CN110495611A - A kind of technique improving sea cucumber nutritional health effect - Google Patents
A kind of technique improving sea cucumber nutritional health effect Download PDFInfo
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- CN110495611A CN110495611A CN201910711437.9A CN201910711437A CN110495611A CN 110495611 A CN110495611 A CN 110495611A CN 201910711437 A CN201910711437 A CN 201910711437A CN 110495611 A CN110495611 A CN 110495611A
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Classifications
-
- 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
- A23L17/00—Food-from-the-sea products; Fish products; Fish meal; Fish-egg substitutes; Preparation or treatment thereof
-
- 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
- A23L33/17—Amino acids, peptides or proteins
- A23L33/18—Peptides; Protein hydrolysates
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P21/00—Preparation of peptides or proteins
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P21/00—Preparation of peptides or proteins
- C12P21/06—Preparation of peptides or proteins produced by the hydrolysis of a peptide bond, e.g. hydrolysate products
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P39/00—Processes involving microorganisms of different genera in the same process, simultaneously
-
- 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
Abstract
The invention discloses a kind of techniques for improving sea cucumber nutritional health effect, belong to field of marine biotechnology.Using sea cucumber deep processing waste sea cucumber internal organ as raw material, integrated modern biotechnology isolation technics, biological enzymolysis technology, mixed bacteria microorganism fermentation technique, the sea cucumber polypeptide biological products of high antioxidant activity in vitro are developed, wherein mixed bacterium aerobic fermentation strain mainly includes saccharomyces cerevisiae, bacillus subtilis and bacillus pumilus.Gained sea cucumber polypeptide has stronger oxidation resistance, and the processing such as extracting solution can be concentrated according to application approach, be adsorbed by drying, alcohol analysis or purifying resin meets in health care, the efficient utilization of food, medicine and other fields.The technology is simple and efficient, products obtained therefrom oxidation resistance and free radical scavenging ability are strong, and product stability is strong, and production cost is low, is easy to industrial-scale production.
Description
Technical field
The present invention relates to a kind of techniques for improving sea cucumber nutritional health effect, especially a kind of to utilize sea cucumber internal organ for raw material
It is raw to belong to ocean for the sea cucumber polypeptide biological products that high anti-oxidation ability and nutritional health function are prepared by microorganism cooperative fermentation
Object technical field.
Background technique
Sea cucumber is a kind of important oceanic invertebrate, fine and tender taste, full of nutrition, is typical high protein, low fat
The food of fat has high nutritive value and medical value, for the patients such as hypertension, coronary heart disease, hepatitis and the elderly
For be diet good food, the immunity and hematopoiesis function of body had not only can be enhanced in it, but also can inhibit a variety of moulds and human carcinomas is thin
Growth and transfer of born of the same parents etc..As soon as there is a habit of beche-de-mer in China before more than thousand years, and sea cucumber is considered as a kind of preciousness
Invigorant is classified as first of " Hai Bazhen ".Sea cucumber is recorded in " new compilation of materia medica " to be had " impotence is treated in sweet, salty, warm, tonifying kidney and benefiting sperm, establishing-Yang "
The effect of, record that " the warm-natured benefit of sea cucumber, sufficient enemy join therefore named sea cucumber in Qing Dynasty's supplementary Amplifications of the Compendium of Materia Medica；Sweet-salty, kidney tonifying warp, benefit
Marrow, dissolving phlegm prolong, take the photograph urine, impotence is treated in establishing-Yang, kills sore worm ".
Currently, the main Leaf-feeding insects of sea cucumber are wall of sea cucumber Stichopus japonicus, therefore to the research of sea cucumber also multi-focus in wall of sea cucumber Stichopus japonicus
On active material.By-product sea cucumber internal organ in Holothurian machining are then discarded more, and utilization rate is low, it is studied both at home and abroad also compared with
Few, developed Related product is even more very little.However sea cucumber internal organ are in addition to containing chemical component similar with body wall, it is also rich
Containing taurine, arginine, phosphatide, unsaturated fatty acid and various high activity microelements, pigment, organized enzyme and function bacterium,
The edible value and medical value of sea cucumber internal organ are higher than wall of sea cucumber Stichopus japonicus in a certain range.Sea cucumber internal organ active material is recognized at present
The shortage and ignorance of knowledge, have resulted in resource serious waste and loss.
