阅读说明：本技术 一种微藻保鲜组合物、保鲜剂及其保存方法 (Microalgae preservation composition, preservative and preservation method thereof ) 是由 黄成潭 潘军 叶蕾 黄敏 于 2021-08-24 设计创作，主要内容包括：本发明涉及微藻技术领域,尤其涉及一种微藻保鲜剂及微藻保存方法。该微藻保鲜组合物包括溶菌酶、肉桂精油和苦皮藤素,在该组合物中进一步添加苯甲酸钠、脱氢乙酸钠、羧甲基纤维素钠和硫酸庆大霉素中的至少一种助剂,获得微藻保鲜剂。实验表明,本发明微藻保鲜剂用于常温保存微藻藻液或藻泥,保存时间长,长达6个月以上微藻活力高,杂菌含量少。(The invention relates to the technical field of microalgae, in particular to a microalgae preservative and a microalgae preservation method. The microalgae preservative composition comprises lysozyme, cinnamon essential oil and celastrus angulatus, and at least one auxiliary agent of sodium benzoate, sodium dehydroacetate, sodium carboxymethylcellulose and gentamicin sulfate is further added into the composition to obtain the microalgae preservative. Experiments show that the microalgae preservative is used for preserving microalgae liquid or algae mud at normal temperature, the preservation time is long, the activity of the microalgae is high for more than 6 months, and the content of mixed bacteria is low.)

1. A composition for keeping microalgae fresh is characterized by comprising lysozyme, cinnamon essential oil and celastrus angulatus.

2. The composition according to claim 1, which comprises the following components in parts by mass:

6-8 parts of lysozyme, 0.25-0.5 part of cinnamon essential oil and 30-40 parts of celastrus angulatus.

3. The composition according to claim 1, which comprises the following components in parts by mass:

6 parts of lysozyme, 0.25 part of cinnamon essential oil and 32 parts of celastrus angulatus.

4. A microalgae preservative, which is characterized by comprising the composition as claimed in any one of claims 1 to 3 and an auxiliary agent; the auxiliary agent comprises at least one of sodium benzoate, sodium dehydroacetate, sodium carboxymethylcellulose and gentamicin sulfate.

5. The microalgae preservative according to claim 4, which is characterized by comprising the following components in parts by mass:

6-8 parts of lysozyme, 0.25-0.5 part of cinnamon essential oil, 30-40 parts of celastrol, 3-5 parts of sodium benzoate, 0.25-0.5 part of sodium benzoate, 200 parts of sodium carboxymethylcellulose and 10-15 parts of gentamycin sulfate.

6. Use of the composition of any one of claims 1 to 3 or the microalgae antistaling agent of claim 4 or 5 in the fresh keeping of microalgae.

7. The use of claim 6, wherein the microalgae comprise Chlorella, Nannochloropsis, oocystis, Chaetoceros or Thalassiosira.

8. A method for preserving microalgae, characterized in that the composition of any one of claims 1 to 3 or the microalgae preservative of claim 4 or 5 is mixed with a microalgae solution and preserved at normal temperature.

9. The method of claim 8, further comprising the step of filtering the microalgae suspension through a 50-100 micron filter before the mixing.

10. The microalgae preserving method as claimed in claim 8, wherein the microalgae preservative accounts for 1-5% of the microalgae solution by mass.

Technical Field

The invention relates to the technical field of microalgae, in particular to a microalgae preservation composition, a preservative and a preservation method thereof.

Background

The main functions of food preservation are to prolong the storage period of food, keep the original flavor of food and reduce energy consumption. The food fresh-keeping is a technology which adopts physical and chemical biological methods to treat the food in the production and circulation processes according to the quality characteristics and the spoilage mechanism of the food, so as to inhibit or delay the spoilage of the fresh food and keep the good freshness and quality of the fresh food. Traditional food storage methods include heat treatment, low temperature, drying, sugar crushing, salt crushing, smoking and the like, and although the methods can prolong the shelf life of the food, the methods have disadvantages, such as high energy consumption caused by a freezing method; the heating, drying and soaking methods are easy to destroy the original flavor of the food; based on the above disadvantages, new food preservation technology is increasingly popular. The food fresh-keeping method mainly comprises three major methods of physical, chemical and biological methods, and each method derives a plurality of new technologies which are respectively based on different fresh-keeping principles. Although the emphasis of various preservation methods is different, the factors playing a key role in the preservation quality are all regulated and controlled. Firstly, the physiological and biochemical change processes of the food are controlled, so that the quality deterioration process is delayed; and secondly, controlling microorganisms, mainly by controlling putrefying bacteria.

