Method for efficiently preparing VB12 from chicken manure and feather

Abstract

The invention discloses a method for efficiently preparing VB12 from chicken manure and feathers, the method comprises the following steps: propionibacterium freudenreichii is inoculated into a chicken manure-glucose culture medium for culture, and the chicken manure-glucose culture medium is prepared from 40-60 g / L of mixed powder of chicken manure and feathers, 5-10 g / L of glucose, 30-40 mg / L of 5, 6-dimethylbenzimidazole, 6-10 mg / L of CoCl2 and water as a solvent. According to the method disclosed by the invention, the yield of VB12 synthesized by propionibacterium freudenreichii can be remarkably improved, and the yield reaches 83.2 mu g / L on the fourth day, so that the quality of chicken manure and feather as animal feed additives is remarkably improved. According to the method disclosed by the invention, sustainable utilization of wastes such as chicken manure and feathers is realized, so that high-quality development of circular economy of poultry breeding industry is effectively promoted, and the method has certain environmental protection benefits and economic benefits.

Description

Technical Field

[0001] The invention belongs to the technical field of organic waste recycling, and specifically relates to a method for efficiently preparing VB using chicken manure and feathers. 12 method. Background Art

[0002] my country is a major poultry breeding and consumption country. Poultry breeding has gradually developed from individual households to large-scale and intensive breeding. However, if the feces and feathers produced by poultry are not treated and utilized, it will cause environmental pollution. Using chicken manure as feed is a sustainable and cost-effective solution. Chicken manure is a nutrient-rich organic material that contains various nutrients necessary for plants and animals. For example, chicken manure is rich in N elements and is an important source of N, mainly in the form of organic nitrogen and NH 3 NH 4 + , urea and other forms. Chicken manure also contains phosphorus (P) in the form of phosphates, potassium (K), calcium (Ca), magnesium (Mg), sulfur (S) and other elements. Not only that, chicken manure also contains a certain amount of essential trace nutrients, such as zinc (Zn), copper (Cu), manganese (Mn), iron (Fe), boron (B), molybdenum (Mo) and chlorine (Cl). In recent years, chicken manure has gradually become a raw material for animal feed and has been applied to animal feed formulas. Chicken manure can be processed and added to poultry feed (chickens, ducks, geese, etc.), ruminant feed (cattle, sheep, horses, etc.), pig feed and fish feed. How to improve the utilization quality of chicken manure in feed additives is a major bottleneck in the sustainable development of chicken manure and feathers.

[0003] Vitamin VB 12 Also known as cobalamin, it is an essential vitamin for animal growth and development. Moreover, higher animals and plants cannot produce vitamin VB. 12 ,VB in nature 12 All are synthesized by microorganisms. 12 It has many functions, such as promoting the development and maturation of red blood cells. It exists in the form of coenzymes, which can increase the utilization of folic acid and promote the metabolism of carbohydrates, fats and proteins. Moreover, it is an essential vitamin for the healthy function of the nervous system and participates in the formation of a lipoprotein in nerve tissue. 12 Can promote the growth and development of poultry, especially young poultry and livestock, with VB 12 Solution treatment of fish eggs and fry can also improve fish tolerance to toxic substances in water such as benzene and heavy metals. 12 It is an important feed additive.

[0004] Therefore, microbial inoculants were used to produce VB using a mixture of chicken manure and feathers as a nitrogen source. 12, which can not only improve the performance of chicken manure as a feed additive, but also reduce the pollution of chicken manure and feathers to the environment. Summary of the invention

[0005] The present invention aims at the sustainable utilization of poultry feces and feathers, and discloses a method for producing vitamin B12 by using a mixture of chicken feces and feathers as a nitrogen source. 12 (VB 12 ) is intended to improve the performance qualities of a mixture of chicken manure and feathers as an animal feed additive.

