CN109706193A - Vitamin K in a kind of enrichment bacillus natto to ferment liquid2Method - Google Patents
Vitamin K in a kind of enrichment bacillus natto to ferment liquid2Method Download PDFInfo
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- CN109706193A CN109706193A CN201910183364.0A CN201910183364A CN109706193A CN 109706193 A CN109706193 A CN 109706193A CN 201910183364 A CN201910183364 A CN 201910183364A CN 109706193 A CN109706193 A CN 109706193A
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- 244000063299 Bacillus subtilis Species 0.000 title claims abstract description 43
- 235000014469 Bacillus subtilis Nutrition 0.000 title claims abstract description 43
- SHUZOJHMOBOZST-UHFFFAOYSA-N phylloquinone Natural products CC(C)CCCCC(C)CCC(C)CCCC(=CCC1=C(C)C(=O)c2ccccc2C1=O)C SHUZOJHMOBOZST-UHFFFAOYSA-N 0.000 title claims abstract description 27
- 229930003448 Vitamin K Natural products 0.000 title claims abstract description 26
- 239000011712 vitamin K Substances 0.000 title claims abstract description 26
- 235000019168 vitamin K Nutrition 0.000 title claims abstract description 26
- 150000003721 vitamin K derivatives Chemical class 0.000 title claims abstract description 26
- 229940046010 vitamin k Drugs 0.000 title claims abstract description 26
- 239000007788 liquid Substances 0.000 claims abstract description 52
- 239000002077 nanosphere Substances 0.000 claims abstract description 35
- 238000000034 method Methods 0.000 claims abstract description 29
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 claims abstract description 29
- 239000002086 nanomaterial Substances 0.000 claims abstract description 24
- 239000006228 supernatant Substances 0.000 claims abstract description 24
- 239000011728 vitamin K2 Substances 0.000 claims abstract description 21
- 238000010521 absorption reaction Methods 0.000 claims abstract description 14
- 239000000696 magnetic material Substances 0.000 claims abstract description 12
- 239000012074 organic phase Substances 0.000 claims abstract description 7
- 238000001914 filtration Methods 0.000 claims abstract description 6
- 238000002604 ultrasonography Methods 0.000 claims abstract description 6
- 239000000706 filtrate Substances 0.000 claims abstract description 5
- SZVJSHCCFOBDDC-UHFFFAOYSA-N iron(II,III) oxide Inorganic materials O=[Fe]O[Fe]O[Fe]=O SZVJSHCCFOBDDC-UHFFFAOYSA-N 0.000 claims description 51
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N silicon dioxide Inorganic materials O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 28
- 229910052681 coesite Inorganic materials 0.000 claims description 20
- 229910052906 cristobalite Inorganic materials 0.000 claims description 20
- 229910052682 stishovite Inorganic materials 0.000 claims description 20
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- 239000000243 solution Substances 0.000 claims description 17
- 238000000855 fermentation Methods 0.000 claims description 15
- 230000004151 fermentation Effects 0.000 claims description 15
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 claims description 14
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 12
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- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 10
- OAKJQQAXSVQMHS-UHFFFAOYSA-N Hydrazine Chemical compound NN OAKJQQAXSVQMHS-UHFFFAOYSA-N 0.000 claims description 10
- 238000001179 sorption measurement Methods 0.000 claims description 9
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- -1 hydrocarbyl succinimide Chemical compound 0.000 claims description 7
- 238000003786 synthesis reaction Methods 0.000 claims description 7
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- 238000002156 mixing Methods 0.000 claims description 6
- 229910052757 nitrogen Inorganic materials 0.000 claims description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 6
- FPQQSJJWHUJYPU-UHFFFAOYSA-N 3-(dimethylamino)propyliminomethylidene-ethylazanium;chloride Chemical compound Cl.CCN=C=NCCCN(C)C FPQQSJJWHUJYPU-UHFFFAOYSA-N 0.000 claims description 5
- BOTDANWDWHJENH-UHFFFAOYSA-N Tetraethyl orthosilicate Chemical compound CCO[Si](OCC)(OCC)OCC BOTDANWDWHJENH-UHFFFAOYSA-N 0.000 claims description 5
- 239000002253 acid Substances 0.000 claims description 5
- 238000005119 centrifugation Methods 0.000 claims description 5
- 239000008367 deionised water Substances 0.000 claims description 5
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- KZNICNPSHKQLFF-UHFFFAOYSA-N dihydromaleimide Natural products O=C1CCC(=O)N1 KZNICNPSHKQLFF-UHFFFAOYSA-N 0.000 claims description 5
- 150000003949 imides Chemical class 0.000 claims description 5
- 239000011259 mixed solution Substances 0.000 claims description 5
- 235000013557 nattō Nutrition 0.000 claims description 5
- 229960002317 succinimide Drugs 0.000 claims description 5
- 238000005406 washing Methods 0.000 claims description 5
- 241000193830 Bacillus <bacterium> Species 0.000 claims description 4
- 238000007605 air drying Methods 0.000 claims description 4
- JJIPIYZWDNKHCN-UHFFFAOYSA-N azane;propan-2-ol;hydrate Chemical compound N.O.CC(C)O JJIPIYZWDNKHCN-UHFFFAOYSA-N 0.000 claims description 4
