CN109971549A - The crude oil obtained using an one-step hydrolysis microbial fermentation, the method for being enriched with PUFAs - Google Patents

The crude oil obtained using an one-step hydrolysis microbial fermentation, the method for being enriched with PUFAs Download PDF

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CN109971549A
CN109971549A CN201910253581.2A CN201910253581A CN109971549A CN 109971549 A CN109971549 A CN 109971549A CN 201910253581 A CN201910253581 A CN 201910253581A CN 109971549 A CN109971549 A CN 109971549A
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fatty acid
temperature
crude oil
purity
polyunsaturated fatty
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谭新
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Hunan Wanquan Yuxiang Biotechnology Co Ltd
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Hunan Wanquan Yuxiang Biotechnology Co Ltd
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    • C—CHEMISTRY; METALLURGY
    • C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
    • C11C—FATTY ACIDS FROM FATS, OILS OR WAXES; CANDLES; FATS, OILS OR FATTY ACIDS BY CHEMICAL MODIFICATION OF FATS, OILS, OR FATTY ACIDS OBTAINED THEREFROM
    • C11C1/00—Preparation of fatty acids from fats, fatty oils, or waxes; Refining the fatty acids
    • C11C1/02—Preparation of fatty acids from fats, fatty oils, or waxes; Refining the fatty acids from fats or fatty oils
    • C11C1/04—Preparation of fatty acids from fats, fatty oils, or waxes; Refining the fatty acids from fats or fatty oils by hydrolysis
    • C—CHEMISTRY; METALLURGY
    • C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
    • C11C—FATTY ACIDS FROM FATS, OILS OR WAXES; CANDLES; FATS, OILS OR FATTY ACIDS BY CHEMICAL MODIFICATION OF FATS, OILS, OR FATTY ACIDS OBTAINED THEREFROM
    • C11C1/00—Preparation of fatty acids from fats, fatty oils, or waxes; Refining the fatty acids
    • C11C1/08—Refining

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  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Microbiology (AREA)
  • General Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Oil, Petroleum & Natural Gas (AREA)
  • Wood Science & Technology (AREA)
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  • Fats And Perfumes (AREA)

Abstract

The present invention relates to the technical field of bioseparation of fatty acid, more particularly to the crude oil obtained using one one-step hydrolysis microbial fermentation of chemical method, it isolates and purifies to obtain the polyunsaturated fatty acid method of high-purity, comprising the following steps: (1) crude oil obtained that will ferment obtains free fatty acid by one one-step hydrolysis of chemical method；(2) fatty acid obtained above is enriched with to obtain the polyunsaturated fatty acid of high-purity by urea clathrate and multiple-grade molecular distillation.The invention using sepiolite as filtering agent, based on isolating and purifying above, the method for the present invention that enrichment coupling polyunsaturated fatty acid is formed obtains crude oil as raw material using microbial fermentation, PUFAs >=96% can be made.The present invention be it is a kind of hydrolyze and be enriched with by efficient chemical method to obtain the polyunsaturated fatty acid method of high-purity, it is creative using crude oil as initial feed, eliminate algae oil refining workshop section, process is simple, at low cost, and greatly improves yield.

Description

The crude oil obtained using an one-step hydrolysis microbial fermentation, the method for being enriched with PUFAs
Technical field
The present invention relates to the crude oils to be fermented using a step chemical method hydrolysising microorganism, obtain free fatty acid, then lead to The method for crossing urea clathrate Yu multiple-grade molecular distillation enriching polyunsaturated fatty acid.
