CN110628634A - Chlorella wall breaking method - Google Patents
Chlorella wall breaking method Download PDFInfo
- Publication number
- CN110628634A CN110628634A CN201910944042.3A CN201910944042A CN110628634A CN 110628634 A CN110628634 A CN 110628634A CN 201910944042 A CN201910944042 A CN 201910944042A CN 110628634 A CN110628634 A CN 110628634A
- Authority
- CN
- China
- Prior art keywords
- chlorella
- wall breaking
- liquid
- wall
- breaking
- 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
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/06—Lysis of microorganisms
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/06—Lysis of microorganisms
- C12N1/066—Lysis of microorganisms by physical methods
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Biotechnology (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Genetics & Genomics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biomedical Technology (AREA)
- Virology (AREA)
- Microbiology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Medicinal Chemistry (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Mycology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The invention relates to a wall breaking method of chlorella, which comprises the following steps: dissolving the chlorella dry powder in water according to a feed-liquid ratio of 10g/L to obtain chlorella liquid; adding cellulase accounting for 1-2% of the mass of the chlorella dry powder into the chlorella liquid, and adjusting the pH of the chlorella liquid to 4-5; adding the algae liquid after pH adjustment into a colloid mill, and grinding at the rotating speed of 8000 r/min-10000 r/min for 1 h; and (3) placing the ground chlorella solution in a water bath at 45-50 ℃, stirring, heating and carrying out enzymolysis for 2-3 h to complete the wall breaking treatment of the chlorella. The method utilizes colloid mill equipment to assist in enzymatic wall breaking, disperses the agglomeration phenomenon of the chlorella powder through the colloid mill, and has grinding and crushing effects on cell walls, thereby improving the wall breaking efficiency, using a small amount of enzyme and a short time to ensure that the chlorella has higher wall breaking rate, and has low cost and simple and convenient operation.
Description
Technical Field
The invention relates to the technical field of algae treatment, in particular to a wall breaking method of chlorella.
Background
Chlorella sp, a kind of unicellular, common green algae, belongs to the family of Oomycetaceae, the class of Chlorophyceae, the phylum Chlorophyceae, and has been widely cultivated. The chlorella grown under good conditions has high application value, the protein content can reach 63.60%, and the chlorella also contains docosahexaenoic acid (DHA). Chlorella has been used as a good food and animal feed additive for over 30 years in the United states and Japan. In recent years, China also pays more attention to the development and utilization of chlorella. However, since chlorella has tough cell walls and cannot be directly eaten, and extraction of its internal functional substances such as growth factor (CGF) is also hindered, the wide use of chlorella is somewhat limited.
At present, the chlorella wall breaking method mainly comprises a physical wall breaking method and a biochemical wall breaking method; the physical wall breaking method comprises a grinding method, a mechanical homogenizing method, an ultrasonic wall breaking method and a repeated freezing and thawing method, the repeated freezing and thawing method needs long time, and the wall breaking efficiency is low; the ultrasonic wall breaking method has high efficiency, but has high requirements on the concentration of wall-broken algae cells, the wall breaking efficiency is greatly reduced under high concentration, and the method needs ice bath and is not suitable for large-scale wall breaking. The enzymolysis wall-breaking method in the biochemical wall-breaking method cannot treat the agglomeration phenomenon of algae cells, and affects the wall-breaking rate, so that a large amount of enzyme needs to be added for treatment or the enzymolysis treatment time needs to be prolonged, the wall-breaking cost is increased, and the efficiency is low.
Therefore, the current wall breaking method for extracting chlorella still needs to be improved.
Disclosure of Invention
The present invention is directed to solving, at least to some extent, one of the technical problems in the related art. Therefore, the invention aims to provide a chlorella wall breaking method. The method can improve wall breaking efficiency of Chlorella, and has simple operation and low cost.