The currently common physical preservation technology mainly utilizes a physical method to preserve food, and comprises ice temperature preservation, modified atmosphere preservation, ultrahigh pressure preservation and ultraviolet irradiation preservation; the chemical fresh-keeping is mainly to utilize a chemical fresh-keeping agent to inhibit or kill microorganisms to achieve the aim of fresh-keeping; biological fresh-keeping is to utilize biological fresh-keeping agent to inhibit the growth of microorganism or harmful bacteria, the biological fresh-keeping agent is extracted from animal and plant and microorganism or is modified by utilizing biological engineering technique, and the mechanism of biological fresh-keeping technique is mainly to utilize the principle of reducing oxidation of organism, isolate organism from air, or exert the bacteriostatic action and bactericidal action of biological fresh-keeping agent to implement fresh-keeping and preservative effect for organism. The biological preservative is a product which is extracted from animals and plants or microorganisms or is obtained by utilizing the modification of a biological engineering technology and has a very safe preservation effect on human bodies. Different biological preservatives have different preservation mechanisms, and some biological preservatives realize preservation by inhibiting or killing organism pathogenic bacteria; some biological preservatives prevent organisms from browning by inhibiting the enzyme activity, so that the organisms have good sensory quality; some biological preservatives prevent fat in organisms from being oxidized and rancid by inducing the organisms to generate defense capacity, and prevent the quality of the organisms from being deteriorated; in addition, by coating organisms of the biological preservative, the loss of water can be reduced, the invasion of microorganisms can be prevented, and free radicals can be effectively eliminated. The biological preservation technology has the advantages that: the biological preservative has the advantages of low treatment cost, easy control of storage conditions, accordance with the requirements of green environmental protection and the like, and generally comprises three technologies of preservation by utilizing microbial cells and metabolites thereof, preservation by natural extracts and preservation by genetic engineering.

In the past decades, people have done a lot of work and accumulated abundant experience in the domestication, breeding and industrial culture of marine chlorella. The seawater chlorella has rich and balanced nutrient substances, and the currently developed seawater chlorella food comprises various seawater chlorella powders, tablets, capsules, instant granules, beverages, oral liquids, health care salts and the like.

The chlorella is a pure natural green biological bait, and the fish and the shrimp are easy to absorb and digest, so the chlorella is an indispensable component link for rockfish hatching and the labeling of the south America white prawns. Meanwhile, the chlorella can improve the dissolved oxygen in the water body, reduce the concentration of pollution indexes such as ammonia nitrogen, sub-salt, phosphorus and the like, effectively improve the water quality of the aquaculture water body, improve the success rate of aquaculture and promote a more environment-friendly culture mode.

In the initial stage of cultivation, the chlorella and the fertilizer are used simultaneously to play a role in quickly fertilizing water. The chlorella can be used independently in a high-temperature period, and can also be used together with bacillus subtilis or EM (effective microorganisms) for splashing, so that the water quality is adjusted, ammonia nitrogen and nitrite are reduced, blue algae is inhibited, and the water environment is improved. Provides a chlorella unicellular source, can be rapidly propagated after entering the culture water body, forms a water body taking the chlorella unicellular as a dominant population, and constructs a good living environment for various aquatic animals such as fish, shrimp, crab, shellfish and the like. The chlorella can provide abundant and balanced natural nutrients, and is rich in proteins, unsaturated fatty acids, vitamins, minerals, chlorophyll, algal polysaccharides, nucleic acids, etc. The nutritional ingredients are helpful for improving disease resistance and resistance. The chlorella has high nutritive value, and can be used as starter feed for young shrimps, crabs and shellfish and direct feed for filter-feeding fishes, to promote growth, reduce cost, and improve survival rate of aquatic animals. Can better carry out photosynthesis, increase dissolved oxygen in water and greatly reduce the possibility of oxygen deficiency floating head.

