Method for producing feed probiotics through fermentation of waste feather degradation products

Abstract

The invention discloses a method for producing feed probiotics through fermentation of waste feather degradation products. Comprising the following steps: (1) collecting waste feathers, cleaning and drying; (2) high-temperature and high-pressure treatment of the waste feathers; (3) adding keratinase for hydrolysis; (4) adding glucose into the feather hydrolysate, and sterilizing to prepare a culture medium; (5) inoculating an activated probiotic seed solution, and culturing in a constant-temperature shaking table; and (6) collecting thalli, and performing spray drying to obtain the probiotic feed. According to the method disclosed by the invention, the waste feather hydrolysate is applied to production of the probiotics capable of producing the vitamin B12, so that the prepared probiotic product has better probiotic potential, the animal immunity is effectively enhanced, and an important guarantee is provided for healthy development of the breeding industry.

Description

Technical Field

[0001] The invention belongs to the field of microbial technology application, and specifically relates to a method for producing probiotic feed by utilizing waste feather degradation products. Background Art

[0002] With the intensive development of poultry farming, poultry is slaughtered in a centralized manner in most parts of the country. The byproducts produced are mainly feathers, among which the output of keratin waste is increasing. However, at present, high-quality keratin resources are quite scarce. Only a small amount of high-quality feathers are used as clothing, handicrafts, etc. The vast majority of the remaining feathers are incinerated or directly discarded as garbage due to the difficulty in handling and the large output. This not only poses a great risk to environmental sanitation, but also is a huge waste of resources.

[0003] The structure of feather keratin is quite stable and degrades very slowly. And due to the complex composition of microorganisms in the environment, it is easy to cause secondary pollution. Feather keratin molecules have a high degree of disulfide cross-linking. Once this chemical structure is broken, it will release rich protein resources, which can be used in the feed industry to solve the problem of protein resource shortage.

[0004] At present, the main treatment methods for discarded feathers include high temperature and high pressure hydrolysis, puffing denaturation, protease hydrolysis and microbial degradation. Taking into account the production cost and operability, the high temperature and high pressure hydrolysis method has become the most widely used treatment method. Under high temperature and high pressure conditions, the chemical structure of the feathers is broken and the protein is hydrolyzed. However, a single high temperature and high pressure method cannot guarantee that the feathers are fully hydrolyzed, and it is necessary to combine the protease hydrolysis method to obtain a complete hydrolyzed protein product. This becomes a good culture medium for microbial culture and can provide sufficient nitrogen source for its growth. This method converts discarded feathers into probiotic protein products with high added value, which can effectively alleviate environmental pressure, reduce resource waste, and bring greater economic benefits. It is a green, environmentally friendly and sustainable application method. Summary of the invention

[0005] The present invention aims to solve the problem of waste of discarded feather resources and provides a high temperature and high pressure combined with enzyme hydrolysis method to prepare feather hydrolyzate, which can be used as a nitrogen source for feed probiotic cultivation, and can convert feather protein into high-quality protein and highly active probiotics, thereby improving the added value of the product.

[0006] To achieve the above object, the present invention provides a method for producing probiotics for feed by fermenting waste feather degradation products, characterized in that it comprises the following steps:

[0007] (1) Collect discarded feathers, wash and dry them;

[0008] (2) High temperature and high pressure treatment of discarded feathers;

[0009] (3) adding keratinase for hydrolysis;

[0010] (4) Add glucose to the feather hydrolysate and sterilize to prepare the culture medium;

[0011] (5) Inoculating the activated probiotic seed solution and culturing in a constant temperature shaking incubator;

[0012] (6) Collect the bacteria and spray-dry them to obtain probiotic feed.

[0013] Preferably, in the hydrolysis system of step (2), the high temperature and high pressure treatment conditions of the waste feathers are: 145-155°C, 0.5-0.6 MPa for 30 min.

[0014] Preferably, in step (3), the solution is dissolved in ultrapure water at a mass-to-volume ratio of 1:20, the pH is adjusted to 9.0, the amount of keratinase added is 15000-20000 U / mL, and the treatment time is 3-5 h.

[0015] Preferably, the glucose in step (4) is added in an amount of 10 g / L glucose, the pH of the system is adjusted to 7.0, and sterilized at 121°C for 20 min.

