CN112831531A - Method for producing alga DHA grease by biological enzyme method wall breaking - Google Patents
Method for producing alga DHA grease by biological enzyme method wall breaking Download PDFInfo
- Publication number
- CN112831531A CN112831531A CN201911161513.XA CN201911161513A CN112831531A CN 112831531 A CN112831531 A CN 112831531A CN 201911161513 A CN201911161513 A CN 201911161513A CN 112831531 A CN112831531 A CN 112831531A
- Authority
- CN
- China
- Prior art keywords
- enzyme
- docosahexaenoic acid
- fermentation liquor
- wall breaking
- oil
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/64—Fats; Fatty oils; Ester-type waxes; Higher fatty acids, i.e. having at least seven carbon atoms in an unbroken chain bound to a carboxyl group; Oxidised oils or fats
- C12P7/6409—Fatty acids
- C12P7/6427—Polyunsaturated fatty acids [PUFA], i.e. having two or more double bonds in their backbone
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Wood Science & Technology (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Oil, Petroleum & Natural Gas (AREA)
- Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
A method for producing alga DHA grease by biological enzyme method wall breaking comprises the following steps: the method comprises the steps of taking Crypthecodinium cohnii as an initial strain, continuously culturing the Crypthecodinium cohnii in a liquid culture medium to obtain seaweed cells, and extracting docosahexaenoic acid grease from the seaweed cells by breaking the walls through a biological enzyme method. The advantages are that: the algae cell wall breaking is complete, the wall breaking rate is more than 95%, drying is not needed, oxidation of crude docosahexaenoic acid oil can be prevented, the peroxide value of the extracted oil is extremely low, the POV value is less than 0.5, the yield is more than 90% of the total oil, and the refining yield is more than 70% of the crude oil.
Description
Technical Field
The invention relates to the technical field of producing docosahexaenoic acid (DHA) grease by using Crypthecodinium cohnii, in particular to a method for producing alga DHA grease by breaking walls by a biological enzyme method.
Background
In recent years, the important role of polyunsaturated fatty acids in human health has been receiving increasing attention. Among them, docosahexaenoic acid is favored because of its important physiological function in the human and animal body. Research has shown that DHA is an essential component of cell membranes in certain tissues of the human body; plays an important role in the development of the visual system and the nervous system of the infant; it also has cholesterol lowering, anticoagulant, anticancer, and senile dementia preventing effects, and is a fatty acid essential for juvenile fish growth. The production of docosahexaenoic acid (DHA) grease by Crypthecodinium cohnii has already formed industrial production, and the production process of docosahexaenoic acid is continuously improved. There are two types of methods for breaking the cell wall of algae in the production of docosahexaenoic acid (DHA) oil by marine microalgae which are now publicly reported. One is physical method wall breaking: mechanical wall breaking such as grinding and squeezing with homogenizer or colloid mill to break cell wall, and drying. The mechanical wall breaking method is adopted to break the cell wall incompletely, and the extraction yield of the organic solvent is 60-70% of the total oil. The drying method consumes time and energy, the peroxidation value of the extracted oil is higher after the heating and ventilating method is adopted for drying, the quality of the crude oil is poorer, the freeze drying cost is higher, and the yield is only 30-50% of the crude oil after the extraction and the refining by the organic solvent after the drying.
Disclosure of Invention
The invention aims to develop a method for producing docosahexaenoic acid grease by using enzyme wall breaking for crypthecodinium cohnii fermentation, which greatly improves the yield of docosahexaenoic acid (DHA for short) and the quality of crude oil.
The invention relates to a method for producing docosahexaenoic acid grease by biological enzyme method wall breaking, which comprises the following steps: the method comprises the steps of taking Crypthecodinium cohnii as an initial strain, continuously culturing the Crypthecodinium cohnii in a liquid culture medium to obtain seaweed cells, and extracting docosahexaenoic acid grease from the seaweed cells by breaking the walls through a biological enzyme method.
The biological enzyme is divided into a main enzyme and an auxiliary enzyme;
the main enzyme is alkaline protease (2709), neutral protease, and papain, and one or more of them can be used, and if several of them are used, the proportion can be used in any proportion. The dosage of the fermentation liquid is 0.12-0.5% of the weight of the fermentation liquid, and if the fermentation liquid is concentrated, the dosage is calculated according to the weight of the fermentation liquid before concentration;
the auxiliary enzyme is trypsin, chymotrypsin, cellulase, glucanase and xylanase, and one or more of the auxiliary enzymes can be used, and if the auxiliary enzymes are more than one, the auxiliary enzymes can be used in any proportion. The dosage of the enzyme is 0 to 0.025 percent of the weight of the fermentation liquor, and if the enzyme is concentrated fermentation liquor, the dosage of the main enzyme and the auxiliary enzyme is calculated according to the weight of the fermentation liquor before concentration;
when used, the wall breaking can be achieved by using the main enzyme alone or the main enzyme and the auxiliary enzyme in combination.