Therefore, in one aspect of the invention, the invention provides a method for breaking the wall of chlorella, which comprises the following steps:
(1) dissolving the chlorella dry powder in water according to a feed-liquid ratio of 10g/L to obtain chlorella liquid;
(2) adding cellulase accounting for 1-2% of the mass of the chlorella dry powder into the chlorella liquid, and adjusting the pH of the chlorella liquid to 4-5;
(3) adding the algae liquid with the pH adjusted in the step (2) into a colloid mill, and grinding at the rotating speed of 8000 r/min-10000 r/min for 1 h;
(4) and (4) placing the algae liquid ground in the step (3) in a water bath at 45-50 ℃, stirring, heating and carrying out enzymolysis for 2-3 h to complete the wall breaking treatment of the chlorella.
According to the chlorella wall breaking method, the colloid mill is used for assisting in carrying out enzymatic wall breaking, the conglomeration phenomenon of chlorella powder is dispersed through the colloid mill, the cell walls have grinding and crushing functions, meanwhile, the colloid mill generates heat during working to heat the chlorella powder, an optimal temperature can be provided for chlorella cell wall enzymolysis during grinding, and therefore the wall breaking efficiency is improved, the chlorella can have a high wall breaking rate in a short time with a small amount of enzyme, the cost is low, and the operation is simple and convenient.
In addition, the method for breaking the wall of the chlorella according to the embodiment of the present invention may further have the following additional technical features:
according to the embodiment of the invention, in the step (3), the temperature in the display screen of the colloid mill is controlled to be 45-50 ℃.
According to an embodiment of the present invention, in the step (3), the model of the colloid mill is an IKA magic LAB type.
According to the embodiment of the invention, the cell count instrument is used for counting the cells before and after the wall breaking of the chlorella to calculate the wall breaking rate of the chlorella.
According to the embodiment of the invention, the cellulase activity is 0.3-1.0 ten thousand U.
Additional aspects and advantages of the invention will be set forth in part in the description which follows and, in part, will be obvious from the description, or may be learned by practice of the invention.
Detailed Description
The technical solution of the present invention is illustrated by specific examples below. It is to be understood that one or more method steps mentioned in the present invention do not exclude the presence of other method steps before or after the combination step or that other method steps may be inserted between the explicitly mentioned steps; it should also be understood that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention. Moreover, unless otherwise indicated, the numbering of the various method steps is merely a convenient tool for identifying the various method steps, and is not intended to limit the order in which the method steps are arranged or the scope of the invention in which the invention may be practiced, and changes or modifications in the relative relationship may be made without substantially changing the technical content.
In order to better understand the above technical solutions, exemplary embodiments of the present invention are described in more detail below. While exemplary embodiments of the invention have been shown, it should be understood that the invention may be embodied in various forms and should not be construed as limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete, and will fully convey the scope of the invention to those skilled in the art.
The test materials adopted by the invention are all common commercial products and can be purchased in the market.
The invention provides a wall breaking method of chlorella, which comprises the following steps:
(1) dissolving the chlorella dry powder in water according to a feed-liquid ratio of 10g/L to obtain chlorella liquid;
(2) adding cellulase accounting for 1-2% of the mass of the chlorella dry powder into the chlorella liquid, and adjusting the pH of the chlorella liquid to 4-5; wherein the activity of the cellulase is 0.3-1.0 ten thousand U.
(3) Adding the algae liquid with the pH adjusted in the step (2) into an IKA magic LAB type colloid mill, and grinding at the rotating speed of 8000 r/min-10000 r/min for 1 h; wherein the temperature in the display screen of the colloid mill is controlled at 45-50 ℃.
(4) And (4) placing the algae liquid ground in the step (3) in a water bath at 45-50 ℃, stirring, heating and carrying out enzymolysis for 2-3 h to complete the wall breaking treatment of the chlorella.
According to the chlorella wall breaking method, the colloid mill is used for assisting in carrying out enzymatic wall breaking, the conglomeration phenomenon of chlorella powder is dispersed through the colloid mill, the cell walls have grinding and crushing functions, meanwhile, the colloid mill generates heat during working to heat the chlorella powder, an optimal temperature can be provided for chlorella cell wall enzymolysis during grinding, and therefore the wall breaking efficiency is improved, the chlorella can have a high wall breaking rate in a short time with a small amount of enzyme, the cost is low, and the operation is simple and convenient.
The invention will now be described with reference to specific examples, which are intended to be illustrative only and not to be limiting in any way.