[0016] Preferably, in step (5), the mixed probiotics are specifically: Bacillus subtilis and vitamin B-producing 12 Propionibacterium freudenreichii (CICC10019), but not limited to common Bacillus.

[0017] Preferably, in step (5), the activation conditions of the probiotics are as follows: Bacillus subtilis is cultured in LB medium at 37°C and 150 rpm for 24 h, Propionibacterium freudenreichii is cultured in RCM medium at 37°C and 150 rpm for 36 h, and the Bacillus subtilis solution and the Propionibacterium freudenreichii solution are mixed in a volume ratio of 5:1 to obtain a probiotic seed solution with a live bacteria concentration of 10 8 CFU / mL.

[0018] Preferably, the spray drying in step (6) is performed using a small spray dryer, and the spray drying conditions are as follows: relative density of the concentrate is 1.10 g / mL, air inlet temperature is 135°C, and pumping speed is 15%.

[0019] Preferably, the feathers include but are not limited to chicken feathers, duck feathers, and goose feathers.

[0020] Compared with the prior art, the present invention has the following advantages:

[0021] 1. Feather degradation products are further utilized by feed probiotics, reducing environmental pollution and resource waste, and increasing the added value of feather products.

[0022] 2. The method of using waste feathers after hydrolysis to cultivate high-quality probiotics provides a rich source of protein for feed production, which is a green, environmentally friendly and sustainable application method.

[0023] 3. The hydrolyzate of discarded feathers is used as a nitrogen source to cultivate probiotics. A high density of active bacteria is retained in the system. It can be used as a feed additive to improve the balance of animal intestinal microorganisms, promote nutrient absorption, and maintain gastrointestinal health.

[0024] 4. The hydrolysis of waste feathers is used to produce probiotics. It is safe, efficient and low-cost. It is an effective means of high-value utilization of waste feathers and has broad application prospects.

[0025] 5. The present invention uses the waste feather hydrolysate to produce vitamin B 12 This makes the prepared probiotic products have better probiotic potential, effectively enhance animal immunity, and provide important guarantees for the healthy development of the breeding industry. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 These are the actual pictures of waste feather hydrolysates. A: Feathers after high temperature and high pressure treatment; B: Feathers hydrolysate after keratinase.

[0027] Figure 2 Waste feather hydrolysate is used for probiotic culture and viable bacteria count. DETAILED DESCRIPTION

[0028] The following examples are provided to further illustrate the present invention, but are not intended to limit the present invention.

[0029] Bacillus subtilis HC2 (Bacillus subtilis ( Bacillus subtilis ) HC2, deposit number GDMCC No: 62466, deposited in Guangdong Provincial Microbiological Culture Collection Center on May 12, 2022, published in Chinese patent application: 202211048781.2, a patent application for a method for preparing animal feed by bioconversion of discarded feathers) was cultured in LB medium at 37°C in a shaking incubator for 24 h. Produces vitamin B 12 Propionibacterium freudenreichii (CICC10019, deposited in the China Industrial Microbiological Culture Collection Center, with the deposit number CICC10019) was cultured in RCM medium at 37°C in a shaking incubator for 36 h. The two bacterial solutions were mixed in a volume ratio of 5:1, and the concentration of live bacteria was 10 8 CFU / mL. Continue to culture in a constant temperature shaker at 37°C, 150 rpm for 24 h to obtain the probiotic seed solution.

[0030] Example 1

[0031] Waste chicken feathers were collected from slaughterhouses, washed and dried. Denatured by treating at 155°C and 0.6 MPa for 30 min. Dissolved in ultrapure water at a mass volume ratio of 1 g:10 ml, adjusted to pH 9.0, and hydrolyzed in a shaker at room temperature for 5 h with 15,000 U / mL keratinase. Feather hydrolysate ( Figure 1 ) 10 g / L glucose was added, the pH of the system was adjusted to 7.0, and the culture medium was prepared by sterilization at 121°C for 20 min. The activated probiotic seed solution (Bacillus subtilis and Lactobacillus plantarum) was inoculated at a density of 2×10 8 CFU / mL. Culture in a 37℃ constant temperature shaker for 2-3 days. Collect the bacteria by centrifugation, spray dry, and store in a sealed container at low temperature. After fermentation, the viable bacteria count is as follows: Figure 2 As shown, Bacillus subtilis and Lactobacillus plantarum were 8.4×10 9 CFU / mL and 3.2×10 9 CFU / mL, the total bacterial density is as high as 1×10 10 CFU / mL and above.