Background technique
Fatty acid is important oil and fat chemical basic material.Polyunsaturated fatty acid (PUFAs) therein to human health, Prevent disease play the role of it is great, especially with docosahexaenoic acid (cis-4,7,10,13,16,19- Docosahexaenoic Acid, DHA) He Ershi carbon two five olefin(e) acid (Docosapentaenoic acid, DPA) effect the most It is prominent.On the one hand, DHA is the important polyunsaturated fatty acid of a kind of pair of human body, has brain tonic and intelligence development, promotes nerve nucleus vision system System development, anticancer, anti-inflammatory, prevention and treatment cardiovascular disease and anti-aging and other effects, in industries such as medicine, food, feeds It has broad application prospects；On the other hand, DPA has effects that anticancer, all has prophylactic function to kinds cancers such as gastric cancers, Also there is anti-inflammatory efficacy, by influencing the expression of cell factor or enzyme, pro-inflammatory cytokine activity is reduced, thus to a certain extent Treat the diseases such as rheumatic arthritis, psoriasis, systemic loupus erythematosus.
The source of traditional PUFA is mainly deep sea fish oil, such as Chinese patent CN101161819A and CN101348807A All disclose and DHA and EPA be enriched with by esterification using fish oil n-3PUFA concentrate and glycerol, above-mentioned two patent with Fish oil is raw material.But since the enrichment of food chain causes in the fish oil extracted containing there are many poisonous and harmful substances, such as two English (Dioxin), toxaphene etc. are disliked, the heavy metal element that the mankind discharge in ocean also can be in the cylinder accumulation of ocean fish, significantly shadow The quality of PUFAs is rung.EPA, pollution-free, environmental-friendly, pure, stability are practically free of with the product that fungi and algae synthesize Well, fishy smell is small, cholesterol-free, and in various algae, Dinophyceae has the PUFAs of high-content, is obtained in recent years with fermentation Algae oil substitutes deep sea fish oil as the source of DHA and DPA and has become a kind of development trend.
PUFAs is enriched with by the algae oil that fermentation obtains, is all using the algae oil of refining as initial feed in traditional handicraft, essential oil is It is obtained by crude oil by workshop sections such as degumming, depickling, decoloration, debrominates.Such as Chinese patent CN104394702B, for being constantly enriched with A kind of method of oil generated by microalgae, with DHA ethyl ester.Algae oil refining process, workshop section's complicated and time consumption, while height can be lost The fatty acid of added value, therefore, the invention using crude oil as raw material, utilize a step chemical method hydrolyze crude oil, process letter Single, the time is short, while can protect the fatty acid of high added value, and percent hydrolysis is high.On the one hand, hydrolysis obtains free fatty acid, On the other hand, reached degumming, depickling, decolorizing effect, meanwhile, debrominate effect is reached by multiple-grade molecular distillation enrichment.Directly Using crude oil as initial feed, the refining workshop section of algae oil is eliminated, process is simple, at low cost.
The hydrolysis of algae oil is enriched in workshop section, requires to be removed by filtration fraction solids and impurity, in traditional workshop section all It is to be filtered using atlapulgite, active carbon and diatomite.The invention cleans by filtering agent of sepiolite, Hai Pao Stone is a kind of magnesia silicates clay compound in the Nature, and chemical formula is Mg8Si12O30(OH)4(OH2)4·8H2O (SEP4H2O, 8H2O), since sepiolite is that have large specific surface area and layered porous by SiO2Tetrahedron and Mg-O eight 3 D stereo special construction made of the vegetative grafting of face, on its surface, there is also more acidity [SiO4] alkalinity [MgO6] center, from And make sepiolite that there is stronger absorption property, adsorption-edulcoration is come with this, is worked well, it is white that traditional activity can be substituted completely Soil, active carbon and diatomite.Selected sepiolite derives from the domestic sepiolite mineral products in xiangtan, hunan province city.
Summary of the invention
The invention belongs to the technical field of bioseparation of fatty acid, the object of the present invention is to provide one kind from microbial fermentation In crude oil obtained, the polyunsaturated fatty acid method of high-purity is obtained by an one-step hydrolysis method and enrichment method, process is simple, It is at low cost, and greatly improve yield.
