shan China typical culture collection center (Wuhan University), postal
Compile：430072.
Embodiment 1
A kind of apple ferment, i.e., with the good apple of fresh mature degree as main material, using Pichia pastorisPichia kudriavzeviiPKD3001 carries out fermentation and is prepared from, and its preparation process specifically includes following steps：
（1）, strain selection
Strain used, is Pichia pastoris subspeciesPichia kudriavzeviiPKD3001 freeze-drying pipes；
（2）, proferment feed liquid preparation
Through drying up, going after the elements in Fuji apple in Shaanxi province running water that fresh, maturity is good, natural ferment content is high is cleaned up
Core, squeezed the juice with juice extractor, obtained pure cider with 100 mesh filtered through gauze, then being mixed with pure water, being obtained raw material mixing
Thing, wherein the ratio of pure cider and pure water, is calculated, pure cider by percent by volume：Pure water is 10%:90%；
It is the glucose of raw mixture gross weight 3% to be added in the raw mixture of above-mentioned gained, and carbon is eaten through 1mol/L
It is 6.0 that acid sodium aqueous solution adjusts pH value, and the 10min that then sterilized in 105 DEG C obtains proferment feed liquid；
（3）, actication of culture
The Pichia pastoris subspecies that the freeze-drying pipe for taking aseptic water dissolves with oese is preservedPichia kudriavzevii
In the flat lining out of potato dextrose agar, 30 DEG C of culture 24h, obtain the ring of PKD3001 strains one in incubator
Flat board activated spawn；
（4）, seed culture
Step is taken with oese（3）The ring of flat board activated spawn one of gained accesses and 50mL fructus hordei germinatus leaching powder meat soup Liquid Cultures is housed
In the triangular flask of the 250mL specifications of base, it is placed in the incubator of 30 DEG C of temperature, then controls rotating speed for 120rpm, it is incubated
36h, obtains nutrient solution；
The rotating speed is controlled to carry out centrifugation 20min for 5000r/min the nutrient solution of above-mentioned gained, the precipitation obtained by centrifugation is used
0.01mol/L sterile phosphate buffer salines solution is cleaned 3 times, then adds 50mL 0.01mol/L aseptic in precipitation after cleaning
PBS shakes resuspended thalline through whirlpool under 500rpm, obtains seed liquor, in the seed liquor of gained, every milliliter of seed
Subspecies containing Pichia pastoris in liquidPichia kudriavzeviiPKD3001 is 1 × 108-1×1010cfu；
（5）, fermented and cultured
In step（2）In the proferment feed liquid of gained, by 1 × 105Cfu/mL inoculum concentrations are by step（4）The Pichia pastoris for obtaining
SubspeciesPichia kudriavzeviiPKD3001 seed liquors are linked into step（2）In the proferment feed liquid of gained, then control
Rotating speed is 120rpm, and temperature carries out fermentation 36h, finally obtains zymotic fluid for 28 DEG C；
（6）, comprehensive allotment
In step（5）In the zymotic fluid of gained, calculated by every liter of zymotic fluid, add sucrose 40g, citric acid successively in zymotic fluid
7g, stabilizer carboxymethyl cellulose sodium 4g, mix after 121 DEG C of sterilization 20min, then canned, obtain final product apple ferment finished product.