- 229910052799 carbon Inorganic materials 0.000 claims description 4
- 229960000935 dehydrated alcohol Drugs 0.000 claims description 4
- 239000008394 flocculating agent Substances 0.000 claims description 4
- 229910021389 graphene Inorganic materials 0.000 claims description 4
- 239000004005 microsphere Substances 0.000 claims description 4
- 229920000620 organic polymer Polymers 0.000 claims description 4
- 229910021642 ultra pure water Inorganic materials 0.000 claims description 4
- 239000012498 ultrapure water Substances 0.000 claims description 4
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- 229920002678 cellulose Polymers 0.000 claims description 3
- 239000010451 perlite Substances 0.000 claims description 3
- 235000019362 perlite Nutrition 0.000 claims description 3
- 241000726221 Gemma Species 0.000 claims description 2
- NLYAJNPCOHFWQQ-UHFFFAOYSA-N kaolin Chemical compound O.O.O=[Al]O[Si](=O)O[Si](=O)O[Al]=O NLYAJNPCOHFWQQ-UHFFFAOYSA-N 0.000 claims description 2
- 230000000694 effects Effects 0.000 abstract description 4
- 239000000463 material Substances 0.000 abstract description 4
- 230000008901 benefit Effects 0.000 abstract description 3
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 15
- 239000003153 chemical reaction reagent Substances 0.000 description 7
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 6
- 238000012545 processing Methods 0.000 description 4
- 238000003756 stirring Methods 0.000 description 4
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 3
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- 238000012986 modification Methods 0.000 description 3
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- 229940088594 vitamin Drugs 0.000 description 3
- 229930003231 vitamin Natural products 0.000 description 3
- 235000013343 vitamin Nutrition 0.000 description 3
- 239000011782 vitamin Substances 0.000 description 3
- 150000003722 vitamin derivatives Chemical class 0.000 description 3
- WYTZZXDRDKSJID-UHFFFAOYSA-N (3-aminopropyl)triethoxysilane Chemical compound CCO[Si](OCC)(OCC)CCCN WYTZZXDRDKSJID-UHFFFAOYSA-N 0.000 description 2
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 description 2
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- LQJBNNIYVWPHFW-UHFFFAOYSA-N 20:1omega9c fatty acid Natural products CCCCCCCCCCC=CCCCCCCCC(O)=O LQJBNNIYVWPHFW-UHFFFAOYSA-N 0.000 description 1
- QCVGEOXPDFCNHA-UHFFFAOYSA-N 5,5-dimethyl-2,4-dioxo-1,3-oxazolidine-3-carboxamide Chemical compound CC1(C)OC(=O)N(C(N)=O)C1=O QCVGEOXPDFCNHA-UHFFFAOYSA-N 0.000 description 1
- QSBYPNXLFMSGKH-UHFFFAOYSA-N 9-Heptadecensaeure Natural products CCCCCCCC=CCCCCCCCC(O)=O QSBYPNXLFMSGKH-UHFFFAOYSA-N 0.000 description 1
- 102000002322 Egg Proteins Human genes 0.000 description 1
- 108010000912 Egg Proteins Proteins 0.000 description 1
- 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 description 1
- 229910021578 Iron(III) chloride Inorganic materials 0.000 description 1
- ABSPRNADVQNDOU-UHFFFAOYSA-N Menaquinone 1 Natural products C1=CC=C2C(=O)C(CC=C(C)C)=C(C)C(=O)C2=C1 ABSPRNADVQNDOU-UHFFFAOYSA-N 0.000 description 1
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- RBTARNINKXHZNM-UHFFFAOYSA-K iron trichloride Chemical compound Cl[Fe](Cl)Cl RBTARNINKXHZNM-UHFFFAOYSA-K 0.000 description 1
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- FRASJONUBLZVQX-UHFFFAOYSA-N naphthoquinone group Chemical group C1(C=CC(C2=CC=CC=C12)=O)=O FRASJONUBLZVQX-UHFFFAOYSA-N 0.000 description 1
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- ZQPPMHVWECSIRJ-KTKRTIGZSA-N oleic acid Chemical compound CCCCCCCC\C=C/CCCCCCCC(O)=O ZQPPMHVWECSIRJ-KTKRTIGZSA-N 0.000 description 1
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- MBWXNTAXLNYFJB-NKFFZRIASA-N phylloquinone Chemical compound C1=CC=C2C(=O)C(C/C=C(C)/CCC[C@H](C)CCC[C@H](C)CCCC(C)C)=C(C)C(=O)C2=C1 MBWXNTAXLNYFJB-NKFFZRIASA-N 0.000 description 1
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Abstract
Vitamin K in a kind of enrichment bacillus natto to ferment liquid2Method, including the following steps: step 1: bacillus natto to ferment liquid is centrifuged, collect supernatant；Step 2: flocculant and filter aid being added into the supernatant that step 1 obtains, is then stirred, is then centrifuged, collects supernatant；Step 3: the Fe being reduced being added into the supernatant of step 23O4@SiO2- G nanosphere adjusts pH value to 6.0-8.0, is then stirred 10min-30min under the conditions of 20 DEG C -35 DEG C, then collects the magnetic material after absorption；Step 4: n-hexane being added into the magnetic material after absorption, carries out ultrasound parsing 5-20min, magnetic Nano material is then collected by additional magnet, organic phase is then poured out into filtering, gained filtrate is to be enriched vitamin K2Hexane solution.The present invention can rapidly and efficiently enrichment bacillus natto to ferment liquid in VK2.Specific steps are simple, material price is cheap and advantage reusable, that concentration effect is efficient.
