CN111466569A - Euphausia superba oil edible nutriment - Google Patents
Euphausia superba oil edible nutriment Download PDFInfo
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Classifications
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/115—Fatty acids or derivatives thereof; Fats or oils
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/56—Materials from animals other than mammals
- A61K35/612—Crustaceans, e.g. crabs, lobsters, shrimps, krill or crayfish; Barnacles
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P39/00—General protective or antinoxious agents
- A61P39/06—Free radical scavengers or antioxidants
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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Abstract
The invention discloses an antarctic krill oil edible nutriment and a preparation method thereof, and belongs to the technical field of marine organisms. The invention uses the leftovers from the processing of the euphausia superba oil as raw materials, integrates the modern bioseparation technology, the biological enzymolysis technology and the mixed microbial fermentation technology, and develops the euphausia superba oil edible nutriment with high in-vitro oxidation resistance, wherein the mixed aerobic fermentation strain mainly comprises aspergillus niger and bacillus subtilis. The obtained antarctic krill oil edible nutriment is rich in antarctic krill polypeptide, has strong oxidation resistance and anti-fatigue effects, and can be widely used for health care and medical rehabilitation of sub-health people. The process technology is simple and efficient, and the obtained product has strong oxidation resistance and free radical scavenging capacity, and has high application value and market prospect.
Description
Technical Field
The invention relates to an antarctic krill oil edible nutriment and a preparation method thereof, in particular to an antarctic krill oil edible nutriment with high oxidation resistance and anti-aging capability prepared by utilizing synergistic fermentation of microorganisms, belonging to the technical field of marine organisms.
Background
Antarctic continents are the only uncontaminated continents on the earth, and the world that has been praised as "clean and clear" is the last piece of "clean soil" on the earth. The ocean around the Antarctic continent is called a rich and clean "marine ranch", and the ocean is filled with living organisms, where there are seaweeds, corals, starfishes and sponges, and there are many small organisms called krill, which is food on which numerous fishes, seabirds, seals, penguins and whales in Antarctic continent live, and plays an important role in the south ocean food chain. Antarctic krill is the fourth largest resource in areas following grains, petroleum, coal. The storage amount of the antarctic krill is quite remarkable, and the nutritional value is extremely high. It is the highest protein-containing organism found today, with a protein content of more than fifty percent, and also contains a great abundance of amino acids and vitamin A essential for human tissues.
At present, Antarctic krill is mainly applied to medicine and food by Antarctic krill oil. Antarctic krill oil is extracted by a patent ultralow temperature extraction method, is natural health food given to human beings by nature, and has the effect that other foods cannot be replaced. The krill oil is clinically proved to have the advantages of ultra-strong blood fat reduction, blood sugar reduction, blood pressure reduction, nutrition passing through a blood brain barrier and activation of brain cells and nerves, and prevention and treatment of cerebrovascular diseases; simultaneously, preventing and treating coronary atherosclerosis and coronary heart disease, and preventing cardiovascular diseases such as myocardial infarction, stent, bypass, secondary recurrence and the like; in addition, the antarctic krill oil can also have multiple effects of resisting bacteria and diminishing inflammation, preventing the occurrence and development of cancers, delaying fatigue, regulating endocrine and the like. With the application and popularization of the Antarctic krill oil, the processing industry develops rapidly, a large amount of Antarctic krill oil processing leftovers are generated, the composition of the leftovers is mainly protein, and therefore the mutual utilization of the resources of the Antarctic krill oil processing leftovers becomes a technical problem to be solved urgently in the field.