Embodiment 2
A kind of apple ferment, i.e., with the good apple of fresh mature degree as main material, using Pichia pastorisPichia kudriavzeviiPKD3001 carries out fermentation and is prepared from, and its preparation process specifically includes following steps：
（1）, strain selection
Strain used, is Pichia pastoris subspeciesPichia kudriavzeviiPKD3001 freeze-drying pipes；
（2）, proferment feed liquid preparation
Through drying up, going after the elements in Fuji apple in Shaanxi province running water that fresh, maturity is good, natural ferment content is high is cleaned up
Core, squeezed the juice with juice extractor, obtained pure cider with 100 mesh filtered through gauze, then being mixed with pure water, being obtained raw material mixing
Thing, wherein the ratio of pure cider and pure water, is calculated, pure cider by percent by volume：Pure water is 10%:90%；
It is the glucose of raw mixture gross weight 3% to be added in the raw mixture of above-mentioned gained, and carbon is eaten through 1mol/L
It is that 6.30,105 DEG C of sterilizing 10min obtain proferment feed liquid that sour sodium adjusts pH value；
（3）, actication of culture
The Pichia pastoris subspecies that the freeze-drying pipe for taking aseptic water dissolves with oese is preservedPichia kudriavzevii
In the flat lining out of potato dextrose agar, 30 DEG C of culture 24h, obtain the ring of PKD3001 strains one in incubator
Flat board activated spawn；
（4）, seed culture
Step is taken with oese（3）The ring of flat board activated spawn one of gained accesses and 50mL fructus hordei germinatus leaching powder meat soup Liquid Cultures is housed
In the triangular flask of the 250mL specifications of base, prepare several pieces, be placed in the incubator of 30 DEG C of temperature, then control the rotating speed to be
120rpm, incubated 36h, obtains nutrient solution；
The rotating speed is controlled to carry out centrifugation 20min for 5000r/min the nutrient solution of above-mentioned gained, the precipitation obtained by centrifugation is used
0.01mol/L sterile phosphate buffer salines solution is cleaned 3 times, then adds 50mL 0.01mol/L aseptic in precipitation after cleaning
PBS shakes resuspended thalline through whirlpool under 500rpm, obtains seed liquor, in the seed liquor of gained, every milliliter of seed
Subspecies containing Pichia pastoris in liquidPichia kudriavzeviiPKD 3001 is 1 × 108-1×1010cfu；
（5）, fermented and cultured
In step（2）In the proferment feed liquid of gained, by 1 × 105Cfu/mL inoculum concentrations are by step（4）The Pichia pastoris for obtaining
SubspeciesPichia kudriavzevii PKD3001 seed liquors are linked into step（2）In the proferment feed liquid of gained, then control
Rotating speed is 120r/min, and temperature carries out fermentation 72h, finally obtains zymotic fluid for 25 DEG C；
（6）, comprehensive allotment
Calculated by every liter of zymotic fluid, in step（5）Sucrose 50g, citric acid 5g, stabilizer carboxylic are added in the zymotic fluid of gained successively
Methyl cellulose sodium 3g, mixes after 121 DEG C of sterilization 20min, then canned, obtains final product apple ferment finished product.
Embodiment 3
A kind of apple ferment, i.e., with the good apple of fresh mature degree as main material, using Pichia pastorisPichia kudriavzeviiPKD3001 carries out fermentation and is prepared from, and its preparation process specifically includes following steps：
（1）, strain selection
Strain used, is Pichia pastoris subspeciesPichia kudriavzeviiPKD3001 freeze-drying pipes；
（2）, proferment feed liquid preparation
Through drying up, going after the elements in Fuji apple in Shaanxi province running water that fresh, maturity is good, natural ferment content is high is cleaned up
Core, squeezed the juice with juice extractor, obtained pure cider with 100 mesh filtered through gauze, then being mixed with pure water, being obtained raw material mixing
Thing, wherein the ratio of pure cider and pure water, is calculated, pure cider by percent by volume：Pure water is 10%:90%；
It is the glucose of raw mixture gross weight 3% to be added in the raw mixture of above-mentioned gained, and carbon is eaten through 1mol/L
It is that 7.0,105 DEG C of sterilizing 10min obtain proferment feed liquid that sour sodium adjusts pH value；
（3）, actication of culture
The Pichia pastoris subspecies that the freeze-drying pipe for taking aseptic water dissolves with oese is preservedPichia kudriavzevii
In the flat lining out of potato dextrose agar, 30 DEG C of culture 24h, obtain the ring of PKD3001 strains one in incubator
Flat board activated spawn；
（4）, seed culture
Step is taken with oese（3）The ring of flat board activated spawn one of gained accesses and 50mL fructus hordei germinatus leaching powder meat soup Liquid Cultures is housed