Although the seawater chlorella is rich in nutrition, the nutrient components of the seawater chlorella are greatly lost and lose vigor in the process of preparing the algae powder and the algae tablets, and the seawater chlorella does not have the functions of regulating water quality and purifying water quality any more. And some substances harmful to the body may be added in the manufacturing process.

The living marine chlorella is easy to age and die in the preservation process, is easy to be polluted by bacteria to cause precipitation and stink, and is not easy to preserve for a long time. Cryopreservation is required, but once frozen, it results in a significant loss of viability and increases the cost of preservation. Because the normal temperature preservation is difficult, the research on the aspect of the normal temperature preservation is less.

Disclosure of Invention

In view of the above, the invention provides a microalgae preservation composition, a preservative and a preservation method thereof. The preservation agent is used for preserving the microalgae at normal temperature, and has long preservation time, high microalgae activity and low content of mixed bacteria.

In order to achieve the above object, the present invention provides the following technical solutions:

the invention provides a composition for keeping microalgae fresh, which comprises lysozyme, cinnamon essential oil and celastrus angulatus.

In some embodiments, the composition comprises the following components in parts by mass:

6-8 parts of lysozyme, 0.25-0.5 part of cinnamon essential oil and 30-40 parts of celastrus angulatus.

In some embodiments, the composition comprises the following components in parts by mass:

6 parts of lysozyme, 0.25 part of cinnamon essential oil and 32 parts of celastrus angulatus.

The invention also provides a microalgae preservative, which comprises the composition and an auxiliary agent; the auxiliary agent comprises at least one of sodium benzoate, sodium dehydroacetate, sodium carboxymethylcellulose and gentamicin sulfate.

In some embodiments, the microalgae preservative comprises the following components in parts by mass:

6-8 parts of lysozyme, 0.25-0.5 part of cinnamon essential oil, 30-40 parts of celastrol, 3-5 parts of sodium benzoate, 0.25-0.5 part of sodium benzoate, 200 parts of sodium carboxymethylcellulose and 10-15 parts of gentamycin sulfate.

The invention also provides the composition and the application of the microalgae preservative in microalgae preservation.

Wherein the microalgae comprises Chlorella, Nannochloropsis, Ascomycetes, Chaetoceros, and Alternaria.

The invention also provides a microalgae preservation method, which comprises the steps of mixing the composition or the microalgae preservative of the invention with microalgae solution, and preserving at normal temperature.

In some embodiments, the method further comprises the step of filtering the microalgae suspension with a 50-100 micron filter membrane prior to the mixing.

In some embodiments, the microalgae preservative is mixed with the microalgae solution or the algae mud in a proportion that the microalgae preservative accounts for 1-5% of the mass of the microalgae solution or the algae mud. In some embodiments, the microalgae preservative accounts for 1%, 1.6%, 2%, 2.4%, 3.2%, or 5% of the microalgae solution by mass. If the addition amount is 1%, the shelf life at normal temperature is relatively short, but not less than 6 months.

The microalgae preservative composition provided by the invention comprises lysozyme, cinnamon essential oil and celastrus angulatus, and at least one auxiliary agent of sodium benzoate, sodium dehydroacetate, sodium carboxymethylcellulose and gentamicin sulfate is further added into the composition to obtain the microalgae preservative. Experiments show that the microalgae preservative can be used for preserving microalgae liquid at normal temperature (20-35 ℃), the preservation time is long, the microalgae activity is high, and the content of mixed bacteria is low. Wherein, the chlorella liquid can still keep more than 90% of activity after being stored for 3 months at normal temperature, and the content of infectious microbes is lower than 5%; the algae mud is stored at normal temperature, the activity and the bacteriostatic ability of the algae mud are obviously improved, the activity is kept at 50% after the algae mud is stored for 3 months, and the content of mixed bacteria is lower than 8%.

