CN115125146A - High-efficiency low-loss microalgae wall breaking and drying method - Google Patents
High-efficiency low-loss microalgae wall breaking and drying method Download PDFInfo
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- 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
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- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/89—Algae ; Processes using algae
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Abstract
The invention discloses a high-efficiency low-loss microalgae wall-breaking drying method, which comprises the following steps: 1) adding a surfactant to the aqueous solution with the enzyme mixture; 2) adding microalgae cells into an aqueous solution containing an enzyme mixture and a surfactant; 3) a freezing process of controlling a freezing rate, freezing the mixture of the aqueous solution with the enzyme mixture and the surfactant and the microalgae cells obtained in the step 2) at a set temperature change rate; 4) and (4) freeze-drying. The method can complete the wall breaking and drying of microalgae cells of any kind by only using a freeze dryer with a heating function and adjustable temperature change rate without using high temperature and strong shearing force. The method has high wall breaking efficiency, simple flow and simple and convenient operation, does not need to replace containers or equipment in the whole process, and can achieve whole-process cleanness; the whole process has the operating temperature not exceeding 40 ℃, and the process hardly loses active ingredients contained in the microalgae.
Description
Technical Field
The invention belongs to a microalgae processing technology, and particularly relates to a high-efficiency low-loss microalgae wall-breaking drying method.
Background
Microalgae refer to microorganisms that are eukaryotic or prokaryotic and can perform photosynthesis. Some microalgae have high protein content and can be used as food, aquatic feed or livestock feed; some microalgae can synthesize a large amount of secondary metabolites such as grease, pigment, polysaccharide and the like under specific conditions, and the products can be used in the fields of functional foods, food additives, pharmacy, biological energy sources and the like and have considerable economic value. In addition, the extraction of microalgae grease from biomass and the conversion of the microalgae grease into biodiesel are considered to be one of the most important ways for carbon sequestration and emission reduction. At present, the microalgae biotechnology has formed a complete industrial chain of enormous scale on a global scale, in which the processing of microalgae, in particular the breaking and drying, is an important process step.
Most microalgae cells can be directly eaten, but the digestion and absorption rate of direct eating is low, and the change of dosage forms cannot be realized, so that the application range of various microalgae active ingredients is limited. Microalgae have a firm cell wall and a flexible cell membrane, and part of the effective components are also enclosed by a layer of cell organelle membrane. To extract effective components from microalgae, it is necessary to destroy the cell wall and membrane structure of the microalgae and remove water from the microalgae. The traditional treatment method is to divide the wall breaking and drying of microalgae into two process flows and use two different devices. When the process is switched, the microalgae needs to be taken out of one device and put into another device, and the pollution is easily caused. In addition, the traditional drying technology has high process temperature, and can also cause adverse effects on heat-sensitive components contained in the microalgae; the traditional wall breaking process breaks cells by using strong shearing force, so that the energy consumption is high, a large amount of heat can be generated in the process, and the adverse effect on components contained in microalgae is generated. In addition, in practical application, the cell breakage rate of a single process is hardly over 80%.
Most of the active ingredients produced by microalgae exist in microalgae cells, and the microalgae cells are protected by cell walls and cell membranes, and in addition, individual active ingredients are secondarily wrapped by organelle membranes in the cells. The wall breaking efficiency of microalgae is closely related to the extraction efficiency of microalgae components, and the extraction efficiency can be directly improved by improving the wall breaking rate, simplifying the extraction process, improving the yield and reducing the production cost.
The composition of the cell walls of different species of microalgae is also different. The cell walls of the microalgae can be only partially destroyed by using the enzymolysis method alone under most conditions, so that the cell walls are softened and weakened, and only a few cell walls of the microalgae can be completely destroyed by using the enzymolysis method; in addition, the enzymatic method only works on the cell wall, and has no influence on the cell membrane and the membrane of the organelle. Therefore, the enzymatic method needs to be matched with other wall breaking methods in most cases. Thus, although the wall breaking efficiency can be improved, the complexity of the process, and the investment of equipment, energy and time are increased. The surfactant alone can completely destroy the cell membrane in only a few cases when the microalgae cells are treated with the surfactant. Particularly for microalgae with cell walls, the cell membranes softened by the surfactant can not be broken due to the supporting effect of the cell walls on the cell membranes. Therefore, the surfactant treatment needs to be used in combination with other methods for breaking the cell wall. For some microalgae, such as the sporozoites of H.pluvialis, even the use of enzymatic hydrolysis in combination with surfactant treatment does not completely disrupt the cell wall and cell membrane. Freezing the cell can not destroy the cell membrane and cell wall in general conditions, but has certain destructive effect on the inner organelle membrane structure of the cell. The reason for this is that the intact cell membrane and cell wall are both flexible, and even if water freezes to ice and expands in volume, the intact cell membrane and cell wall will expand without being damaged, and the organelles in the cell will be partially damaged by being squeezed.