In the triangular flask of the 250mL specifications of base, prepare several pieces, be placed in the incubator of 30 DEG C of temperature, then control the rotating speed to be
120rpm, incubated 36h, obtains nutrient solution；
The rotating speed is controlled to carry out centrifugation 20min for 5000r/min the nutrient solution of above-mentioned gained, the precipitation obtained by centrifugation is used
0.01mol/L sterile phosphate buffer salines solution is cleaned 3 times, and the aseptic phosphoric acid buffers of 50mL are then added in precipitation after cleaning
Salting liquid shakes resuspended thalline through whirlpool under 500rpm, obtains seed liquor, in the seed liquor of gained, containing complete in every milliliter of seed liquor
Red yeast subspeciesPichia kudriavzevii PKD3001 is 1 × 108-1×1010cfu；
（5）, fermented and cultured
In step（2）In the proferment feed liquid of gained, by 1 × 105Cfu/mL inoculum concentrations are by step（4）The Pichia pastoris for obtaining
SubspeciesPichia kudriavzevii PKD3001 seed liquors are linked into step（2）In the proferment feed liquid of gained, then control
Rotating speed is 120r/min, and temperature carries out fermentation 24h, finally obtains zymotic fluid for 33 DEG C；
（6）, comprehensive allotment
Calculated by every liter of zymotic fluid, in step（5）Sucrose 80g, citric acid 8g, stabilizer carboxylic are added in the zymotic fluid of gained successively
Methyl cellulose sodium 2g, mixes after 121 DEG C of sterilization 20min, then canned, obtains final product apple ferment finished product.
Embodiment 4
A kind of apple ferment, i.e., with the good apple of fresh mature degree as main material, apple ferment is prepared using Pichia pastoris fermentation,
Its preparation process specifically includes following steps：
（1）, strain selection
Strain used, is Pichia pastoris subspeciesPichia kudriavzeviiPKD3001；
（2）, proferment feed liquid preparation
Through drying up, going after the elements in Fuji apple in Shaanxi province running water that fresh, maturity is good, natural ferment content is high is cleaned up
Core, squeezed the juice with juice extractor, obtained pure cider with 100 mesh filtered through gauze, then being mixed with pure water, being obtained raw material mixing
Thing, wherein the ratio of pure cider and pure water, is calculated, pure cider by percent by volume：Pure water is 10%:90%；
It is the glucose of raw mixture gross weight 3% to be added in the raw mixture of above-mentioned gained, and carbon is eaten through 1mol/L
It is that 6.7,105 DEG C of sterilizing 10min obtain proferment feed liquid that sour sodium adjusts pH value；
（3）, actication of culture
The Pichia pastoris subspecies that the freeze-drying pipe for taking aseptic water dissolves with oese is preservedPichia kudriavzevii
In the flat lining out of potato dextrose agar, 30 DEG C of culture 24h, obtain the ring of PKD3001 strains one in incubator
Flat board activated spawn；
（4）, seed culture
Step is taken with oese（3）The ring of flat board activated spawn one of gained accesses and 50mL fructus hordei germinatus leaching powder meat soup Liquid Cultures is housed
In the triangular flask of the 250mL specifications of base, prepare several pieces, be placed in the incubator of 30 DEG C of temperature, then control the rotating speed to be
120rpm, incubated 36h, obtains nutrient solution；
The rotating speed is controlled to carry out centrifugation 20min for 5000r/min the nutrient solution of above-mentioned gained, the precipitation obtained by centrifugation is used
0.01mol/L sterile phosphate buffer salines solution is cleaned 3 times, and the aseptic phosphoric acid buffers of 50mL are then added in precipitation after cleaning
Salting liquid shakes resuspended thalline through whirlpool under 500rpm, obtains seed liquor, in the seed liquor of gained, containing complete in every milliliter of seed liquor
Red yeast subspeciesPichia kudriavzevii PKD3001 is 1 × 108-1×1010cfu；
（5）, fermented and cultured
In step（2）In the proferment feed liquid of gained, by 1 × 105Cfu/mL inoculum concentrations are by step（4）The Pichia pastoris for obtaining
SubspeciesPichia kudriavzevii PKD3001 seed liquors are linked into step（2）In the proferment feed liquid of gained, then control
Rotating speed is 120r/min, and temperature carries out fermentation 60h, finally obtains zymotic fluid for 35 DEG C；
（6）, comprehensive allotment
Calculated by every liter of zymotic fluid, in step（5）Sucrose 70g, citric acid 10g, stabilizer are added in the zymotic fluid of gained successively
Carboxymethyl cellulose sodium 2.5g, mixes after 121 DEG C of sterilization 20min, then canned, obtains final product apple ferment finished product.
