CN109182023A - A kind of fermentation fructus lycii slag and flavouring Lycium chinense wine - Google Patents
A kind of fermentation fructus lycii slag and flavouring Lycium chinense wine Download PDFInfo
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Abstract
The present invention relates to a kind of fermentation fructus lycii slags, and are related to the flavouring Lycium chinense wine using the fermentation fructus lycii slag Titian, belong to fructus lycii deep process technology field.The fermentation fructus lycii slag is prepared by the following method: Ku Teshi bacillus (Kurthia sp) NXUGQ15 that 4-6% deposit number is CCTCC NO:M2017524 is added in fructus lycii slag, semi-solid ferment 10-15 hours at 35-37 DEG C, 130r/min, must ferment fructus lycii slag.It can be applied to the multiple fields such as feed, cosmetics, food, being especially applied to fermentation Lycium chinense wine being capable of significant Titian.The mutagenic obtained Ku Teshi bacillus strain NXUGQ15 of the present invention, the ability for carotenoid of degrading is strong, and the production carotenoid degrading enzyme time of fermenting is short, and enzyme activity is high, the enzyme activity 8.87U/mL of crude enzyme liquid, and producing enzyme is preferable.Fructus lycii slag can be turned waste into wealth, and make full use of the carotenoid in fructus lycii slag.
Description
Technical field
The present invention relates to a kind of fermentation fructus lycii slags, and are related to the flavouring Lycium chinense wine using the fermentation fructus lycii slag Titian, belong to
Fructus lycii deep process technology field.
Background technique
It is well known that fructus lycii has nourishing liver and kidney, benefiting shrewd head, soreness of waist and knee joint, dizziness and tinnitus, Heat Diabetes, blood are treated
Empty sallow effect, therefore, Lycium chinense wine is made as health liquor in fructus lycii by people.Lycium chinense wine preparation method has infusion method and fermentation at present
Two kinds of method.Infusion method is generally formed whole grain fructus lycii white wine or yellow rice wine are brewed, and wine degree is higher, and nutritional ingredient can not be abundant
Dissolution.Fermentation method Lycium chinense wine is also less to have oxygen participation because not being related to high-temperature heating process, not only maintains in fructus lycii substantially
Natural nutrition ingredient, and be more conducive to absorption of human body through everfermentation, be a kind of extraordinary health-care nutritive fruit wine.
But fermented type Lycium chinense wine, due to being influenced by factors such as raw material and processing technologys, quality is irregular, the mouthfeel of product is fragrant
Gas is generally insufficient, needs to improve.
Publication number: 101323823 invention " brewing method of Lycium chinense wine " discloses a kind of brewing method of Lycium chinense wine, will
Diastatic fermentation distiller's yeast and distiller's yeast is added in the fructus lycii of broken deseeding and the starchy material after curing, is sent out in the fermenter
Ferment；Then the mash fermented is squeezed, wine juice is clarified, sterilization, ageing, blends and bottles after filtering.Using shallow lake
Carbon source of the silty raw material as fermentation, brews pure fermentation Lycium chinense wine, mainly solves the problems, such as are as follows: reduces be produced into the maximum extent
This.
Publication number: 1513970 invention " a kind of fermented wolfberry fruit grape and brew method " discloses a kind of fermentation fructus lycii
Wine and fermentating wine blend preparation method, and this method is separately fermented using fructus lycii and grape as raw material, then carry out preparation tune
The process matched.It is with fresh fructus lycii or dry wolfberry are broken pulp (fruit juice) fermentation to be made fermentation Lycium chinense wine is made；White grape or
Red grape is made fruit juice (pulp) fermentation and is brewed into fermentating wine through broken.Ferment Lycium chinense wine and fermentating wine according to
(1%-99%): the ratio of (99%-1%) is tuned into different types of pol after blending proportion, through Overheating Treatment, cold treatment, mistake
Filter, bottling, sterilization, are made fermented wolfberry fruit grape.The fermented wolfberry fruit grape that the method is brewed, Lycium berry and grape fragrance is harmonious,
The plentiful coordination of mouthfeel, fructus lycii taste is strong, and typicalness is prominent.
Publication number: a kind of 1104248 invention " brew method of Lycium chinense wine " discloses a kind of brew method of Lycium chinense wine,
This method includes featured raw material, squeezing, fermentation, filtering, sterilizing, it is characterised in that fresh fructus lycii is featured, squeezes into wolfberry juice
Afterwards, after sodium sulfite vulcanization is added, yeast is added, after adding glucose to pol to be 22 ° of BX, sealing hair in investment pressurized tank
Ferment, fermentation temperature are (20-25) DEG C, and the time is after 5-8 days, and adding glucose to pol again is 18 ° of BX, is carried out again close
Seal ferment, fermentation temperature are (15-20) DEG C, and when fermentation to tank bottom precipitating has pomace and yeast, wine liquid tentatively to clarify, pol is
It after 5 ° of BX or less, after medlar liquid pomace, yeast mixt are stirred, heat in tank, then is cooled down, by fructus lycii mixture
Suitable sodium sulfite is added in filtering resulting medlar liquid and adds honey clarification, filters again, filtered fluid is store for filtering
In in bucket, changes bucket and once carry out filling pasteurization afterwards within storage 2-3 months under being 10 DEG C or so in temperature, that is, be brewed into fructus lycii
The medlar liquid of above-mentioned brew is put into a certain amount of fructus lycii soak, is configured to the Lycium chinense wine of different wine degree by wine.
Publication number: the brew method of Lycium chinense wine disclosed in a kind of 1782060 invention " brew method of Lycium chinense wine ": fresh Chinese holly
Qi or dry wolfberry are sorted, clean, if dry wolfberry is also impregnated 12-24 hours with the water of 2-5 times of its weight, are then crushed
At wolfberry fruit syrup, alcoholic strength is adjusted to 20-25% (v/v) with the deodorizing alcohol of 95% (v/v), is impregnated 10-20 days, during immersion often
It beat circulation 1-4 times, after isolate soaking wine；Fresh fructus lycii pomace after separation adds the soft water of 0.5-2 times of fresh fructus lycii weight；
Dry wolfberry pomace adds the soft water of 2-5 times of dry wolfberry weight, with sucrose sugar addition to 210-230g/L, with citric acid or tartaric acid
Total acid is adjusted to 6.0-8.0g/L, adjusts SO2To 80-100mg/L, add pectase 0.2-0.5g/L, adds dry ferment 0.15-
0.3g/L, control temperature are fermented at 18-30 DEG C, the time 5-7 days, when specific gravity is down to 1000g/L or less, analyze residual sugar < 4g/L
Isolate fermented wine；Soaking wine and fermented wine are mixed, then be through ageing, allotment, lower glue, filtering, degerming, bottling, packaging
Finished product.