Description
Technical field
The present invention relates to vitamin Ks in a kind of enrichment bacillus natto to ferment liquid2Method, belong to bioengineering and life
Object separation engineering field.
Background technique
Vitamin K2It is a series of isoprene containing 2- methyl-1,4-naphthaquinone parent nucleus and C3 with number not etc.
The general designation of the terpenes side chain compound of structural unit can be divided into K2 (10), K2 according to the number of carbon on terpenes side chain
(20), K2 (35), K2 (40) etc..Vitamin K2It is a kind of liposoluble vitamin, the naphthoquinones group with phylloquinone bioactivity
Derivative, be one of indispensable important vitamin in human body.It with the research to its function, has been demonstrated that, vitamin
K2Have the function of the diseases such as pre- preventing bone rarefaction, angiocarpy, Parkinson, liver cancer.Therefore, functional food, medical products, makeup
The fields such as product are to vitamin K2Demand increasingly increase, good market prospect.
Vitamin K2With chiral structure, trans-vitamin K2With bioactivity, cis vitamin K2It is living without biology
Property.Vitamin K made from chemical synthesis2Contain cis and trans simultaneously；In contrast, microbe fermentation method, it is available
Alltrans vitamin K2.Therefore, it has provided in the world, for the vitamin K in functional food2Microbial fermentation must be derived from.
Currently, bacillus natto to ferment prepares vitamin K2It is a kind of microbial fermentation production vitamin K generally used2Method.
Bacillus natto to ferment prepares vitamin K2In, the metabolite of bafillus natto is more, simultaneously because its
Aqtocytolysis can generate plurality of impurities, such as foreign protein, nucleic acid, grease, cause to obtain High Purity vitamin K2Difficulty compared with
Greatly.
Currently, organic solvent, which is widely used, extracts vitamin K from bacillus natto to ferment liquid2, this method need into
Pretreatment operations, the subsequent purification process such as the crushing of row thallus, dehydration and drying also need to carry out macroreticular resin and reverse phase silica gel etc.
Processing, integrated artistic is complicated, and expensive, recovery rate is lower.Extraction purification technology, it has also become bacillus natto to ferment preparation
Vitamin K2Bottleneck problem.
Summary of the invention
The object of the invention is that providing vitamin K in a kind of enrichment bacillus natto to ferment liquid2Method.
In order to achieve the above objects and other related objects, present invention provide the technical scheme that a kind of enrichment natto gemma
Vitamin K in bacillus fermentation liquid2Method, including the following steps:
Step 1: bacillus natto to ferment liquid being centrifuged, supernatant is collected；
Step 2: flocculant and filter aid are added into the supernatant that step 1 obtains, is then stirred, is then centrifuged,
Collect supernatant；
Step 3: the Fe being reduced being added into the supernatant of step 23O4@SiO2- G nanosphere, adjusting pH value to 6.0-8.0,
Then it is stirred 10min-30min under the conditions of 20 DEG C -35 DEG C, then collects the magnetic material after absorption；The quilt
The Fe of reduction3O4@SiO2The preparation method of-G nanosphere includes the following steps:
S1, by Fe3O4Magnetic microsphere is added to the mixed solution being made of deionized water, isopropanol ammonium hydroxide and ethyl orthosilicate
In, it reacts 10-20h at room temperature after mixing evenly, collects the nanosphere of synthesis, obtain after washing by SiO2Package
Fe3O4@SiO2-NH2；
S2, Xiang You SiO2The Fe of package3O4@SiO2-NH2Middle addition graphene oxide, dimethylformamide, 1-(3- dimethylamino
Propyl) -3- ethyl-carbodiimide hydrochloride and hydrocarbyl succinimide acid imide, pH value is controlled in 4-6, is reacted at room temperature
1-3h obtains Fe3O4@SiO2-GO；Then by Fe3O4@SiO2- GO with hydrazine reacts 1-3h at 75-85 DEG C to be reduced
Fe3O4@SiO2- G nanosphere；
Step 4: n-hexane being added into the magnetic material after absorption, carries out ultrasound parsing 5-20min, then passes through additional magnet
Magnetic Nano material is collected, organic phase is then poured out into filtering, gained filtrate is to be enriched vitamin K2Hexane solution.