Comparative examples 1
Apple ferment is prepared using aging process, with the good apple of fresh mature degree as main material, by aging process
Into its preparation process specifically includes following steps：
（1）, proferment feed liquid preparation
Through drying up, going after the elements in Fuji apple in Shaanxi province running water that fresh, maturity is good, natural ferment content is high is cleaned up
Core, squeezed the juice with juice extractor, obtained pure cider with 100 mesh filtered through gauze, then being mixed with pure water, being obtained raw material mixing
Thing, wherein the ratio of pure cider and pure water, is calculated, pure cider by percent by volume：Pure water is 10%:90%, as
Proferment feed liquid；
（2）, fermented and cultured
Proferment feed liquid obtained above is dispensed into vial, special lid is added, room temperature is placed and carries out nature hair in 20 days
Ferment, obtains zymotic fluid；
（3）, comprehensive allotment
Calculated by every liter of zymotic fluid, in step（2）Sucrose 40g, citric acid 7g, stabilizer carboxylic are added in the zymotic fluid of gained successively
Methyl cellulose sodium 4g, mixes after 121 DEG C of sterilization 20min, then canned, obtains final product apple ferment finished product.
Effect example 1
Using alcohol meter method[1]Respectively to embodiment 1,2,3,4 by adding Pichia pastoris subspeciesPichia kudriavzevii
PKD3001 fermented obtained by apple ferment as obtained by the method for spontaneous fermentation of apple ferment sample and comparative examples 1
Alcoholic strength in sample is detected that its step is as follows：
The apple of the control of the apple ferment sample and the gained of comparative examples 1 of the gained of 100mL embodiments 1,2,3,4 is taken respectively
Ferment sample is in 200mL cucurbits, then adds several cleaned glass pearls, connects condenser, and receiver is made with 100mL volumetric flasks
（Additional ice bath）, condensed water is opened, slow heating distillation is collected distillate close to scale, 30min is incubated in 20 DEG C of water-baths,
Benefit adds water to scale, mixes, and pours into clean, dry 100mL graduated cylinders, and static several minutes, after after wherein bubble collapse, are put into
Clean, dry alcohol meter（Uoe, Wuqiang County absolute sincerity instrucment and meter plant）, then gently click, must not exposure barrel, while insertion
Thermometer, balances 5min, and the scale indicating value with meniscus the tangent is read in level observation, while temperature is recorded, according to what is measured
Alcohol meter indicating value and temperature, look into alcohol meter temperature concentration conversion table, alcoholic strength when being converted into 20 DEG C.Each sample parallel determination 3
Secondary, acquired results are represented to a decimal.
Experimental result is shown in accompanying drawing 1, and the apple ferment obtained by preparation method of the present invention is can be seen that from accompanying drawing 1, and its alcohol is produced
Amount is less than 1%.
Effect example 2
TTC (2,3,5 monochlor(in)ate phenyl tetrazole) is a kind of developer, and its metabolite to yeast occurs color reaction, leads to
The size that it may determine that breathing enzyme activity in saccharomycete is crossed, i.e. saccharomycete produces the height of alcohol ability.On solid medium
Different colours can be shown after one layer of TTC developer is covered on different yeast colonies, it is in peony to produce the strong yeast colony of alcohol ability,
It is pink to take second place.Fermentation is tested with metabolism of yeasts product and TTC developers generation chromogenic reaction to be produced with Pichia yeast
Alcohol ability is strong and weak, and the chromogenic reaction of fermentation Pichia yeast, other saccharomycete and TTC is determined respectively.
Picking fermentation Pichia yeast and other saccharomycete are distinguished in potato dextrose agar with sterilized toothpick
On culture medium, fermentation Pichia yeast black marking pen in culture dish bottom marker, after 30 DEG C of culture 36h, in culture medium
On cover one layer of TTC upper stratas culture medium again, lucifuge stands half a day.Experimental result is evident that fermentation Pichia yeast in white
Color, other saccharomycete are in peony, pink colour, are indicated above Pichia pastoris subspecies usedPichia kudriavzevii
It is low that PKD3001 produces alcohol amount.