In the prior art, the preparation process of sea cucumber internal organ active peptides focuses primarily upon conbined enzymolysis or composite fermentation
Method, if patent CN201811098258.4 discloses a kind of preparation method of sea cucumber internal organ polypeptide, step includes: by sea cucumber internal organ
Frozen goods are successively thawed, are digested, is centrifuged, is concentrated, drying steps, and complex enzyme is used in the enzymolysis step, described compound
Enzyme is lipase and proteinase combination.The complex enzyme is the combination of Collagenase, papain and lipase；Patent
CN201810512423.X discloses a kind of flavouring and preparation method thereof using the production of sea cucumber internal organ, by weight, including
20-30 parts of clean sea cucumber internal organ, 20-40 parts of soy meal, 10-15 parts of salt, 1-2 parts of composite fermentation strain；By what is usually abandoned
Sea cucumber internal organ make full use of, and by raw material by processes such as homogenate, sterilization, fermentations, big egg can be made under the action of microorganism
White matter molecule is decomposed into the flavor substances such as water-soluble small peptide, polysaccharide, saponin(e.But on how to improve the anti-of sea cucumber internal organ active peptides
The research of oxidisability and nutritive effect is relatively fewer, is especially for sea cucumber internal organ polypeptide OH free radical scavenging ability is improved
The key technology drawback of this field.
Summary of the invention
In order to overcome the above-mentioned deficiencies of the prior art, meet waste sea cucumber processing object high efficiency extraction and higher value application, this
Application provides a kind of technique for improving sea cucumber nutritional health effect, integrates modern biotechnology isolation technics, biological enzymolysis technology, mixed bacterium
Microbial fermentation technology develops the sea cucumber polypeptide biological products of high antioxidant activity in vitro, meets health care, medical treatment, food
The fields such as product to the application demand of sea cucumber polypeptide, which is simple and efficient, products obtained therefrom oxidation resistance and
Free radical scavenging ability is strong, and production cost is low, is easy to industrial-scale production.
The present invention is achieved through the following technical solutions above-mentioned technical effect:
A kind of technique improving sea cucumber nutritional health effect, it is characterised in that preparation method includes the following steps:
(1) it cleans: the new fresh sea cucumber internal organ of processing is cleaned up with tap water, then clean drip with natural mineral water or purified water
It is dry stand-by；
(2) it is homogenized: high-speed homogenization machine is added in sea cucumber internal organ obtained by step (1), natural mineral water or purified water is added, carry out high
Speed homogenate, until can all pass through the aperture 2mm rotary strainer；
(3) it sterilizes: the sea cucumber internal organ after homogenate is sterilized by autoclave, it is stand-by after cooling；
(4) complex enzyme hydrolysis: the complex enzyme formulation of mass ratio 2% is added into the slurries after sterilizing, is stirred under the conditions of 40~60 DEG C
Digest 30min；
(5) aerobic fermentation: the complex micro organism fungicide of enzymolysis liquid mass ratio 3% is added into the enzymolysis liquid after enzymatic hydrolysis, 28~35
Aerobic fermentation 8h under the conditions of DEG C；
(6) 2% Bacillus acidi lactici powder, anaerobic fermentation 12h under the conditions of 28~35 DEG C anaerobic fermentation: are added after aerobic fermentation；
(7) it filters: carrying out coarse filtration and remove fermentation thalli；
(8) enzyme deactivation: enzyme deactivation 10min under the conditions of 100 DEG C；
(9) be centrifuged: with the centrifuge separation of 10000r/min rate, supernatant is sea cucumber polypeptide extracting solution.
Preferably, the enteromorpha oligosaccharide of sea cucumber internal organ mass ratio 5% is added in sea cucumber internal organ homogenization process.
It is further preferred that the molecular weight of the enteromorpha oligosaccharide is 3000-5000D.
