CN107189848A - A kind of method that kettle algae oil fat is split in extraction of highly effective and safe environmental protection - Google Patents
A kind of method that kettle algae oil fat is split in extraction of highly effective and safe environmental protection Download PDFInfo
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- CN107189848A CN107189848A CN201710352848.4A CN201710352848A CN107189848A CN 107189848 A CN107189848 A CN 107189848A CN 201710352848 A CN201710352848 A CN 201710352848A CN 107189848 A CN107189848 A CN 107189848A
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- kettle algae
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- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11B—PRODUCING, e.g. BY PRESSING RAW MATERIALS OR BY EXTRACTION FROM WASTE MATERIALS, REFINING OR PRESERVING FATS, FATTY SUBSTANCES, e.g. LANOLIN, FATTY OILS OR WAXES; ESSENTIAL OILS; PERFUMES
- C11B1/00—Production of fats or fatty oils from raw materials
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- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11B—PRODUCING, e.g. BY PRESSING RAW MATERIALS OR BY EXTRACTION FROM WASTE MATERIALS, REFINING OR PRESERVING FATS, FATTY SUBSTANCES, e.g. LANOLIN, FATTY OILS OR WAXES; ESSENTIAL OILS; PERFUMES
- C11B1/00—Production of fats or fatty oils from raw materials
- C11B1/02—Pretreatment
- C11B1/04—Pretreatment of vegetable raw material
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- Chemical Kinetics & Catalysis (AREA)
- Oil, Petroleum & Natural Gas (AREA)
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- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Fats And Perfumes (AREA)
Abstract
The invention discloses the method that kettle algae oil fat is split in a kind of extraction of highly effective and safe environmental protection, comprise the steps：1）It will split and homogenization is carried out after kettle algae powder dilutes through sodium citrate buffer solution, then obtained homogenizing fluid will be ground, alkali carries, neutral protease enzymolysis, alkali protease enzymolysis processing are then carried out successively to the kettle algae powder that splits after treatment, enzymolysis liquid is obtained；2）Enzymolysis liquid is centrifuged, free oil is obtained, the free oil is to split kettle algae oil fat.Based on the technique that this method splits kettle algae oil by aqueous enzymatic extraction, counterincision kettle algae powder carries out enzymolysis and obtains enzymolysis liquid, and enzymolysis liquid can be obtained to the greatest extent by centrifugation splits kettle algae oil fat, and oil extracting rate is up to 91.21%.
Description
Technical field
The present invention relates to the method that kettle algae oil fat is split in a kind of extraction of highly effective and safe environmental protection.
Background technology
Microalgae is widely distributed, and their figure can be found in land, ocean.Microalgae has growth cycle short, numerous
The features such as speed is fast, nutritional need is simple is grown, therefore people have been studied by biological engineering method and control culture bar at present
Part realizes the high density heterotrophic fermentation of microalgae.Correlative study comparative maturity and carry out having for industrialized production split kettle algae, Yin Jia
Algae, chlorella etc..Microalgae is nutritious, rich in protein, fat, carbohydrate and amino acid and polyunsaturated fatty acid
（PUFAs）Etc. a variety of nutriments, it is widely used in the fields such as food, medicine, cosmetics.The fat content of part microalgae compared with
Height, such as splits kettle algae, hidden dinoflagellate, and its fat content can reach more than the 40% of dry cell weight.These oil-producing microalgaes are not rich in more
Saturated fatty acid, some polyunsaturated fatty acids such as DHA, EPA content are up to more than the 10% of dry cell weight；And how unsaturated fat
The species of fat acid is fairly simple, and the separating-purifying for carrying out a certain specific polyunsaturated fatty acid is relatively easy.Therefore utilize
Microalgae has very wide prospect to produce polyunsaturated fatty acid.Because the existence form of grease in microalgae is and other big point
Son（Such as protein, carbohydrate etc.）It is combined to form " lipopolysaccharides ", " lipoprotein " complex, so needing the thin of microalgae
Born of the same parents' structure and grease compounded body are sufficiently destroyed, and just can discharge grease therein, are further extraction how unsaturated
Aliphatic acid is prepared.
