CN111748478B - Microalgae wall-breaking processing method and application thereof - Google Patents
Microalgae wall-breaking processing method and application thereof Download PDFInfo
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Abstract
The invention relates to a microalgae wall-breaking processing method, which comprises the following steps: performing screw extrusion treatment on microalgae with humidity of 25-75%, drying the extruded material, and pulverizing to obtain wall-broken microalgae powder. The wall breaking method for microalgae provided by the invention has the advantages of high wall breaking rate, high speed, low cost and low energy consumption; after microalgae with high protein content such as chlorella pyrenoidosa is treated, the protein dissolution rate is remarkably improved to 92% at most, and the high-level structure of the protein is fully dissociated, so that the protein is easier to be absorbed by a human body; after microalgae with high oil content such as chlorella vulgaris is treated, the oil leaching rate is remarkably improved and can reach 92% at most.
Description
Technical Field
The invention belongs to the technical field of microalgae wall breaking, and particularly relates to a microalgae wall breaking processing method and application thereof.
Background
Microalgae are one of the earliest lives on the earth 20 billion years ago, are ubiquitous and exist in oceans, lakes, lands, deserts and even high-altitude dust, are main contributors to oxygen of the earth and are main primary producers of biomass on the earth. Microalgae such as chlorella, spirulina, etc. contain many edible high-value substances such as polyunsaturated fatty acids, proteins, carotenoids, etc.
However, many microalgae have cell walls, such as chlorella, spirulina, and especially chlorella has a thick cell wall that is not digestible when eaten directly. Therefore, as a food raw material, microalgae with cell walls need to be subjected to wall breaking treatment to ensure that a human body can obtain functional components in the microalgae. At present, the mainstream wall breaking methods include a grinding method, a high-pressure homogenization method, an ultrasonic method, a repeated freeze-thaw method, an enzyme hydrolysis method, a cavitation method and the like, and have limited effects on plant materials and microorganisms.
CN109609255A discloses a method for extracting microalgae oil by bio-enzyme catalysis wall breaking, which comprises four steps of microalgae culture centrifugal screening, protease treatment and boiling, ultrasonic treatment after complex enzyme treatment and distillation extraction; through changing the adding sequence of the enzyme and controlling the temperature of the solution, the activity of the enzyme is maximized, the using amount of the enzyme is reduced, finally, the decomposition of the whole microalgae cells is accelerated through short-time ultrasonic treatment, meanwhile, the decomposition products are separated from the grease on the microalgae, and the extraction of the follow-up grease is facilitated.
CN101817738A discloses a method for breaking the walls of algae and fungi cells and extracting the intracellular lipid product DHA by physical method without the assistance of any organic solvent and chemical drugs. Separating and collecting cells of fermented microalgae or fungus fermentation liquor by a separation system, adjusting pH of the bacterial sludge to 2.0-4.0 with acid, controlling the temperature of the bacterial sludge to 10-20 deg.C, adding antioxidant, and performing high-pressure homogenization and wall breaking by a high-pressure homogenizer; and adding water into the bacterial sludge after wall breaking, stirring, and separating the feed liquid by a three-phase separator to obtain DHA grease.
CN102051330A discloses a method for rapidly crushing microalgae cells, mixing the microalgae to be crushed with liquid nitrogen for 0.1-1h, mechanically grinding and crushing to obtain the microalgae with crushed cells, and illuminating for 0.5-2h for extracting oil.
However, the above-mentioned wall-breaking technique has the following disadvantages: the equipment is complex and the cost is high; the energy consumption is too high, and the waste water discharge is large; the efficiency is too low, and the method is not suitable for industrial production, so that the development of the microalgae wall-breaking processing method which has high wall-breaking rate, high speed, low cost and low energy consumption and is suitable for industrial production is very meaningful.
Disclosure of Invention
Aiming at the defects of the prior art, the invention aims to provide a microalgae wall-breaking processing method and application thereof.
