阅读说明：本技术 一种提高几丁聚糖功能食品营养价值的方法 (Method for improving nutritive value of chitosan functional food ) 是由 马轩 王冠丹 孙倩 王晓瞳 于 2020-05-26 设计创作，主要内容包括：本发明公开了一种提高几丁聚糖功能食品营养价值的方法,属于海洋生物技术领域。以几丁聚糖为原料,集成现代生物分离技术、生物酶解技术、混菌微生物发酵技术,研制开发出高体外抗氧化能力的几丁聚糖,其中混菌好氧发酵菌种主要包括酿酒酵母、地衣芽孢杆菌和凝结芽孢杆菌。所得几丁聚糖具有较强的抗氧化能力,可满足在保健、食品、医药等领域的高效利用。该工艺技术简单高效,所得产品抗氧化能力和自由基清除能力强,产品稳定性好,具有较强的市场应用前景。(The invention discloses a method for improving the nutritive value of chitosan functional food, belonging to the technical field of marine organisms. The chitosan with high in vitro oxidation resistance is developed by taking chitosan as a raw material and integrating a modern bioseparation technology, a biological enzymolysis technology and a mixed bacteria microbial fermentation technology, wherein the mixed bacteria aerobic fermentation strain mainly comprises saccharomyces cerevisiae, bacillus licheniformis and bacillus coagulans. The obtained chitosan has strong oxidation resistance, and can be efficiently utilized in the fields of health care, food, medicine and the like. The process technology is simple and efficient, and the obtained product has strong oxidation resistance and free radical scavenging capacity, good product stability and stronger market application prospect.)

1. A method for improving the nutritive value of chitosan functional food is characterized by comprising the following steps:

(1) crushing: crushing chitosan, sieving with a 100-mesh sieve, adding purified water in an amount which is 3-5 times that of the chitosan, and stirring for 10 min;

(2) compound enzymolysis: adding a complex enzyme preparation with the mass ratio of 2% into the mixed solution, and stirring and performing enzymolysis for 30min at the temperature of 40-60 ℃;

(3) aerobic fermentation: adding a compound microbial agent with the mass ratio of 3% of the enzymolysis liquid into the enzymolysis liquid after enzymolysis, and carrying out aerobic fermentation for 8h at the temperature of 28-35 ℃;

(4) anaerobic fermentation: after the aerobic fermentation is finished, 2% of lactobacillus casei powder is added, and anaerobic fermentation is carried out for 12 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) concentrating and drying: and (4) carrying out spray drying on the concentrated fermentation liquor.

2. The method for improving the nutritional value of the chitosan functional food as claimed in claim 1, wherein mannan is added in an amount of 3-5% by mass of chitosan during the enzymatic hydrolysis in the step (2).

3. The method as claimed in claim 2, wherein the molecular weight of the mannan is 5000-1000 Da.

4. The method for improving the nutritional value of the chitosan functional food as claimed in claim 1, wherein the complex enzyme preparation in the step (4) is a combination of mannanase, alpha-1, 6 glycosidase and chitosanase, and the mass ratio is 2:2: 1.

5. The method for improving the nutritional value of the chitosan functional food as claimed in claim 1, wherein the compound microbial agent in the step (5) is a mixed microbial agent of saccharomyces cerevisiae, bacillus licheniformis and bacillus coagulans, and the mass ratio of the compound microbial agent to the bacillus coagulans is 2:3: 1.

6. The method of claim 1, wherein the aerobic fermentation in step (5) is performed in a fermentation tank at a ventilation rate of 0.2-0.3 m³/m³.min。

7. The method for improving the nutritional value of chitosan functional food as claimed in claim 1, wherein the Saccharomyces cerevisiae seed solution is prepared by the following steps: under the aseptic operation condition, two rings of strains are picked from the inclined plane of the yeast YDP by using an inoculating ring, inoculated into a 500mL triangular flask containing 300mLPDA liquid culture medium, subjected to shaking culture at the temperature of between 28 and 32 ℃ and at the speed of 140r/min for 24 hours, and then inoculated into a seed tank for expanded culture according to the process to prepare the yeast seed liquid, wherein the effective viable count of the seed liquid is required to be more than or equal to 10⁹cfu/mL。