Publication number: the brew method of Lycium chinense wine disclosed in a kind of 1265420 invention " brew method of Lycium chinense wine ": will divide
Blanching in the fructus lycii merging citric acid and sodium sulfite mixed liquor chosen, the fructus lycii after blanching first after crushing, add soft water
It impregnates, fructus lycii and soft water are stood after adjusting composition with 7:3 (volume ratio) tinning with sulfur dioxide and pectinase treatment, inoculation hair
Ferment isolates fermented wine stored for future use, sorted broken 95% deodorization edible wine of fructus lycii merging after primary fermentation and post-fermentation
It impregnates 30 days or so and separates in essence, soaking wine is made, soaking wine and fermented wine are blent into allotment, storage one by 1:4 (volume ratio)
After the section time, filters through lower glue, bottle after aseptic filtration.
Publication number: the preparation method of fermentation Chinese wolfberry fruit wine disclosed in 1077744 invention " process for preparation of fermentation Chinese wolfberry fruit wine ": will
Fructus lycii is rinsed through impurity elimination, and hot dipping mentions, and is crushed, and filtering adds auxiliary material, sterilizes, cooling, prior fermentation, purified treatment, filtering, ageing,
Altar processed, allotment, filtering etc. processes and be made fermentation Chinese wolfberry fruit wine, specifically comprises the processes of: a: fructus lycii impurity elimination is rinsed, with 65 DEG C -75
DEG C hot water extraction 1.5-2.5 hours, its residue is placed again into 65 DEG C of -75 DEG C of hot water after crushing filtering, extracts 0.8-1.2
It carries out filtering for second after hour, b: the residue after a secondary filters is crushed, its filter residue is abandoned after filtering again, c: in b
Filtrate in 20% white sugar auxiliary material is added, d: solution made from c is sterilized 20-25 seconds at a temperature of 95 DEG C -100 DEG C, cold
But to 65 DEG C -70 DEG C, it is put into wine jar, continues to be cooled to 30 DEG C -32 DEG C, e: by solution made from d under the conditions of 28 DEG C -30 DEG C
It carries out prior fermentation 6-8 days, then progress later stage fermentation 9-11 days at room temperature, f: solution made from e is purified
Gelatin (being added in the ratio of 10mL solution 0.008g gelatin), filtering in lower glue 14-16 days, g: will be molten made from f is added in processing
Liquid ageing 3-5 months, altar, placed 10-20 days under the conditions of 50 DEG C -60 DEG C, places 5-7 under the conditions of subzero 2 DEG C -44 DEG C
It, filtering, filtrate is stored 1 month, h: flavoring agent is added in filtrate made from g and tonic is deployed, i: is obtained by h
Solution filtering, sterilizing, as finished product.
Although Catotenoids From Lycium Barbarum content is very rich, after traditional handicraft undergoes microbial fermentation, in Lycium chinense wine
Carotenoid content is not high, not soluble in water mainly due to carotenoid, greatly when fermentation ends pomace separates
It is removed, another part is degraded in brewing process and is lost.
Oneself has centuries for the time-honored Traditional Brand of lycium barbarum, cultural advantage history, and Zhongning County is State Council's name
" township of Chinese fructus lycii ", lycium barbarum is classified as the homologous food of medicine food by " Chinese Pharmacopoeia ", this be other any provinces and regions all without
Method analogy.
National No. 1 file clearly proposes that " reinforcing new raw-food material, integration of drinking and medicinal herbs food development and application " " adds within 2017
Strong modern biotechnology and fortification technical research, excavating exploitation has the function of the food of health care ", it is called in response to country,
Development function health food develops Ningxia advantage characteristic fructus lycii resource, promotes the quality of health lycium chinense, find a kind of fermentation Chinese holly
The preparation method of Qi slag is the technical problems to be solved by the invention.
Summary of the invention
To solve the above problems, the present invention provides a kind of fermentation fructus lycii slag, dropped using one plant of isolated carotenoid
Bacterium is solved, the pomace being filtered to remove in Lycium chinense wine manufacturing process, light, oxygen, heat, biological enzyme unstable using carotenoid are handled
Etc. the degradable characteristic for generating drop isoprenoid volatile compound, it is made to generate a large amount of C9-、C10-、C13And C15It drops different
Pentadiene class compound (norisoprenoids), these substances are that have volatile aromatic compound, these compounds because
It is lower for sense organ threshold value, there is positive contribution to food.The carotenoid that fructus lycii is rich in mainly includes beta carotene, corn
Flavine, luteole acid dipalmitate, and wherein most with luteole acid dipalmitate, about carotenoid total amount
77.5%.It is most of to be lost in fructus lycii slag due to not soluble in water but in brewed wine, cause very big waste.The present invention
Using microbial degradation, fructus lycii slag is utilized, makes contained a large amount of carotenoid degradations not soluble in water, degradation in fructus lycii
Fermentation fructus lycii slag afterwards is put back to again in the Lycium chinense wine of fermentation, is played the role of increasing wine aroma, is improved the quality of Lycium chinense wine, is made
Flavouring fructus lycii is spilt.
Traditional zymotechnique, since Catotenoids From Lycium Barbarum is not soluble in water, a large amount of carotenoid is by fructus lycii slag band
It walks, the present invention is made using carotenoid not soluble in water in isolated one plant of carotenoid degradation bacteria degradation fructus lycii slag
Drop isoprenoid aroma substance is generated, preparation fermentation fructus lycii slag can be used for the preparation of Lycium chinense wine, can be improved aroma, improves
Fructus lycii wine aroma improves Lycium chinense wine quality, innovates fructus lycii wine brewage technology.