Preferred technical solution are as follows: the vitamin K of the bacillus natto to ferment liquid2Content is 30-100mg/L, residual
Sugared content is 0.01-2g/L, residual nitrogen content is 3-8g/L, surface tension 18-24Mn/m.
Preferred technical solution are as follows: in step 1, the revolving speed of centrifugation is 5000-8000rpm, and the time of centrifugally operated is 10-
20min, centrifugally operated carry out in the 2h after the bacillus natto to ferment liquid is made.
Preferred technical solution are as follows: the flocculant is at least one in inorganic flocculating agent and organic polymer coargulator
Kind, the dosage of the flocculant is the 3-30% of bacillus natto to ferment liquid quality.
Preferred technical solution are as follows: the flocculant is chitosan.
Preferred technical solution are as follows: the filter aid is diatomite, perlite, cellulose, graphite powder, active carbon and acidity
At least one of carclazyte, the dosage of the filter aid are the 2.5-25% of bacillus natto to ferment liquid quality.
Preferred technical solution are as follows: the filter aid is diatomite.
Preferred technical solution are as follows: the magnetic Nano material that step 4 is collected first uses ultrapure water, then is rushed with dehydrated alcohol
It washes, then obtains the reusable Fe being reduced with 60-65 DEG C of heated-air drying3O4@SiO2Then-G nanosphere can weigh
It is multiplexed in vitamin K2Specific adsorption operation；It reuses 10-15 times.
Since above-mentioned technical proposal is used, the present invention has the advantage, that compared with prior art
1, the present invention can greatly increase the effect of later period magnetic Nano material absorption first with the impurity in flocculant removal fermentation liquid
Rate and specificity, reduce enrichment time and material used.
2, the present invention uses the Fe by modification3O4@SiO2- G magnetic Nano material chemical property is stablized, and magnetic stability can
With duplicate use, G is able to achieve by π-π static buildup to the XK2 specific adsorption in fermentation liquid, also, magnetic Nano material
Expect good dispersion, the efficiency of enrichment is rapidly and efficiently.
3, this method rate of departure is fast.Used magnet in the separation link of magnetic material, magnet close to when, it is powerful
Magnetic fields make magnetic Nano material rapid aggregation, so that isolated process is almost just to complete moment.With other
Method it is incomparable it is quick with it is efficient
4, flocculant chitosan or Fe either used in this method3O4@SiO2- G magnetic Nano material will not pollute ring
Border, and property is stablized, and raw material are cheap and easily-available, and may be reused, and greatly reduce the cost of enrichment.
5, the present invention greatly reduces the dosage of organic reagent, reduces the dirt of environment compared with traditional liquid-liquid extraction
Dye.
Detailed description of the invention
Fig. 1 is the process flow chart of magnetite gathering of the present invention.
Fig. 2 is the influence for investigating the dosage that different magnetic Nano materials are added to adsorbance.
Fig. 3 is the influence for investigating magnetic Nano material adsorption time to adsorbance.
Fig. 4 is the influence for investigating organic reagent dosage to parsing.
Specific embodiment
Embodiments of the present invention are illustrated by particular specific embodiment below, those skilled in the art can be by this explanation
Content disclosed by book is understood other advantages and efficacy of the present invention easily.
Please refer to Fig. 1-Fig. 4.It should be clear that this specification structure depicted in this specification institute accompanying drawings, ratio, size etc., only to
Cooperate the revealed content of specification, so that those skilled in the art understands and reads, being not intended to limit the invention can be real
The qualifications applied, therefore do not have technical essential meaning, the tune of the modification of any structure, the change of proportionate relationship or size
It is whole, in the case where not influencing the effect of present invention can be generated and the purpose that can reach, it should all still fall in disclosed skill
Art content obtains in the range of capable of covering.Meanwhile in this specification it is cited as "upper", "lower", "left", "right", " centre " and
The term of " one " etc. is merely convenient to being illustrated for narration, rather than to limit the scope of the invention, relativeness
It is altered or modified, under the content of no substantial changes in technology, when being also considered as the enforceable scope of the present invention.
A kind of embodiment 1: vitamin K in enrichment bacillus natto to ferment liquid2Method
(1) acquisition of fermented liquid supernatant
It is 69.6g/L, glucose 34.5g/L, K that fermentation liquid handled by the present embodiment, which is by glycerol,2HPO4For 4.0g/L, egg
White peptone is 20g/L, and yeast leachate is culture medium composed by 25g/L, and 250rpm cultivates five days at 37 DEG C, obtains natto bud
Spore bacillus fermentation liquid.