8. The method of claim 1, wherein the bacillus licheniformis and bacillus coagulans seed solution is prepared by the following steps: under the aseptic condition, LB culture medium is adopted to culture for 24 hours at the temperature of 28-32 ℃ to prepare liquid bacillus licheniformis and bacillus coagulans seed liquid, and the effective viable count of the seed liquid is required to be more than or equal to 10⁹cfu/mL。

9. Chitosan produced by the process of claims 1-8.

10. The chitosan of claim 9, for use in health care, food, and medicine.

Technical Field

The invention relates to a method for improving the nutritive value of chitosan functional food, in particular to a chitosan biological product with high oxidation resistance and nutrition and health care effects prepared by using chitosan as a raw material through microbial synergistic fermentation, belonging to the technical field of marine organisms.

Background

Chitin is called the last renewable resource on the earth, and with the continuous and deep research, the potential value of the chitin is continuously explored, and particularly, the chitosan is gradually a new favorite in the field of health care. Chitosan, also known as chitin, chitosan, chitosamine, deacetylated chitin, and radical polyacid, is an edible cellulose with positive charges, cations and alkalinity; chitosan has a variety of health care functions, and is known as the sixth vital element behind protein, fat, carbohydrate, vitamins and mineral elements by nutriologists. The nutrition and medical effects of chitosan are summarized to have all-round health care functions of three-regulation (regulating immunity, regulating pH value and hormone secretion), three-reduction (reducing cholesterol, reducing blood sugar and blood pressure), three-reduction (discharging redundant harmful cholesterol, discharging heavy metal ions and removing toxins), three-inhibition (inhibiting cancer cell growth, inhibiting cancer metastasis and inhibiting toxins released by cancer cell proliferation) and the like. Therefore, chitosan has a good development prospect in the field of functional foods.

At present, relatively few researches on the degradation and the improvement of the antioxidant capacity of chitosan exist, and especially the improvement of the chitosan-OH free radical scavenging capacity is a key technical disadvantage in the field, which restricts the application of the chitosan-OH free radical scavenging capacity in functional foods.

Disclosure of Invention

In order to overcome the defects of the prior art and meet the requirements of efficient extraction and high-value utilization of chitosan, the application provides a method for improving the nutritional value of chitosan functional food, integrates a modern bioseparation technology, a biological enzymolysis technology and a mixed-bacterium microbial fermentation technology, develops and develops a chitosan biological product with high in-vitro oxidation resistance, meets the application requirements of fields such as health care, medical treatment, food and the like on chitosan, and has the advantages of simple and efficient process technology, strong oxidation resistance and free radical scavenging capacity of the obtained product, strong product stability and strong market application prospect.

The invention realizes the technical effects through the following technical scheme:

a method for improving the nutritive value of chitosan functional food is characterized by comprising the following steps:

(1) crushing: crushing chitosan, sieving with a 100-mesh sieve, adding purified water in an amount which is 3-5 times that of the chitosan, and stirring for 10 min;

(2) compound enzymolysis: adding a complex enzyme preparation with the mass ratio of 2% into the mixed solution, and stirring and performing enzymolysis for 30min at the temperature of 40-60 ℃;

(3) aerobic fermentation: adding a compound microbial agent with the mass ratio of 3% of the enzymolysis liquid into the enzymolysis liquid after enzymolysis, and carrying out aerobic fermentation for 8h at the temperature of 28-35 ℃;

(4) anaerobic fermentation: after the aerobic fermentation is finished, 2% of lactobacillus casei powder is added, and anaerobic fermentation is carried out for 12 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) concentrating and drying: and (4) carrying out spray drying on the concentrated fermentation liquor.

Preferably, mannan with the mass ratio of chitosan being 3-5% is added in the enzymolysis process in the step (2).

Preferably, the mannan has a molecular weight of 5000-1000 Da.

Preferably, the complex enzyme preparation in the step (4) is a combination of mannanase, alpha-1, 6 glycosidase and chitosanase, and the mass ratio of the mannanase, the alpha-1, 6 glycosidase and the chitosanase is 2:2: 1.