Drop isoprenoid compound (norisoprenoids) is a kind of special by having for carotenoid degradation generation
The substance of fragrance, and the substance have lower odor threshold, thus on a small quantity exist can to the flavor of food generate compared with
Big organoleptic effects.Drop isoprenoid compound containing 9,10,11 and 13 carbon atoms in food is mostly by carotenoid
Degradation generates, such as dorinone, alpha, beta-lonone, dihydroactinidiolide, all has good flavor.Isoprenoid drops
Compound is one of main aroma-producing substance of the freshs fruit of vegetables such as grape, and influences the important compound of fruit wine flavouring essence quality.Fruit
There is explored drop isoprenoid compound in wine aroma ingredient: alpha, beta-lonone, isophorone, limonene, safranal,
Geranyl acetone, dihydro-β-ionone, dorinone, dihydroactinidiolide, methyl heptenone, β-cyclocitral, 2- heptan
Olefine aldehydr, dihydro jasmone, α-cyclocitral, isogeraniol, 2,5,6- trimethyl -4- teracrylic acid -one, 2,4- nonadienal, 3-
Decanone, 2- nonenyl aldehyde, 2,19 kinds of drop pentadiene class compounds such as 2,6- trimethyl annulenones, odor characteristic are shown in Table 3.
The present invention makes full use of fructus lycii slag, using the degradation of carotenoid, increases the flavor in fructus lycii slag
Fructus lycii slag prepared by the method for the present invention is used for fructus lycii liquor brewing, increases drop isoprenoid in Lycium chinense wine after testing by matter
Compounds content and type, they are mainly alpha, beta-lonone, 2,2,6- trimethylcyclohexanones (TCH), isophorone, spiceleaf
Benzylacetone, dihydro-β-ionone, geraniol, safranal, β-cyclocitral, limonene, pseudoionone, dihydro macaque
Peach lactone.And aldehyde C-9, amyl valerate, certain herbaceous plants with big flowers acetoacetic ester.The taste threshold value of these substances is low, plays very for Lycium chinense wine sensory evaluation
Good effect.
The Lycium chinense wine carbonyl compound content increases, and relative peak area accounts for the 5.78% of total ingredient, higher than old technology
4.23%；It is 3- undecyl ketone, dorinone, alpha, beta-lonone, β-cyclocitral, safranal and two that wherein content is higher
Hydrogen-alpha, beta-lonone, the higher alpha, beta-lonone 3.136% of relative amount, β-cyclocitral 3.615%, it is special that they have
Fruit aroma.
Ester type compound relative peak area be 14.08%, higher than the 11.03% of prior art, be in Lycium chinense wine content compared with
More aromatic substances.The 13.68% of total ingredient is accounted for, higher than the 10.73% of prior art, larger, certain herbaceous plants with big flowers is contributed to fructus lycii wine aroma
Acetoacetic ester and amyl valerate are also important catabolite, the fragrance with pears perfume (or spice) and banana；Dihydroactinidiolide, acetone are fragrant
Leaf ester is one of intermediate product and the higher esters of content of carotenoid degradation, and fruit and flowery odour are prominent.
A kind of fermentation fructus lycii slag is prepared by the method included the following steps:
Ku Teshi bacillus (Kurthia sp) is expanded and is cultivated, Ku Teshi bacillus bacterium solution is obtained；
Fresh fructus lycii mashing, juicing, filtering, obtain wolfberry juice and fructus lycii slag；
Ku Teshi bacillus bacterium solution is added in fructus lycii slag, ferment to obtain fermentation fructus lycii slag；
Fermentation fructus lycii slag is played back in Lycium chinense wine, dipping, dissolves in catabolite drop isoprenoid in wine, filter
Wine is taken, then through low temperature ageing, the fragrance of fermented type Lycium chinense wine can be significantly improved, obtains flavouring Lycium chinense wine；
Preferably, the Ku Teshi bacillus is NXUGQ15 (Kurthia sp), and deposit number is CCTCC NO:
M2017524；
The fermentation fructus lycii slag is prepared by the method included the following steps:
The Ku Teshi bacillus bacterium solution that will have been spread cultivation accesses in fructus lycii slag according to the inoculum concentration of 4-6% mass percent,
35-37 DEG C, semi-solid ferment 10-15 hours under 130r/min, primary every stirring in 2 hours, must ferment fructus lycii slag, rich in dropping
Isoprenoid compound；
The preparation method of the Ku Teshi bacillus bacterium solution includes the following steps:
(1) prepared by culture medium
Fluid nutrient medium (g/L): sodium nitrate 3, dipotassium hydrogen phosphate 1, magnesium sulfate 0.5, potassium chloride 0.5, ferric sulfate 0.01, sugarcane
Sugared 30, YNB synthetic media (no amino yeast nitrogen culture medium) 6.7,15'-dioxygenase；PH=3.2.
(2) expand culture
Take slant tube Ku Teshi bacillus (Kurthia sp) 1 ring of NXUGQ15 → 35-37 DEG C of 10mL fluid nutrient medium,
It is cultivated under 130r/min and cultivates 10-12h → 3000mL liquid under 35-37 DEG C of 10-12h → 100mL fluid nutrient medium, 130r/min
10-12h → bacterial concentration is cultivated under 35-37 DEG C of culture medium, 130r/min reaches 106Cfu/ml → obtain Ku Teshi bacillus bacterium solution；
Present invention simultaneously provides it is described fermentation fructus lycii slag application, the fermentation fructus lycii slag can be applied to feed, cosmetics,
The multiple fields such as food, such as can be used for improving the fragrance of fermented type Lycium chinense wine；Such as fermentation fructus lycii slag is put into Lycium chinense wine,
Dipping dissolves in catabolite drop isoprenoid in wine, filters to take wine, then through low temperature ageing, can significantly improve hair
The fragrance of ferment type Lycium chinense wine, obtains flavouring Lycium chinense wine.
Present invention simultaneously provides a kind of flavouring Lycium chinense wines.
The separation and acquisition of the Ku Teshi bacillus strain
The strain of the isolated carotenoid that can degrade from wolfberry juice, and go out Ku Teshi bar through Uv-induced screening
Bacteria strain NXUGQ15 (Kurthia sp), using the carotenoid degrading enzyme of this plant of bacterium generation, the class in wolfberry fruit syrup of degrading is recklessly
Radish element, improves fructus lycii wine brewage technology, improves Lycium chinense wine quality.The bacterium starting strain is by Zhang Huiling from Ningxia Berry source fructus lycii
It is isolated in the wolfberry juice of limited liability company.
Ku Teshi bacillus strain NXUGQ15 (Kurthia sp), deposit number CCTCC NO:M2017524, in 2017 9
The moon is preserved in China typical culture collection center (Wuhan) on the 21st, address: the Chinese Wuhan Wuhan University.The bacterium can degrade
Beta carotene, optimum growth temp are 35-37 DEG C, pH=2-3；Degrading enzyme tolerable temperature caused by the bacterium is 70-90 DEG C,
Best enzyme reaction pH=1-3.The bacterium produces that the carotenoid degrading enzyme time is short, and enzyme activity is high, the enzyme activity 8.87U/m L of crude enzyme liquid, excellent
In bacterium germination GQ-16 out.