By above-mentioned bacillus natto to ferment liquid, centrifugal filtration removes thallus and sediment, obtains supernatant.
(2) flocculant purifying fermentation liquor
Acetic acid is first added into water, is configured to the acetic acid solution of 0.18%wt, then chitosan is added into 0.18%wt acetic acid solution,
It is configured to the chitosan acetic acid solution of 0.4%wt.
The supernatant of the above-mentioned removal thallus of 100m is taken, the above-mentioned chitosan acetic acid solution of 9g is added thereto, stirs 30min,
It is eventually adding filter aid, precipitating is abandoned after centrifugation, obtains supernatant.The filter aid is cellulose.
(3) magnetic Fe3O4@SiO2The preparation of-G nano material
Chemical precipitation method prepares Fe3O4.At room temperature, to equipped with blender, condenser pipe, N2The three-necked flask of the 500ml of protection
In, the ammonium hydroxide of 112ml 12.5% is added, leads to nitrogen, 0.4mol/L FeCl is added in the case of stirring2And 0.8mol/L
FeCl3, pH to 6 is then adjusted, is heated rapidly to 80 DEG C, adds oleic acid, constant temperature stirs 1h, is then down to room temperature, anhydrous second
Alcohol washs repeatedly, magnet separation, dry, obtains the Fe of Nano grade3O4。
Fe3O4/SiO2The preparation of nanosphere.Take the Fe of the above-mentioned synthesis of 500mg3O4It is added to by 20ml deionized water,
In the mixed solution of 80ml isopropanol, 1ml ammonium hydroxide and 20ml ethyl orthosilicate, 12h, Zhi Houyong are reacted at room temperature after mixing evenly
The nanosphere that magnet collects synthesis three times with ethanol washing obtains Fe3O4/SiO2Nanosphere is scattered in isopropanol
In solution；100ml is taken to contain Fe3O4/SiO2Aqueous isopropanol, the APTES(3- aminopropyl-triethoxy silicon of 0.2ml is added
Alkane), ultrasound 30min, is stirred to react 6h at 70 DEG C later under the protection of nitrogen, obtains the Fe being modified3O4/SiO2It receives
Meter Wei Qiu i.e. Fe3O4/SiO2-NH2Nanosphere, after ethyl alcohol repeated flushing, vacuum drying.
Fe3O4@SiO2The preparation of-G nanosphere.To Fe3O4/SiO2-NH2GO, DMF(dimethyl are added in nanosphere
Formamide), EDC(1-(3- dimethylamino-propyl) -3- ethyl-carbodiimide hydrochloride), NHS(hydrocarbyl succinimide acid imide) and, by PH
Control reacts 2h at room temperature and obtains Fe in 4-63O4@SiO2- GO nanosphere.Fe3O4@SiO2- GO and hydrazine exist
2h is reacted at 80 DEG C, the Fe being just reduced3O4@SiO2- G nanosphere.
(4) optimize the amount of magnetic Nano material
The Fe of 300mg-800mg is separately added into the fermentation liquid after processing3O4@SiO2- G nanosphere, the present embodiment
Specific addition 500mg.At room temperature after stirring and adsorbing 1h, additional magnet recycles Fe3O4@SiO2- G nanosphere, outwells liquid, to
The n-hexane for completing to be added 30ml in the nano material of enrichment carries out ultrasonic parsing, time 5min-30min, the present embodiment tool
The body time is 20min.Organic phase is filtered later and carries out HPLC detection, determines the amount for completing the magnetic Nano material of enrichment.
As shown in Fig. 2, the increase of the amount of magnetic material with addition, bioaccumulation efficiency are consequently increased, when material therefor is super
When crossing 500mg, accumulation rate tends towards stability, and illustrates that the magnetic Nano material of 500mg can complete most of VK2 in fermentation liquid
Absorption, therefore select the material of 500mg as optimal additive amount.
(5) magnetic Nano material adsorption time
The Fe of 500mg is separately added into the fermentation liquid after processing3O4@SiO2- G nanosphere, is stirred respectively at room temperature
5min-60min is adsorbed, optimizes the time of absorption, its adsorption rate of result such as Fig. 3,20min has just reached 88%, illustrated its absorption
Process it is fast quickly, this and Fe3O4@SiO2- G nanosphere good dispersion, it is related by force to the specific adsorption of VK2.
(6) influence of the organic reagent dosage to resolution factor
The Fe of 500mg is separately added into the fermentation liquid after processing3O4@SiO2- G nanosphere, is stirred respectively at room temperature
20min is adsorbed, additional magnet recycles Fe3O4@SiO2- G nanosphere, outwells liquid, respectively into the nano material for completing enrichment
The n-hexane that 10ml-50ml is added carries out ultrasonic parsing, and organic phase is filtered carry out HPLC detection later, really by time 20min
The amount of the fixed magnetic Nano material for completing enrichment.