[0006] To achieve the above-mentioned purpose, the present invention adopts the following technical solutions:

[0007] A method for efficiently preparing VB using chicken manure and feathers 12 The method comprises the following steps: inoculating Propionibacterium freudenreichii into a chicken manure-glucose medium for cultivation, wherein the formula of the chicken manure-glucose medium is: 40-60 g / L of a mixed powder of chicken manure and feathers, 5-10 g / L of glucose, 30-40 mg / L of 5,6-dimethylbenzimidazole (DMB), and CoCl 2 6~10mg / L, the solvent is water.

[0008] Preferably, the formula of the chicken manure-glucose medium is: 50 g / L of mixed powder of chicken manure and feathers, 5 g / L of glucose, 35.56 mg / L of 5,6-dimethylbenzimidazole, and 1.5 g / L of CoCl. 2 8.01mg / L, the solvent is water.

[0009] Preferably, the Propionibacterium freudenreichii is Propionibacterium freudenreichii CICC 10019, which is deposited in the China Industrial Microbiological Culture Collection Center (CICC), with the strain number: CICC 10019, and the address: Building 6, No. 24, Jiuxianqiao Middle Road, Chaoyang District, Beijing (100015).

[0010] Preferably, the culture conditions are culturing at 30° C. and 200 rpm for 4 days.

[0011] On the other hand, the present invention also provides the above method for preparing VB 12 Purpose.

[0012] The present invention has the following beneficial effects: the present invention uses a mixture of chicken manure and chicken feathers as a nitrogen source for microbial culture, supplements a certain amount of glucose as a carbon source, and adds an appropriate amount of DMB and CoCl 2 , suitable for preparing VB by fermentation of Propionibacterium freudenreichii 12 Chicken manure feather-glucose medium can significantly increase the synthesis of VB by Propionibacterium freudenreichii12 The chicken manure feather-glucose medium was used to culture Propionibacterium freudenreichii to produce VB 12 , the yield reached 83.2 μg / L on the 4th day, significantly improving the quality of chicken manure and feathers as animal feed additives. The method of the present invention realizes the sustainable utilization of waste such as chicken manure and feathers, which will effectively promote the high-quality development of the circular economy of the poultry farming industry and has certain environmental and economic benefits. BRIEF DESCRIPTION OF THE DRAWINGS

[0013] Figure 1 Propionibacterium freudenreichii produces VB in chicken manure-glucose medium 12 Yield diagram of different treatments. Different capital letters indicate significant differences among treatments at the p≤0.05 level. DETAILED DESCRIPTION

[0014] The following examples are provided to further illustrate the present invention, rather than to limit the present invention.

[0015] Example 1

[0016] Detecting VB 12 The method was high performance liquid chromatography, and the detection conditions were: C18-250A chromatographic column (4.6mm×150mm, 5μm), the mobile phase was methanol:water=28:72 (volume ratio), isocratic elution, the elution flow rate was 0.8mL / min, the absorption wavelength was 361nm, the column temperature was 38℃, and the injection volume was 10μL.

[0017] The experimental control was Reinforced Clostridium Medium (RCM). The formula of RCM medium was as follows: 10.0 g beef extract powder, 5.0 g peptone, 3.0 g yeast extract powder, 5.0 g glucose, 1.0 g soluble starch, 5.0 g NaCl, 3.0 g NaAc, 0.5 g L-cysteine ​​hydrochloride, and 1000 mL distilled water.

[0018] The experiment was divided into five treatments: culturing Propionibacterium freudenreichii to produce VB 12 Treatment 1: 5 g chicken manure and feather powder was added to 100 mL distilled water (5% chicken manure); Treatment 2: 5% chicken manure + 1% glucose; Treatment 3: 5% chicken manure + 1% glucose + DMB; Treatment 4: 5% chicken manure + 0.5% glucose + DMB + CoCl 2 ; Treatment 5: Culture using RCM medium.

[0019] The preparation steps of the treatment medium are as follows: crush the mixture of chicken manure and chicken feathers into powder using a grinder, take 5g of the mixed powder of chicken manure and feathers and add it to 100mL of distilled water, stir and mix, and sterilize at high temperature and high pressure to obtain the obtained medium.