The identification of the Ku Teshi bacillus strain
Preliminary Identification: after fluid nutrient medium culture 48h, carrying out microscopy using microscope, in oily its form under the microscope.
The result shows that bacterial strain NXUGQ15 is Gram-positive bacillus, no gemma.
Molecular biology identification: it is analyzed using 16S-23S rDNA ISR polymorphism and sequence, constructs its systematic evolution tree
See attached drawing 2.By systematic evolution tree it is found that be based on 16rDNA region sequence phylogenetic tree, bacterial strain NXUGQ15 (Kurthia sp) with
Zuo Shi Al Kut Salmonella is got together, and illustrates bacterial strain NXUGQ15 (Kurthia sp) and Zuo Shi Al Kut Salmonella is same species, i.e. bacterium
Strain NXUGQ15 (Kurthia sp) is Al Kut Salmonella.
Degradation of the Ku Teshi bacillus to carotenoid
Bacterial strain NXUGQ15 (Kurthia sp) is inoculated into fluid nutrient medium, the β-that luteole is sole carbon source respectively
Carrotene is the fluid nutrient medium of sole carbon source, luteole acid dipalmitate is to carry out in the fluid nutrient medium of sole carbon source
Breeding metabolism.It takes 5ml fermentation liquid in 20ml ml headspace bottle after 48h, carries out solid-phase headspace microextraction and GC-MS measurement.Determination condition
Are as follows:
The solid phase microextraction of sample
It takes sample Lycium chinense wine 8ml in 20ml ml headspace bottle, 2.0g sodium chloride is added, and 8 μ L sec-n-octyl alcohol solution are added, it is permanent
40 DEG C of balance 10min on warm magnetic stirring apparatus, insertion CAR/DVB/PDMS fiber head 40 DEG C of absorption 15min, GC desorb 5min, use
It is analyzed in GC-MS.
Gaschromatographic mass spectrometry operating condition
Chromatographic condition are as follows: chromatographic column is DB-5MS (30m × 0.25mm × 0.25 μm), and carrier gas He, volume flow is
1mL/min, injector temperature are 250 DEG C.Temperature programming: 40 DEG C of holding 3min rise to 120 DEG C with the heating rate of 5 DEG C/min,
230 DEG C are risen to the heating rate of 8 DEG C/min again, keeps 10min.Filament flow is 0.20mA.Mass Spectrometry Conditions are as follows: EI ionization
Source, electron energy 70eV, detector voltage 350V.Scanning range is 20~450AMU, and ion source temperature is 200 DEG C.
Beneficial effect
The degradation bacteria Ku Teshi bacillus strain NXUGQ15 of the mutagenic obtained medlar carotenoid that can degrade of the present invention
(Kurthia sp) (deposit number CCTCC NO:M2017524).Compared to the ability that bacterium germination out improves degradation carotenoid, and
The fermentation production carotenoid degrading enzyme time is short, and enzyme activity is high, the enzyme activity 8.87U/mL of crude enzyme liquid, and producing enzyme is preferable.
Fructus lycii slag is turned waste into wealth, and makes full use of the carotenoid in fructus lycii slag
The present invention has innovated the zymotechnique of fructus lycii slag, handles fructus lycii slag using preservation of bacteria strain, recklessly to the class in fructus lycii slag
Radish element is degraded, and fructus lycii slag is turned waste into wealth, and makes full use of the carotenoid in fructus lycii slag, and carotenoid is made to degrade,
The aroma substance in fructus lycii slag is increased, can be applied to the multiple fields such as feed, cosmetics, food, is especially used for sending out
Ferment Lycium chinense wine can increase the volatilization fragrance component in Lycium chinense wine, and increase nine Carotenoids aroma substances, Neng Gougai
The quality of kind Lycium chinense wine, improves the market competitiveness of Lycium chinense wine.Reduce the waste that fermentation Lycium chinense wine generates simultaneously, reduces
Waste, reduces the difficulty of subsequent processing, energy conservation and environmental protection has great popularization value.
Detailed description of the invention
Fig. 1-the present invention ferment fructus lycii slag preparation method and present invention fermentation fructus lycii slag is used for Lycium chinense wine Titian work
Skill flow chart；
Fig. 2-Ku Teshi bacillus phyletic evolution tree graph of the present invention；
The sensory evaluation result figure of Fig. 3-three kinds of method for increasing aroma flavouring Lycium chinense wines；
In figure: wine sample 1 is that present invention fermentation fructus lycii slag is put back to gained flavouring Lycium chinense wine in Lycium chinense wine；Wine sample 2 is to use
High pressure degradation fructus lycii slag puts back to gained flavouring Lycium chinense wine in Lycium chinense wine；Wine sample 3 is the carotenoid enzyme produced using Ku Teshi bacillus
The flavouring Lycium chinense wine of assistant degradation preparation.
Specific embodiment
The present invention is described below by specific embodiment.Unless stated otherwise, technological means used in the present invention
It is method known in those skilled in the art.In addition, embodiment is interpreted as illustrative, it is not intended to limit the present invention
Range, the spirit and scope of the invention are limited only by the claims that follow.To those skilled in the art, without departing substantially from this
Under the premise of invention spirit and scope, to the various changes or change of material component and dosage progress in these embodiments
It belongs to the scope of protection of the present invention.
Embodiment 1
The separation and acquisition of Ku Teshi bacillus strain
The strain of the isolated carotenoid that can degrade from wolfberry juice, and go out Ku Teshi bar through Uv-induced screening
Bacteria strain NXUGQ15 (Kurthia sp), using the carotenoid degrading enzyme of this plant of bacterium generation, the class in wolfberry fruit syrup of degrading is recklessly
Radish element, improves fructus lycii wine brewage technology, improves Lycium chinense wine quality.The bacterium starting strain is by Zhang Huiling from Ningxia Berry source fructus lycii
It is isolated in the wolfberry juice of limited liability company.
Ku Teshi bacillus strain NXUGQ15 (Kurthia sp), deposit number CCTCC NO:M2017524, in 2017 9
The moon is preserved in China typical culture collection center (Wuhan) on the 21st, address: the Chinese Wuhan Wuhan University.The bacterium can degrade
Beta carotene, optimum growth temp are 35-37 DEG C, pH=2-3；Degrading enzyme tolerable temperature caused by the bacterium is 70-90 DEG C,
Best enzyme reaction pH=1-3.The bacterium produces that the carotenoid degrading enzyme time is short, and enzyme activity is high, the enzyme activity 8.87U/m L of crude enzyme liquid, excellent
In bacterium germination GQ-16 out.