Influence of the dosage of organic reagent to parsing effect when n-hexane volume used is 20ml as shown in figure 4, parse
Rate has reached 92%, and with continuing growing for organic reagent dosage, resolution factor is not greatly improved, therefore selects 20ml just
Volume of the hexane as parsing.Compared with traditional organic reagent extracts, organic reagent used in such method greatly reduces.
Vitamin K in a kind of enrichment bacillus natto to ferment liquid of embodiment 22Method
Vitamin K in a kind of enrichment bacillus natto to ferment liquid2Method, including the following steps:
Step 1: bacillus natto to ferment liquid being centrifuged, supernatant is collected；
Step 2: flocculant and filter aid are added into the supernatant that step 1 obtains, is then stirred, is then centrifuged,
Collect supernatant；
Step 3: the Fe being reduced being added into the supernatant of step 23O4@SiO2- G nanosphere, adjusting pH value to 6.0, then
It is stirred 10minmin at 20 °C, then collects the magnetic material after absorption；The Fe being reduced3O4@
SiO2The preparation method of-G nanosphere includes the following steps:
S1, by Fe3O4Magnetic microsphere is added to the mixed solution being made of deionized water, isopropanol ammonium hydroxide and ethyl orthosilicate
In, it reacts 10h at room temperature after mixing evenly, collects the nanosphere of synthesis, obtain after washing by SiO2The Fe of package3O4@
SiO2-NH2；
S2, Xiang You SiO2The Fe of package3O4@SiO2-NH2Middle addition graphene oxide, dimethylformamide, 1-(3- dimethylamino
Propyl) -3- ethyl-carbodiimide hydrochloride and hydrocarbyl succinimide acid imide, pH value is controlled 4, reacts 1h at room temperature
Obtain Fe3O4@SiO2-GO；Then by Fe3O4@SiO2- GO reacts 1h at 75 DEG C to get the Fe being reduced with hydrazine3O4@
SiO2- G nanosphere；
Step 4: n-hexane being added into the magnetic material after absorption, carries out ultrasound parsing 5min, is then received by additional magnet
Collect magnetic Nano material, organic phase is then poured out into filtering, gained filtrate is to be enriched vitamin K2Hexane solution.
Preferred embodiment are as follows: the vitamin K of the bacillus natto to ferment liquid2Content is 30mg/L, residual sugar contains
Amount is 0.01g/L, residual nitrogen content is 3g/L, surface tension 18Mn/m.
Preferred embodiment are as follows: in step 1, the revolving speed of centrifugation is 5000rpm, and the time of centrifugally operated is 10min, from
Heart operation carries out in the 2h after the bacillus natto to ferment liquid is made.
Preferred embodiment are as follows: the flocculant is at least one in inorganic flocculating agent and organic polymer coargulator
Kind, the dosage of the flocculant is the 3% of bacillus natto to ferment liquid quality.
Preferred embodiment are as follows: the flocculant is chitosan.
Preferred embodiment are as follows: the mixing that the filter aid is diatomite, perlite is constituted according to the mass ratio of 1:1
Object, the dosage of the filter aid are the 2.5% of bacillus natto to ferment liquid quality.
Preferred embodiment are as follows: the filter aid is diatomite.
Preferred technical solution are as follows: the magnetic Nano material that step 4 is collected first uses ultrapure water, then is rushed with dehydrated alcohol
It washes, then obtains the reusable Fe being reduced with 60 DEG C of heated-air dryings3O4@SiO2Then-G nanosphere is repeated and is used
In vitamin K2Specific adsorption operation；It reuses 10 times.
A kind of embodiment 3: vitamin K in enrichment bacillus natto to ferment liquid2Method
Vitamin K in a kind of enrichment bacillus natto to ferment liquid2Method, including the following steps:
Step 1: bacillus natto to ferment liquid being centrifuged, supernatant is collected；
Step 2: flocculant and filter aid are added into the supernatant that step 1 obtains, is then stirred, is then centrifuged,
Collect supernatant；
Step 3: the Fe being reduced being added into the supernatant of step 23O4@SiO2- G nanosphere, adjusting pH value to 8.0, then
It is stirred 30min under the conditions of 35 DEG C, then collects the magnetic material after absorption；The Fe being reduced3O4@SiO2-
The preparation method of G nanosphere includes the following steps:
S1, by Fe3O4Magnetic microsphere is added to the mixed solution being made of deionized water, isopropanol ammonium hydroxide and ethyl orthosilicate
In, it reacts 20h at room temperature after mixing evenly, collects the nanosphere of synthesis, obtain after washing by SiO2The Fe of package3O4@
SiO2-NH2；
S2, Xiang You SiO2The Fe of package3O4@SiO2-NH2Middle addition graphene oxide, dimethylformamide, 1-(3- dimethylamino
Propyl) -3- ethyl-carbodiimide hydrochloride and hydrocarbyl succinimide acid imide, pH value is controlled 6, reacts 3h at room temperature
Obtain Fe3O4@SiO2-GO；Then by Fe3O4@SiO2- GO reacts 3h at 85 DEG C to get the Fe being reduced with hydrazine3O4@
SiO2- G nanosphere；
Step 4: n-hexane being added into the magnetic material after absorption, carries out ultrasound parsing 20min, is then received by additional magnet
Collect magnetic Nano material, organic phase is then poured out into filtering, gained filtrate is to be enriched vitamin K2Hexane solution.