In order to achieve the purpose, the invention adopts the following technical scheme:
in one aspect, the invention provides a microalgae wall-breaking processing method, which comprises the following steps: performing screw extrusion treatment on microalgae with humidity of 25-75%, drying the extruded material, and pulverizing to obtain wall-broken microalgae powder.
The humidity of the microalgae can be 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70% or 75%, and any specific point value in the above numerical range can be selected, and is not described in detail herein.
And the screw extrusion treatment adopts a single-screw extruder or a double-screw extruder for treatment. The outer side of the machine barrel of the extruder is provided with a heating device for heating the material in the machine barrel.
The wall breaking method of the microalgae related by the invention has high wall breaking rate, high speed and low cost, and can perform extrusion wall breaking treatment without complete drying after the microalgae is collected and cleaned because the humidity of the microalgae needs to be controlled within the range of 25-75 percent, thereby greatly saving energy; the invention creatively discovers that the wall breaking rate of the microalgae can be better improved by controlling the humidity of the microalgae in the range, the protein dissolution rate of the microalgae with high protein content, such as chlorella pyrenoidosa, is obviously improved and can reach 92 percent at most after the microalgae with high protein content is treated, and the high-level structure of the protein is fully dissociated, so that the protein is easier to be absorbed by a human body; after microalgae with high oil content such as chlorella vulgaris is treated, the oil leaching rate is remarkably improved and can reach 92% at most.
Preferably, the humidity of the microalgae is 55-65%. The wall breaking rate of the microalgae can be remarkably improved by controlling the humidity of the microalgae to be within 55-65%.
Preferably, the screw extrusion process has a starting temperature T 0 The temperature is 80-130 ℃, such as 80 ℃, 85 ℃, 90 ℃, 95 ℃, 100 ℃, 105 ℃, 110 ℃, 115 ℃, 120 ℃, 125 ℃ or 130 ℃, any specific point value in the above numerical value range can be selected, and the description is omitted.
The starting temperature T of the screw extrusion process according to the invention 0 The specific selection is in the range of 80-130 deg.C, because excessive temperature can cause back end heating and paste, and excessive temperature can cause insufficient preheating and low wall breaking rate.
Preferably, the said snailStarting temperature T of the rod extrusion process 0 Is 85-125 ℃.
Preferably, the terminal temperature T of the screw extrusion process 1 Is 130-220 ℃, such as 130 ℃, 140 ℃, 145 ℃, 150 ℃, 160 ℃, 170 ℃, 180 ℃, 190 ℃, 195 ℃, 200 ℃, 210 ℃ or 220 ℃ and the like, and any specific value in the numerical value range can be selected, which is not repeated herein.
End point temperature T of screw extrusion processing according to the present invention 1 The specific selection is 130-220 deg.C, because the temperature is too high, which causes the phenomena of paste or swelling, and too low, which causes insufficient heating and low wall breaking rate.
Preferably, the terminal temperature T of the screw extrusion process 1 The temperature is 140-195 ℃.
Preferably, the total length of time t of the screw extrusion process 1 Is 10-200s, such as 10s, 20s, 50s, 80s, 100s, 120s, 150s, 180s, or 200s, any specific point value within the above numerical range can be selected, and is not repeated herein.
The total duration t of the screw extrusion process 1 The specific selection is in the range of 10-200s, because the algae powder is pasted and knotted, nutrient elements are destroyed and oil oxidation peculiar smell is generated due to too long treatment time, and insufficient heating and low wall breaking rate are caused due to too short treatment time.
Preferably, the total length of time t of the screw extrusion process 1 Is 20-150s.
As a preferred technical scheme of the invention, the microalgae wall-breaking processing method comprises the following steps:
carrying out screw extrusion treatment on microalgae with the humidity of 25-75%, wherein the initial temperature T of the screw extrusion treatment 0 80-130 ℃, and the terminal temperature T of screw extrusion treatment 1 At 130-220 deg.C, the total duration t of screw extrusion treatment 1 Is 10-200s; drying the extruded material to reduce the humidity below 10%, and pulverizing to obtain wall-broken microalgae powder.