Therefore, a high-efficiency and low-loss microalgae wall-breaking drying method is urgently needed to be proposed.
Disclosure of Invention
In order to overcome the defects in the prior art, the invention provides a high-efficiency low-loss microalgae wall-breaking drying method.
In order to solve the technical problems, the invention provides the following technical scheme:
the invention provides a high-efficiency low-loss microalgae wall breaking and drying method, which comprises the following steps:
1) adding a surfactant to the aqueous solution with the enzyme mixture;
2) adding microalgae cells into the aqueous solution with the enzyme mixture and the surfactant obtained in the step 1); the step carries out a cell wall enzymolysis process and a surfactant treatment process, wherein the cell wall enzymolysis process uses an aqueous solution with an enzyme mixture to be mixed with microalgae cells, and the microalgae cell walls are hydrolyzed by using the action of enzyme; in the surfactant treatment process, the microalgae cell walls and cell membranes are treated by using a surfactant, and the microalgae cell walls and cell membranes are softened and weakened under the action of the surfactant;
3) a freezing process of controlling a freezing rate, freezing the mixture of the aqueous solution with the enzyme mixture and the surfactant and the microalgae cells obtained in the step 2) at a set temperature change rate;
4) freeze-drying, namely drying the mixture of the aqueous solution with the enzyme mixture and the surfactant and the microalgae cells frozen in the step 3) in a frozen state through sublimation.
Further, the enzyme is at least two of cellulase, pectinase, glucanase, mannanase, chitinase, phospholipase and cholesterol enzyme.
Further, the weight ratio of the enzyme mixture to water is 0.01-0.1; the pH of the aqueous solution containing the enzyme mixture is 3 to 7.
Further, the weight ratio of the aqueous solution with the enzyme mixture to the microalgae cells is 0.5-5, the hydrolysis temperature is 20-40 ℃, and the hydrolysis time is 1-24 hours.
Further, the surfactant is an aqueous solution containing one or more of hydrolyzed lecithin, sodium lauryl sulfate, ammonium lauryl sulfate, poloxamer, sucrose ester, PEG and its esterified product, and polyglycerol ester.
Further, the weight ratio of the surfactant to the aqueous solution with the enzyme mixture is 0.001-0.1.
Further, the cell wall enzymolysis process and the surfactant treatment process are carried out simultaneously.
Further, in the step 3), the temperature is reduced from the initial temperature to below-50 ℃ by using the temperature change rate of 1-50 ℃/h for freezing.
Compared with the prior art, the invention has the following beneficial effects:
the invention can hydrolyze microalgae cell walls with different components through the synergistic effect of different enzymes. Firstly softening and weakening cell walls through the hydrolysis of enzyme; meanwhile, the incompletely hydrolyzed cell wall is softened and weakened again through the action of the surfactant, and the structure of the microalgae cell membrane is weakened and locally damaged. In addition, the invention promotes the water inside the microalgae cells to form an ice crystal structure through the freezing process of controlling the freezing rate, and makes the ice crystals sharper. Through the formation of ice crystals and the characteristic that the volume of water after freezing into ice becomes large, the method not only can destroy the membrane structure of organelles in cells, but also can completely destroy the weakened and softened cell walls and cell membrane structures from the inside, so that effective components contained in the microalgae cells are released, and the preparation is prepared for the subsequent extraction process.