In the prior art, the preparation methods of antarctic krill polypeptides are relatively few, and mainly include an enzymatic hydrolysis method, for example, patent 201810373432.5 discloses a method for degrading defatted antarctic krill powder by a combinatorial enzymatic method to recover polypeptide intermediate products and prepare N-acetylglucosamine; firstly, pretreatment of the degreased antarctic krill powder is carried out, and polypeptides in the degreased antarctic krill powder are recovered through pretreatment of alkaline protease, so that raw materials are fully utilized, and the relative content of chitin in the degreased antarctic krill powder is improved. Compared with the documents, the complex enzymatic hydrolysis method is mostly adopted for the preparation of the antarctic krill polypeptide, the key technology for resource utilization of the processing leftovers of the antarctic krill oil by the fermentation of complex microorganisms and the application thereof are lacked, and especially the research for improving the nutrition of the antarctic krill polypeptide becomes the key bottleneck for restricting the industrial development.
Disclosure of Invention
In order to overcome the defects of the prior art and meet the requirement of high-valued comprehensive utilization of leftovers in the processing of the euphausia superba oil, the invention provides the euphausia superba oil edible nutriment, integrates the modern bioseparation technology, the biological enzymolysis technology and the mixed microbial fermentation technology, develops and develops the euphausia superba oil edible nutriment with high in-vitro antioxidant capacity and anti-aging capacity, meets the application requirements of fields such as health care, medical treatment, food and the like on functional polypeptides of the euphausia superba, and has the advantages of simple and efficient process, strong antioxidant capacity and free radical scavenging capacity of the obtained product, low production cost and easiness in industrial scale production.
The invention realizes the technical effects through the following technical scheme:
An antarctic krill oil edible nutriment is characterized by comprising the following steps:
(1) Dissolving: crushing the leftovers of the euphausia superba oil processing, adding water and stirring until the leftovers are completely dissolved;
(2) Enzymolysis: adding 2-3% of compound protease and 0.5% of naringinase in mass ratio to the protein of the leftovers of the Antarctic krill oil processing after dissolving, and carrying out enzymolysis for 4-8 h under the condition of controlling the temperature to be 40-60 ℃;
(3) Aerobic fermentation: adding a compound microbial agent with the mass ratio of 2-3% of the enzymolysis liquid into the enzymolysis liquid after enzymolysis, and performing aerobic fermentation for 6-12 h at the temperature of 28-35 ℃;
(4) Anaerobic fermentation: after the aerobic fermentation is finished, adding 1-2% of lactobacillus acidophilus powder, and carrying out anaerobic fermentation for 8-16 h at the temperature of 28-35 ℃;
(5) And (3) filtering: performing coarse filtration to remove fermentation thalli;
(6) Enzyme deactivation: inactivating enzyme at 100 deg.C for 10 min;
(7) Centrifuging: centrifuging at 10000r/min to obtain supernatant as antarctic krill oil nutriment.
The optimal water adding proportion in the step (1) is the tailings of the euphausia superba oil processing: water =1: 8.
Preferably, astaxanthin with the protein mass ratio of 2-3% of the tailings of the Antarctic krill oil processing is added in the tailings dissolving process of the Antarctic krill oil processing.
The compound protease in the step (2) is a combination of papain and flavourzyme, and the mass ratio of the compound protease to the flavourzyme is 2: 1.
The compound microbial agent in the step (3) is a mixed microbial agent of aspergillus niger and bacillus subtilis, and the mass ratio of the compound microbial agent to the bacillus subtilis is 2: 3.
The ventilation rate is 0.1-0.2 m when the aerobic fermentation process in the step (3) is carried out in a fermentation tank 3/m3And min, and controlling the stirring speed to be 30-60 r/min.
The preparation method of the aspergillus niger seed liquid comprises the following steps: culturing for 24h at 35-40 deg.C in potato culture medium under aseptic condition to obtain Aspergillus niger seed liquid with effective viable count not less than 10 9cfu/mL。
the preparation method of the bacillus subtilis seed solution comprises the steps of adopting L B culture medium to culture for 24 hours at 28-32 ℃ under aseptic condition to prepare the liquid bacillus subtilis seed solution, wherein the effective viable count of the seed solution is required to be more than or equal to 10 9cfu/mL。
The invention provides an antarctic krill oil edible nutrient which is rich in antarctic krill polypeptide and has strong antioxidant capacity and anti-aging effect, and the extract can be subjected to concentration, adsorption drying, alcohol precipitation, resin purification and other treatments according to application approaches, so that the high-efficiency utilization in the fields of health care, food, medical treatment and the like is met.