To achieve the above object, the invention adopts the following technical scheme:
From crude oil made from microbial fermentation, the polyunsaturated fatty acid method for obtaining high-purity is isolated and purified, fermentation is made The crude oil obtained hydrolyzes to obtain free fatty acid by chemical method, and process is simple, and at low cost, percent hydrolysis reaches 95% or more；It will be upper The fatty acid stated is enriched with to obtain the polyunsaturated fatty acid of high-purity by urea clathrate and multiple-grade molecular distillation.Water Solution is enriched with reaction solution obtained in workshop section using sepiolite as filtering agent and obtains crude product, selected sepiolite content 25%, 40%, 55%, 75%, 90%, preferential 75% content.Itself specific steps are as follows:
(1) hydrolysis: mCrude oil:V Sodium hydroxide-ethanol water=1:5-10,200 rpm/min-500 rpm/min of mixing speed, reaction It 40 DEG C -80 DEG C of temperature, is condensed back, 50 min-120 min of reaction time, process is led to nitrogen and all hydrolyzed, nitrogen envelope pressure 0.02 Mpa-0.05 Mpa.Reaction end is acidified, stands, and extracts, and washes, water removal, and sepiolite filters and evaporation, obtains free-fat Acid.
(2) urea clathrate: fatty acid: urea: ethyl alcohol=1:1-5:5-15(w:w:v), 2 DEG C -12 DEG C of temperature are included, inclusion 10 h-24 h of time.It is acidified after inclusion, it filters, extracts, water removal, sepiolite filtering rotates and obtains PUFAs crude product.
(3) molecular distillation is enriched with: the crude product in step (2) being distilled in molecular distillation apparatus, level-one distillation, evaporator Temperature: 50 DEG C -85 DEG C, cryosurface temperature: 30 DEG C -50 DEG C, vacuum degree: 2.5-4.5*10-1mbar；Retain first order molecular distillation Heavy constituent afterwards carries out secondary molecules distillation, evaporator temperature: 85 DEG C -115 DEG C, cryosurface temperature: and 45 DEG C -75 DEG C, vacuum degree: 2.5-3.5*10-2 mbar；Heavy constituent after retaining secondary molecules distillation carries out three-level molecular distillation, evaporator temperature: 115 DEG C -140 DEG C, cryosurface temperature: 55 DEG C -90 DEG C, vacuum degree: 2.5-3.5*10-2 mbar, the weight after retaining three-level molecular distillation Component carries out quaternary molecule distillation, evaporator temperature: 140 DEG C -170 DEG C, cryosurface temperature: and 65 DEG C -90 DEG C, vacuum degree: 2.5- 3.5*10-3 mbar, the heavy constituent after retaining quaternary molecule distillation carry out Pyatyi molecular distillation, evaporator temperature: 170 DEG C -200 DEG C, cryosurface temperature: 65 DEG C -90 DEG C, vacuum degree: 1.5-2.5*10-3 mbar retains light component, and sepiolite adsorption filtration obtains To the polyunsaturated fatty acid of high-purity.
Compared with prior art, the invention has the following beneficial effects:
The present invention is to obtain the polyunsaturated fatty acid method of high-purity by an one-step hydrolysis method and enrichment, and process is simple, cost It is low, and greatly improve yield.
(1) chemical method hydrolyzes, and reaction condition is mild, and the reaction time is short, simple process, and algae oil percent hydrolysis reaches 95% or more；
(2) the invention using crude oil as initial feed, eliminate the refining workshop section of algae oil, process is simple, environmental-friendly, Greatly reduce cost of material；
(3) the invention using sepiolite as filtering agent, take full advantage of regional advantages, it is at low cost, work well；
(4) the invention is with urea clathrate and the coupled enriched PUFAs of multiple-grade molecular distillation, it will be apparent that it is pure to improve PUFAs Degree, PUFAs content reach 95-99%.