The breeding of Ku Teshi bacillus strain
(1) inventor separated and identifies from wolfberry juice in January, 2016, had obtained the 1 plant of carotenoid that can degrade
Strain is Ku Teshi bacillus strain GQ-16.In order to improve the degradation capability of its carotenoid of degrading, which is lured
Become, see below:
(2) mutagenesis Al Kut Salmonella GQ-16
Using Al Kut Salmonella GQ-16 as starting strain, mutagenic and breeding is carried out, when purpose improves inulinase-producing activity shortening fermentation
Between.
It tests as follows:
Ultraviolet mutagenesis:
One ring of starting strain is taken, is linked into fluid nutrient medium, cultivates 5-6h under the conditions of 35 DEG C, until logarithmic phase mid-term.It will
It is 10 that bacteria suspension, which is diluted to concentration,5CFU/m L.With lethality 80% be condition by preliminary experiment, obtain best irradiation time with
Irradiation distance.Bacteria suspension is spread evenly across on fluid nutrient medium, distance 25cm under the ultraviolet lamp of 30W is placed in, irradiates 5min,
After radiation treatment, with black cloth package culture medium under the conditions of 35 DEG C -37 DEG C, 8-12h is cultivated, screening grows fast and colonial morphology
Good 20 plants of bacterial strain, then pass through fermenting property test experiments, and excellent 5 plants of bacterial strain of screenability are respectively designated as
NXUGQ15, NXUGQ46, NXUGQ7, NXUGQ18, NXUGQ39 extract each enzyme respectively and carry out fermentation of medlar wine, pass through measurement
Fermentation time come identify its fermenting property height.Fermentation time is shorter, and indicating that fermenting property is better the results are shown in Table 1.
Microwave irradiation:
5m L bacteria suspension is taken in the culture dish of diameter 9cm, with lethality 80% is condition by preliminary experiment, selects maximum
Power 700W, pulse power 2450MHz household microwave oven are irradiated 5s, are quickly cooled down 5s on ice, repeat this step, take
It states treatment fluid 0.5m L to be spread evenly across on solid medium, with black cloth package culture medium under the conditions of 35 DEG C -37 DEG C, cultivates 8-
12h.Screening grows fast and good colonial morphology 20 plants of bacterial strain, then passes through fermenting property test experiments, and screenability is excellent
5 plants of bacterial strain, be respectively designated as W-NXUGQ-5, W-NXUGQ-26, W-NXUGQ-1, W-NXUGQ-18, W-NXUGQ-35, respectively
It extracts each enzyme and carries out fermentation of medlar wine, its fermenting property height is identified by measurement fermentation time.Fermentation time is shorter, table
Showing that fermenting property is better the results are shown in Table 2.
Enzyme activity definition: under the conditions of 50 DEG C, p H 3.5, the amount that 1h decomposes carotenoid is one for 1g enzyme powder or 1m L enzyme solution
A enzyme-activity unit (U).The enzyme activity calculation formula of crude enzyme liquid is as follows:
In formula: Y is the quality .mg of enzyme effect degradation carotenoid；N is sample extension rate；2 be when measuring enzyme activity
The 1/2 of reaction solution is taken；T is to react time .h used.
The influence result of strain enzyme-producing (carotenoid degrading enzyme) is distinguished in ultraviolet mutagenesis processing and microwave irradiation processing
It is shown in Table 1 and table 2.
The processing of 1 ultraviolet mutagenesis of table
| Number | NXUGQ 15 | NXUGQ46 | NXUGQ 7 | NXUGQ 18 | NXUGQ39 | Bacterium germination GQ-16 out |
| Enzyme activity U/mL | 8.87 | 7.90 | 8.46 | 8.84 | 8.12 | 6.32 |
| Fermentation time | 8h | 13h | 10h | 12h | 11h | 10h |
The processing of 2 microwave irradiation of table
| Number | W-NXUGQ-5 | W-NXUGQ-26 | W-NXUGQ-1 | W-NXUGQ-18 | W-NXUGQ-35 | Bacterium germination GQ-16 out |
| Enzyme activity U/mL | 7.87 | 7.89 | 7.46 | 8.05 | 8.12 | 6.32 |
| Fermentation time | 11h | 8h | 10h | 10h | 14h | 10h |
As can be seen from the above two tables, preferably, microwave irradiation handles 5 plants to NXUGQ 15 in 5 plants of bacterium of ultraviolet mutagenesis processing
W-NXUGQ-5 is best in bacterium,
It is carried out stability test 15 times with two plants of bacterium respectively, by comparing finding that NXUGQ 15 is more stable, compared to setting out
Bacterium improves the ability of degradation carotenoid, and the production carotenoid degrading enzyme time of fermenting is short, and enzyme activity is high, the enzyme activity of crude enzyme liquid
8.87U/mL, producing enzyme are preferable.The strain is selected, NXUGQ15, i.e. Ku Teshi bacillus strain NXUGQ15 are finally named as
(Kurthia sp), and carry out culture presevation.
Embodiment 2
A kind of preparation method for the fructus lycii slag that ferments, includes the following steps:
By Ku Teshi bacillus NXUGQ15 (Kurthia sp), deposit number is CCTCC NO:M2017524, expands culture,
Obtain Ku Teshi bacillus bacterium solution；
Fresh fructus lycii mashing, juicing, filtering, obtain wolfberry juice and fructus lycii slag；
The Ku Teshi bacillus bacterium solution that will have been spread cultivation accesses in fructus lycii slag according to 5% inoculum concentration, in 35-37 DEG C, 130r/
Semi-solid ferment 12 hours under min, primary every stirring in 2 hours, must ferment fructus lycii slag；
The preparation method of the Ku Teshi bacillus bacterium solution includes the following steps:
(1) prepared by culture medium
Fluid nutrient medium (g/L): sodium nitrate 3, dipotassium hydrogen phosphate 1, magnesium sulfate 0.5, potassium chloride 0.5, ferric sulfate 0.01, sugarcane
Sugared 30, YNB synthetic media (no amino yeast nitrogen culture medium) 6.7,15'-dioxygenase；PH=3.2.