At present, the extracting method of grease is mainly milling process, extraction method and CO2Supercritical extract etc..Milling process
Grease yield is low, labor intensity is big, cost is high, power consumption is big, and the investment of extraction method is big, the crude oil composition extracted
Complexity is, it is necessary to strict refining treatment；Extraction fado uses organic solvent so that technique becomes complicated, and danger coefficient is high, together
When organic solvent use easily cause environmental pollution；CO2It is higher that supercritical extract is limited to cost, it is impossible to carries out extensive work
Industry metaplasia is produced.Aqueous enzymatic method carries oil because it has the features such as less energy consumption, pollution are small, the few quality of the oily impurity that extracts is high, gradually
Become the study hotspot of people.The principle that aqueous enzymatic method carries oil is that utilization can drop on the basis of using Mechanical Crushing oil plant
Oil plant cell membrane is solved, or there is the enzyme of destruction to complexs such as lipoprotein, lipopolysaccharides（Such as cellulase, pectase, starch
Enzyme, protease etc.）, oil plant is acted on, the grease in oil plant is discharged.Enzyme used except can degrade oil plant cell membrane and
Destroy that lipopolysaccharides, lipoprotein etc. are compound external, can also destroy that oil plant produces in process of lapping is wrapped in oil droplets
Lipoprotein membrane, it is further to emulsion to destroy to improve oil yield.Not only grease extraction efficiency is high for aqueous enzymatic extraction microalgae oil, and
The quality better of oil.
Research related on aqueous enzymatic extraction grease at present has been directed to various plants, such as maize germ, soybean, flower
Raw, tea seed etc..The research of aqueous enzymatic extraction microalgae oil also compares many at present, but is all to crack microalgae cell using enzyme process
Wall, is then extracted using organic solvent.Such technique is complex, has used the organic solvent that there is potential safety hazard, no
Meet the requirement of current environmental protection, and grease extraction efficiency is not very high, including the present inventor's early stage patent ZL
201110111271.0 wait.
The content of the invention
It is an object of the invention to provide the method that kettle algae oil fat is split in a kind of extraction of highly effective and safe environmental protection.This method is with water enzyme
Method extraction is split based on the technique of kettle algae oil, and counterincision kettle algae powder carries out enzymolysis and obtains enzymolysis liquid, and enzymolysis liquid can be obtained to the greatest extent by centrifuging
Kettle algae oil fat must be split, oil extracting rate is up to 91.21%.
The method that kettle algae oil fat is split in the extraction of highly effective and safe environmental protection provided by the present invention, comprises the steps：
1）It will split and homogenization is carried out after kettle algae powder dilutes through sodium citrate buffer solution, then obtained homogenizing fluid will be ground,
Then alkali carries, neutral protease enzymolysis, alkali protease enzymolysis processing are carried out successively to the kettle algae powder that splits after treatment, obtained
Enzymolysis liquid；
2）Enzymolysis liquid is centrifuged, free oil is obtained, the free oil is to split kettle algae oil fat.
Step 1 of the present invention) in, the condition of the alkali carries is：PH value 9.0-9.6,60-65 DEG C of temperature extracts 1-3 h.Institute
The condition for stating neutral protease enzymolysis is：Enzyme concentration is that 0.03-0.13 g/g split kettle algae powder, pH value 6.3-6.8, temperature 43-46
DEG C, enzymolysis time 2-5 h, neutral proteinase used is concretely：Jie Neng sections bacterium neutral proteinase Protex 7L, enzyme activity
Power is 1600 AU/g.The condition of alkali protease enzymolysis is：Enzyme concentration is that 0.05-0.15 g/g split kettle algae powder, pH value
9.2-9.7,63-67 DEG C of temperature, enzymolysis time 2-7h.Alkali protease used is concretely：Jie Neng sections alkaline bacterial albumen
Enzyme Protex 6L, enzyme activity is 580000 DU/g.