Example 1
A wall breaking method of chlorella comprises the following steps:
(1) weighing 20g of chlorella dry powder, and adding 2L of water for dissolving to obtain chlorella liquid.
(2) Adding 0.4g of cellulase with the enzyme activity of 0.5 ten thousand U into the algae liquid, and adjusting the pH value of the algae liquid to be 4 by using a citric acid solution.
(3) Adding the algae liquid with the pH adjusted in the step (2) into an IKA magic LAB type colloid mill, and grinding at the rotating speed of 10000r/min for 1 h; wherein the temperature in the display screen of the colloid mill is controlled at 45-50 ℃.
(4) And (4) placing the algae liquid ground in the step (3) in a water bath at 50 ℃, stirring, heating and carrying out enzymolysis for 3 hours to complete enzymolysis, so as to complete the wall breaking treatment of the chlorella.
In the embodiment, a cell counter is adopted to count the cells before and after wall breaking, and the wall breaking rate (%) of the chlorella is (the number of the chlorella before wall breaking-the number of the chlorella after wall breaking)/the number of the chlorella before wall breaking; the wall breaking rate of the chlorella is calculated to be 84.5%.
Example 2
A wall breaking method of chlorella comprises the following steps:
(1) weighing 20g of chlorella dry powder, and adding 2L of water for dissolving to obtain chlorella liquid.
(2) Adding 0.2g of cellulase with the enzyme activity of 1.0 ten thousand U into the algae liquid, and adjusting the pH value of the algae liquid to be 5 by using a citric acid solution.
(3) Adding the algae liquid with the pH adjusted in the step (2) into an IKA magic LAB type colloid mill, and grinding at the rotating speed of 8000r/min for 1 h; wherein the temperature in the display screen of the colloid mill is controlled at 45-50 ℃.
(4) And (4) placing the algae liquid ground in the step (3) in a water bath at 45 ℃, stirring, heating and carrying out enzymolysis for 2 hours to complete enzymolysis, so as to complete the wall breaking treatment of the chlorella.
In the embodiment, a cell counter is adopted to count the cells before and after wall breaking, and the wall breaking rate (%) of the chlorella is (the number of the chlorella before wall breaking-the number of the chlorella after wall breaking)/the number of the chlorella before wall breaking; the wall breaking rate of the chlorella is calculated to be 82.7%.
Example 3
A wall breaking method of chlorella comprises the following steps:
(1) weighing 40g of chlorella dry powder, and adding 4L of water for dissolving to obtain chlorella liquid.
(2) Adding 0.8g of cellulase with the enzyme activity of 0.7 ten thousand U into the algae liquid, and adjusting the pH value of the algae liquid to be 4 by using a citric acid solution.
(3) Adding the algae liquid with the pH adjusted in the step (2) into an IKA magic LAB type colloid mill, and grinding at the rotating speed of 10000r/min for 1 h; wherein the temperature in the display screen of the colloid mill is controlled at 45-50 ℃.
(4) And (4) placing the algae liquid ground in the step (3) in a water bath at 50 ℃, stirring, heating and carrying out enzymolysis for 3 hours to complete enzymolysis, so as to complete the wall breaking treatment of the chlorella.
In the embodiment, a cell counter is adopted to count the cells before and after wall breaking, and the wall breaking rate (%) of the chlorella is (the number of the chlorella before wall breaking-the number of the chlorella after wall breaking)/the number of the chlorella before wall breaking; the wall breaking rate of the chlorella is calculated to be 86.3%.
In conclusion, according to the chlorella wall breaking method provided by the invention, the colloid mill is used for assisting in carrying out enzymatic wall breaking, the conglomeration phenomenon of the chlorella powder is dispersed through the colloid mill, the cell walls have grinding and crushing functions, meanwhile, the colloid mill generates heat during working to heat the chlorella powder, an optimal temperature can be provided for the enzymolysis of the chlorella cell walls during grinding, the optimal temperature and the synergistic effect of the two can be realized, so that the wall breaking efficiency is improved, the chlorella can have a high wall breaking rate by using a small amount of enzyme in a short time, the wall breaking rate can reach 86.3%, the cost is low, and the operation is simple and convenient.