(2) expand culture
Take slant tube Ku Teshi bacillus (Kurthia sp) 1 ring of NXUGQ15 → 35-37 DEG C of 10mL fluid nutrient medium,
It is cultivated under 130r/min and cultivates 11h → 3000mL fluid nutrient medium under 35-37 DEG C of 11h → 100mL fluid nutrient medium, 130r/min
35-37 DEG C, culture 11h → bacterial concentration reaches 10 under 130r/min6Cfu/ml → obtain Ku Teshi bacillus bacterium solution；
Fermentation fructus lycii slag is played back in Lycium chinense wine, 1h is impregnated, dissolves in catabolite drop isoprenoid in wine, mistake
Leaching wine, then through low temperature ageing, the fragrance of fermented type Lycium chinense wine can be significantly improved, obtain flavouring Lycium chinense wine；
Flavor substance and sensory evaluation are measured, the results are shown in Table 3, table 4 and attached drawing 3.
Embodiment 3
A kind of preparation method for the fructus lycii slag that ferments, includes the following steps:
Fresh fructus lycii mashing, juicing, filtering, obtain wolfberry juice and fructus lycii slag；
By Ku Teshi bacillus NXUGQ15 (Kurthia sp), deposit number is CCTCC NO:M2017524, expands culture,
Obtain Ku Teshi bacillus bacterium solution；
The Ku Teshi bacillus bacterium solution that will have been spread cultivation accesses in fructus lycii slag according to 4% inoculum concentration, in 35-37 DEG C, 130r/
Semi-solid ferment 10 hours under min, primary every stirring in 2 hours, must ferment fructus lycii slag；
The preparation method of the Ku Teshi bacillus bacterium solution includes the following steps:
(1) prepared by culture medium
Fluid nutrient medium (g/L): sodium nitrate 3, dipotassium hydrogen phosphate 1, magnesium sulfate 0.5, potassium chloride 0.5, ferric sulfate 0.01, sugarcane
Sugared 30, YNB synthetic media (no amino yeast nitrogen culture medium) 6.7,15'-dioxygenase；PH=3.2.
(2) expand culture
Take slant tube Ku Teshi bacillus (Kurthia sp) 1 ring of NXUGQ15 → 35-37 DEG C of 10mL fluid nutrient medium,
It is cultivated under 130r/min and cultivates 10h → 3000mL fluid nutrient medium under 35-37 DEG C of 10h → 100mL fluid nutrient medium, 130r/min
35-37 DEG C, culture 10h → bacterial concentration reaches 10 under 130r/min6Cfu/ml → obtain Ku Teshi bacillus bacterium solution.
Embodiment 4
A kind of preparation method for the fructus lycii slag that ferments, includes the following steps:
By Ku Teshi bacillus NXUGQ15 (Kurthia sp), deposit number is CCTCC NO:M2017524, expands culture,
Obtain Ku Teshi bacillus bacterium solution；
Fresh fructus lycii mashing, juicing, filtering, obtain wolfberry juice and fructus lycii slag；
The Ku Teshi bacillus bacterium solution that will have been spread cultivation accesses in fructus lycii slag according to 6% inoculum concentration, in 35-37 DEG C, 130r/
Semi-solid ferment 15 hours under min, primary every stirring in 2 hours, must ferment fructus lycii slag；
The preparation method of the Ku Teshi bacillus bacterium solution includes the following steps:
(1) prepared by culture medium
Fluid nutrient medium (g/L): sodium nitrate 3, dipotassium hydrogen phosphate 1, magnesium sulfate 0.5, potassium chloride 0.5, ferric sulfate 0.01, sugarcane
Sugared 30, YNB synthetic media (no amino yeast nitrogen culture medium) 6.7,15'-dioxygenase；PH=3.2.
(2) expand culture
Take slant tube Ku Teshi bacillus (Kurthia sp) 1 ring of NXUGQ15 → 35-37 DEG C of 10mL fluid nutrient medium,
It is cultivated under 130r/min and cultivates 12h → 3000mL fluid nutrient medium under 35-37 DEG C of 12h → 100mL fluid nutrient medium, 130r/min
35-37 DEG C, culture 12h → bacterial concentration reaches 10 under 130r/min6Cfu/ml → obtain Ku Teshi bacillus bacterium solution.
Test example
Flavor substance measurement
It drops the extraction of isoprenoid compound: carrying out fragrance enrichment using headspace solid-phase microextraction (HS-SPME) method, take
One 20mL ml headspace bottle, with Sterile pipette plus 8mL wolfberry juice to be analyzed or Lycium chinense wine sample and 2.0g NaCl, in 40 DEG C of perseverances
10min is balanced on warm magnetic stirring apparatus, 40 DEG C of absorption 15min, GC desorption 5min of insertion CAR/DVB/PDMS fiber head are used for
GC-MS analysis.
Gaschromatographic mass spectrometry operating condition
Chromatographic condition are as follows: chromatographic column be DB-5MS (30m × 0.25mm × 0.25 μm), temperature programming: 40 DEG C of holding 3min,
120 DEG C are risen to the heating rate of 5 DEG C/min, then rises to 230 DEG C with the heating rate of 8 DEG C/min, keeps 10min.Carrier gas is
He, volume flow 1mL/min, injector temperature are 250 DEG C.Mass Spectrometry Conditions are as follows: EI ionization source, electron energy 70eV, lamp
Silk flow is 0.20mA.Detector voltage is 350V.Scanning range is 20~450AMU, and ion source temperature is 200 DEG C.
Analysis method:
(1) analysis method of carotenoid
It is calculated before processing by standard curve respectively and the content of each single factor test treated carotenoid.Substitute into following formula
In calculate the degradation rates of all kinds of carrotene.
Degradation rate (%)=[(content after content-processing before handling)/content before handling] × 100%
(2) analysis method of isoprenoid drops
According to the structure feature of compound, it is compared with the map of NIST spectrum library Plays compound, according to ion stream
Figure determines its relative amount.
By the present invention fermentation fructus lycii slag be used to ferment Lycium chinense wine Titian and other two method preparation Lycium chinense wine volatilize fragrance
Comparison of ingredients, the Lycium chinense wine with traditional handicraft fermentation is control:
Method I: the method for the present invention uses NXUGQ15 (Kurthia sp) bacterium direct fermentation fructus lycii slag, drops carotenoid
Solution plays back slag in Lycium chinense wine, dissolves in catabolite drop isoprenoid in wine, filters to take wine.
Method II: using NXUGQ15 (Kurthia sp) bacterium, by fermented and cultured bacterium solution, collects the class of bacterium generation recklessly
Degrading enzyme is added in wolfberry fruit syrup and digests by radish element degrading enzyme, and then fermentation prepares Lycium chinense wine, and catabolite is made to drop isoamyl
Diolefin compound dissolves in wine, filters to take wine.