Preferably, the compound microbial agent in the step (5) is a mixed microbial agent of saccharomyces cerevisiae, bacillus licheniformis and bacillus coagulans, and the mass ratio of the compound microbial agent to the bacillus coagulans is 2:3: 1.

Preferably, the ventilation rate of the aerobic fermentation process in the step (5) is 0.2-0.3 m³/m³.min。

The preparation method of the saccharomyces cerevisiae seed liquid comprises the following steps: under the aseptic operation condition, two rings of strains are picked from the inclined plane of the yeast YDP by using an inoculating ring, inoculated into a 500mL triangular flask containing 300mLPDA liquid culture medium, subjected to shaking culture at the temperature of between 28 and 32 ℃ and at the speed of 140r/min for 24 hours, and then inoculated into a seed tank for expanded culture according to the process to prepare the yeast seed liquid, wherein the effective viable count of the seed liquid is required to be more than or equal to 10⁹cfu/mL。

The preparation method of the bacillus licheniformis and bacillus coagulans seed solution comprises the following steps: under the aseptic condition, LB culture medium is adopted to culture for 24 hours at the temperature of 28-32 ℃ to prepare liquid bacillus licheniformis and bacillus coagulans seed liquid, and the effective viable count of the seed liquid is required to be more than or equal to 10⁹cfu/mL。

The invention provides a method for improving the nutritive value of chitosan functional food, and the obtained chitosan has stronger oxidation resistance and can meet the high-efficiency utilization in the fields of health care, food, medicine and the like.

Compared with the prior art, the method for improving the nutritive value of the chitosan functional food has the following remarkable advantages:

(1) the chitosan is used as a raw material, and a modern bioseparation technology, a biological enzymolysis technology and a mixed-bacterium microbial fermentation technology are integrated, so that the oxidation resistance of the chitosan is remarkably improved; more importantly, the application creatively discovers that the antioxidant capacity of the chitosan prepared by the enzymolysis and fermentation method can be obviously improved by adding a certain proportion of mannan into a chitosan dissolving solution; meanwhile, test results show that the addition of mannan has obvious influence on the improvement of the efficacy of mannan;

(2) this application utilizes aerobic bacteria (saccharomyces cerevisiae, bacillus licheniformis, bacillus coagulans) to ferment chitosan jointly and draws chitosan, and the synergistic action between the make full use of microorganism promotes chitosan's extraction rate and external antioxidant capacity, wherein: the saccharomyces cerevisiae can generate various enzymes for degrading cell walls in the fermentation process, so that more functional active substances such as polypeptides and the like are released from the cell walls, and the saccharomyces cerevisiae can generate the enzymes capable of releasing conjugated phenolic acid, thereby being beneficial to improving the in-vitro antioxidant capacity of chitosan; the addition of the bacillus licheniformis and the bacillus coagulans can obviously improve the chitosan-OH free radical scavenging capability and has obvious biocatalysis effect.

Detailed Description

In the present application, the activation method of saccharomyces cerevisiae, bacillus licheniformis and bacillus coagulans is as follows:

(1) the preparation method of the saccharomyces cerevisiae seed liquid comprises the following steps: under the aseptic operation condition, two rings of strains are picked from the inclined plane of the yeast YDP by using an inoculating ring, inoculated into a 500mL triangular flask containing 300mLPDA liquid culture medium, subjected to shaking culture at the temperature of between 28 and 32 ℃ and at the speed of 140r/min for 24 hours, and then inoculated into a seed tank for expanded culture according to the process to prepare the yeast seed liquid, wherein the effective viable count of the seed liquid is required to be more than or equal to 10⁹cfu/mL；

(2) The preparation method of the bacillus licheniformis and bacillus coagulans seed solution comprises the following steps: under the aseptic condition, LB culture medium is adopted to culture for 24 hours at the temperature of 28-32 ℃ to prepare liquid bacillus licheniformis and bacillus coagulans seed liquid, and the effective viable count of the seed liquid is required to be more than or equal to 10⁹cfu/mL。