Step 1 of the present invention) in, the concentration of the sodium citrate buffer solution can be that 0.05-0.2 mol/L, pH value are 4.6-
5.2；During dilution, the solid-liquid ratio for splitting kettle algae powder and sodium citrate buffer solution can be 1g ﹕ 6-9 ml；The condition being fully ground
For：Utilize grinding 120-180 s under the conditions of tissue grinder instrument 50-60 Hz.
The step 2) in the rotating speed that is centrifuged to enzymolysis liquid can be 1000-6000 rpm.
Beneficial effects of the present invention：Advantage of the aqueous enzymatic extraction compared with traditional handicraft is mainly reflected in economy, ring
Protect and safety in terms of.Have the advantage that as follows：
1）Technique is simple, safe operation, is adapted to large-scale production；
2）Make clasmatosis complete using homogeneous and the processing of tissue grinder instrument；
3）Without using organic solvent, green safety pollution is small；
4）Enzymolysis liquid after centrifugation gained upper strata free oil be carried split kettle algae oil, the collection method of grease is simple.
Embodiment
The method of the present invention is illustrated below by specific embodiment, but the invention is not limited in this.
Experimental method described in following embodiments, is conventional method unless otherwise specified；The reagent and material, such as
Without specified otherwise, commercially obtain.
Neutral proteinase used is：Jie Neng sections bacterium neutral proteinase Protex 7L, enzyme activity is 1600 AU/g.
Alkali protease used is：Jie Neng sections bacterial alkaline protease Protex 6L, enzyme activity is 580000 DU/g.
Split the measure of kettle algae fat content：Using soxhlet extraction methods (GB5009.6-85).
Oil extracting rate：Oil extracting rate=（The fat content of free oil quality/sample）×100%.
Embodiment 1
The g of kettle algae powder 20 is split in weighing（Containing the g of grease about 8.34）, the pH value for adding 0.1 mol/L is 4.8 sodium citrate buffer solution
120 ml, homogeneous, using grinding 120 s under the conditions of the Hz of tissue grinder instrument 55.It is 9.5 with NaOH solution regulation homogenizing fluid pH value,
60 DEG C of temperature, at the uniform velocity stirs the h of alkali carries 2.After alkali carries terminate, kettle algae powder weight is split in regulation system pH value 6.5,45 DEG C of temperature, addition
7% (g/g) neutral proteinase is measured, 3 h are at the uniform velocity digested；PH value 9.5 is adjusted again, and kettle algae powder weight 10% is split in 65 DEG C of temperature, addition
(g/g) alkali protease, at the uniform velocity digests 6 h, 4800 rpm centrifugations while hot, 10 mim, take upper strata free oil after enzymolysis terminates,
Quality is 7.61 g, and final oil extracting rate is 91.21%.
Embodiment 2
The g of kettle algae powder 20 is split in weighing（Containing the g of grease about 8.34）, the pH value for adding 0.1 mol/L is 4.8 sodium citrate buffer solution
120 ml, homogeneous, using grinding 120 s under the conditions of the Hz of tissue grinder instrument 55.With NaOH solution regulation homogenizing fluid pH value 9.5, temperature
60 DEG C of degree, at the uniform velocity stirs the h of alkali carries 2.After alkali carries terminate, kettle algae powder weight is split in regulation system pH value 6.5,45 DEG C of temperature, addition
7% (g/g) neutral proteinase, at the uniform velocity digests 3 h；PH value 9.5 is adjusted again, and (the g/ of kettle algae powder weight 8% is split in 65 DEG C of temperature, addition
G) alkali protease, at the uniform velocity digests 6 h, 4800 rpm centrifugations while hot, 10 mim, take upper strata free oil, quality after enzymolysis terminates
For 7.57 g, final oil extracting rate is 90.85%.
