阅读说明：本技术 可得然胶在制备饲料添加剂中的应用 (Application of curdlan in preparation of feed additive ) 是由 何珣 周勇 万欣 卢宗梅 邴狄祥 郭世堂 陈可泉 叶震 佟毅 于 2021-09-10 设计创作，主要内容包括：本发明公开了可得然胶在制备饲料添加剂中的应用。通过可得然胶改性处理的织物乳杆菌,能够提高植物乳杆菌细胞的制备和保存过程的活菌率。换句话说就是：利用本发明的菌剂,能够在环境条件下维持较高活菌率,使得植物乳杆菌在常温条件下也可以延长保存期,在农业生产上有着广泛的用途,对保护环境,减少抗生素的使用量,促进农业生产可持续发展有着重要的技术价值和社会价值,本发明改性工艺简单,原料广泛,降低了生产成本,所得添加剂的稳定性大大提高,性能优异；也便于运输和储存,更适宜产业化推广。(The invention discloses an application of curdlan in preparing a feed additive. The fabric lactobacillus modified by curdlan can improve the viable bacteria rate of lactobacillus plantarum cells in the preparation and preservation processes. In other words, it is: the microbial inoculum can maintain higher viable bacteria rate under environmental conditions, so that the storage life of the lactobacillus plantarum can be prolonged under normal temperature conditions, the lactobacillus plantarum has wide application in agricultural production, has important technical and social values in protecting environment, reducing the use amount of antibiotics and promoting sustainable development of agricultural production, and has the advantages of simple modification process, wide raw materials, reduced production cost, greatly improved stability of the obtained additive and excellent performance; is also convenient for transportation and storage, and is more suitable for industrialized popularization.)

1. Application of curdlan in preparing feed additive is provided.

2. Use of curdlan according to claim 1 for the preparation of a feed additive, characterized in that it comprises the following steps:

step 1, preparing an MRS agar plate culture medium and an MRS liquid culture medium, sterilizing at 121 ℃ for 20 minutes, and cooling to room temperature for later use;

step 2, selecting lactobacillus plantarum to grow on an MRS agar culture medium plate for 24 hours at 37 ℃;

step 3, picking the cultured strains from the flat plate to an MRS liquid culture medium, and carrying out shake culture in a nitrogen shaker at 37 ℃ to the middle stage of logarithmic phase of growth;

step 4, inoculating the strain into a 50L fermentation tank from a shake flask according to the inoculation amount of 10%, fermenting and culturing for 72 hours at 37 ℃, and centrifugally collecting thalli after fermentation;

and 4, after counting the number of the viable bacteria, uniformly mixing the curdlan and the lactobacillus plantarum, and freeze-drying to obtain the lactobacillus plantarum packaged by the curdlan, namely the feed additive.

3. The use according to claim 2, wherein the Lactobacillus plantarum CGMCC 15013, China general microbiological culture Collection center, is used in step 2.

4. The use as claimed in claim 2, wherein the mass ratio of curdlan to lactobacillus plantarum in step 4 is 0.2-1.5: 100.

5. The use of claim 2, wherein step 4 further comprises mixing one or more of skimmed milk powder, sucrose, molasses, starch, whey protein isolate, or alginate.

6. Use of a feed additive according to any one of claims 1 to 4 for the preparation of an animal feed.

7. An animal feed comprising the feed additive of any one of claims 1 to 5.

Technical Field

The invention relates to the technical field of preparation of animal feed additives, in particular to application of curdlan in preparation of feed additives.

Background

The microbial exopolysaccharide has the advantage of relatively low toxicity, which makes it widely used in the health care, food and cosmetic industries. Meanwhile, the food and drug administration (U.S.) has legally approved curdlan for use as a food additive.

Curdlan (Curdlan) is a high molecular polymer produced by fermentation of a microorganism such as Agrobacterium sp using sugars such as glucose and sucrose as raw materials. It is a straight-chain polysaccharide formed by connecting glucose structural units by beta-1, 3-glycosidic bonds. It is also known as thermal gel, known as curdlan gum in taiwan china due to its unique property of forming a gel under heating. It is safe and non-toxic to human body, and is insoluble in water but soluble in alkaline solution to form viscous solution. Curdlan as food additive is widely applied to food industry, and can improve the water holding capacity, viscoelasticity and stability of the product and improve the taste of the product. In addition, the compound has good application prospect in the fields of feed, medicine, building materials, petroleum development and the like.