Compound protease described in above-mentioned steps (4) is the combination of trypsase, bromelain and papain,
Mass ratio is 2:2:1.
Complex micro organism fungicide described in above-mentioned steps (5) is saccharomyces cerevisiae, bacillus subtilis and bacillus pumilus
Mix bacterium agent, mass ratio group become 2:3:1.
Ventilation quantity is 0.2~0.3m when aerobic fermentation process carries out in the fermenter in above-mentioned steps (5)3/m3.min。
Above-mentioned saccharomyces cerevisiae seed liquor from the inclined-plane saccharomycete YDP the preparation method comprises the following steps: under aseptic technique, use inoculation
Two ring strain of ring picking is inoculated into the triangular flask of 500mL dress 300mLPDA fluid nutrient medium, 28 DEG C~32 DEG C, 140r/min
Shaken cultivation for 24 hours after, then according to above-mentioned technique inoculation seeding tank expand culture preparation saccharomycete seed liquor, it is desirable that seed liquor
Living bacteria count >=109cfu/mL。
Above-mentioned bacillus subtilis and bacillus pumilus seed liquor using LB the preparation method comprises the following steps: aseptically, trained
It supports base to cultivate under the conditions of 28~32 DEG C for 24 hours, liquid bacillus subtilis and bacillus pumilus seed liquor is made, it is desirable that seed
Living bacteria count >=10 of liquid9cfu/mL。
The present invention provides a kind of technique for improving sea cucumber nutritional health effect, and gained sea cucumber polypeptide has stronger anti-oxidant
Ability, the processing such as extracting solution can be concentrated according to application approach, be adsorbed by drying, alcohol analysis or purifying resin, meets in health care, food
The efficient utilization of product, medicine and other fields.
The present invention provides a kind of technique for improving sea cucumber nutritional health effect, compared with prior art, has following significant
Advantage:
(1) the application integrates modern biotechnology isolation technics, biological enzymolysis technology, the mixed micro- life of bacterium using sea cucumber visceral protein as raw material
Object fermentation technique has been obviously improved sea cucumber visceral protein oxidation resistance；Particular, it is important that the discovery of the application creativeness,
A certain proportion of enteromorpha oligosaccharide is added in sea cucumber internal organ lysate can significantly improve the sea cucumber polypeptide of acidic hydrolysis preparation
Oxidation resistance；Meanwhile test result show the molecular weight of enteromorpha oligosaccharide its effect is promoted influence it is significant, when molecular weight not
Do not have above-mentioned effect then in herein described range；
(2) the application utilizes aerobic bacteria (saccharomyces cerevisiae, bacillus subtilis, bacillus pumilus) co-fermentation sea cucumber internal organ egg
White extraction sea cucumber polypeptide, makes full use of the synergistic effect between microorganism, promotes the recovery rate and antioxidation in vitro energy of sea cucumber polypeptide
Power, in which: saccharomyces cerevisiae can generate the enzyme of various degradation cell walls during the fermentation, so that the functions such as more polypeptides are living
Property substance released from cell wall, and yeast is to generate that the enzyme of reference state phenolic acid can be discharged, and is conducive to promote sea cucumber
The antioxidant activity in vitro of polypeptide；Bacillus subtilis can kill the pathogenic bacteria in fermentation liquid during hot fermentation, and
With abilities such as stronger production amylase, protease, macromolecules starch, protein substance can be degraded to Institute of Micro-biology's utilization,
To improve the oxidation resistance and nutritional health function of active peptide；The addition of bacillus pumilus can then be obviously improved sea cucumber
Polypeptide OH free radical scavenging ability；
(3), the present invention carries out anaerobic fermentation using Bacillus acidi lactici, improves the stability of sea cucumber polypeptide, test result shows In
Within the scope of 20~80 DEG C, temperature and time influences less, to resist when temperature is more than 100 DEG C on sea cucumber internal organ range of hydrolysed peptides inoxidizability
Oxidisability starts to reduce with the increase of temperature and time, and stability is higher than the technical level reported at present；
(4) liquid fermentation is used, the growth rate of thallus is improved, shortens fermentation time, improves production efficiency fermentation work
Skill mild condition, production cost is low, and is easy to large-scale production, and applying to promotion industrial economy benefit and large-scale promotion has
Important function.