Effect example 3
Using DPPH methods[2]Embodiment 1,2,3,4 is determined respectively by adding Pichia pastoris subspeciesPichia kudriavzevii
PKD 3001 fermented obtained by apple ferment as obtained by the method for spontaneous fermentation of apple ferment sample and comparative examples 1
The antioxidation activity of sample, i.e., with 1,1- diphenyl -2- trinitrophenyl-hydrazines（DPPH）Removing vigor is detected that it is walked for index
It is rapid as follows：
In 5 test tubes of 10 mL, the apple ferment sample and comparative examples of the gained of 2mL embodiments 1,2,3,4 are separately added into
The apple ferment sample of 1 gained, is then separately added into 2mL 0.1mmol/L DPPH solution in each test tube, shake up, respectively
Dark place stands 30min at room temperature, with absolute ethyl alcohol as blank, absorbance of the sample under 517nm in each test tube is determined respectively,
It is denoted asA sample.2mL DPPH solution and the mixed absorbance of 2mL ethanol are determined simultaneously, are denoted asA control, and 2mL hairs
Bright sample solution and the mixed absorbance of 2mL absolute ethyl alcoholsA blank.Then the apple ferment of the gained of embodiment 1,2,3,4 is calculated
The radical scavenging activity of the apple ferment sample of sample and the gained of comparative examples 1, i.e. clearance rate, each sample parallel determination
3 times, its computational methods is as follows：
Clearance rate shows that more greatly the Scavenging activity of free radical is stronger, is indicated above its oxidation resistance stronger.
The above-mentioned experimental result for measuring is shown in accompanying drawing 2, figure it is seen that by the present invention by adding Pichia yeast
SubspeciesPichia kudriavzevii PKD3001 fermented obtained by apple ferment sample to DPPH free radical scavenging activities ratio
At least more than 75% is improved using the apple ferment obtained by natural fermentating method in comparative examples, is shown preferably anti-oxidant
Vigor.
In sum, a kind of apple ferment that the present invention is provided, remains the originally nutritional ingredient in apple, by addition
Pichia yeast subspeciesPichia kudriavzevii PKD3001 is fermented, and not only increases new local flavor, is also improved
The antioxidation activity of apple ferment, and product alcohol amount is low, is suitably applied beverage and cosmetic field.
The above is only the citing of embodiments of the present invention, is not intended to limit the invention, for the art
Those of ordinary skill for, on the premise of the technology of the present invention principle is not departed from, some improvement and modification can also be made, this
A little improvement also should be regarded as protection scope of the present invention with modification.
Claims (7)
1. a kind of preparation method of apple ferment, it is characterised in that the preparation method, i.e., based on the good apple of fresh mature degree
Material, uses Pichia pastoris subspeciesPichia kudriavzevii PKD 3001 carries out fermentation and is prepared from, its preparation process tool
Body comprises the following steps：
（1）, strain selection
Strain used, is September in 2016 29 in China typical culture collection center preservation, and its deposit number is
CCTCC NO:The Pichia pastoris subspecies of M 2016537Pichia kudriavzevii The freeze-drying pipes of PKD 3001；
（2）, proferment feed liquid preparation
After the apple running water that fresh, maturity is good, natural ferment content is high is cleaned up through drying, stoning, with squeezing the juice
Machine is squeezed the juice, is obtained pure cider with 100 mesh filtered through gauze, is then mixed with pure water, raw mixture is obtained, wherein pure
The ratio of cider and pure water, is calculated, pure cider by percent by volume：Pure water is 10%:90%；
The glucose of raw mixture gross weight 3% is added in the raw mixture of above-mentioned gained, carbonic acid is eaten through 1mol/L
It is 6.0-7.0 that sodium water solution adjusts pH value, and the 10min that then sterilized in 105 DEG C obtains proferment feed liquid；
（3）, actication of culture
The Pichia pastoris subspecies that the freeze-drying pipe for taking aseptic water dissolves with oese is preservedPichia kudriavzevii
In the flat lining out of potato dextrose agar, 30 DEG C of culture 24h, obtain the ring of PKD3001 strains one in incubator