Specific embodiment
Embodiment 1
(1) will ferment 280 g(DHA+DPA >=55% of crude oil obtained), 160 g of sodium hydroxide, 1550 mL ethyl alcohol (95%) solution Mixing, 300 rpm/min of mixing speed, are condensed back, 60 min of reaction time by 50 DEG C of reaction temperature, and it is whole that process leads to nitrogen Hydrolysis, 0.03 Mpa of nitrogen envelope pressure.Reaction terminates, and percent hydrolysis reaches 96.1%, and reaction solution is acidified, stands, and extracts, and washing removes Water, the filtering of 75% sepiolite and evaporation, obtain free fatty acid.
(2) it is poured into round-bottomed flask after first mixing 200 g urea with 1600 g ethyl alcohol (95%), is placed in 60 DEG C of water-baths It is condensed back reaction, until solution is clarified, 100 g of free fatty acid of step (1) is taken to be added in round-bottomed flask, logical nitrogen protection, 20 min are reacted, are cooled to room temperature, is placed in after 5 DEG C of standings include 16 h and takes out.Hydrated sheath is acidified to acid with 4 mol/L sulfuric acid Property, it is placed in separatory funnel, three times with n-hexane extraction, retains organic phase (upper phase), be washed with deionized water three times.It is added appropriate Anhydrous sodium sulfate stands water removal, the filtering of 75% sepiolite, and 40 DEG C of revolvings remove n-hexane, obtain polyunsaturated fatty acid concentrate.
(3) step (2) crude product is isolated and purified into molecular distillation apparatus, level-one distillation, evaporator temperature: 60 DEG C, cryosurface temperature: 35 DEG C, vacuum degree: 3.0*10-1 Mbar, 0.2 kg/h of sample rate；Retain the weight of first order molecular distillation Component carries out secondary molecules distillation, evaporator temperature: 90 DEG C, cryosurface temperature: and 50 DEG C, vacuum degree: 2.5*10-2 Mbar, sample introduction 0.2 kg/h of rate；The heavy constituent for retaining secondary molecules distillation carries out three-level molecular distillation, evaporator temperature: 120 DEG C, cryosurface Temperature: 60 DEG C, vacuum degree: 2.5*10-2 Mbar, 0.2 kg/h of sample rate；The heavy constituent for retaining three-level molecular distillation carries out four Grade molecular distillation, evaporator temperature: 140 DEG C, cryosurface temperature: 70 DEG C, vacuum degree: 3.5*10-3 Mbar, sample rate 0.2 kg/h；The heavy constituent for retaining quaternary molecule distillation carries out Pyatyi molecular distillation, evaporator temperature: 190 DEG C, cryosurface temperature: 80 DEG C, vacuum degree: 1.5*10-3 Mbar, 0.2 kg/h of sample rate retain light component, and after being filtered with sepiolite, centrifuge separation is obtained To the polyunsaturated fatty acid of high-purity, after esterification, through gas chromatographic detection, obtaining DHA content is that 84.3%, DPA content is 12.9%, it the results are shown in Table one.
Embodiment 2
(1) will ferment 300 g(DHA+DPA >=55% of crude oil obtained), 170 g of sodium hydroxide, 1650 mL ethyl alcohol (95%) solution Mixing, 300 rpm/min of mixing speed, are condensed back, 70 min of reaction time by 50 DEG C of reaction temperature, and it is whole that process leads to nitrogen Hydrolysis, 0.03 Mpa of nitrogen envelope pressure.Reaction terminates, and percent hydrolysis reaches 97.4%, and reaction solution is acidified, stands, and extracts, and washing removes Water, the filtering of 75% sepiolite and evaporation, obtain free fatty acid.