In another aspect, the invention provides an application of the microalgae wall-breaking processing method in preparation of microalgae food.
Compared with the prior art, the invention has the following beneficial effects:
the wall breaking method of the microalgae related by the invention has high wall breaking rate, high speed and low cost, and can perform extrusion wall breaking treatment without complete drying after the microalgae is collected and cleaned because the humidity of the microalgae needs to be controlled within the range of 25-75 percent, thereby greatly saving energy; the invention creatively discovers that the wall breaking rate of the microalgae can be better improved by controlling the humidity of the microalgae in the range, the protein dissolution rate of the microalgae with high protein content, such as chlorella pyrenoidosa, is obviously improved and can reach 92 percent at most after the microalgae with high protein content is treated, and the high-level structure of the protein is fully dissociated, so that the protein is easier to be absorbed by a human body; the oil leaching rate of microalgae with high oil content such as chlorella vulgaris is remarkably improved to 92% at most after the microalgae are treated.
Drawings
FIG. 1 is the polyacrylamide gel electrophoresis chart of the wall-broken protein Chlorella powder prepared in example 1.
Detailed Description
The technical solution of the present invention is further explained by the following embodiments. It should be understood by those skilled in the art that the examples are only for the understanding of the present invention and should not be construed as the specific limitations of the present invention.
Example 1
This example provides a method for breaking the wall of Chlorella pyrenoidosa (Chlorella pyrenoidosa), which comprises the following steps:
concentrating Chlorella pyrenoidosa to humidity of 65%, and screw extruding in a twin-screw extruder at rotation speed of 90rpm and initial temperature T 0 Set to 115 ℃ and end point temperature T 1 Set to 165 ℃, total duration t of screw extrusion treatment 1 Is 100s, and the specific heating mode is as follows: uniformly heating the temperature of the first section from 115 ℃ to 130 ℃ for 20s, and uniformly heating the temperature of the second section from 130 ℃ to 165 ℃ for 80s; and drying the extruded material by hot air to reduce the humidity to 5%, and crushing to obtain the wall-broken egg white nodule chlorella powder.
Example 2
This example provides a method for breaking the cell wall of Chlorella pyrenoidosa (Chlorella pyrenoidosa), which comprises the following steps:
concentrating Chlorella pyrenoidosa to 55% humidity, and screw extruding in single screw extruder at 90rpm and initial temperature T 0 Set at 125 ℃ and end point temperature T 1 Set to 160 ℃, total duration t of screw extrusion treatment 1 Is 65s, and the specific heating mode is as follows: uniformly heating the temperature of one section from 125 ℃ to 130 ℃ for 15s, uniformly heating the temperature of the other section from 130 ℃ to 150 ℃ for 20s, uniformly heating the temperature of the other section from 150 ℃ to 170 ℃ for 20s, and uniformly cooling the temperature of the other section from 170 ℃ to 160 ℃ for 10s; and drying the extruded material by hot air to reduce the humidity to 5%, and crushing to obtain the wall-broken egg white nodule chlorella powder.
Example 3
This example provides a method for breaking the cell wall of Chlorella pyrenoidosa (Chlorella pyrenoidosa) which differs from example 1 only in that Chlorella pyrenoidosa is concentrated to a moisture level of 75% and all other conditions are maintained.
Example 4
This example provides a method for breaking the cell wall of Chlorella pyrenoidosa (Chlorella pyrenoidosa) which differs from example 1 only in that Chlorella pyrenoidosa is concentrated to a moisture level of 45% and all other conditions are maintained.
Example 5
This example provides a method for breaking the cell wall of Chlorella pyrenoidosa (Chlorella pyrenoidosa) which differs from example 1 only in that Chlorella pyrenoidosa is concentrated to a moisture content of 25% and all other conditions are maintained.