Preferred embodiment are as follows: the vitamin K of the bacillus natto to ferment liquid2Content is 100mg/L, residual sugar contains
Amount is 2g/L, residual nitrogen content is 8g/L, surface tension 24Mn/m.
Preferred embodiment are as follows: in step 1, the revolving speed of centrifugation is 8000rpm, and the time of centrifugally operated is 20min, from
Heart operation carries out in the 2h after the bacillus natto to ferment liquid is made.
Preferred embodiment are as follows: the flocculant is at least one in inorganic flocculating agent and organic polymer coargulator
Kind, the dosage of the flocculant is the 30% of bacillus natto to ferment liquid quality.
Preferred embodiment are as follows: the flocculant is chitosan.
Preferred embodiment are as follows: the filter aid is that active carbon and Emathlite are constituted according to the mass ratio of 1:2
Mixture, the dosage of the filter aid are the 25% of bacillus natto to ferment liquid quality.
Preferred embodiment are as follows: the filter aid is diatomite.
Preferred embodiment are as follows: the magnetic Nano material that step 4 is collected first uses ultrapure water, then is rushed with dehydrated alcohol
It washes, then obtains the reusable Fe being reduced with 65 DEG C of heated-air dryings3O4@SiO2Then-G nanosphere is repeated and is used
In vitamin K2Specific adsorption operation；It reuses 15 times.
As described above is only to be not intended to tool to explain the preferred embodiments of the invention to do any shape to the present invention
Limitation in formula should all wrap therefore all have any modification or change for making the related present invention under identical spirit
It includes in the scope that the invention is intended to protect.
Claims (8)
1. vitamin K in a kind of enrichment bacillus natto to ferment liquid2Method, it is characterised in that: include the following steps:
Step 1: bacillus natto to ferment liquid being centrifuged, supernatant is collected；
Step 2: flocculant and filter aid are added into the supernatant that step 1 obtains, is then stirred, is then centrifuged,
Collect supernatant；
Step 3: the Fe being reduced being added into the supernatant of step 23O4@SiO2- G nanosphere, adjusting pH value to 6.0-8.0,
Then it is stirred 10min-30min under the conditions of 20 DEG C -35 DEG C, then collects the magnetic material after absorption；The quilt
The Fe of reduction3O4@SiO2The preparation method of-G nanosphere includes the following steps:
S1, by Fe3O4Magnetic microsphere is added in the mixed solution being made of deionized water, isopropanol ammonium hydroxide and ethyl orthosilicate,
It reacts 10-20h at room temperature after mixing evenly, collects the nanosphere of synthesis, obtain after washing by SiO2The Fe of package3O4@
SiO2-NH2；
S2, Xiang You SiO2The Fe of package3O4@SiO2-NH2Middle addition graphene oxide, dimethylformamide, 1-(3- dimethylamino
Propyl) -3- ethyl-carbodiimide hydrochloride and hydrocarbyl succinimide acid imide, pH value is controlled in 4-6, is reacted at room temperature
1-3h obtains Fe3O4@SiO2-GO；Then by Fe3O4@SiO2- GO with hydrazine reacts 1-3h at 75-85 DEG C to be reduced
Fe3O4@SiO2- G nanosphere；
Step 4: n-hexane being added into the magnetic material after absorption, carries out ultrasound parsing 5-20min, then passes through additional magnet
Magnetic Nano material is collected, organic phase is then poured out into filtering, gained filtrate is to be enriched vitamin K2Hexane solution.
2. vitamin K in enrichment bacillus natto to ferment liquid according to claim 12Method, it is characterised in that: institute
State the vitamin K of bacillus natto to ferment liquid2Content is 30-100mg/L, residual sugar content is 0.01-2g/L, residual nitrogen content is
3-8g/L, surface tension 18-24Mn/m.
3. vitamin K in enrichment bacillus natto to ferment liquid according to claim 12Method, it is characterised in that: step
In rapid 1, the revolving speed of centrifugation is 5000-8000rpm, and the time of centrifugally operated is 10-20min, and centrifugally operated is in the natto bud
It is carried out in 2h after spore bacillus fermentation liquid is obtained.
4. vitamin K in enrichment bacillus natto to ferment liquid according to claim 12Method, it is characterised in that: institute
Stating flocculant is at least one of inorganic flocculating agent and organic polymer coargulator, and the dosage of the flocculant is natto gemma
The 3-30% of bacillus fermentation liquid quality.