(2) it is poured into round-bottomed flask after first mixing 200 g urea with 1600 g ethyl alcohol (95%), is placed in 60 DEG C of water-baths It is condensed back reaction, until solution is clarified, 100 g of free fatty acid of step (1) is taken to be added in round-bottomed flask, logical nitrogen protection, 20 min are reacted, are cooled to room temperature, is placed in after 4 DEG C of standings include 20 h and takes out.Hydrated sheath is acidified to acid with 4 mol/L sulfuric acid Property, it is placed in separatory funnel, three times with n-hexane extraction, retains organic phase (upper phase), be washed with deionized water three times.It is added appropriate Anhydrous sodium sulfate stands water removal, the filtering of 75% sepiolite, and 40 DEG C of revolvings remove n-hexane, obtain polyunsaturated fatty acid concentrate.
(3) step (2) crude product is isolated and purified into molecular distillation apparatus, level-one distillation, evaporator temperature: 65 DEG C, cryosurface temperature: 35 DEG C, vacuum degree: 2.5*10-1 Mbar, 0.2 kg/h of sample rate；Retain the weight of first order molecular distillation Component carries out secondary molecules distillation, evaporator temperature: 100 DEG C, cryosurface temperature: and 50 DEG C, vacuum degree: 2.5*10-2 Mbar, into 0.2 kg/h of sample rate；The heavy constituent for retaining secondary molecules distillation carries out three-level molecular distillation, evaporator temperature: 125 DEG C, condensation Face temperature: 60 DEG C, vacuum degree: 2.5*10-2 Mbar, 0.2 kg/h of sample rate；The heavy constituent for retaining three-level molecular distillation carries out Quaternary molecule distillation, evaporator temperature: 145 DEG C, cryosurface temperature: 70 DEG C, vacuum degree: 3.5*10-3 Mbar, sample rate 0.2 kg/h；The heavy constituent for retaining quaternary molecule distillation carries out Pyatyi molecular distillation, evaporator temperature: 195 DEG C, cryosurface temperature: 80 DEG C, vacuum degree: 1.5*10-3 Mbar, 0.2 kg/h of sample rate retain light component, and after being filtered with sepiolite, centrifuge separation is obtained To the polyunsaturated fatty acid of high-purity, after esterification, through gas chromatographic detection, obtaining DHA content is that 85.6%, DPA content is 12.6%, it the results are shown in Table one.
Embodiment 3
(1) will ferment 300 g(DHA+DPA >=55% of crude oil obtained), sodium hydroxide 175g, 1700 mL ethyl alcohol (95%) solution Mixing, 300 rpm/min of mixing speed, are condensed back, 75 min of reaction time by 55 DEG C of reaction temperature, and it is whole that process leads to nitrogen Hydrolysis, 0.03 Mpa of nitrogen envelope pressure.Reaction terminates, and percent hydrolysis reaches 98.7%, and reaction solution is acidified, stands, and extracts, and washing removes Water, the filtering of 75% sepiolite and evaporation, obtain free fatty acid.
(2) it is poured into round-bottomed flask after first mixing 200 g urea with 1600 g ethyl alcohol (95%), is placed in 60 DEG C of water-baths It is condensed back reaction, until solution is clarified, 100 g of free fatty acid of step (1) is taken to be added in round-bottomed flask, logical nitrogen protection, 30 min are reacted, are cooled to room temperature, is placed in after 4 DEG C of standings include 18 h and takes out.Hydrated sheath is acidified to acid with 4 mol/L sulfuric acid Property, it is placed in separatory funnel, three times with n-hexane extraction, retains organic phase (upper phase), be washed with deionized water three times.It is added appropriate Anhydrous sodium sulfate stands water removal, the filtering of 75% sepiolite, and 45 DEG C of revolvings remove n-hexane, obtain polyunsaturated fatty acid concentrate.