Specific embodiment
In this application, saccharomyces cerevisiae, bacillus subtilis and the activation method of bacillus pumilus are as follows:
(1) saccharomyces cerevisiae seed liquor from the inclined-plane saccharomycete YDP with oese the preparation method comprises the following steps: under aseptic technique, chosen
Two ring strains are taken, are inoculated into the triangular flask of 500mL dress 300mLPDA fluid nutrient medium, 28 DEG C~32 DEG C, 140r/min oscillation
Culture for 24 hours after, then according to above-mentioned technique inoculation seeding tank expand culture preparation saccharomycete seed liquor, it is desirable that seed liquor it is effective
Viable count >=109cfu/mL；
(2) above-mentioned bacillus subtilis and bacillus pumilus seed liquor using LB the preparation method comprises the following steps: aseptically, cultivated
Base is cultivated for 24 hours under the conditions of 28~32 DEG C, liquid bacillus subtilis and bacillus pumilus seed liquor is made, it is desirable that seed liquor
Living bacteria count >=109cfu/mL。
Embodiment 1
A kind of technique improving sea cucumber nutritional health effect, specific process step include:
(1) it cleans: the new fresh sea cucumber internal organ of processing is cleaned up with tap water, then clean drip with natural mineral water or purified water
It is dry stand-by；
(2) it is homogenized: high-speed homogenization machine is added in sea cucumber internal organ obtained by step (1), natural mineral water or purified water is added, carry out high
Speed homogenate, until can all pass through the aperture 2mm rotary strainer；
(3) it sterilizes: the sea cucumber internal organ after homogenate is sterilized by autoclave, it is stand-by after cooling；
(4) complex enzyme hydrolysis: the complex enzyme formulation of mass ratio 2% is added into the slurries after sterilizing, is stirred under the conditions of 40~60 DEG C
Digest 30min；Combination of the compound protease for trypsase, bromelain and papain, mass ratio 2:
2:1；
(5) aerobic fermentation: the complex micro organism fungicide of enzymolysis liquid mass ratio 3% is added into the enzymolysis liquid after enzymatic hydrolysis, 28~35
Aerobic fermentation 8h under the conditions of DEG C；The complex micro organism fungicide is saccharomyces cerevisiae, bacillus subtilis and bacillus pumilus
Mix bacterium agent, mass ratio group become 2:3:1；Ventilation quantity is 0.2~0.3m during aerobic fermentation3/m3.min；
(6) 2% Bacillus acidi lactici powder, anaerobic fermentation 12h under the conditions of 28~35 DEG C anaerobic fermentation: are added after aerobic fermentation；
(7) it filters: carrying out coarse filtration and remove fermentation thalli；
(8) enzyme deactivation: enzyme deactivation 10min under the conditions of 100 DEG C；
(9) be centrifuged: with the centrifuge separation of 10000r/min rate, supernatant is sea cucumber polypeptide extracting solution.
The oxidation resistance of sea cucumber polypeptide extracting solution in embodiment 1 is measured, wherein the measurement ginseng of OH free radical scavenging activity
According to method described in Shao Xiuzhi etc. " Food Chemistry test "；The measurement of DPPH free radical scavenging activity is referring to Lin Lianzhu etc. " when reaction
Between to DPPH method, ABTS method evaluation inoxidizability result influence " described in method；Measurement result is as follows:
| Test group | OH clearance rate (%) | DPPH free radical scavenging activity (%) |
| 1 test group of embodiment | 3.28 | 80.12 |
| Compare 1 group | 2.09 | 52.76 |
| Compare 2 groups | 2.77 | 70.02 |
| Compare 3 groups | 2.03 | 65.47 |
1 group is compareed in above-mentioned experiment as the sea cucumber polypeptide product after step (2) enzymolysis liquid enzyme deactivation centrifugation in embodiment 1；Compare 2 groups
The sea cucumber polypeptide product after bacillus pumilus is fermented is substituted for bacillus subtilis in aerobic fermentation in embodiment 1,
Remaining same embodiment 1；3 groups are compareed to ferment for bacillus pumilus substitution bacillus subtilis in aerobic fermentation in embodiment 1
Sea cucumber polypeptide product afterwards, remaining is the same as embodiment 1.