The lactobacillus plantarum is a gram-positive lactobacillus which widely exists in nature, is anaerobic or facultative anaerobic, has a straight or bent rod-shaped strain, is single or sometimes paired or chained, has the optimal pH value of about 6.5, and belongs to homofermentation lactobacillus. However, the survival rate of the existing lactobacillus plantarum at normal temperature is low, and the application of the existing lactobacillus plantarum is limited. Further modification is needed to improve survival rate and facilitate application.

Disclosure of Invention

Aiming at the defects of the prior art, the invention aims to provide the application of curdlan in preparing the feed additive, the feed additive utilizes the curdlan to modify lactobacillus plantarum, improves the survival capability of the lactobacillus plantarum at normal temperature, can prolong the shelf life when used in feed, has low cost and safe components, and is suitable for industrial application.

In order to solve the problems of the prior art, the invention adopts the technical scheme that:

application of curdlan in preparing feed additive is provided.

The application comprises the following steps:

step 1, preparing an MRS agar plate culture medium and an MRS liquid culture medium, sterilizing at 121 ℃ for 20 minutes, and cooling to room temperature for later use;

step 2, selecting lactobacillus plantarum to grow on an MRS agar culture medium plate for 24 hours at 37 ℃;

step 3, picking the cultured strains from the flat plate to an MRS liquid culture medium, and carrying out shake culture in a nitrogen shaker at 37 ℃ to the middle stage of logarithmic phase of growth;

step 4, inoculating the strain into a 50L fermentation tank from a shake flask according to the inoculation amount of 10%, fermenting and culturing for 72 hours at 37 ℃, and centrifugally collecting thalli after fermentation;

and 4, after counting the number of the viable bacteria, uniformly mixing the curdlan and the lactobacillus plantarum, and freeze-drying to obtain the lactobacillus plantarum packaged by the curdlan, namely the feed additive.

The improvement is that the Lactobacillus plantarum CGMCC 15013, China general microbiological culture collection center, is adopted in the step 2.

The improvement is that the mass ratio of curdlan to lactobacillus plantarum in the step 4 is 0.2-1.5: 100.

The improvement is that the step 4 also comprises one or more of skimmed milk powder, sucrose, molasses, starch, isolated whey protein or alginate.

The feed additive is applied to preparing animal feed.

An animal feed contains the feed additive.

Has the advantages that:

compared with the prior art, the application of curdlan in preparing the feed additive has the following advantages:

1. the fabric lactobacillus modified by curdlan can improve the viable bacteria rate of lactobacillus plantarum cells in the preparation and preservation processes. In other words, it is: the microbial inoculum of the invention can maintain higher viable bacteria rate under environmental conditions, so that the lactobacillus plantarum can prolong the storage life under normal temperature conditions, has wide application in agricultural production, and has important technical and social values for protecting environment, reducing the use amount of antibiotics and promoting sustainable development of agricultural production.

2. The modification process is simple, the raw materials are wide, the production cost is reduced, the stability of the obtained additive is greatly improved, and the performance is excellent; is also convenient for transportation and storage, and is more suitable for industrialized popularization.

Drawings

FIG. 1 is a picture of freeze-dried Lactobacillus plantarum used in the present invention.

FIG. 2 is a picture of the Lactobacillus plantarum encapsulated with curdlan according to the invention, stored at 4 ℃.

FIG. 3 is a picture of the Lactobacillus plantarum encapsulated with curdlan according to the invention, stored at 25 ℃.

Detailed Description

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although only preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention. All documents mentioned in this specification are incorporated by reference herein for the purpose of disclosing and describing the methods and/or materials associated with the documents. In case of conflict with any incorporated document, the present specification will control.

The Lactobacillus plantarum used in the invention is Lactobacillus plantarum CGMCC 15013, China general microbiological culture Collection center.

Isolated whey protein was purchased from Beijing Congbit sports science and technology, Inc.;

the starch is soluble starch, and the common corn starch is used in the embodiment of the invention;

MRS liquid medium formula: 10.0 g/L of peptone, 8.0 g/L of beef extract powder, 4.0 g/L of yeast extract powder, 20.0 g/L of glucose, 2.0 g/L of dipotassium phosphate, 2.0 g/L of diammonium hydrogen citrate, 5.0 g/L of sodium acetate, 0.2 g/L of magnesium sulfate, 0.04 g/L of manganese sulfate and 801.0 g/L of tween;

MRS solid medium formula: 10.0 g/L of peptone, 5.0 g/L of beef extract powder, 4.0 g/L of yeast extract powder, 20.0 g/L of glucose, 2.0 g/L of dipotassium phosphate, 2.0 g/L of triammonium citrate, 5.0 g/L of sodium acetate, 0.2 g/L of magnesium sulfate, 0.05 g/L of manganese sulfate, 801 mL of tween and 10 g/L of agar powder.