5. vitamin K in enrichment bacillus natto to ferment liquid according to claim 42Method, it is characterised in that: institute
Stating flocculant is chitosan.
6. vitamin K in enrichment bacillus natto to ferment liquid according to claim 12Method, it is characterised in that: institute
Stating filter aid is at least one of diatomite, perlite, cellulose, graphite powder, active carbon and Emathlite, the filter aid
Dosage be bacillus natto to ferment liquid quality 2.5-25%.
7. vitamin K in enrichment bacillus natto to ferment liquid according to claim 62Method, it is characterised in that: institute
Stating filter aid is diatomite.
8. vitamin K in enrichment bacillus natto to ferment liquid according to claim 12Method, it is characterised in that: step
Rapid 4 magnetic Nano materials collected first use ultrapure water, then are rinsed with dehydrated alcohol, are then obtained with 60-65 DEG C of heated-air drying
To the reusable Fe being reduced3O4@SiO2- G nanosphere, then can be repeated for vitamin K2Specific adsorption
Operation；It reuses 10-15 times.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110129234A (en) * | 2019-05-27 | 2019-08-16 | 沈阳农业大学 | The bacillus subtilis strain of high yield Agua-Mephyton 2 through mutagenesis and its application |
| CN110652968A (en) * | 2019-09-29 | 2020-01-07 | 中国农业科学院油料作物研究所 | Magnetic bacterium ball composite material loaded with reduced graphene oxide and preparation method thereof |
| CN113403348A (en) * | 2021-06-18 | 2021-09-17 | 山东润德生物科技有限公司 | Preparation method of vitamin K2 |
| CN114538589A (en) * | 2022-02-25 | 2022-05-27 | 中国科学院合肥物质科学研究院 | Method for enriching terpenoid in fermentation liquor and cellulose-based flocculant used by method |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20010046697A1 (en) * | 2000-04-21 | 2001-11-29 | Shinsaku Takaoka | Bacillus natto culture extract |
| US20060057688A1 (en) * | 2004-08-30 | 2006-03-16 | Chisso Corporation | Method for separating vitamin K2 from Bacillus bacterium culture |
| US20060105445A1 (en) * | 2002-07-29 | 2006-05-18 | Klaus Godl | Medium and method for enriching, purifying or depleting atp binding proteins from a pool of proteins |
| CN109402187A (en) * | 2018-11-15 | 2019-03-01 | 沈阳农业大学 | Agua-Mephyton 2 and thrombus dissolving enzyme co-production |
-
2019
- 2019-03-12 CN CN201910183364.0A patent/CN109706193A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20010046697A1 (en) * | 2000-04-21 | 2001-11-29 | Shinsaku Takaoka | Bacillus natto culture extract |
| US20060105445A1 (en) * | 2002-07-29 | 2006-05-18 | Klaus Godl | Medium and method for enriching, purifying or depleting atp binding proteins from a pool of proteins |
| US20060057688A1 (en) * | 2004-08-30 | 2006-03-16 | Chisso Corporation | Method for separating vitamin K2 from Bacillus bacterium culture |
| CN109402187A (en) * | 2018-11-15 | 2019-03-01 | 沈阳农业大学 | Agua-Mephyton 2 and thrombus dissolving enzyme co-production |
Non-Patent Citations (1)
| Title |
|---|
| 李亚娟等: "磁性还原石墨烯的制备及其对抗生素的吸附", 《环境工程学报》 * |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110129234A (en) * | 2019-05-27 | 2019-08-16 | 沈阳农业大学 | The bacillus subtilis strain of high yield Agua-Mephyton 2 through mutagenesis and its application |
| CN110652968A (en) * | 2019-09-29 | 2020-01-07 | 中国农业科学院油料作物研究所 | Magnetic bacterium ball composite material loaded with reduced graphene oxide and preparation method thereof |
| CN110652968B (en) * | 2019-09-29 | 2023-08-18 | 中国农业科学院油料作物研究所 | Magnetic fungus ball composite material loaded with reduced graphene oxide and preparation method thereof |
| CN113403348A (en) * | 2021-06-18 | 2021-09-17 | 山东润德生物科技有限公司 | Preparation method of vitamin K2 |
| CN113403348B (en) * | 2021-06-18 | 2022-03-22 | 山东润德生物科技有限公司 | Preparation method of vitamin K2 |
| CN114538589A (en) * | 2022-02-25 | 2022-05-27 | 中国科学院合肥物质科学研究院 | Method for enriching terpenoid in fermentation liquor and cellulose-based flocculant used by method |
| CN114538589B (en) * | 2022-02-25 | 2023-08-25 | 中国科学院合肥物质科学研究院 | Method for enriching terpenoid in fermentation liquor and cellulose-based flocculant used by same |
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