Method III: using in 121 DEG C of sterilizing 20min HIGH PRESSURE TREATMENTs fructus lycii slag, and carotenoid in fructus lycii slag is made to degrade,
Then it is dipped in 0.5-1h in Lycium chinense wine again, makes to drop in isoprenoid dissolution wine, filters to take wine.
Lycium chinense wine with traditional handicraft fermentation is control, and method I, method II, method III is respectively adopted and carries out Lycium chinense wine
Flavouring, Lycium chinense wine obtained mainly volatilizees fragrance component and relative amount comparison result is shown in Table 3:
Mainly volatilize fragrance component and relative amount in 3 different process Lycium chinense wine of table
Note: " nd " expression is not detected.
By upper table it can be concluded that mainly being waved with the Lycium chinense wine after the fermentation fructus lycii slag flavouring of the method for the present invention preparation
Hair fragrance component type and content is much higher than the Lycium chinense wine of traditional handicraft preparation, and increases nine Carotenoids fragrance objects
Matter.
Lycium chinense wine carbonyl compound content of the present invention increases, and relative peak area accounts for the 5.78% of total ingredient, is higher than traditional work
The 4.23% of skill；Wherein content it is higher be 3- undecyl ketone, dorinone, alpha, beta-lonone, β-cyclocitral, safranal and
Dihydro-β-ionone, the higher alpha, beta-lonone of relative amount 4.22%, β-cyclocitral at least higher than traditional handicraft is at least
High by 1.17%, they have special fruit aroma.
Ester type compound relative peak area be 14.08%, higher than the 11.03% of prior art, be in Lycium chinense wine content compared with
More aromatic substances.The 13.68% of total ingredient is accounted for, higher than the 10.73% of prior art, larger, certain herbaceous plants with big flowers is contributed to fructus lycii wine aroma
Acetoacetic ester and amyl valerate are also important catabolite, the fragrance with pears perfume (or spice) and banana；Dihydroactinidiolide, acetone are fragrant
Leaf ester is one of intermediate product and the higher esters of content of carotenoid degradation, and fruit and flowery odour are prominent.
Drop isoprene kind compound content compares in the Lycium chinense wine of three kinds of methods preparation
Lycium chinense wine with traditional handicraft fermentation is control, and method I, method II, method III is respectively adopted and carries out Lycium chinense wine
Flavouring, drop isoprenoid compound is compared in Lycium chinense wine obtained, the results are shown in Table 4
Drop isoprenoid compound compares (peak area) in the Lycium chinense wine of 4 different process of table preparation
Such as drawn a conclusion by table 4 is available: fermentation fructus lycii slag prepared by the present invention is added in Lycium chinense wine after dipping, drops isoamyl
Diolefinic compounds content is much higher than the Lycium chinense wine of traditional handicraft preparation, also above the Lycium chinense wine of high-pressure treatment process preparation.This
Invention, which increases, drops isoprene kind compound content and type in Lycium chinense wine, they are mainly alpha, beta-lonone, 2,2,6-
Trimethylcyclohexanone (TCH), isophorone, geranyl acetone, dihydro-β-ionone, geraniol, safranal, β-ring lemon
Lemon aldehyde, limonene, pseudoionone, dihydroactinidiolide.And aldehyde C-9, amyl valerate, certain herbaceous plants with big flowers acetoacetic ester.The taste of these substances
Threshold value is low, plays a good role for Lycium chinense wine sensory evaluation.
Fragrance control is referring to table:
The odor characteristic of isoprenoid compound partially drops in 5 Lycium chinense wine of table
According to the sensory evaluation method of following table, prepared by III 3 kinds of the method for the present invention II, method I, method distinct methods
Lycium chinense wine carries out sensory evaluation, judges result and draws sensory evaluation wind rose, sees attached drawing 3
6 fermentation Chinese wolfberry fruit wine sensory evaluation scores table of table
By finding prepared by the present invention to the measurement for dropping pentadiene class compound in three kinds of wine samples and in conjunction with sensory evaluation
The fructus lycii slag that ferments is used for Lycium chinense wine Titian, and fragrance and mouthfeel are preferable.Fructus lycii slag, first one side are fermented not using Al Kut Salmonella
The degradation rate of carotenoid only can be improved, to generate more drop pentadiene fragrance components.Secondly, fermentation temperature is low, keep away
Exempt to cause bad flavor to wine body because of high-temperature heating.
The present invention carries out degradation treatment using Al Kut Salmonella during fructus lycii liquor brewing, by fructus lycii slag, can improve
The brewage process of Lycium chinense wine improves fructus lycii wine aroma.And can appropriateness carry out high pressure sterilization, or using Al Kut Salmonella produce
Raw carotenoid enzyme aid in treatment wolfberry fruit syrup can significantly improve fermentation fructus lycii so that several method for increasing aroma are combined
The fragrance of wine.
The embodiments described above only express several embodiments of the present invention, and the description thereof is more specific and detailed, but simultaneously
The limitation to the scope of the patents therefore cannot be interpreted as.It should be pointed out that for those of ordinary skill in the art,
Under the premise of not departing from present inventive concept, the respective embodiments described above can also make combination and improvement, naturally it is also possible to send out this
Bright disclosed several preparation methods are reconfigured and the increase and decrease of processing step, recombination, these belong to guarantor of the invention
Protect range.
Claims (10)
1. a kind of fermentation fructus lycii slag is prepared by the method included the following steps:
Fresh fructus lycii mashing, juicing, filtering, obtain fructus lycii slag；
Ku Teshi bacillus liquid is added in fructus lycii slag, ferment to obtain fermentation fructus lycii slag；
The Ku Teshi bacillus liquid is that Ku Teshi bacillus (Kurthia sp) is expanded to culture gained.
2. fermentation fructus lycii slag according to claim 1, which is characterized in that the Ku Teshi bacillus is (Kurthia sp)
NXUGQ15, deposit number are CCTCC NO:M2017524.
3. fermentation fructus lycii slag according to claim 1 or 2, which is characterized in that the preparation method includes the following steps:
The Ku Teshi bacillus liquid that will have been spread cultivation, according to 4-6% mass percent inoculum concentration access fructus lycii slag in, 35-37 DEG C,
Semi-solid ferment 10-15 hours under 130r/min, primary every stirring in 2 hours, must ferment fructus lycii slag.