Compared with the prior art, the euphausia superba oil edible nutriment provided by the invention has the following remarkable advantages:
(1) The processing leftovers of the euphausia superba oil are used as raw materials, and the modern bioseparation technology, the biological enzymolysis technology and the mixed microbial fermentation technology are integrated, so that the oxidation resistance and the anti-aging effect of the processing leftovers of the euphausia superba oil are obviously improved; more importantly, the application creatively discovers that the antioxidant capacity of the antarctic krill polypeptide prepared by the enzymolysis fermentation method can be remarkably improved by adding astaxanthin with a certain proportion into the protein solution of the leftovers after the antarctic krill oil processing, the analysis reason is probably that the complex chemical combination reaction between the astaxanthin and the antarctic krill polypeptide plays a role in improving the antioxidant capacity, and the chitosan oligosaccharide can promote the fermentation of microorganisms, so that the antioxidant capacity of the product is further improved; meanwhile, the test result shows that the change of the composition of the microorganism can obviously nourish the synergistic effect of the astaxanthin;
(2) This application utilizes aerobic bacteria (aspergillus niger, bacillus subtilis) to ferment antarctic krill oil processing leftover bits and pieces jointly and draws functional activity material such as antarctic krill polypeptide, and the synergism between the make full use of microorganism promotes the extraction rate and the external antioxidant capacity of antarctic krill polypeptide, wherein: aspergillus niger can generate various enzymes for degrading cell walls in the fermentation process, so that more functional active substances such as polypeptide and the like are released from the cell walls, and the in-vitro antioxidant capacity of the euphausia superba polypeptide is improved; the bacillus subtilis can kill pathogenic bacteria in fermentation liquor in the high-temperature fermentation process, has strong capabilities of producing amylase, protease and the like, and can degrade macromolecular starch and protein substances into microorganisms for utilization; the synergistic effect of the aspergillus niger and the bacillus subtilis obviously improves the oxidation resistance and the health care effect of the product;
(3) According to the invention, lactobacillus acidophilus is adopted for anaerobic fermentation, so that the stability of antarctic krill polypeptide is improved, test results show that the temperature and time have little influence on the oxidation resistance of the protein hydrolysis peptide of the leftovers after the processing of the antarctic krill oil at the temperature of 20-80 ℃, the oxidation resistance begins to be reduced along with the increase of the temperature and time when the temperature exceeds 100 ℃, and the stability is higher than the technical level reported at present;
(4) According to the invention, naringinase is added in the composite enzymolysis process, so that the nutrition taste of the product is improved, and the obvious effects of improving the oxidation resistance and the health care effect of the product are achieved.