[0020] The preparation steps of the treatment 2 culture medium are as follows: crush the mixture of chicken manure and chicken feathers into powder using a grinder, take 5g of the mixed powder of chicken manure and feathers and 1g of glucose and add them to 100mL of distilled water, stir and mix, and sterilize at high temperature and high pressure to obtain the culture medium.

[0021] The preparation steps of treatment three culture medium are as follows: grind the mixture of chicken manure and chicken feathers into powder using a grinder, take 5g of the mixed powder of chicken manure and feathers, 1g of glucose, and 3.556mg of DMB and add them to 100mL of distilled water, stir and mix, and sterilize at high temperature and high pressure to obtain.

[0022] The preparation steps of treatment 4 culture medium are as follows: the mixture of chicken manure and chicken feathers is crushed into powder using a grinder, 5g of the mixed powder of chicken manure and feathers, 0.5g of glucose, 3.556mg of DMB, 0.801mg of CoCl 2 Add into 100 mL of distilled water, stir to mix, and sterilize under high temperature and high pressure.

[0023] Propionibacterium freudenreichii purchased from the China Industrial Microbiological Culture Collection (CICC) was streaked on RCM solid medium and cultured in an anaerobic bag at 30°C for 24 hours to activate the strain. A single colony was picked and transferred to RCM liquid medium and cultured at 30°C and 200rpm for 16 hours. Then it was inoculated into the above medium at a volume ratio of 5%, and cultured in the above five culture matrices for 4 days. The RCM medium was used as a control to measure the production of VB under the culture medium. 12 situation.

[0024] Figure 1 It was shown that VB could be produced by fermentation with Propionibacterium freudenreichii using a mixture of chicken manure and feathers as a nitrogen source and glucose as a carbon source. 12 The yield on the 4th day reached 6.2 μg / L. At the same time, appropriate amounts of DMB (35.56 mg / L) and CoCl 2 (8.01mg / L) can significantly increase VB 12 The yield reached 83.2 μg / L on the 4th day, which was significantly higher than that of VB produced by RCM medium. 12 These results indicate that chicken manure can be used as a nitrogen source for microorganisms to produce vitamin B 12 , which will further improve the quality of chicken manure as a feed additive.

[0025] The above are only preferred embodiments of the present invention. It should be noted that the above preferred embodiments should not be regarded as limiting the present invention, and the protection scope of the present invention should be based on the scope defined by the claims. For ordinary technicians in this technical field, several improvements and modifications can be made without departing from the spirit and scope of the present invention, and these improvements and modifications should also be regarded as the protection scope of the present invention.

Claims

1. A method for efficiently preparing VB using chicken manure and feathers 12 The method is characterized in that The following steps are involved: The method is prepared by inoculating Propionibacterium freudenreichii into a chicken manure-glucose culture medium for cultivation. The formula of the chicken manure-glucose culture medium is: 40-60 g / L of mixed powder of chicken manure and feathers, 5-10 g / L of glucose, 30-40 mg / L of 5,6-dimethylbenzimidazole, 6-10 mg / L of CoCl2, and the solvent is water.

2. The method for efficiently preparing VB using chicken manure and feathers according to claim 1 12 The method is characterized in that The formula of the chicken manure-glucose culture medium is: 50 g / L of mixed powder of chicken manure and feathers, 5 g / L of glucose, 35.56 mg / L of 5,6-dimethylbenzimidazole, 8.01 mg / L of CoCl2, and the solvent is water.

3. The method for efficiently preparing VB using chicken manure and feathers according to claim 1 12 The method is characterized in that The Propionibacterium freudenreichii is Propionibacterium freudenreichii CICC 10019.

4. The method for efficiently preparing VB using chicken manure and feathers according to claim 1 12 The method is characterized in that The culture conditions are culturing at 30° C. and 200 rpm for 4 days.

5. The method according to any one of claims 1 to 4 for preparing VB 12 Purpose.

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