(3) step (2) crude product is isolated and purified into molecular distillation apparatus, level-one distillation, evaporator temperature: 70 DEG C, cryosurface temperature: 40 DEG C, vacuum degree: 2.5*10-1 Mbar, 0.2 kg/h of sample rate；Retain the weight of first order molecular distillation Component carries out secondary molecules distillation, evaporator temperature: 105 DEG C, cryosurface temperature: and 55 DEG C, vacuum degree: 2.5*10-2 Mbar, into 0.2 kg/h of sample rate；The heavy constituent for retaining secondary molecules distillation carries out three-level molecular distillation, evaporator temperature: 125 DEG C, condensation Face temperature: 65 DEG C, vacuum degree: 2.5*10-2 Mbar, 0.2 kg/h of sample rate；The heavy constituent for retaining three-level molecular distillation carries out Quaternary molecule distillation, evaporator temperature: 150 DEG C, cryosurface temperature: 70 DEG C, vacuum degree: 3.5*10-3 Mbar, sample rate 0.2 kg/h；The heavy constituent for retaining quaternary molecule distillation carries out Pyatyi molecular distillation, evaporator temperature: 195 DEG C, cryosurface temperature: 85 DEG C, vacuum degree: 1.5*10-3 Mbar, 0.2 kg/h of sample rate retain light component, and after being filtered with sepiolite, centrifuge separation is obtained To the polyunsaturated fatty acid of high-purity, after esterification, through gas chromatographic detection, obtaining DHA content is that 85.9%, DPA content is 12.1%, it the results are shown in Table one.
Embodiment 4
(1) will ferment 330 g(DHA+DPA >=55% of crude oil obtained), sodium hydroxide 185g, 1850 mL ethyl alcohol (95%) solution Mixing, 300 rpm/min of mixing speed, are condensed back, 90 min of reaction time by 75 DEG C of reaction temperature, and it is whole that process leads to nitrogen Hydrolysis, 0.03 Mpa of nitrogen envelope pressure.Reaction terminates, and percent hydrolysis reaches 99.1%, and reaction solution is acidified, stands, and extracts, and washing removes Water, the filtering of 75% sepiolite and evaporation, obtain free fatty acid.
(2) it is poured into round-bottomed flask after first mixing 200 g urea with 1600 g ethyl alcohol (95%), is placed in 60 DEG C of water-baths It is condensed back reaction, until solution is clarified, 100 g of free fatty acid of step (1) is taken to be added in round-bottomed flask, logical nitrogen protection, 30 min are reacted, are cooled to room temperature, is placed in after 4 DEG C of standings include 18 h and takes out.Hydrated sheath is acidified to acid with 4 mol/L sulfuric acid Property, it is placed in separatory funnel, three times with n-hexane extraction, retains organic phase (upper phase), be washed with deionized water three times.It is added appropriate Anhydrous sodium sulfate stands water removal, the filtering of 75% sepiolite, and 45 DEG C of revolvings remove n-hexane, obtain polyunsaturated fatty acid concentrate.
(3) step (2) crude product is isolated and purified into molecular distillation apparatus, level-one distillation, evaporator temperature: 75 DEG C, cryosurface temperature: 40 DEG C, vacuum degree: 2.5*10-1 Mbar, 0.2 kg/h of sample rate；Retain the weight of first order molecular distillation Component carries out secondary molecules distillation, evaporator temperature: 110 DEG C, cryosurface temperature: and 55 DEG C, vacuum degree: 2.5*10-2 Mbar, into 0.2 kg/h of sample rate；The heavy constituent for retaining secondary molecules distillation carries out three-level molecular distillation, evaporator temperature: 130 DEG C, condensation Face temperature: 65 DEG C, vacuum degree: 2.5*10-2 Mbar, 0.2 kg/h of sample rate；The heavy constituent for retaining three-level molecular distillation carries out Quaternary molecule distillation, evaporator temperature: 155 DEG C, cryosurface temperature: 65 DEG C, vacuum degree: 3.0*10-3 Mbar, sample rate 0.2 kg/h；The heavy constituent for retaining quaternary molecule distillation carries out Pyatyi molecular distillation, evaporator temperature: 195 DEG C, cryosurface temperature: 80 DEG C, vacuum degree: 1.5*10-3 Mbar, 0.2 kg/h of sample rate retain light component, and after being filtered with sepiolite, centrifuge separation is obtained To the polyunsaturated fatty acid of high-purity, after esterification, through gas chromatographic detection, obtaining DHA content is that 86.8%, DPA content is 11.8%, it the results are shown in Table one.