Flat board activated spawn；
（4）, seed culture
Step is taken with oese（3）The ring of flat board activated spawn one of gained accesses and 50mL fructus hordei germinatus leaching powder meat soup Liquid Cultures is housed
In the triangular flask of the 250mL specifications of base, it is placed in the incubator of 30 DEG C of temperature, then controls rotating speed for 120rpm, it is incubated
36h, obtains nutrient solution；
The rotating speed is controlled to carry out centrifugation 20min for 5000r/min the nutrient solution of above-mentioned gained, the precipitation obtained by centrifugation is used
0.01mol/L sterile phosphate buffer salines solution is cleaned 3 times, then adds 50mL 0.01mol/L aseptic in precipitation after cleaning
PBS shakes resuspended thalline through whirlpool under 500rpm, obtains seed liquor, in the seed liquor of gained, every milliliter of seed
Subspecies containing Pichia pastoris in liquidPichia kudriavzevii PKD 3001 is 1 × 108-1×1010cfu；
（5）, fermented and cultured
In step（2）In the proferment feed liquid of gained, by 1 × 105Cfu/mL inoculum concentrations are by step（4）The seed liquor for obtaining is accessed
Step（2）In the proferment feed liquid of gained, then control rotating speed for 120r/min, temperature carries out fermentation 24-72h for 25-35 DEG C,
Obtain zymotic fluid；
（6）, comprehensive allotment
In step（5）In the zymotic fluid of gained, calculated by every liter of zymotic fluid, add sucrose 40-80g, lemon successively in zymotic fluid
Lemon acid 5-10g, stabilizer carboxymethyl cellulose sodium 2-4g, mix after 121 DEG C of sterilization 20min, then canned, obtain final product apple ferment
Finished product.
2. a kind of preparation method of apple ferment as claimed in claim 1, it is characterised in that step（2）The system of proferment feed liquid
In standby：It is the glucose of raw mixture gross weight 3% to be added in the raw mixture of above-mentioned gained, edible through 1mol/L
It is 6.0 that aqueous sodium carbonate adjusts pH value, and the 10min that then sterilized in 105 DEG C obtains proferment feed liquid；
Step（5）In fermented and cultured：Control rotating speed for 120rpm, temperature carries out fermentation 36h for 28 DEG C；
Step（6）In comprehensive allotment：Calculated by every liter of zymotic fluid, in step（5）Sucrose is added successively in the zymotic fluid of gained
40g, citric acid 7g, stabilizer carboxymethyl cellulose sodium 4g are mixed after 121 DEG C of sterilization 20min, then canned, obtain final product apple ferment
Plain finished product.
3. a kind of preparation method of apple ferment as claimed in claim 1, it is characterised in that step（2）The system of proferment feed liquid
In standby：It is the glucose of raw mixture gross weight 3% to be added in the raw mixture of above-mentioned gained, edible through 1mol/L
It is that 6.30,105 DEG C of sterilizing 10min obtain proferment feed liquid that sodium carbonate adjusts pH value；
Step（5）In fermented and cultured：Control rotating speed for 120r/min, temperature carries out fermentation 72h, finally obtains fermentation for 25 DEG C
Liquid；
Step（6）In comprehensive allotment：Calculated by every liter of zymotic fluid, in step（5）Sucrose is added successively in the zymotic fluid of gained
50g, citric acid 5g, stabilizer carboxymethyl cellulose sodium 3g are mixed after 121 DEG C of sterilization 20min, then canned, obtain final product apple ferment
Plain finished product.
4. a kind of preparation method of apple ferment as claimed in claim 1, it is characterised in that step（2）The system of proferment feed liquid
In standby：It is the glucose of raw mixture gross weight 3% to be added in the raw mixture of above-mentioned gained, edible through 1mol/L
It is that 7.0,105 DEG C of sterilizing 10min obtain proferment feed liquid that sodium carbonate adjusts pH value；
Step（5）In fermented and cultured：Control rotating speed for 120r/min, temperature carries out fermentation 24h, finally obtains fermentation for 33 DEG C
Liquid；
Step（6）In comprehensive allotment：Calculated by every liter of zymotic fluid, in step（5）Sucrose is added successively in the zymotic fluid of gained
80g, citric acid 8g, stabilizer carboxymethyl cellulose sodium 2g are mixed after 121 DEG C of sterilization 20min, then canned, obtain final product apple ferment
Plain finished product.