The cell wall breaking rate of the method of the invention is over 99 percent, and the wall breaking and drying of microalgae cells of any kind can be completed only by using one freeze dryer with heating function and adjustable temperature change rate. In the whole process, equipment does not need to be replaced, the microalgae to be treated does not need to be directly operated, the loss caused by equipment replacement and transportation is completely avoided, and the pollution possibly caused by equipment replacement is completely avoided. Compared with a plurality of independent devices and a plurality of processes for drying and wall breaking, the invention only uses one device, thereby reducing the device cost; in addition, the method only needs one flow, is simple and convenient to operate, does not need to replace containers or equipment in the whole process, reduces the personnel cost, saves the operation time, and can achieve the whole-process cleanness of the process. In addition, the whole process of the invention has the operating temperature not exceeding 40 ℃, and the process hardly loses active ingredients contained in the microalgae.
Detailed Description
The preferred embodiments of the present invention are described below, and it should be understood that the preferred embodiments described herein are only for illustrating and explaining the present invention and are not to be construed as limiting the present invention.
Example 1
Mixing cellulase, pectinase, glucanase and mannase with water according to the weight ratio of 0.05; after dissolution, the pH of the solution was adjusted to 5 using 1M hydrochloric acid solution. Hydrolyzed lecithin, sodium lauryl sulfate, and ammonium lauryl sulfate were added to the above aqueous solution with the enzyme mixture in a weight ratio of 0.005. After complete dissolution, the above solution was mixed with 500g of fresh haematococcus pluvialis sporophyte cells in a weight ratio of 1. The mixture was charged into a freeze dryer of Tianfeng brand (Shanghai) model TF-HFD-4. The temperature inside the freeze dryer was adjusted to 37 ℃ and the above mixture was allowed to stand for 4 hours. And (3) cooling the mixture in the freeze dryer from room temperature at the speed of 12 ℃/hour, starting a vacuum pump of the freeze dryer to start freeze drying after the mixture is completely frozen and the temperature reaches-56 ℃, and finishing the process drying after 4 hours.
Comparative example 1
Mixing cellulase, pectinase, glucanase and mannase with water according to the weight ratio of 0.05; after dissolution, the pH of the solution was adjusted to 5 using 1M hydrochloric acid solution. After complete dissolution, the above solution was mixed with 500g of fresh haematococcus pluvialis sporophyte cells in a weight ratio of 1. The mixture was charged into a freeze dryer of Tianfeng brand (Shanghai) model TF-HFD-4. The temperature inside the freeze dryer was adjusted to 37 ℃ and the above mixture was allowed to stand for 4 hours. And (3) cooling the mixture in the freeze dryer from room temperature at the speed of 12 ℃/hour, starting a vacuum pump of the freeze dryer to start freeze drying after the mixture is completely frozen and the temperature reaches-56 ℃, and finishing the process drying after 4 hours.
Comparative example 2
The hydrolyzed lecithin, sodium lauryl sulfate, and ammonium lauryl sulfate were added to water in a weight ratio of 0.005. After complete dissolution, the above solution was mixed with 500g of fresh haematococcus pluvialis sporophyte cells in a weight ratio of 1. Loading the mixture into freeze-drying machine of Tianfeng brand (Shanghai) model TF-HFD-4. The mixture was charged into a freeze dryer of Tianfeng brand (Shanghai) model TF-HFD-4. The temperature inside the freeze dryer was adjusted to 37 ℃ and the above mixture was allowed to stand for 4 hours. And (3) cooling the mixture in the freeze dryer from room temperature at the speed of 12 ℃/hour, starting a vacuum pump of the freeze dryer to start freeze drying after the mixture is completely frozen and the temperature reaches-56 ℃, and finishing the process drying after 4 hours.
Comparative example 3
Mixing cellulase, pectinase, glucanase and mannase with water according to the weight ratio of 0.05; after dissolution, the pH of the solution was adjusted to 5 using 1M hydrochloric acid solution. Hydrolyzed lecithin, sodium lauryl sulfate, and ammonium lauryl sulfate were added to the above aqueous solution with the enzyme mixture in a weight ratio of 0.005. After complete dissolution, the above solution was mixed with 500g of fresh haematococcus pluvialis sporophyte cells in a weight ratio of 1. Loading the mixture into freeze-drying machine of Tianfeng brand (Shanghai) model TF-HFD-4. The temperature inside the freeze dryer was adjusted to 37 ℃ and the above mixture was allowed to stand for 4 hours. Freezing the mixture in the freeze dryer at the highest speed, starting a vacuum pump of the freeze dryer to start freeze drying after the mixture is completely frozen and the temperature reaches-56 ℃, and finishing the process drying after 4 hours.
