阅读说明：本技术 一种发酵榨油粕类制备活性多肽的方法 (Method for preparing active polypeptide by fermenting oil pressing meals ) 是由 李清玉 刘希军 于 2020-06-17 设计创作，主要内容包括：本发明提供了一种发酵榨油粕类制备活性多肽的方法,其包括以下的步骤：(1)配置发酵培养基;(2)接种复合菌剂5-10%,在200rpm的摇床中摇床培养30-40h；(3)8000rpm离心,20min；(4)发酵液采用10kDa,5kDa,3kDa,1kDa的卷式滤膜进行截留；(5)对5-10kDa的组分进行硫酸铵沉淀；(6)将硫酸铵沉淀采用PBS缓冲液溶解并在PBS溶液中进行透析；(7)将透析所得5-10kDa以及3-5kDa,1-3kDa的组分进行冷冻干燥即得活性多肽。本发明将豆粕等转化为高附加值的活性多肽,该活性多肽并未检出黄曲霉毒素,安全可靠,其具有促进动物生长,提高机体免疫力的功效。(The invention provides a method for preparing active polypeptide by fermenting oil cakes, which comprises the following steps: (1) inoculating 5-10% of the compound microbial inoculum, and performing shake culture in a shaker at 200rpm for 30-40 h; (3) centrifuging at 8000rpm for 20 min; (4) the fermentation liquor is intercepted by a 10kDa, 5kDa, 3kDa and 1kDa roll-up filter membrane; (5) performing ammonium sulfate precipitation on the 5-10kDa fraction; (6) dissolving the ammonium sulfate precipitate by adopting PBS buffer solution and dialyzing in the PBS solution; (7) freeze drying the 5-10kDa and 3-5kDa, 1-3kDa fractions obtained by dialysis to obtain active polypeptide. The invention converts the soybean meal and the like into the active polypeptide with high added value, the active polypeptide does not detect aflatoxin, is safe and reliable, and has the effects of promoting the growth of animals and improving the immunity of organisms.)

1. A method for preparing active polypeptide by fermenting oil pressing meals is characterized in that: which comprises the following steps:

(1) preparing a fermentation culture medium;

(2) inoculating 5-10% of the compound microbial inoculum, and performing shake culture in a shaking table at 200rpm for 30-40 h;

(3) centrifuging at 8000rpm for 20 min;

(4) the fermentation liquor is intercepted by a 10kDa, 5kDa, 3kDa and 1kDa roll-up filter membrane;

(5) performing ammonium sulfate precipitation on the 5-10kDa fraction;

(6) dissolving the ammonium sulfate precipitate by adopting PBS buffer solution and dialyzing in the PBS solution;

(7) freeze drying the 5-10kDa and 3-5kDa, 1-3kDa fractions obtained by dialysis to obtain active polypeptide.

2. The method for preparing active polypeptide by fermenting oil cakes according to claim 1, which is characterized in that: in the step (1), the specific method is as follows: dissolving 60-80% of oil cakes, 20-30% of starch, 1-5% of NaCl, K2HPO40.2-0.4% and MnCl20.3-0.5% in 1000mL of distilled water, and sterilizing at 121 ℃ for 20 min.

3. The method for preparing active polypeptide by fermenting oil meals according to claim 2, characterized in that: the oil pressing meals are peanut meals: bean pulp: rapeseed meal =1-1.5: 1-1.5: 2-3.

4. The method for preparing active polypeptide by fermenting oil cakes according to claim 1, which is characterized in that: the compound microbial inoculum adopted in the step (2) is trichoderma viride: b, bacillus licheniformis: bacillus pumilus: b, bacillus amyloliquefaciens: bifidobacteria: lactobacillus wenselii =1:1:1:4:1: 2.

5. The method for preparing active polypeptide by fermenting oil cakes according to claim 1, which is characterized in that: the concentration of the water saturated ammonium sulfate precipitated by the ammonium sulfate in the step (5) is 30-60%.

6. The method for preparing active polypeptide by fermenting oil cakes according to claim 1, which is characterized in that: the number of the viable bacteria in the composite microbial inoculum is more than 108 cfu/mL.

Technical Field

The invention belongs to the field of fermentation preparation of active products, and particularly relates to a method for preparing active polypeptide by fermenting oil pressing cakes.

Background

Bean pulp, peanut pulp, rapeseed pulp and the like are byproducts obtained after oil pressing. The oil is extracted only from oil, and the dregs contain many nutritive substances such as protein. For example, the rapeseed meal has a crude protein content of 32% or more and a crude fiber content of 12% or less. The peanut meal is rich in vegetable protein and has good taste. Meals are therefore often used directly in the feed sector. However, a large amount of nutrient components contained in meals are not easy to be directly absorbed by animals, which causes resource waste and has important significance for further extracting and utilizing nutrient substances in meals.

The bioactive polypeptide is prepared from meals by a microbial fermentation method, so that the utilization value of the meals can be improved. Meanwhile, meal residues left by microbial fermentation are easier to absorb by animals, some polysaccharides which are difficult to absorb and utilize by the animals can be degraded into active substances such as oligosaccharides and the like by the microorganisms, and the contained probiotics can also promote the intestinal health of the animals.

The preparation of bioactive polypeptides from meals by a microbial fermentation method and the realization of high-value utilization of the meals become problems to be solved urgently.

Disclosure of Invention

The invention aims to solve the problem of preparing bioactive polypeptide from meals by using a microbial fermentation method and realize high-value utilization of the meals.

The purpose of the invention is realized by the following technical scheme:

a method for preparing active polypeptide by fermenting oil cakes comprises the following steps:

(1) preparing a fermentation culture medium;

(2) inoculating 5-10% of the compound microbial inoculum, and performing shake culture in a shaking table at 200rpm for 30-40 h;

(3) centrifuging at 8000rpm for 20 min;

(4) the fermentation liquor is intercepted by a 10kDa, 5kDa, 3kDa and 1kDa roll-up filter membrane;

(5) performing ammonium sulfate precipitation on the 5-10kDa fraction;

(6) dissolving the ammonium sulfate precipitate by adopting PBS buffer solution and dialyzing in the PBS solution;

(7) freeze drying the 5-10kDa and 3-5kDa, 1-3kDa fractions obtained by dialysis to obtain active polypeptide.

Further, in the step (1), the specific method is as follows: dissolving 60-80% of oil cakes, 20-30% of starch, 1-5% of NaCl, K2HPO40.2-0.4% and MnCl20.3-0.5% in 1000mL of distilled water, and sterilizing at 121 ℃ for 20 min.

Further, the oil pressing meals are peanut meals: bean pulp: rapeseed dregs are 1-1.5:1-1.5: 2-3.

Further, the compound microbial inoculum adopted in the step (2) is trichoderma viride: b, bacillus licheniformis: bacillus pumilus: b, bacillus amyloliquefaciens: bifidobacteria: lactobacillus weissei is 1:1:1:4:1: 2.

Further, the concentration of the water saturated ammonium sulfate precipitated by the ammonium sulfate in the step (5) is 30-60%.

Further, the number of viable bacteria in the composite microbial inoculum is more than 108 cfu/mL.

Compared with the prior art, the invention has the following advantages:

1. the invention belongs to high-value utilization of oil pressing meals, and converts soybean meal and the like which can only be used for feed into active polypeptide with high added value.

2. The active polypeptide obtained by the invention has the effects of promoting the growth of animals and improving the immunity of organisms.

3. Aflatoxin is not detected in the active polypeptide treated by the method, and the method is safe and reliable.

Detailed Description

The present invention will be described in detail with reference to examples, experimental examples and tables.