Example 1

Preparing MRS agar plate culture medium and MRS liquid culture medium, sterilizing at 121 deg.C for 20 min, and cooling to room temperature. Wherein the content of the first and second substances,

selecting lactobacillus plantarum to grow on an MRS agar culture medium plate for 24 hours at 37 ℃;

selecting the cultured strain to a shake flask filled with MRS liquid culture medium, and shake-culturing in a nitrogen shaker at 37 ℃ to the middle stage of logarithmic phase;

inoculating the strain into a 50L seeding tank according to the inoculation amount of 10%, fermenting and culturing for 72 hours at 37 ℃, and after the fermentation is finished, transferring the fermentation liquid from the fermentation tank to a rotary drum type centrifuge under pressure to collect the thallus of the lactobacillus plantarum.

Mixing curdlan and lactobacillus plantarum fully according to the mass ratio of 1:100, and remembering the lactobacillus plantarum packaged by the curdlan.

Quick freezing at-40 deg.C, and drying.

Plates were coated by serial dilution and viable count was calculated.

Under the condition of 4 ℃, the viable count is reduced by 5 percent after the mushroom is stored for six months, and the distribution uniformity of thalli is not changed.

Plates were coated by serial dilution and viable count was calculated. Reference may be made specifically to the literature: yuanlihong. microbiological experiment [ M ]. Beijing: chemical industry publishers, 2010: 66-67.

Under the condition of 4 ℃, the viable count is reduced by 5 percent after the mushroom is stored for six months, and the distribution uniformity of thalli is not changed.

Comparative example 1

The same procedure as in example 1 was repeated, except that curdlan was replaced with whey protein isolate.

Under the condition of 4 ℃, the viable count is reduced by 23 percent after the culture medium is stored for six months, and the distribution uniformity of thalli is not changed.

Example 2

The same as example 1 was repeated, except that curdlan and lactobacillus plantarum were replaced by mixing curdlan, starch, whey protein isolate and lactobacillus plantarum at a mass ratio of 1:20:15: 100.

Under the condition of 25 ℃, the viable count is reduced by 45 percent after the mushroom is stored for six months, and the distribution uniformity of thalli is not changed.

Comparative example 2

The same procedure as in example 1 was repeated, except that curdlan and lactobacillus plantarum were mixed and replaced with starch, whey protein isolate and lactobacillus plantarum at a mass ratio of 20:15: 100.

Quick freezing at-40 deg.C, and drying.

Under the condition of 25 ℃, the viable count is reduced by 68 percent after the strain is stored for six months, and the distribution uniformity of the strain is unchanged.

Example 3

The same as example 1 was repeated, except that curdlan and lactobacillus plantarum were mixed and replaced with curdlan, starch, whey protein isolate, and lactobacillus plantarum at a mass ratio of 1:10:15: 100.

Under the condition of 25 ℃, the viable count is reduced by 54 percent after the mushroom is stored for six months, and the distribution uniformity of thalli is not changed.

In the above examples, it should be noted that the viable count mentioned herein refers to the number of viable Lactobacillus plantarum.

The living microbial cells are stored at low temperature, so that the activity is easy to ensure, and the living microbial cells are easy to die at high temperature for a long time. The invention improves the normal temperature survival ability after the modification treatment of the lactobacillus plantarum agent. As can be seen from fig. 1, 2 and 3, curdlan can encapsulate microorganisms, while the embedded structure can be maintained for a longer time at room temperature, and thus, the cell death rate is maintained to be low, and the viable count during storage is increased. In particular, fig. 3, it is very intuitive to reflect that the encapsulated (embedded) state remains after storage at room temperature, and the embedded structure is not destroyed.

The above description is only a preferred embodiment of the present invention, and the scope of the present invention is not limited thereto, and any simple modifications or equivalent substitutions of the technical solutions that can be obviously obtained by those skilled in the art within the technical scope of the present invention are within the scope of the present invention.