Example 6
This example provides a method for breaking the cell wall of Chlorella vulgaris (Chlorella vulgaris), which comprises the following steps:
concentrating Chlorella vulgaris to 55% humidity, and performing screw extrusion treatment in a twin-screw extruder at rotation speed of 90rpm and initial temperature T 0 Set to 120 ℃ and end point temperature T 1 Set to 190 ℃, total duration t of screw extrusion treatment 1 Is 110s, and the specific heating mode is as follows: uniformly heating the temperature of the first section from 120 ℃ to 150 ℃ for 60s, and uniformly heating the temperature of the second section from 150 ℃ to 190 ℃ for 50s; drying the extruded material with hot air to reduce humidity to 5%, and pulverizing to obtain wall-broken common Chlorella powder.
Example 7
This example provides a method for breaking the cell wall of Chlorella vulgaris (Chlorella vulgaris) which differs from example 6 only in that the Chlorella vulgaris is concentrated to a moisture of 40% and all other conditions are kept constant.
Example 8
This example provides a method for breaking the cell wall of Chlorella vulgaris (Chlorella vulgaris) which differs from example 6 only in that Chlorella vulgaris is concentrated to 70% humidity, and all other conditions are kept constant.
Comparative example 1
This comparative example provides a method of breaking the cell wall of Chlorella pyrenoidosa (Chlorella pyrenoidosa) which differs from example 1 only in that Chlorella pyrenoidosa is concentrated to 85% moisture and all other conditions are maintained.
Comparative example 2
This comparative example provides a method for breaking the cell wall of Chlorella pyrenoidosa (Chlorella pyrenoidosa) which is different from that of example 1 only in that Chlorella pyrenoidosa is concentrated to a humidity of 15%, and all other conditions are maintained.
Evaluation test:
(1) Evaluation of dissolution rate of Water-soluble protein:
the wall breaking rate of the methods described in examples 1-5 and comparative examples 1-2 was evaluated by the dissolution rate of water-soluble protein, and the BCA colorimetric method was used to determine when the BCA working reagent bound to the protein, which bound Cu in the protein, under alkaline conditions 2+ Reduction to Cu + The working reagent is changed from original apple green to purple compound, and the light absorption intensity of the working reagent is in direct proportion to the protein concentration at 562 nm. Preparing standard protein solution by using kit operation, and making standard yeastAnd line, calculating the protein concentration of the sample to be detected according to the regression equation of the standard curve.
Taking a small amount of chlorella pyrenoidosa with the humidity of 5%, fully grinding and breaking the wall by using a mortar, weighing 2g of wall-broken algae powder, adding 20mL of distilled water, shaking for 1min, standing for 10min, and repeating for three times. Centrifuging at 5000rpm for 15min, collecting supernatant, and determining protein content A by BCA method 0 % of the total weight of the composition. Then 2g of the wall-broken chlorella protein powder obtained in examples 1-8 is taken, water-soluble protein is extracted according to the steps, supernatant is taken, and the protein content is A according to the BCA method 1 % of the total weight of the composition. The wall-breaking rate Y can be calculated by the following formula: y = A 1 ×100％/A 0 . The results are shown in table 1:
TABLE 1
| Group of | Wall breaking rate |
| Example 1 | 89％ |
| Example 2 | 92％ |
| Example 3 | 62％ |
| Example 4 | 69％ |
| Example 5 | 44％ |
| Comparative example 1 | 33％ |
| Comparative example 2 | 38％ |
From the results in Table 1, it can be seen that: the wall breaking method for microalgae related by the invention has high wall breaking rate, high speed and low cost, and can perform extrusion wall breaking treatment without completely drying the microalgae after harvesting and cleaning, thereby greatly saving energy; compared with a comparative example, the invention can better improve the wall breaking rate of the microalgae by controlling the humidity of the microalgae to be within the range of 25-75%, and the protein dissolution rate of the chlorella pyrenoidosa is obviously improved to be up to 92% after the chlorella pyrenoidosa is treated.