Detailed Description
In the present application, the activation method of Aspergillus niger and Bacillus subtilis is as follows:
(1) The preparation method of the aspergillus niger seed liquid comprises the following steps: culturing for 24h at 35-40 deg.C in potato culture medium under aseptic condition to obtain Aspergillus niger seed liquid with effective viable count not less than 10 9cfu/mL；
(2) the preparation method of the bacillus subtilis seed solution comprises the steps of adopting L B culture medium to culture for 24 hours at 28-32 ℃ under aseptic condition to prepare the liquid bacillus subtilis seed solution, wherein the effective viable count of the seed solution is required to be more than or equal to 10 9cfu/mL。
Example 1
An antarctic krill oil edible nutriment specifically comprises the following process steps:
(1) Dissolving: crushing the leftovers of the euphausia superba oil processing, adding water and stirring until the leftovers are completely dissolved; the water adding proportion is the processing leftovers of the euphausia superba oil: water =1: 8;
(2) Enzymolysis: adding compound protease accounting for 3% of the protein mass ratio of the leftovers of the Antarctic krill oil processing and 0.5% of naringinase after dissolving, and performing enzymolysis for 6 hours at the temperature of 40-60 ℃; the compound protease is a combination of papain and flavourzyme, and the mass ratio of the compound protease to the flavourzyme is 2: 1;
(3) Aerobic fermentation: adding a compound microbial agent with the mass ratio of 2% of the enzymolysis liquid into the enzymolysis liquid after enzymolysis, and carrying out aerobic fermentation for 12h at the temperature of 28-35 ℃; the composite microbial agent is a mixed microbial agent of aspergillus niger and bacillus subtilis, and the mass ratio of the composite microbial agent to the bacillus subtilis is 2: 3; the ventilation rate is 0.1-0.2 m when the aerobic fermentation process is carried out in a fermentation tank 3/m3Min, controlling the stirring speed to be 30-60 r/min;
(4) Anaerobic fermentation: after the aerobic fermentation is finished, 2% of lactobacillus acidophilus powder is added, and anaerobic fermentation is carried out for 8 hours at the temperature of 28-35 ℃;
(5) And (3) filtering: performing coarse filtration to remove fermentation thalli;
(6) Enzyme deactivation: inactivating enzyme at 100 deg.C for 10 min;
(7) Centrifuging: centrifuging at 10000r/min to obtain supernatant as antarctic krill oil nutriment.
The antioxidant capacity of the krill oil nutriment of example 1 was determined, wherein the OH radical clearance was determined according to the method described in "food chemistry test" of Shao shou et al; ABTS free radical clearance and DPPH free radical clearance are measured by referring to the method in the method of 'influence of reaction time on oxidation resistance results evaluated by DPPH method and ABTS method' of Linlian bamboo and the like; the results of the measurement were as follows:
The control group 1 in the above experiment is the antarctic krill oil nutriment without aerobic fermentation process in example 1; the control 2 group is the antarctic krill oil nutriment obtained by adding only aspergillus niger for fermentation in the aerobic fermentation in the example 1, and the rest is the same as the example 1; the control group 3 is the antarctic krill oil nutriment obtained by adding only bacillus subtilis to the aerobic fermentation in the example 1 for fermentation; the control group 4 is an antarctic krill oil nutriment without naringinase in the enzymolysis process; the control group 5 is an antarctic krill oil nutriment only containing naringinase in the enzymolysis process; the control group 6 was a Euphausia superba oil nutriment without enzymatic hydrolysis process during the preparation process; the rest is the same as example 1.
The experimental results show that the antioxidant capacity of the euphausia superba oil nutriment in the embodiment 1 of the invention is remarkably improved, and the mixed fermentation between aspergillus niger and bacillus subtilis in the aerobic fermentation process plays a remarkable synergistic effect.
Example 2
An antarctic krill oil edible nutriment specifically comprises the following process steps:
(1) Dissolving: crushing the tailings of the Euphausia superba oil processing, adding water, stirring until the tailings are completely dissolved, and adding astaxanthin with the protein mass ratio of 2% to the tailings of the Euphausia superba oil processing; the water adding proportion is the processing leftovers of the euphausia superba oil: water =1: 8;
(2) Enzymolysis: adding compound protease accounting for 3% of the protein mass ratio of the leftovers of the Antarctic krill oil processing and 0.5% of naringinase after dissolving, and performing enzymolysis for 6 hours at the temperature of 40-60 ℃; the compound protease is a combination of papain and flavourzyme, and the mass ratio of the compound protease to the flavourzyme is 2: 1;
(3) Aerobic fermentation: adding a compound microbial agent with the mass ratio of 2% of the enzymolysis liquid into the enzymolysis liquid after enzymolysis, and carrying out aerobic fermentation for 12h at the temperature of 28-35 ℃; the composite microbial agent is a mixed microbial agent of aspergillus niger and bacillus subtilis, and the mass ratio of the composite microbial agent to the bacillus subtilis is 2: 3; the ventilation rate is 0.1-0.2 m when the aerobic fermentation process is carried out in a fermentation tank 3/m3Min, controlling the stirring speed to be 30-60 r/min;
(4) Anaerobic fermentation: after the aerobic fermentation is finished, 2% of lactobacillus acidophilus powder is added, and anaerobic fermentation is carried out for 8 hours at the temperature of 28-35 ℃;
(5) And (3) filtering: performing coarse filtration to remove fermentation thalli;
(6) Enzyme deactivation: inactivating enzyme at 100 deg.C for 10 min;
(7) Centrifuging: centrifuging at 10000r/min to obtain supernatant as antarctic krill oil nutriment.