Embodiment 5
(1) will ferment 350 g(DHA+DPA >=55% of crude oil obtained), sodium hydroxide 190g, 1900 mL ethyl alcohol (95%) solution Mixing, 300 rpm/min of mixing speed, are condensed back, 120 min of reaction time by 80 DEG C of reaction temperature, and it is whole that process leads to nitrogen Hydrolysis, 0.03 Mpa of nitrogen envelope pressure.Reaction terminates, and percent hydrolysis reaches 99.5%, and reaction solution is acidified, stands, and extracts, and washing removes Water, the filtering of 75% sepiolite and evaporation, obtain free fatty acid.
(2) it is poured into round-bottomed flask after first mixing 200 g urea with 1600 g ethyl alcohol (95%), is placed in 60 DEG C of water-baths It is condensed back reaction, until solution is clarified, 100 g of free fatty acid of step (1) is taken to be added in round-bottomed flask, logical nitrogen protection, 30 min are reacted, are cooled to room temperature, is placed in after 4 DEG C of standings include 18 h and takes out.Hydrated sheath is acidified to acid with 4 mol/L sulfuric acid Property, it is placed in separatory funnel, three times with n-hexane extraction, retains organic phase (upper phase), be washed with deionized water three times.It is added appropriate Anhydrous sodium sulfate stands water removal, the filtering of 75% sepiolite, and 45 DEG C of revolvings remove n-hexane, obtain polyunsaturated fatty acid concentrate.
(3) step (2) crude product is isolated and purified into molecular distillation apparatus, level-one distillation, evaporator temperature: 85 DEG C, cryosurface temperature: 45 DEG C, vacuum degree: 2.5*10-1 Mbar, 0.2 kg/h of sample rate；Retain the weight of first order molecular distillation Component carries out secondary molecules distillation, evaporator temperature: 115 DEG C, cryosurface temperature: and 50 DEG C, vacuum degree: 2.5*10-2 Mbar, into 0.2 kg/h of sample rate；The heavy constituent for retaining secondary molecules distillation carries out three-level molecular distillation, evaporator temperature: 140 DEG C, condensation Face temperature: 60 DEG C, vacuum degree: 2.5*10-2 Mbar, 0.2 kg/h of sample rate；The heavy constituent for retaining three-level molecular distillation carries out Quaternary molecule distillation, evaporator temperature: 165 DEG C, cryosurface temperature: 70 DEG C, vacuum degree: 3.0*10-3 Mbar, sample rate 0.2 kg/h；The heavy constituent for retaining quaternary molecule distillation carries out Pyatyi molecular distillation, evaporator temperature: 200 DEG C, cryosurface temperature: 70 DEG C, vacuum degree: 1.5*10-3 Mbar, 0.2 kg/h of sample rate retain light component, and after being filtered with sepiolite, centrifuge separation is obtained To the polyunsaturated fatty acid of high-purity, after esterification, through gas chromatographic detection, obtaining DHA content is that 87.1%, DPA content is 11.0%, it the results are shown in Table one.
Table one

Claims (7)

1. the crude oil obtained using one one-step hydrolysis microbial fermentation of chemical method, isolates and purifies the polyunsaturated fat for obtaining high-purity Sour method, the crude oil obtained that will ferment obtain free fatty acid by one one-step hydrolysis of chemical method, and crude oil and water are by a certain percentage It is blended under the catalysis of alkali, reaction condition is mild, and process is simple, and at low cost, percent hydrolysis reaches 95% or more, after reaction will Reaction solution is filtered with sepiolite, centrifuge separation；Obtained fatty acid is enriched with to obtain by urea clathrate and multiple-grade molecular distillation The polyunsaturated fatty acid of high-purity.