Embodiment 2
A kind of technique improving sea cucumber nutritional health effect, specific process step include:
(1) it cleans: the new fresh sea cucumber internal organ of processing is cleaned up with tap water, then clean drip with natural mineral water or purified water
It is dry stand-by；
(2) it is homogenized: high-speed homogenization machine is added in sea cucumber internal organ obtained by step (1), natural mineral water or purified water is added, carry out high
Speed homogenate, until can all pass through the aperture 2mm rotary strainer；The enteromorpha oligosaccharide of sea cucumber internal organ mass ratio 5% is added, the Enteromorpha is few
The molecular weight of sugar is 3000D.
(3) it sterilizes: the sea cucumber internal organ after homogenate is sterilized by autoclave, it is stand-by after cooling；
(4) complex enzyme hydrolysis: the complex enzyme formulation of mass ratio 2% is added into the slurries after sterilizing, is stirred under the conditions of 40~60 DEG C
Digest 30min；Combination of the compound protease for trypsase, bromelain and papain, mass ratio 2:
2:1；
(5) aerobic fermentation: the complex micro organism fungicide of enzymolysis liquid mass ratio 3% is added into the enzymolysis liquid after enzymatic hydrolysis, 28~35
Aerobic fermentation 8h under the conditions of DEG C；The complex micro organism fungicide is saccharomyces cerevisiae, bacillus subtilis and bacillus pumilus
Mix bacterium agent, mass ratio group become 2:3:1；Ventilation quantity is 0.2~0.3m during aerobic fermentation3/m3.min；
(6) 2% Bacillus acidi lactici powder, anaerobic fermentation 12h under the conditions of 28~35 DEG C anaerobic fermentation: are added after aerobic fermentation；
(7) it filters: carrying out coarse filtration and remove fermentation thalli；
(8) enzyme deactivation: enzyme deactivation 10min under the conditions of 100 DEG C；
(9) be centrifuged: with the centrifuge separation of 10000r/min rate, supernatant is sea cucumber polypeptide extracting solution.
The oxidation resistance of sea cucumber polypeptide extracting solution in embodiment 2 is measured, measuring method is the same as antioxygen described in embodiment 1
Change performance measurement method.Test result is as follows:
| Test group | OH clearance rate (%) | DPPH free radical scavenging activity (%) |
| 2 test group of embodiment | 51.93 | 90.62 |
| Compare 1 group | 8.72 | 59.17 |
| Compare 2 groups | 21.96 | 80.35 |
| Compare 3 groups | 28.94 | 74.65 |
1 group is compareed in above-mentioned experiment as the sea cucumber polypeptide product after step (2) enzymolysis liquid enzyme deactivation centrifugation in embodiment 2；Compare 2 groups
The sea cucumber polypeptide product after bacillus pumilus is fermented is substituted for bacillus subtilis in aerobic fermentation in embodiment 2,
Remaining same embodiment 2；3 groups are compareed to ferment for bacillus pumilus substitution bacillus subtilis in aerobic fermentation in embodiment 2
Sea cucumber polypeptide product afterwards, remaining is the same as embodiment 2.
Embodiment 3
A kind of technique improving sea cucumber nutritional health effect, specific process step include:
(1) it cleans: the new fresh sea cucumber internal organ of processing is cleaned up with tap water, then clean drip with natural mineral water or purified water
It is dry stand-by；
(2) it is homogenized: high-speed homogenization machine is added in sea cucumber internal organ obtained by step (1), natural mineral water or purified water is added, carry out high
Speed homogenate, until can all pass through the aperture 2mm rotary strainer；The enteromorpha oligosaccharide of sea cucumber internal organ mass ratio 5% is added, the Enteromorpha is few
The molecular weight of sugar is 5000D.