The antioxidant capacity of the krill oil nutriment of example 2 was measured according to the same method as the antioxidant capacity measurement method described in example 1. The test results are as follows:
The control 1 group in the above experiment was the antarctic krill oil nutriment without aerobic fermentation in example 2; the control 2 group is the antarctic krill oil nutriment obtained by adding only aspergillus niger for fermentation in the aerobic fermentation in the example 2, and the rest is the same as the example 2; the control 3 group was the antarctic krill oil nutriment obtained by fermenting the example 2 aerobic fermentation with only bacillus subtilis, and the rest was the same as the example 2.
The experimental results show that the antioxidant capacity of the euphausia superba oil nutriment in the embodiment 1 of the invention is remarkably improved, the mixed fermentation between aspergillus niger and bacillus subtilis in the aerobic fermentation process plays a remarkable synergistic effect, and the technical effect of 1+1>2 is achieved. Example 2 is the best embodiment of the invention.
Example 3
An antarctic krill oil edible nutriment specifically comprises the following process steps:
(1) Dissolving: crushing the tailings of the Euphausia superba oil processing, adding water, stirring until the tailings are completely dissolved, and adding astaxanthin with the protein mass ratio of 3% to the tailings of the Euphausia superba oil processing; the water adding proportion is the processing leftovers of the euphausia superba oil: water =1: 8;
(2) Enzymolysis: adding compound protease accounting for 3% of the protein mass ratio of the leftovers of the Antarctic krill oil processing and 0.5% of naringinase after dissolving, and performing enzymolysis for 6 hours at the temperature of 40-60 ℃; the compound protease is a combination of papain and flavourzyme, and the mass ratio of the compound protease to the flavourzyme is 2: 1;
(3) Aerobic fermentation: adding a compound microbial agent with the mass ratio of 2% of the enzymolysis liquid into the enzymolysis liquid after enzymolysis, and carrying out aerobic fermentation for 12h at the temperature of 28-35 ℃; the composite microbial agent is a mixed microbial agent of aspergillus niger and bacillus subtilis, and the mass ratio of the composite microbial agent to the bacillus subtilis is 2: 3; the ventilation rate is 0.1-0.2 m when the aerobic fermentation process is carried out in a fermentation tank 3/m3Min, controlling the stirring speed to be 30-60 r/min;
(4) Anaerobic fermentation: after the aerobic fermentation is finished, 2% of lactobacillus acidophilus powder is added, and anaerobic fermentation is carried out for 8 hours at the temperature of 28-35 ℃;
(5) And (3) filtering: performing coarse filtration to remove fermentation thalli;
(6) Enzyme deactivation: inactivating enzyme at 100 deg.C for 10 min;
(7) Centrifuging: centrifuging at 10000r/min to obtain supernatant as antarctic krill oil nutriment.
The antioxidant capacity of the krill oil nutriment of example 3 was measured according to the same method as the antioxidant capacity measurement method described in example 1. The test results are as follows:
The control 1 group in the above experiment was the antarctic krill oil nutriment without aerobic fermentation in example 3; the control group 2 is the antarctic krill oil nutriment obtained by adding only aspergillus niger for fermentation in the aerobic fermentation in the example 3, and the rest is the same as the example 3; the control 3 group was the antarctic krill oil nutriment obtained by fermenting the example 3 aerobic fermentation with only bacillus subtilis, and the rest was the same as the example 3.