In the description herein, references to the description of the term "one embodiment," "some embodiments," "an example," "a specific example," or "some examples," etc., mean that a particular feature, structure, material, or characteristic described in connection with the embodiment or example is included in at least one embodiment or example of the invention. In this specification, the schematic representations of the terms used above should not be understood to necessarily refer to the same embodiment or example. Furthermore, the particular features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples. Furthermore, various embodiments or examples described in this specification can be combined and combined by those skilled in the art.
Although embodiments of the present invention have been shown and described above, it is understood that the above embodiments are exemplary and should not be construed as limiting the present invention, and that variations, modifications, substitutions and alterations can be made to the above embodiments by those of ordinary skill in the art within the scope of the present invention.
Claims (5)
1. A wall breaking method of chlorella is characterized by comprising the following steps:
(1) dissolving the chlorella dry powder in water according to a feed-liquid ratio of 10g/L to obtain chlorella liquid;
(2) adding cellulase accounting for 1-2% of the mass of the chlorella dry powder into the chlorella liquid, and adjusting the pH of the chlorella liquid to 4-5;
(3) adding the algae liquid with the pH adjusted in the step (2) into a colloid mill, and grinding at the rotating speed of 8000 r/min-10000 r/min for 1 h;
(4) and (4) placing the algae liquid ground in the step (3) in a water bath at 45-50 ℃, stirring, heating and carrying out enzymolysis for 2-3 h to complete the wall breaking treatment of the chlorella.
2. A method for breaking cell wall of Chlorella vulgaris according to claim 1 wherein in said step (3), the temperature in the display screen of the colloid mill is controlled to be 45-50 ℃.
3. The method for breaking cell wall of chlorella according to claim 1, wherein in the step (3), the model of the colloid mill is IKA magic LAB type.
4. A wall-breaking method of chlorella according to claim 1, wherein the cell count before and after the wall-breaking of chlorella is carried out by a cell counter to calculate the wall-breaking rate of chlorella.
5. The method for breaking a wall of chlorella according to claim 1, wherein the cellulase activity is 0.3 to 1.0 ten thousand U.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910944042.3A CN110628634A (en) | 2019-09-30 | 2019-09-30 | Chlorella wall breaking method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910944042.3A CN110628634A (en) | 2019-09-30 | 2019-09-30 | Chlorella wall breaking method |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN110628634A true CN110628634A (en) | 2019-12-31 |
Family
ID=68974916
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910944042.3A Pending CN110628634A (en) | 2019-09-30 | 2019-09-30 | Chlorella wall breaking method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110628634A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114432348A (en) * | 2022-02-21 | 2022-05-06 | 日照职业技术学院 | Method for extracting growth factor from microalgae |
| CN115074250A (en) * | 2022-03-27 | 2022-09-20 | 中科技术物理青岛研究院 | Efficient chlorella cell wall breaking method and application |
Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101429467A (en) * | 2008-12-24 | 2009-05-13 | 青岛生物能源与过程研究所 | Method for simultaneously extracting lipid and protein from microalgae |
| CN101756300A (en) * | 2009-12-30 | 2010-06-30 | 张炳泉 | Chlorella cell wall disruption method |
| WO2012002447A1 (en) * | 2010-06-30 | 2012-01-05 | 武田薬品工業株式会社 | Method for detection of mutated polynucleotide |
| CN103555582A (en) * | 2013-10-22 | 2014-02-05 | 渤海大学 | Method for breaking wall of chlorella by using compound enzyme |
| CN104312720A (en) * | 2014-11-11 | 2015-01-28 | 甘肃德福生物科技有限公司 | Method for extracting oil from chlorella |
| CN104432318A (en) * | 2014-12-03 | 2015-03-25 | 甘肃德福生物科技有限公司 | Preparation method of chlorella beverage |