(3) it sterilizes: the sea cucumber internal organ after homogenate is sterilized by autoclave, it is stand-by after cooling；
(4) complex enzyme hydrolysis: the complex enzyme formulation of mass ratio 2% is added into the slurries after sterilizing, is stirred under the conditions of 40~60 DEG C
Digest 30min；Combination of the compound protease for trypsase, bromelain and papain, mass ratio 2:
2:1；
(5) aerobic fermentation: the complex micro organism fungicide of enzymolysis liquid mass ratio 3% is added into the enzymolysis liquid after enzymatic hydrolysis, 28~35
Aerobic fermentation 8h under the conditions of DEG C；The complex micro organism fungicide is saccharomyces cerevisiae, bacillus subtilis and bacillus pumilus
Mix bacterium agent, mass ratio group become 2:3:1；Ventilation quantity is 0.2~0.3m during aerobic fermentation3/m3.min；
(6) 2% Bacillus acidi lactici powder, anaerobic fermentation 12h under the conditions of 28~35 DEG C anaerobic fermentation: are added after aerobic fermentation；
(7) it filters: carrying out coarse filtration and remove fermentation thalli；
(8) enzyme deactivation: enzyme deactivation 10min under the conditions of 100 DEG C；
(9) be centrifuged: with the centrifuge separation of 10000r/min rate, supernatant is sea cucumber polypeptide extracting solution.
The oxidation resistance of sea cucumber polypeptide extracting solution in embodiment 3 is measured, measuring method is the same as antioxygen described in embodiment 1
Change performance measurement method.Test result is as follows:
| Test group | OH clearance rate (%) | DPPH free radical scavenging activity (%) |
| 3 test group of embodiment | 31.38 | 88.21 |
| Compare 1 group | 9.21 | 63.02 |
| Compare 2 groups | 18.89 | 79.39 |
| Compare 3 groups | 22.31 | 72.01 |
1 group is compareed in above-mentioned experiment as the sea cucumber polypeptide product after step (2) enzymolysis liquid enzyme deactivation centrifugation in embodiment 3；Compare 2 groups
The sea cucumber polypeptide product after bacillus pumilus is fermented is substituted for bacillus subtilis in aerobic fermentation in embodiment 3,
Remaining same embodiment 3；3 groups are compareed to ferment for bacillus pumilus substitution bacillus subtilis in aerobic fermentation in embodiment 3
Sea cucumber polypeptide product afterwards, remaining is the same as embodiment 3.
The above examples are only used to illustrate the technical scheme of the present invention, rather than is limited；Although referring to aforementioned reality
It applies example to be described in detail to by invention, but for those of ordinary skill in the art, it still can be to aforementioned reality
Technical solution documented by example is applied to modify or equivalent replacement of some of the technical features；And to these modifications
Or replacement, the spirit and scope for claimed technical solution of the invention that it does not separate the essence of the corresponding technical solution.
Claims (10)
1. a kind of technique for improving sea cucumber nutritional health effect, it is characterised in that preparation method includes the following steps:
(1) it cleans: the new fresh sea cucumber internal organ of processing is cleaned up with tap water, then clean drip with natural mineral water or purified water
It is dry stand-by；
(2) it is homogenized: high-speed homogenization machine is added in sea cucumber internal organ obtained by step (1), natural mineral water or purified water is added, carry out high
Speed homogenate, until can all pass through the aperture 2mm rotary strainer；
(3) it sterilizes: the sea cucumber internal organ after homogenate is sterilized by autoclave, it is stand-by after cooling；
(4) complex enzyme hydrolysis: the complex enzyme formulation of mass ratio 2% is added into the slurries after sterilizing, is stirred under the conditions of 40~60 DEG C
Digest 30min；
(5) aerobic fermentation: the complex micro organism fungicide of enzymolysis liquid mass ratio 3% is added into the enzymolysis liquid after enzymatic hydrolysis, 28~35
Aerobic fermentation 8h under the conditions of DEG C；
(6) 2% Bacillus acidi lactici powder, anaerobic fermentation 12h under the conditions of 28~35 DEG C anaerobic fermentation: are added after aerobic fermentation；
(7) it filters: carrying out coarse filtration and remove fermentation thalli；
(8) enzyme deactivation: enzyme deactivation 10min under the conditions of 100 DEG C；
(9) be centrifuged: with the centrifuge separation of 10000r/min rate, supernatant is sea cucumber polypeptide extracting solution.