The experimental results show that the antioxidant capacity of the euphausia superba oil nutriment in the embodiment 1 of the invention is remarkably improved, and the mixed fermentation between aspergillus niger and bacillus subtilis in the aerobic fermentation process plays a remarkable synergistic effect.
The above embodiments are only used for illustrating the technical solution of the present invention, and not for limiting the same; although the invention has been described in detail with reference to the foregoing embodiments, it will be apparent to those skilled in the art that various changes may be made and equivalents may be substituted for elements thereof; such modifications and substitutions do not depart from the spirit and scope of the present invention as set forth in the appended claims.
Claims (10)
1. An antarctic krill oil edible nutriment is characterized by comprising the following steps:
Dissolving: crushing the leftovers of the euphausia superba oil processing, adding water and stirring until the leftovers are completely dissolved;
Enzymolysis: adding 2-3% of compound protease and 0.5% of naringinase in mass ratio to the protein of the leftovers of the Antarctic krill oil processing after dissolving, and carrying out enzymolysis for 4-8 h under the condition of controlling the temperature to be 40-60 ℃;
Aerobic fermentation: adding a compound microbial agent with the mass ratio of 2-3% of the enzymolysis liquid into the enzymolysis liquid after enzymolysis, and performing aerobic fermentation for 6-12 h at the temperature of 28-35 ℃;
Anaerobic fermentation: after the aerobic fermentation is finished, adding 1-2% of lactobacillus acidophilus powder, and carrying out anaerobic fermentation for 8-16 h at the temperature of 28-35 ℃;
And (3) filtering: performing coarse filtration to remove fermentation thalli;
Enzyme deactivation: inactivating enzyme at 100 deg.C for 10 min;
Centrifuging: centrifuging at 10000r/min to obtain supernatant as antarctic krill oil nutriment.
2. The antarctic krill oil edible nutrient according to claim 1, characterized in that the optimal water addition ratio in step (1) is the antarctic krill oil processing leftovers: water =1: 8.
3. The antarctic krill oil edible nutritional product according to claim 1, wherein astaxanthin with a protein mass ratio of 2-3% is added to the antarctic krill oil processing leftovers in the process of dissolving the antarctic krill oil processing leftovers.
4. The euphausia superba oil edible nutritional product according to claim 1, wherein the compound protease in step (2) is a combination of papain and flavourzyme, and the mass ratio of the papain to the flavourzyme is 2: 1.
5. The antarctic krill oil edible nutrient according to claim 1, characterized in that the compound microbial agent in the step (3) is a mixed microbial agent of aspergillus niger and bacillus subtilis, and the mass ratio of the compound microbial agent to the bacillus subtilis is 2: 3.
6. The euphausia superba oil edible nutrient according to claim 1, wherein the ventilation rate of the aerobic fermentation process in step (3) is 0.1-0.2 m when the aerobic fermentation process is performed in a fermentation tank 3/m3And min, and controlling the stirring speed to be 30-60 r/min.
7. The antarctic krill oil edible nutrient according to claim 1, wherein the preparation method of the aspergillus niger seed liquid comprises: culturing in potato culture medium at 35-40 deg.C for 24 hr under aseptic condition to obtain Aspergillus niger seed liquid The effective viable count of the liquid is more than or equal to 10 9cfu/mL。
8. the euphausia superba oil edible nutritional product as claimed in claim 1, wherein the preparation method of the bacillus subtilis seed liquid comprises the steps of culturing the euphausia superba oil edible nutritional product for 24 hours at 28-32 ℃ by using L B culture medium under aseptic conditions to obtain the liquid bacillus subtilis seed liquid, wherein the effective viable count of the seed liquid is not less than 10 9cfu/mL。
9. Euphausia superba oil comestible nutritional product prepared by the process of claims 1-8.
10. The antarctic krill oil edible nutritional product of claim 9 for use in health care, food, medical treatment.
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