The invention relates to a method for producing docosahexaenoic acid grease by biological enzyme method wall breaking, which comprises the following steps: adding main enzyme into the fermentation liquor or the filtered and concentrated fermentation liquor, wherein the dosage of the main enzyme is 0.12-0.5% of the weight of the fermentation liquor, and if the main enzyme is the concentrated fermentation liquor, the dosage is calculated according to the weight of the fermentation liquor before concentration; and (3) auxiliary enzyme, the dosage of which is 0-0.025% of the weight of the fermentation broth, if the fermentation broth is concentrated, the dosage of the main enzyme and the auxiliary enzyme is calculated according to the weight of the fermentation broth before concentration, the temperature is increased to 50-70 ℃, the heat preservation and the stirring are carried out for 3-9 hours, 95% ethanol is added, the dosage of the ethanol is 30-150% of the concentrated fermentation broth of the fermentation broth, and the dosage of the organic solvent is 1.5-4.5 times of the weight of the fermentation broth or the concentrated fermentation broth, so as to extract the docosahexaenoic acid crude oil. The refined docosahexaenoic acid oil is obtained by hydrating, alkali refining, decolorizing and deodorizing the crude docosahexaenoic acid oil.
Wherein the organic solvent is n-hexane or chloroform, etc.
The addition of the main enzyme and the auxiliary enzyme in the concentrated fermentation liquor is calculated according to the fermentation liquor amount before the fermentation liquor is filtered and concentrated.
The method for producing the docosahexaenoic acid grease by using the biological enzyme method wall breaking has the advantages that: the algae cell wall breaking is complete, the wall breaking rate is more than 95%, drying is not needed, oxidation of crude docosahexaenoic acid oil can be prevented, the peroxide value of the extracted oil is extremely low, the POV value is less than 0.5, the yield is more than 90% of the total oil, and the refining yield is more than 70% of the crude oil.
Detailed Description
The first embodiment is as follows:
a method for producing alga DHA grease by breaking walls by a biological enzyme method comprises the following steps: performing liquid shake culture, liquid shake flask amplification culture, primary seed culture, secondary seed culture and tertiary fermentation culture on Crypthecodinium cohnii slant strain to obtain fermentation liquor. Adding alkaline protease (2709)0.2kg into fermentation broth 100kg, heating to 50-70 deg.C, stirring for 3-9 hr, adding 95% ethanol 30kg, adding n-hexane 150kg, and extracting to obtain docosahexaenoic acid crude oil 3 kg. The refined docosahexaenoic acid oil 2.34kg is obtained by hydrating, alkali refining, decolorizing and deodorizing the crude docosahexaenoic acid oil.
Example two:
a method for producing alga DHA grease by breaking walls by a biological enzyme method comprises the following steps: performing liquid shake culture, liquid shake flask amplification culture, primary seed culture, secondary seed culture and tertiary fermentation culture on Crypthecodinium cohnii slant strain to obtain fermentation liquor. Adding neutral protease 0.35kg into fermentation broth 100kg, heating to 50-70 deg.C, stirring for 3-9 hr, adding 95% ethanol 30kg, adding n-hexane 150kg, and extracting to obtain docosahexaenoic acid crude oil 3 kg. The refined docosahexaenoic acid oil 2.34kg is obtained by hydrating, alkali refining, decolorizing and deodorizing the crude docosahexaenoic acid oil.
Example three:
a method for producing alga DHA grease by breaking walls by a biological enzyme method comprises the following steps: performing liquid shake culture, liquid shake flask amplification culture, primary seed culture, secondary seed culture and tertiary fermentation culture on Crypthecodinium cohnii slant strain to obtain fermentation liquor. Adding papain 0.45kg into fermentation broth 100kg, heating to 50-70 deg.C, stirring for 3-9 hr, adding 95% ethanol 30kg, adding n-hexane 150kg, and extracting to obtain docosahexaenoic acid crude oil 3 kg. The refined docosahexaenoic acid oil 2.34kg is obtained by hydrating, alkali refining, decolorizing and deodorizing the crude docosahexaenoic acid oil.