Detailed Description

The invention provides a microalgae preservation composition, a preservative and a preservation method thereof. Those skilled in the art can modify the process parameters appropriately to achieve the desired results with reference to the disclosure herein. It is expressly intended that all such similar substitutes and modifications which would be obvious to one skilled in the art are deemed to be included in the invention. While the methods and applications of this invention have been described in terms of preferred embodiments, it will be apparent to those of ordinary skill in the art that variations and modifications in the methods and applications described herein, as well as other suitable variations and combinations, may be made to implement and use the techniques of this invention without departing from the spirit and scope of the invention.

The test materials adopted by the invention are all common commercial products and can be purchased in the market.

Example 1 the microalgae preservative composition of the invention

TABLE 1 microalgae preservative composition of the invention (Material A)

Serial number	Name of material	Weight (D)
			01	Lysozyme	0.06g
02	Cinnamon essential oil	2.5mg
			03	Celastrus angulatus extract	0.32g

Example 2 the microalgae antistaling agent of the invention

Consisting of the preservative composition of example 1 (material a) and the adjuvant (material B):

TABLE 2 adjuvant (Material B) composition

The configuration method comprises the following steps:

1. taking two high-temperature resistant plastic containers (or glass containers), forbidding metal containers, and sterilizing (if sterilization is better) for standby;

2. respectively adding 3-4L of sterile water (or boiling water to accelerate the dissolution rate) into high-temperature resistant plastic containers, slowly pouring the material A and the material B into the two containers respectively, and fully stirring until the materials are uniformly mixed (a small amount of precipitate is normal), and adding the sterile water to 5L when the mixed solution in a single container is less than 5L;

3. and after the water in the two containers is cooled at the temperature, respectively pouring the liquid in the two containers into a third container for mixing to obtain 10L of mixed liquid, uniformly stirring, and sealing for later use, thus obtaining the normal-temperature microalgae preservative.

Example 3 Normal temperature preservation of fresh Chlorella species

The storage method comprises the following steps:

1. filtering cultured Chlorella vulgaris with 50 μm filter membrane, and removing large granule precipitate.

2. Before subpackaging the chlorella liquid in seawater, uniformly shaking the microalgae preservative, uniformly pouring the microalgae preservative into the chlorella liquid, uniformly stirring, and after subpackaging the chlorella liquid, wherein a small amount of precipitate in a bottle is normal;

3. example 2 when preparing a microalgae preservative (10L), the addition amount of the microalgae preservative accounts for 1.6-3.2% (w/w) of the algae liquid,

4. after the algae liquid added with the microalgae preservative is bottled, the algae liquid is filled in a paper box to keep a lightless state and is stored at normal temperature.

The fresh-collected marine chlorella (the density of the chlorella is 500-.

The control group does not add the normal temperature antistaling agent of the marine chlorella, and other preservation processes are the same as above.

TABLE 3

Therefore, the seawater chlorella preserved for 3 months at normal temperature still keeps more than 90% of activity after the preservative is added, and the contrast group is basically died.

Example 4 preservation of concentrated algae with addition of preservative

The collected marine chlorella is centrifugally concentrated to form paste algae mud, and then 5% of preservative is added. The control group 1 was not added with preservative, and the control group 2 was added with 1% (w/w) sodium benzoate chemical preservative. Storing the three groups of algae mud in a refrigerator at 4 ℃, and detecting the activity, color, character and amount of mixed bacteria of algae every week.

TABLE 4

The concentrated algae mud is rich in a large amount of impurities, algae in a control group without the preservative are killed in a large amount, and the content of infectious microbes is high.

The foregoing is only a preferred embodiment of the present invention, and it should be noted that it is obvious to those skilled in the art that various modifications and improvements can be made without departing from the principle of the present invention, and these modifications and improvements should also be considered as the protection scope of the present invention.