2. the crude oil obtained according to claim 1 using one one-step hydrolysis microbial fermentation of chemical method, isolates and purifies to obtain height The polyunsaturated fatty acid method of purity, it is characterised in that: by one one-step hydrolysis of chemical method, urea clathrate and many-level molecule steam It evaporates enrichment and obtains the polyunsaturated fatty acid of high-purity.
3. isolating and purifying to obtain high-purity according to the crude oil for using one one-step hydrolysis microbial fermentation of chemical method to obtain described in claim 1 The polyunsaturated fatty acid method of degree, it is NaOH or KOH that chemical method, which hydrolyzes the alkali,.
4. one one-step hydrolysis of chemical method answers mild condition, and process is simple, and at low cost, percent hydrolysis reaches according to claim 3 the method To 95% or more.
5. isolating and purifying to obtain high-purity according to the crude oil for using one one-step hydrolysis microbial fermentation of chemical method to obtain described in claim 1 The polyunsaturated fatty acid method of degree, in urea clathrate, fatty acid: urea: ethyl alcohol=1:1-5:5-15(w:w:v), include temperature 2 DEG C -12 DEG C, 10 h-24 h of time, acidified, suction filtration after inclusion are included, extraction removes water, and sepiolite filtering rotates To PUFAs crude product.
6. isolating and purifying to obtain high-purity according to the crude oil for using one one-step hydrolysis microbial fermentation of chemical method to obtain described in claim 1 The polyunsaturated fatty acid method of degree is distilled in molecular distillation apparatus, level-one distillation, evaporator temperature: 50 DEG C -85 DEG C, condensation Face temperature: 30 DEG C -50 DEG C, vacuum degree: 2.5-4.5*10-1mbar；Heavy constituent after retaining first order molecular distillation carries out two fractions Son distillation, evaporator temperature: 85 DEG C -115 DEG C, cryosurface temperature: 45 DEG C -75 DEG C, vacuum degree: 2.5-3.5*10-2mbar；It protects Heavy constituent after staying secondary molecules to distill carries out three-level molecular distillation, evaporator temperature: 115 DEG C -140 DEG C, cryosurface temperature: 55 DEG C -90 DEG C, vacuum degree: 2.5-3.5*10-2Mbar, the heavy constituent after retaining three-level molecular distillation carry out quaternary molecule distillation, steam Hair device temperature: 140 DEG C -170 DEG C, cryosurface temperature: 65 DEG C -90 DEG C, vacuum degree: 2.5-3.5*10-3Mbar retains four fractions Heavy constituent after son distillation carries out Pyatyi molecular distillation, evaporator temperature: 170 DEG C -200 DEG C, cryosurface temperature: and 65 DEG C -90 DEG C, Vacuum degree: 1.5-2.5*10-3Mbar, retains light component, and sepiolite adsorption filtration obtains the polyunsaturated fatty acid of high-purity.
7. being connected according to claim 6 the method using multiple-grade molecular distillation, it will be apparent that improve PUFAs purity, PUFAs contains Amount reaches 95-99%, and molecular distillation enrichment, temperature is low, heated time is short, separation degree is high, energy consumption is low, environmental-friendly, separates Journey is physical process, does not use organic solvent in separation process, separated object matter can be protected well not contaminated, obtained pure The product of safety, and molecular distillation carries debrominate function, obtained polyunsaturated fatty acid is obviously reduced compared to former algae oil taste.
CN201910253581.2A 2019-03-30 2019-03-30 The crude oil obtained using an one-step hydrolysis microbial fermentation, the method for being enriched with PUFAs Pending CN109971549A (en)

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傅红，等: "分子蒸馏法制备鱼油多不饱和脂肪酸", 《无锡轻工大学学报》 *
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Application publication date: 20190705