4. fermentation fructus lycii slag according to claim 3, which is characterized in that the preparation method of the Ku Teshi bacillus liquid includes
Following steps:
(1) prepared by culture medium
Fluid nutrient medium (g/L): sodium nitrate 3, dipotassium hydrogen phosphate 1, magnesium sulfate 0.5, potassium chloride 0.5, ferric sulfate 0.01, sucrose
30, YNB synthetic media 6.7,15'-dioxygenase；PH=3.2；
(2) expand culture
Take slant tube Ku Teshi bacillus (Kurthia sp) NXUGQ151 ring → 35-37 DEG C of 10mL fluid nutrient medium, 130r/
It is cultivated under min and cultivates 10-12h → 3000mL Liquid Culture under 35-37 DEG C of 10-12h → 100mL fluid nutrient medium, 130r/min
10-12h → bacterial concentration is cultivated under 35-37 DEG C of base, 130r/min reaches 106Cfu/ml → obtain Ku Teshi bacillus liquid.
5. fermentation fructus lycii slag according to claim 4, which is characterized in that the preparation method includes the following steps:
By Ku Teshi bacillus NXUGQ15 (Kurthia sp), deposit number is CCTCC NO:M2017524, expands culture, obtains library
Te Shi bacillus bacterium solution；
Fresh fructus lycii mashing, juicing, filtering, obtain wolfberry juice and fructus lycii slag；
The Ku Teshi bacillus bacterium solution that will have been spread cultivation, according to 5% mass percent inoculum concentration access fructus lycii slag in, 35-37 DEG C,
Semi-solid ferment 12 hours under 130r/min, primary every stirring in 2 hours, must ferment fructus lycii slag；
The preparation method of the Ku Teshi bacillus bacterium solution includes the following steps:
(1) prepared by culture medium
Fluid nutrient medium (g/L): sodium nitrate 3, dipotassium hydrogen phosphate 1, magnesium sulfate 0.5, potassium chloride 0.5, ferric sulfate 0.01, sucrose
30, YNB synthetic media 6.7,15'-dioxygenase；PH=3.2；
(2) expand culture
Take slant tube Ku Teshi bacillus (Kurthia sp) 1 ring of NXUGQ15 → 35-37 DEG C of 10mL fluid nutrient medium, 130r/
It is cultivated under min and cultivates 11h → 3000mL fluid nutrient medium 35-37 under 35-37 DEG C of 11h → 100mL fluid nutrient medium, 130r/min
DEG C, culture 11h → bacterial concentration reaches 10 under 130r/min6Cfu/ml → obtain Ku Teshi bacillus bacterium solution.
6. fermentation fructus lycii slag according to claim 4, which is characterized in that the preparation method includes the following steps:
Fresh fructus lycii mashing, juicing, filtering, obtain wolfberry juice and fructus lycii slag；
By Ku Teshi bacillus NXUGQ15 (Kurthia sp), deposit number is CCTCC NO:M2017524, expands culture, obtains library
Te Shi bacillus bacterium solution；
The Ku Teshi bacillus bacterium solution that will have been spread cultivation accesses in fructus lycii slag according to 4% inoculum concentration, at 35-37 DEG C, 130r/min
Semi-solid ferment 10 hours, primary every stirring in 2 hours, must ferment fructus lycii slag；
The preparation method of the Ku Teshi bacillus bacterium solution includes the following steps:
(1) prepared by culture medium
Fluid nutrient medium (g/L): sodium nitrate 3, dipotassium hydrogen phosphate 1, magnesium sulfate 0.5, potassium chloride 0.5, ferric sulfate 0.01, sucrose
30, YNB synthetic media 6.7,15'-dioxygenase；PH=3.2；
(2) expand culture
Take slant tube Ku Teshi bacillus (Kurthia sp) 1 ring of NXUGQ15 → 35-37 DEG C of 10mL fluid nutrient medium, 130r/
It is cultivated under min and cultivates 10h → 3000mL fluid nutrient medium 35-37 under 35-37 DEG C of 10h → 100mL fluid nutrient medium, 130r/min
DEG C, culture 10h → bacterial concentration reaches 10 under 130r/min6Cfu/ml → obtain Ku Teshi bacillus bacterium solution.
7. fermentation fructus lycii slag according to claim 4, which is characterized in that the preparation method includes the following steps:
By Ku Teshi bacillus NXUGQ15 (Kurthia sp), deposit number is CCTCC NO:M2017524, expands culture, obtains library
Te Shi bacillus bacterium solution；
Fresh fructus lycii mashing, juicing, filtering, obtain wolfberry juice and fructus lycii slag；
The Ku Teshi bacillus bacterium solution that will have been spread cultivation accesses in fructus lycii slag according to 6% inoculum concentration, at 35-37 DEG C, 130r/min
Semi-solid ferment 15 hours, primary every stirring in 2 hours, must ferment fructus lycii slag；
The preparation method of the Ku Teshi bacillus bacterium solution includes the following steps:
(1) prepared by culture medium
Fluid nutrient medium (g/L): sodium nitrate 3, dipotassium hydrogen phosphate 1, magnesium sulfate 0.5, potassium chloride 0.5, ferric sulfate 0.01, sucrose
30, YNB synthetic media 6.7,15'-dioxygenase；PH=3.2；
(2) expand culture
Take slant tube Ku Teshi bacillus (Kurthia sp) 1 ring of NXUGQ15 → 35-37 DEG C of 10mL fluid nutrient medium, 130r/
It is cultivated under min and cultivates 12h → 3000mL fluid nutrient medium 35-37 under 35-37 DEG C of 12h → 100mL fluid nutrient medium, 130r/min
DEG C, culture 12h → bacterial concentration reaches 10 under 130r/min6Cfu/ml → obtain Ku Teshi bacillus bacterium solution.
8. application of any fermentation fructus lycii slag of claim 1-7 in feed or cosmetics or food.
9. application of any fermentation fructus lycii slag of claim 1-7 in fermentation Lycium chinense wine: fermentation fructus lycii slag is put into fructus lycii
In wine, wine is filtered to take after dipping, then through low temperature ageing, obtain flavouring Lycium chinense wine.
10. the flavouring Lycium chinense wine that application obtains according to claim 9.
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| 吴秀玲: "《微生物制药技术》", 31 January 2015, 中国轻工业出版社 * |
| 李勇等: "《枸杞皮渣的综合利用》", 《中国酿造》 * |
| 李金鹏等: "一株降解β-胡萝卜素细菌的分离鉴定及产酶条件优化", 《中国酿造》 * |
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