[0032] Example 2

[0033] Collect discarded duck feathers, clean them and dry them. Treat them at 145 ℃ and 0.50 MPa for 0.5 h. Then, dissolve the feathers in water at a mass volume ratio of 1g:20ml, adjust the pH of the system to 9.0, add 20000 U / mL keratinase and hydrolyze them on a shaker at room temperature for 3 h. Add 10 g / L glucose to the feather hydrolysate, adjust the pH of the system to 7.0, and sterilize at 121℃ for 20 min to prepare the culture medium. Inoculate the activated probiotic seed liquid (Bacillus subtilis and Bacillus amyloliquefaciens) with an inoculation density of 1.5×10 8 CFU / mL. Cultured in a 37℃ constant temperature shaker for 1-3 days. Collected the bacteria by centrifugation, spray dried and sealed for storage. After the production was completed, samples were taken for one week, one month and three months respectively, and used to determine the changes in the quality of probiotic feed as shown in Table 1. The results showed that the probiotics were stable in activity and could maintain a high density of viable bacteria after three months of storage. LB medium: 5 g yeast powder, 10 g tryptone, 10 g sodium chloride, dilute to 1000 mL with ultrapure water, adjust pH to 7.0. Sterilize at 121°C for 20 min after mixing well. 15 g agar powder should be added to LB agar medium.

[0034] MRS medium: 10 g peptone, 5 g beef extract powder, 4 g yeast extract powder, 20 g glucose, 2 g triammonium citrate, 4 g K2HP04, 5 g sodium acetate, 0.4 g MgS04, 0.05 g MnS04, 0.1% Tween 80, ultrapure water to 1000 mL, adjust pH to 6.2. Sterilize at 121°C for 20 min. MRS agar medium needs to add 15 g agar powder.

Claims

1. A method for producing probiotics for feed by fermenting waste feather degradation products, characterized in that: The following steps are involved: (1) Collect discarded feathers, clean and dry them; (2) High temperature and high pressure treatment of discarded feathers; (3) adding keratinase for hydrolysis; (4) adding glucose to the feather hydrolysate and sterilizing it to prepare a culture medium; (5) inoculating activated probiotic seed solution and culturing in a constant temperature shaking incubator; (6) Collect the bacteria and spray-dry them to obtain probiotic feed.

2. The method according to claim 1, characterized in that In the hydrolysis system of step (2), the high temperature and high pressure treatment conditions of the waste feathers are: 145-155° C., 0.5-0.6 MPa, for 30 min.

3. The method according to claim 1, characterized in that As described in step (3), the solution is dissolved in ultrapure water at a mass volume ratio of 1:20, the pH is adjusted to 9.0, the amount of keratinase added is 15000-20000U / mL, and the treatment time is 3-5h.

4. The method according to claim 1, characterized in that The glucose described in step (4) is added in an amount of 10 g / L glucose, the pH of the system is adjusted to 7.0, and sterilized at 121° C. for 20 min.

5. The method according to claim 1, characterized in that The mixed probiotics described in step (5) are specifically: Bacillus subtilis and vitamin B 12 freudenreichii, but not limited to common Bacillus.

6. The method according to claim 1, characterized in that The activation conditions of the probiotics in step (5) are as follows: Bacillus subtilis is cultured in LB medium at 37°C and 150 rpm for 24 h, Propionibacterium freudenreichii is cultured in RCM medium at 37°C and 150 rpm for 36 h, and the Bacillus subtilis solution and the Propionibacterium freudenreichii solution are mixed in a volume ratio of 5:1 to obtain a probiotic seed solution with a live bacteria concentration of 10 8 CFU / mL.

7. The method according to claim 1, characterized in that The spray drying in step (6) is carried out using a small spray drying apparatus, and the spray drying conditions are as follows: relative density of the concentrated liquid is 1.10 g / mL, air inlet temperature is 135° C., and pumping speed is 15%.

8. The method according to claim 1, characterized in that: The feathers include but are not limited to chicken feathers, duck feathers, and goose feathers.

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