2. a kind of technique for improving sea cucumber nutritional health effect according to claim 1, it is characterised in that the step (2)
The enteromorpha oligosaccharide of sea cucumber internal organ mass ratio 5% is added in homogenization process.
3. a kind of technique for improving sea cucumber nutritional health effect according to claim 2, it is characterised in that the Enteromorpha is few
The molecular weight of sugar is 3000-5000D.
4. a kind of technique for improving sea cucumber nutritional health effect according to claim 1, it is characterised in that the step (4)
Described in compound protease be trypsase, bromelain and papain combination, mass ratio 2:2:1.
5. a kind of technique for improving sea cucumber nutritional health effect according to claim 1, it is characterised in that the step (5)
Described in complex micro organism fungicide be saccharomyces cerevisiae, bacillus subtilis and the mix bacterium agent of bacillus pumilus, mass ratio
Group becomes 2:3:1.
6. a kind of technique for improving sea cucumber nutritional health effect according to claim 1, it is characterised in that the step (5)
Ventilation quantity is 0.2~0.3m when middle aerobic fermentation process carries out in the fermenter3/m3.min。
7. a kind of technique for improving sea cucumber nutritional health effect according to claim 1, it is characterised in that the wine brewing ferment
The sub- liquid and preparation method thereof of parent species are as follows: under aseptic technique, two ring strain of oese picking is used from the inclined-plane saccharomycete YDP, is connect
Kind to 500mL dress 300mLPDA fluid nutrient medium triangular flask in, 28 DEG C~32 DEG C, 140r/min shaken cultivation for 24 hours after, then
Expand culture preparation saccharomycete seed liquor according to above-mentioned technique inoculation seeding tank, it is desirable that the living bacteria count of seed liquor >=
109cfu/mL。
8. a kind of technique for improving sea cucumber nutritional health effect according to claim 1, it is characterised in that the withered grass bud
Spore bacillus and bacillus pumilus seed liquor are the preparation method comprises the following steps: aseptically, using LB culture medium in 28~32 DEG C of conditions
For 24 hours, liquid bacillus subtilis and bacillus pumilus seed liquor is made in lower culture, it is desirable that and the living bacteria count of seed liquor >=
109cfu/mL。
9. by the sea cucumber polypeptide of the preparation of technique described in claim 1-8.
10. sea cucumber polypeptide described in claim 9 is in the application in health care, food, medicine.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110973342A (en) * | 2020-03-05 | 2020-04-10 | 中国科学院烟台海岸带研究所 | Apostichopus japonicus oligopeptide and preparation method and application thereof |
| CN112370369A (en) * | 2020-10-28 | 2021-02-19 | 梁志勇 | Marine protein skin moisturizer for resisting skin aging |
| CN112690387A (en) * | 2020-12-24 | 2021-04-23 | 大连鑫澜生物技术有限公司 | Sea cucumber intestine extract probiotic drink capable of strengthening kidney and strengthening body |
| CN114504083A (en) * | 2020-11-16 | 2022-05-17 | 烟台东宇海珍品有限公司 | Processing method of sea cucumber sausages |
-
2019
- 2019-08-02 CN CN201910711437.9A patent/CN110495611A/en not_active Withdrawn
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110973342A (en) * | 2020-03-05 | 2020-04-10 | 中国科学院烟台海岸带研究所 | Apostichopus japonicus oligopeptide and preparation method and application thereof |
| CN112370369A (en) * | 2020-10-28 | 2021-02-19 | 梁志勇 | Marine protein skin moisturizer for resisting skin aging |
| CN114504083A (en) * | 2020-11-16 | 2022-05-17 | 烟台东宇海珍品有限公司 | Processing method of sea cucumber sausages |
| CN112690387A (en) * | 2020-12-24 | 2021-04-23 | 大连鑫澜生物技术有限公司 | Sea cucumber intestine extract probiotic drink capable of strengthening kidney and strengthening body |
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