Comparative example 4
Quickly freezing 500g of fresh haematococcus pluvialis sporophyte cells by using liquid nitrogen, then filling the frozen haematococcus pluvialis sporophyte cells into a precooled Tianfeng brand (Shanghai) freeze dryer with the model of TF-HFD-4, starting a vacuum pump of the freeze dryer to freeze and dry, and finishing the process drying after 4 hours.
Comparative example 5
The water was mixed with 500g of fresh haematococcus pluvialis sporophyte cells in a weight ratio of 1. The above mixture was ground using a Tianxiao powder (Changsha) brand planetary ball mill at 500rpm for 4 hours using stainless steel balls. After grinding, separating the stainless steel balls from the water containing the microalgae by using a screen, filling the water containing the microalgae into a freeze dryer with the model of Tianfeng brand (Shanghai) TF-HFD-4, cooling to-56 ℃ at the highest speed, starting a vacuum pump of the freeze dryer to perform freeze drying, and drying after 4 hours.
After the drying of example 1 and comparative examples 1, 2, 3, 4 and 5 was completed, 0.05mg of sample was accurately weighed from each sample, 6 parallel samples were taken, each sample was suspended with 500uL of pure water, 50uL of sample was added to a cell counting slide after shaking was uniform, and the number of cells that were not wall-broken was recorded by observing through a microscope, and the wall-breaking rate was calculated based on comparative example 4 (0% wall-broken). The results obtained are shown in the following table:
| example 1 | Comparative example 1 | Comparative example 2 | Comparative example 3 | Comparative example 5 | |
| Wall breaking rate | 99.6％±0.2％ | 2.3％±1.2％ | 1.1％±0.9％ | 73.2％±8.1％ | 78.2％±11.2％ |
Finally, it should be noted that: although the present invention has been described in detail with reference to the foregoing embodiments, it will be apparent to those skilled in the art that changes may be made in the embodiments and/or equivalents thereof without departing from the spirit and scope of the invention. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (8)
1. A high-efficiency low-loss microalgae wall breaking and drying method is characterized by comprising the following steps:
1) adding a surfactant to the aqueous solution with the enzyme mixture;
2) adding microalgae cells into the aqueous solution with the enzyme mixture and the surfactant obtained in the step 1); the step carries out a cell wall enzymolysis process and a surfactant treatment process, wherein the cell wall enzymolysis process uses an aqueous solution with an enzyme mixture to be mixed with microalgae cells, and the microalgae cell walls are hydrolyzed by the action of enzyme; in the surfactant treatment process, the microalgae cell walls and cell membranes are treated by using a surfactant, and are softened and weakened under the action of the surfactant;
3) a freezing process of controlling a freezing rate, freezing the mixture of the aqueous solution with the enzyme mixture and the surfactant and the microalgae cells obtained in the step 2) at a set temperature change rate;
4) freeze-drying, namely drying the mixture of the aqueous solution with the enzyme mixture and the surfactant and the microalgae cells frozen in the step 3) in a frozen state through sublimation.
2. The method for breaking wall and drying microalgae according to claim 1, wherein the enzyme is at least two of cellulase, pectinase, glucanase, mannanase, chitinase, phospholipase and cholesterol enzyme.
3. The method for breaking the wall and drying the microalgae according to claim 1, wherein the weight ratio of the enzyme mixture to water in step 1) is 0.01-0.1, and the pH of the aqueous solution containing the enzyme mixture is 3-7.
4. The method for breaking the wall of microalgae according to claim 1, wherein the weight ratio of the aqueous solution containing the enzyme mixture to the microalgae cells is 0.5-5, the hydrolysis temperature is 20-40 ℃, and the hydrolysis time is 1-24 hours.
5. The method for breaking the wall and drying the microalgae according to claim 1, wherein the surfactant is an aqueous solution containing one or more of hydrolyzed lecithin, sodium lauryl sulfate, ammonium lauryl sulfate, poloxamer, sucrose esters, PEG and its esters, and polyglycerol esters.
6. The method for breaking the wall and drying the microalgae according to claim 1, wherein the weight ratio of the surfactant to the aqueous solution containing the enzyme mixture is 0.001-0.1.