Claims (1)
1. A method for producing alga DHA grease by biological enzyme method wall breaking is characterized in that: the method comprises the following steps: continuously culturing Crypthecodinium cohnii in a liquid culture medium by using Crypthecodinium cohnii as an initial strain to obtain algal cells, and extracting docosahexaenoic acid oil from the algal cells by biological enzyme method wall breaking; adding a main enzyme and an auxiliary enzyme into fermentation liquor or filtered and concentrated fermentation liquor, heating to 50-70 ℃, keeping the temperature and stirring for 3-9 hours, adding 95% of ethanol, wherein the ethanol accounts for 30% -150% of the fermentation liquor, and adding an organic solvent for extraction to obtain docosahexaenoic acid crude oil; hydrating, alkali refining, decolorizing and deodorizing the crude docosahexaenoic acid oil to obtain refined docosahexaenoic acid oil; the main enzyme is one or more of alkaline protease 2709, neutral protease and papain, and if the main enzyme is several, the main enzyme can be used in any proportion; the auxiliary enzyme is one or more of trypsin, chymotrypsin, cellulase, glucanase and xylanase, and if the auxiliary enzyme is more than one, the auxiliary enzyme is used in any proportion; the dosage of the main enzyme is 0.12 to 0.5 percent of the weight of the fermentation liquor; the dosage of the auxiliary enzyme is 0-0.025% of the weight of the fermentation liquor, and if the fermentation liquor is concentrated, the dosage of the main enzyme and the auxiliary enzyme is calculated according to the weight of the fermentation liquor before concentration.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201911161513.XA CN112831531A (en) | 2019-11-25 | 2019-11-25 | Method for producing alga DHA grease by biological enzyme method wall breaking |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201911161513.XA CN112831531A (en) | 2019-11-25 | 2019-11-25 | Method for producing alga DHA grease by biological enzyme method wall breaking |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN112831531A true CN112831531A (en) | 2021-05-25 |
Family
ID=75921902
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201911161513.XA Pending CN112831531A (en) | 2019-11-25 | 2019-11-25 | Method for producing alga DHA grease by biological enzyme method wall breaking |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN112831531A (en) |
-
2019
- 2019-11-25 CN CN201911161513.XA patent/CN112831531A/en active Pending
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101307341B (en) | Production process of docosahexenoic acid grease by bioenzyme method wall-breaking | |
| EP3626806B1 (en) | Methods of recovering oil from microorganisms | |
| CN104394702B (en) | For being constantly enriched with a kind of method of oil generated by microalgae, that there is DHA ethyl ester | |
| CN109777607B (en) | Method for purifying DHA (docosahexaenoic acid) crude oil | |
| CN102199482B (en) | Method for extracting grease from oleaginous microorganisms | |
| WO2004020647A1 (en) | Process for producing and recovering mannosylerythritol lipids from culture medium containing the same | |
| Wan et al. | Production of exopolysaccharide by Ganoderma lucidum in a repeated-batch fermentation | |
| CN106977582A (en) | A kind of method of Hydrolysis kinetics phytosterol in deodorization distillate | |
| CN107418982B (en) | Low-chloropropanol microbial oil and preparation method thereof | |
| EP3083974A1 (en) | Methods of recovering oil from microorganisms | |
| CN109956856A (en) | The method of Co-Q10 and phosphatide is extracted from Co-Q10 zymophyte powder | |
| CN107099561A (en) | Solvent-free extraction method of docosahexaenoic acid-containing oil | |
| CN101870915B (en) | Process for extracting arachidonic acid grease by aqueous enzymatic method | |
| CN112159825A (en) | Method for extracting DHA (docosahexaenoic acid) by fermentation | |
| CN107267282A (en) | Immobilization composite phospholipid enzyme enzymatic degumming technique | |
| CN111690462A (en) | Method for demulsifying and extracting oil from oil-containing algae or fungus cell wall-broken liquid | |
| CN110628840B (en) | Method for extracting myricetin by microbial fermentation | |
| CN112831531A (en) | Method for producing alga DHA grease by biological enzyme method wall breaking | |
| CN109266698B (en) | Method for adjusting fatty acid composition components in Mortierella microbial oil | |
| CN105368885A (en) | Method for enhancing production of alpha-linolenic acid from rhodosporidium toruloides | |
| CN104164411A (en) | Method for preparing glucose oxidase | |
| CN108641802A (en) | A method of extracting peanut oil using microbial fermentation | |
| CN114774485A (en) | Production method of DHA algae oil rich in Sn-2-position palmitic acid | |
| KR101856849B1 (en) | Method of producing glucose from chinese cabbage wastes and algae culture media containing glucose | |
| Nair et al. | Production of lipase of Aspergillus foetidus in a batch stirred reactor |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20210525 |
|
| WD01 | Invention patent application deemed withdrawn after publication |