Embodiment 3
The g of kettle algae powder 20 is split in weighing（Containing the g of grease about 8.34）, the pH value for adding 0.1 mol/L is 4.8 sodium citrate buffer solution
120 ml, homogeneous, using grinding 120 s under the conditions of the Hz of tissue grinder instrument 55.With NaOH solution regulation homogenizing fluid pH value 9.0, temperature
60 DEG C of degree, at the uniform velocity stirs the h of alkali carries 2.After alkali carries terminate, kettle algae powder weight is split in regulation system pH value 6.5,45 DEG C of temperature, addition
5% (g/g) neutral proteinase, at the uniform velocity digests 3 h；PH value 9.5 is adjusted again, and (the g/ of kettle algae powder weight 5% is split in 65 DEG C of temperature, addition
G) alkali protease, at the uniform velocity digests 6 h, 4800 rpm centrifugations while hot, 10 mim, take upper strata free oil, quality after enzymolysis terminates
For 6.24 g, final oil extracting rate is 74.82%.
Embodiment 4
The g of kettle algae powder 20 is split in weighing（Containing the g of grease about 8.34）, the pH value for adding 0.1 mol/L is 4.8 sodium citrate buffer solution
120 ml, homogeneous, using grinding 120 s under the conditions of the Hz of tissue grinder instrument 55.With NaOH solution regulation homogenizing fluid pH value 9.5, temperature
60 DEG C of degree, at the uniform velocity stirs the h of alkali carries 2.After alkali carries terminate, kettle algal gel dry weight is split in regulation system pH value 6.5,45 DEG C of temperature, addition
7% (g/g) neutral proteinase, at the uniform velocity digests 3 h；PH value 9.5 is adjusted again, and (the g/ of kettle algae powder weight 6% is split in 65 DEG C of temperature, addition
G) alkali protease, at the uniform velocity digests 3 h, 4800 rpm centrifugations while hot, 10 mim, take upper strata free oil, quality after enzymolysis terminates
For 5.73 g, final oil extracting rate is 68.71%.
Embodiment 5
The g of kettle algae powder 20 is split in weighing（Containing the g of grease about 8.34）, the pH value for adding 0.1 mol/L is 4.8 sodium citrate buffer solution
120 ml, homogeneous, using grinding 120 s under the conditions of the Hz of tissue grinder instrument 55.With NaOH solution regulation homogenizing fluid pH value 9.5, temperature
60 DEG C of degree, at the uniform velocity stirs the h of alkali carries 2.After alkali carries terminate, kettle algae powder weight is split in regulation system pH value 6.5,45 DEG C of temperature, addition
3% (g/g) neutral proteinase, at the uniform velocity digests 2 h；PH value 9.5 is adjusted again, and kettle algal gel dry weight 10% is split in 65 DEG C of temperature, addition
(g/g) alkali protease, at the uniform velocity digests 6 h, 4800 rpm centrifugations while hot, 10 mim, take upper strata free oil, matter after enzymolysis terminates
Measure as 6.38 g, final oil extracting rate is 76.50%.
Embodiment 6
The g of kettle algae powder 20 is split in weighing（Containing the g of grease about 8.34）, the pH value for adding 0.1 mol/L is 4.8 sodium citrate buffer solution
120 ml, homogeneous, using grinding 120 s under the conditions of the Hz of tissue grinder instrument 55.With NaOH solution regulation homogenizing fluid pH value 9.5, temperature
60 DEG C of degree, at the uniform velocity stirs the h of alkali carries 2.After alkali carries terminate, kettle algae powder weight is split in regulation system pH value 6.5,45 DEG C of temperature, addition
13% (g/g) neutral proteinase, at the uniform velocity digests 5 h；PH value 9.5 is adjusted again, and kettle algal gel dry weight 15% is split in 60 DEG C of temperature, addition
(g/g) alkali protease, at the uniform velocity digests 7 h, 4800 rpm centrifugations while hot, 10 mim, take upper strata free oil, matter after enzymolysis terminates
Measure as 7.21 g, final oil extracting rate is 86.45%.