(2) Evaluating the oil separation rate:
the wall breaking rate of the method described in example 6-8 was evaluated by using the oil extraction rate, and 2g (based on dry basis) of the wall-broken common chlorella powder prepared in example 6-8 was weighed, added with 12mL of n-hexane in a centrifuge tube, shaken for 30s, left to stand for 10min, and repeated three times. Then, the mixture was centrifuged at 8000rpm for 10min, and the upper organic layer was aspirated by a pipette and transferred to a new centrifuge tube (previously dried and accurately weighed). The n-hexane was then removed with a nitrogen blower and then weighed. Weighing 2g of common chlorella powder with humidity of 5% (calculated on dry basis), grinding with mortar to break cell wall, extracting oil according to the above steps, and weighing. The oil extraction rate = sample oil weight × 100%/crude algae powder oil weight. The results are shown in table 2:
TABLE 2
| Group of | Wall breaking rate |
| Example 6 | 92％ |
| Example 7 | 65％ |
| Example 8 | 84％ |
From the results of table 2, it can be seen that: the wall breaking processing method of microalgae related by the invention has high wall breaking rate to common chlorella powder, and after treatment, the oil precipitation rate is obviously improved and can reach 92% to the maximum; and the initial humidity of the microalgae has a remarkable influence on the extraction rate of the final oil.
(3) Evaluation of the effect of dissociation of the higher structure of the protein:
the wall-broken chlorella pyrenoidosa powder prepared in example 1 is taken, polyacrylamide gel electrophoresis (SDS-PAGE) electrophoresis is adopted to characterize the influence on the distribution of protein molecules after extrusion wall breaking, and the result is shown in figure 1 by taking chlorella pyrenoidosa raw powder with the humidity of 5% as a control. Therefore, the wall-breaking processing method can obviously make protein molecules migrate to a low molecular weight region, so that high-order structures of the protein are dissociated, and partial macromolecules are decomposed.
(4) Evaluation of Sterilization Effect
The total number of microorganisms was measured by taking the products prepared in examples 1 and 3 to 5, and the total number was measured by the method of GB4789.2 to 2016 food safety national standard food microbiology inspection colony count determination, and Chlorella pyrenoidosa raw powder with 5% humidity was used as a control. The results are shown in Table 3. It can be seen that each treatment group meets the national limit requirements, and the total number of colonies is significantly reduced when the humidity is 45-65%.
TABLE 3
The applicant states that the present invention is illustrated by the above examples to describe a microalgae wall-breaking method and its application, but the present invention is not limited to the above examples, which does not mean that the present invention is implemented by relying on the above examples. It should be understood by those skilled in the art that any modifications of the present invention, equivalent substitutions of the raw materials of the product of the present invention, and the addition of auxiliary components, selection of specific modes, etc., are within the scope and disclosure of the present invention.
The preferred embodiments of the present invention have been described in detail, however, the present invention is not limited to the specific details of the above embodiments, and various simple modifications may be made to the technical solution of the present invention within the technical idea of the present invention, and these simple modifications are within the protective scope of the present invention.
It should be noted that, in the above embodiments, the various features described in the above embodiments may be combined in any suitable manner, and in order to avoid unnecessary repetition, the present invention does not separately describe various possible combinations.
Claims (2)
1. A microalgae wall-breaking processing method is characterized by comprising the following steps: performing screw extrusion treatment on Chlorella pyrenoidosa or Chlorella vulgaris with humidity of 55-65%, drying the extruded material, and pulverizing to obtain wall-broken Chlorella pyrenoidosa powder or wall-broken Chlorella vulgaris powder;
the initial temperature T0 of the screw extrusion treatment is 115-125 ℃, the final temperature T1 of the screw extrusion treatment is 160-190 ℃, and the total time T1 of the screw extrusion heat treatment is 65-110s.
2. Use of the method of breaking the walls of microalgae according to claim 1 in the preparation of microalgae food.
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