5. a kind of preparation method of apple ferment as claimed in claim 1, it is characterised in that step（2）The system of proferment feed liquid
In standby：It is the glucose of raw mixture gross weight 3% to be added in the raw mixture of above-mentioned gained, edible through 1mol/L
It is that 6.7,105 DEG C of sterilizing 10min obtain proferment feed liquid that sodium carbonate adjusts pH value；
Step（5）In fermented and cultured：Control rotating speed for 120r/min, temperature carries out fermentation 60h, finally obtains fermentation for 35 DEG C
Liquid；
Step（6）In comprehensive allotment：Calculated by every liter of zymotic fluid, in step（5）Sucrose is added successively in the zymotic fluid of gained
70g, citric acid 10g, stabilizer carboxymethyl cellulose sodium 2.5g are mixed after 121 DEG C of sterilization 20min, then canned, obtain final product apple
Ferment finished product.
6. a kind of apple ferment as obtained by the preparation method of claim 1,2,3,4 or 5.
7. a kind of apple ferment as obtained by the preparation method of claim 1,2,3,4 or 5 is in propionibacterium acnes is suppressed
Using.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN107573941A (en) * | 2017-08-30 | 2018-01-12 | 覃祖桓 | Application and its method of a kind of storehouse Delhi A Ziweishi Pichia yeasts in soil |
| CN108094790A (en) * | 2017-12-22 | 2018-06-01 | 光明乳业股份有限公司 | A kind of fermented apple juice and preparation method thereof |
| CN108968039A (en) * | 2018-06-19 | 2018-12-11 | 上海应用技术大学 | A kind of tomato ferment and its preparation method and application |
| CN108968038A (en) * | 2018-06-19 | 2018-12-11 | 上海应用技术大学 | A kind of cherry ferment and preparation method thereof |
| CN111659275A (en) * | 2020-05-23 | 2020-09-15 | 常州市武进区环境监测站 | Preparation process and device of environment-friendly enzyme |
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| CN104872681A (en) * | 2015-06-30 | 2015-09-02 | 卞洪锋 | Apple microorganism ferment and preparation method thereof |
| CN105029600A (en) * | 2015-05-25 | 2015-11-11 | 江南大学 | Apple enzyme beverage production method |
| CN105995349A (en) * | 2016-06-13 | 2016-10-12 | 江南大学 | Fast making method of apple enzyme beverage |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105029600A (en) * | 2015-05-25 | 2015-11-11 | 江南大学 | Apple enzyme beverage production method |
| CN104872681A (en) * | 2015-06-30 | 2015-09-02 | 卞洪锋 | Apple microorganism ferment and preparation method thereof |
| CN105995349A (en) * | 2016-06-13 | 2016-10-12 | 江南大学 | Fast making method of apple enzyme beverage |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107573941A (en) * | 2017-08-30 | 2018-01-12 | 覃祖桓 | Application and its method of a kind of storehouse Delhi A Ziweishi Pichia yeasts in soil |
| CN108094790A (en) * | 2017-12-22 | 2018-06-01 | 光明乳业股份有限公司 | A kind of fermented apple juice and preparation method thereof |
| CN108968039A (en) * | 2018-06-19 | 2018-12-11 | 上海应用技术大学 | A kind of tomato ferment and its preparation method and application |
| CN108968038A (en) * | 2018-06-19 | 2018-12-11 | 上海应用技术大学 | A kind of cherry ferment and preparation method thereof |
| CN111659275A (en) * | 2020-05-23 | 2020-09-15 | 常州市武进区环境监测站 | Preparation process and device of environment-friendly enzyme |
| CN111659275B (en) * | 2020-05-23 | 2022-05-31 | 常州市武进区环境监测站 | Preparation process and device of environment-friendly enzyme |
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