Claims (8)
1. a kind of method that kettle algae oil fat is split in extraction of highly effective and safe environmental protection, it is characterized in that, comprise the steps：
1）It will split and homogenization is carried out after kettle algae powder dilutes through sodium citrate buffer solution, then obtained homogenizing fluid will be ground,
Then alkali carries, neutral protease enzymolysis, alkali protease enzymolysis processing are carried out successively to the kettle algae powder that splits after treatment, obtained
Enzymolysis liquid；
2）Enzymolysis liquid is centrifuged, free oil is obtained, the free oil is to split kettle algae oil fat.
2. the method that kettle algae oil fat is split in the extraction of highly effective and safe environmental protection according to claim 1, it is characterized in that, step 1)
In, the concentration of the sodium citrate buffer solution is that 0.05-0.2 mol/L, pH value are 4.6-5.2；It is described to split kettle algae powder during dilution
Solid-liquid ratio with sodium citrate buffer solution is 1g ﹕ 6-9 ml；The condition of the grinding is：Utilize tissue grinder instrument 50-60 Hz bars
120-180 s are ground under part.
3. the method for kettle algae oil fat is split according to the extraction of the highly effective and safe of claim 1 or 2 environmental protection, it is characterized in that, the step 1)
In, the condition of the alkali carries is：PH value 9.0-9.6,60-65 DEG C of temperature extracts 1-3 h.
4. the method that kettle algae oil fat is split in the extraction of highly effective and safe environmental protection according to claim 3, it is characterized in that, the neutrality
The condition of protease hydrolyzed is：Enzyme concentration is that 0.03-0.13 g/g split kettle algae powder, pH value 6.3-6.8,43-46 DEG C of temperature, enzyme
Solution time 2-5 h.
5. the method that kettle algae oil fat is split in the extraction of highly effective and safe according to claim 4 environmental protection, it is characterized in that, it is described in
Property protease is：Jie Neng sections bacterium neutral proteinase Protex 7L, enzyme activity is 1600 AU/g.
6. the method that kettle algae oil fat is split in the extraction of highly effective and safe environmental protection according to claim 4, it is characterized in that, the alkalescence
The condition of protease hydrolyzed is：Enzyme concentration is that 0.05-0.15 g/g split kettle algae powder, pH value 9.2-9.7,63-67 DEG C of temperature, enzyme
Solution time 2-7h.
7. the method that kettle algae oil fat is split in the extraction of highly effective and safe environmental protection according to claim 6, it is characterized in that, the alkalescence
Protease is：Jie Neng sections bacterial alkaline protease Protex 6L, enzyme activity is 580000 DU/g.
8. the method that kettle algae oil fat is split in the extraction of highly effective and safe environmental protection according to claim 1, it is characterized in that, the step
2) rotating speed centrifuged in enzymolysis liquid is 1000-6000 rpm.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113142394A (en) * | 2021-03-15 | 2021-07-23 | 湖南加农正和生物技术有限公司 | Feed additive, feed and preparation method and application thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20100261918A1 (en) * | 2009-04-13 | 2010-10-14 | Board Of Regents, The University Of Texas System | Novel Process for Separating Lipids From a Biomass |
| CN102199485A (en) * | 2011-04-29 | 2011-09-28 | 国家海洋局第三海洋研究所 | Method for extracting schizochytrium aggregatum oil |
-
2017
- 2017-05-18 CN CN201710352848.4A patent/CN107189848A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20100261918A1 (en) * | 2009-04-13 | 2010-10-14 | Board Of Regents, The University Of Texas System | Novel Process for Separating Lipids From a Biomass |
| CN102199485A (en) * | 2011-04-29 | 2011-09-28 | 国家海洋局第三海洋研究所 | Method for extracting schizochytrium aggregatum oil |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113142394A (en) * | 2021-03-15 | 2021-07-23 | 湖南加农正和生物技术有限公司 | Feed additive, feed and preparation method and application thereof |
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