7. The method for breaking the wall and drying the microalgae according to claim 1, wherein the temperature change rate of the microalgae in the step 3) is reduced from the initial temperature to below-50 ℃ per hour for freezing.
8. The method for breaking the wall and drying the microalgae with high efficiency and low loss as claimed in claim 1, wherein the equipment used in the method is a freeze dryer with heating function and adjustable temperature change rate.
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Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101352249A (en) * | 2008-09-04 | 2009-01-28 | 蔡志武 | Production method for fully using oil-containing micro-algae |
| CN101756300A (en) * | 2009-12-30 | 2010-06-30 | 张炳泉 | Chlorella cell wall disruption method |
| CN102504999A (en) * | 2011-12-22 | 2012-06-20 | 新奥科技发展有限公司 | Treatment method of microalgae |
| CN103396951A (en) * | 2013-08-12 | 2013-11-20 | 云南爱尔康生物技术有限公司 | Method for cultivating haematococcus pluvialis in large scale and producing natural astaxanthin seasoning packet by haematococcus pluvialis |
| CN104928183A (en) * | 2015-06-26 | 2015-09-23 | 新奥科技发展有限公司 | Wall breaking method for microalgae cell walls and method for extracting protein from microalgae |
| CN106029624A (en) * | 2013-12-20 | 2016-10-12 | 帝斯曼知识产权资产管理有限公司 | Processes for obtaining microbial oil from microbial cells |
| US20160324167A1 (en) * | 2008-10-14 | 2016-11-10 | Terravia Holdings, Inc. | Novel microalgal food compositions |
| CN107460129A (en) * | 2017-08-21 | 2017-12-12 | 李彤 | The industrialization microalgae culture method that biomass separates with nutrient solution |
| CN109142549A (en) * | 2018-08-27 | 2019-01-04 | 睿藻生物科技（苏州）有限公司 | A kind of microalgae metabolic rate measurement chamber for mass spectrometer |
-
2022
- 2022-07-15 CN CN202210839702.3A patent/CN115125146A/en active Pending
Patent Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101352249A (en) * | 2008-09-04 | 2009-01-28 | 蔡志武 | Production method for fully using oil-containing micro-algae |
| US20160324167A1 (en) * | 2008-10-14 | 2016-11-10 | Terravia Holdings, Inc. | Novel microalgal food compositions |
| CN101756300A (en) * | 2009-12-30 | 2010-06-30 | 张炳泉 | Chlorella cell wall disruption method |
| CN102504999A (en) * | 2011-12-22 | 2012-06-20 | 新奥科技发展有限公司 | Treatment method of microalgae |
| CN103396951A (en) * | 2013-08-12 | 2013-11-20 | 云南爱尔康生物技术有限公司 | Method for cultivating haematococcus pluvialis in large scale and producing natural astaxanthin seasoning packet by haematococcus pluvialis |
| CN106029624A (en) * | 2013-12-20 | 2016-10-12 | 帝斯曼知识产权资产管理有限公司 | Processes for obtaining microbial oil from microbial cells |
| CN104928183A (en) * | 2015-06-26 | 2015-09-23 | 新奥科技发展有限公司 | Wall breaking method for microalgae cell walls and method for extracting protein from microalgae |
| CN107460129A (en) * | 2017-08-21 | 2017-12-12 | 李彤 | The industrialization microalgae culture method that biomass separates with nutrient solution |
| CN109142549A (en) * | 2018-08-27 | 2019-01-04 | 睿藻生物科技（苏州）有限公司 | A kind of microalgae metabolic rate measurement chamber for mass spectrometer |
Non-Patent Citations (4)
| Title |
|---|
| BENJAMIN SCHMID等: "Drying Microalgae Using an Industrial Solar Dryer: A Biomass Quality Assessment", 《FOODS》, vol. 11, no. 13, pages 1873 * |
| 周湘池等: "雨生红球藻(Haematococcus pluvialis)破壁方法对虾青素提取率的影响", 《海洋与湖沼》, no. 05, pages 424 - 429 * |
| 唐佳芮等: "微藻油加工技术研究进展", 《粮食与食品工业》, vol. 26, no. 01, pages 10 - 12 * |
| 梁英妹等: "小球藻食品工艺中的关键技术", 《科技创新导报》, vol. 15, no. 10, pages 247 - 250 * |
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