| WO2016009146A1 (en) * | 2014-07-18 | 2016-01-21 | Roquette Freres | Method for extracting soluble proteins from microalgal biomass |
-
2019
- 2019-09-30 CN CN201910944042.3A patent/CN110628634A/en active Pending
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101429467A (en) * | 2008-12-24 | 2009-05-13 | 青岛生物能源与过程研究所 | Method for simultaneously extracting lipid and protein from microalgae |
| CN101756300A (en) * | 2009-12-30 | 2010-06-30 | 张炳泉 | Chlorella cell wall disruption method |
| WO2012002447A1 (en) * | 2010-06-30 | 2012-01-05 | 武田薬品工業株式会社 | Method for detection of mutated polynucleotide |
| CN103555582A (en) * | 2013-10-22 | 2014-02-05 | 渤海大学 | Method for breaking wall of chlorella by using compound enzyme |
| WO2016009146A1 (en) * | 2014-07-18 | 2016-01-21 | Roquette Freres | Method for extracting soluble proteins from microalgal biomass |
| CN104312720A (en) * | 2014-11-11 | 2015-01-28 | 甘肃德福生物科技有限公司 | Method for extracting oil from chlorella |
| CN104432318A (en) * | 2014-12-03 | 2015-03-25 | 甘肃德福生物科技有限公司 | Preparation method of chlorella beverage |
Non-Patent Citations (3)
| Title |
|---|
| ELKE BRAUN 等: "Enzymatic Degradation of the Cell Wall of Chlorella", 《PLANTA》 * |
| 梁英妹 等: "小球藻食品工艺中的关键技术", 《科技创新导报》 * |
| 钟韵山 等: "小球藻破壁技术研究进展", 《食品研究与开发》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114432348A (en) * | 2022-02-21 | 2022-05-06 | 日照职业技术学院 | Method for extracting growth factor from microalgae |
| CN115074250A (en) * | 2022-03-27 | 2022-09-20 | 中科技术物理青岛研究院 | Efficient chlorella cell wall breaking method and application |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102276301B (en) | Straw decomposing agent and production method thereof | |
| CN108013213A (en) | A kind of moist microorganism particle fermented feed for aquatic livestock and preparation method thereof | |
| CN103555556B (en) | Method for preparing ginseng vinegar by virtue of fermentation with corncobs and corn ears as raw materials | |
| CN101974386A (en) | Method for preparing novel small yeast | |
| CN110628634A (en) | Chlorella wall breaking method | |
| CN103555582B (en) | Method for breaking wall of chlorella by using compound enzyme | |
| CN105661013B (en) | A kind of method that fermentation of Chinese herbal medicine prepares compound feed additive for promoting growth | |
| CN104830346B (en) | Agricultural wastes are as the screening of acid soil improving agent and Application way | |
| CN102887732A (en) | Method for producing amino acid liquid fertilizer by processing wastewater via tilapias | |
| CN112352870A (en) | Preparation method of marine fish peptide suitable for piglet daily feed | |
| CN105533289A (en) | Preparation method of pelteobagrus fulvidraco feed | |
| CN104171406A (en) | Selenium-enriched traditional Chinese herbal medicine fermented broiler feed | |
| CN102337225B (en) | Preparation method of high-nitrogen fresh yeast and extract | |
| CN104016779B (en) | A kind of Rape Straw is culture medium for golden mushroom of raw material and preparation method thereof | |
| CN104140302A (en) | Special fertilizer for carrots and production method thereof | |
| CN106146143A (en) | A kind of oily nutrient fluid with high efficiency special with Paeonia suffruticosa and preparation method thereof | |
| CN102206567A (en) | Method for producing rice-koji for spirits | |
| CN101525564B (en) | Method for improving beer wort nutrition and malt culms nutrient used thereof | |
| CN101864401B (en) | Liquid complex enzyme for use in brewing red wine and use thereof | |
| CN105503392A (en) | Nutrient solution for improving growth environment of aquatic plants | |
| CN108378230A (en) | A kind of production method of feed for goat | |
| CN107573146A (en) | A kind of fertilizer for family planting vegetables | |
| CN101695290B (en) | Method for pearl culture | |
| CN100497209C (en) | Preparation of probiotics bacterium and method for rehabilitation of aquaculture environment | |
| CN105746886B (en) | A method of compound green feed additive for promoting growth is prepared using aquatic product protein and Chinese herbal medicine fermentation |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination |