阅读说明：本技术 一种功能性混合饲料添加剂及其制备的方法 (Functional mixed feed additive and preparation method thereof ) 是由 常艺海 于 2019-10-18 设计创作，主要内容包括：本发明提出了一种功能性混合饲料添加剂,添加剂为蛹虫草菌先对玉米芯进行第一阶段发酵后,再利用红法夫酵母菌对发酵后的玉米芯进行第二阶段发酵的产物；其中添加剂中蛹虫草素不少于100mg/kg,虾青素不少于300mg/kg。本发明充分结合了生物工程发酵技术中不同菌种的特性,根据蛹虫草菌和红法夫酵母菌的生理特性,结合固体发酵技术的应用,制备出含有虫草素和虾青素的新型发酵玉米芯功能性饲料添加剂。(The invention provides a functional mixed feed additive, which is a product obtained by first-stage fermentation of corncobs by cordyceps militaris and then second-stage fermentation of the fermented corncobs by using phaffia rhodozyma; wherein the additive contains Cordyceps militaris essence not less than 100mg/kg, and astaxanthin not less than 300 mg/kg. The invention fully combines the characteristics of different strains in the bioengineering fermentation technology, prepares the novel fermented corncob functional feed additive containing cordycepin and astaxanthin according to the physiological characteristics of cordyceps militaris and phaffia rhodozyma and by combining the application of the solid fermentation technology.)

1. A functional mixed feed additive is characterized in that the additive is a product obtained by first-stage fermentation of corncobs by cordyceps militaris and then second-stage fermentation of the fermented corncobs by using phaffia rhodozyma; wherein the cordycepin in the additive is not less than 100mg/kg, and the astaxanthin is not less than 300 mg/kg.

2. The use of the functional mixed feed additive according to claim 1 in livestock and poultry farming.

3. A process for the preparation of the additive according to claim 1 or 2, comprising the steps of:

(1) mixing cordyceps militaris bacterium liquid and corncobs according to the ratio of 1: 1.3-1.5, and performing solid fermentation at 25-28 ℃;

(2) ventilating every 2-4 h; stirring and turning materials every 1-2 h; completing the fermentation in the first stage after 120-144 h;

(3) mixing the phaffia rhodozyma bacterial liquid and the fermented corncobs in the step (2) according to the ratio of 1: mixing at a mass ratio of 0.4-0.6, and performing solid fermentation at 26-30 deg.C;

(4) ventilating every 4-6 h; stirring and turning materials every 30-40 min; completing the second stage fermentation after 48-60 h;

(5) and (4) sterilizing the fermentation product obtained in the step (4) to obtain an additive finished product.

4. The method for preparing the additive according to claim 3, wherein the cordyceps militaris bacterial liquid is prepared by inoculating cordyceps militaris in a seed culture medium, and performing shake culture at 25-27 ℃ and 150-180 rpm in a dark environment for 3-5 d; inoculating the seed liquid into a fermentation culture medium according to the volume percentage of 10-15%, wherein the seed liquid is inoculated for 96-120 h at the rotating speed of 160-220 rpm; when the pH value is 3.5, the fermentation is stopped.

5. The method for preparing the additive as claimed in claim 4, wherein the Cordyceps militaris seed medium comprises 130g of potato 110-.

6. The method for preparing the additive according to claim 4, wherein the fermentation medium of the cordyceps militaris is 5% -10% of soybean cake powder, 3% -6% of sucrose, 4% -8% of silkworm chrysalis defatted powder, 0.2% -0.5% of ammonium sulfate, 0.1% -0.3% of sodium carbonate and 0.005% of vitamin B10,003% -0.005%.

7. The method for preparing the additive according to claim 3, wherein the Phaffia rhodozyma strain solution is prepared by inoculating Phaffia rhodozyma in a seed culture medium, and shake-culturing at 26-28 ℃ and 130-160 rpm for 1d-2 d; inoculating the seed liquid into a fermentation culture medium according to the volume percentage of 8-18%, wherein the seed liquid is inoculated for 36-48 h, and the rotating speed is 130-180 rpm; when the pH value is 4.5, the fermentation is stopped.

8. The method for preparing the additive according to claim 7, wherein the Phaffia rhodozyma seed medium comprises malt extract powder 10-12g, glucose 10-15g, peptone 8-12g, sucrose 10-20g, magnesium sulfate 1.1-1.7g, sodium chloride 1.1-1.8g, and pH 7.0-7.5.

9. The method for preparing the additive as claimed in claim 7, wherein the fermentation medium of Phaffia rhodozyma is yeast extract 6-10%, fructose 4-7%, apple raw vinegar 4-8%, ammonium sulfate 0.1-0.3%, and potassium dihydrogen phosphate 0.2-0.4%.

Technical Field

The invention relates to the technical field of feed additives, in particular to a functional mixed feed additive and a preparation method thereof.

Background

China is a big agricultural country and the corn yield is extremely rich. The corn yield is 1.1-1.3 million tons every year, and 2000 million tons of corncobs can be produced as byproducts, however, the corncobs as agricultural wastes cannot be widely used for a long time, and most of the corncobs as farmyard fuel are burnt off in vain, so that great waste is caused. In recent years, with the continuous enhancement of scientific and technological strength of China, the field of industrial deep processing of the corncobs is continuously expanded, and a series of high-value-added products such as furfural, xylose, xylitol, xylo-oligosaccharide and the like are successively produced industrially, so that the corncob resources are fully utilized.

However, the main components of corncobs are cellulose, hemicellulose and lignin. Wherein the cellulose accounts for 32-36 percent, the pentosan accounts for 35-40 percent, the lignin accounts for 25 percent, and a small amount of ash and the like are also contained. Cellulose and hemicellulose are difficult to be absorbed and utilized by livestock and poultry, and can be only added in a small amount in feed, so that the feed utilization range of the corncobs is seriously influenced.

Provides a novel feed additive, i.e. the cordyceps militaris can utilize saprophytic organisms such as cellulose, lignin and the like, can metabolize to generate bioactive substances such as cordycepin, cordyceps polysaccharide, cordycepic acid and the like, and in addition, the substances such as sugar alcohol and the like are generated by decomposition, and the special fragrance of the cordyceps militaris can improve the palatability of livestock and poultry. The phaffia rhodozyma can perform secondary fermentation on corncobs fermented by cordyceps militaris, can produce a large amount of astaxanthin, and is used for the livestock feed additive. The problem of corncob comprehensive utilization is solved, the types of novel green and environment-friendly feed additives are increased, huge economic benefits can be generated, and the healthy development of animal husbandry is better served. The novel feed additive contains various beneficial bioactive substances such as cordycepin, cordyceps polysaccharide, cordycepic acid, astaxanthin and the like, and realizes the solution of the comprehensive utilization and the feed application of corncobs.

Disclosure of Invention

The invention provides a functional mixed feed additive and a preparation method thereof based on at least one of the technical problems, and the invention fully combines the characteristics of different strains in the bioengineering fermentation technology, and combines the physiological characteristics of cordyceps militaris and phaffia rhodozyma and the application of the solid fermentation technology to prepare the novel fermented corn cob functional feed additive containing cordycepin and astaxanthin. The content of bioactive substances can be remarkably improved by adopting a staged fermentation mode, wherein cordycepin is more than or equal to 100mg/kg, and astaxanthin is more than or equal to 300 mg/kg.

In view of the above, the invention provides a functional mixed feed additive, which is a product obtained by first-stage fermentation of corncobs by cordyceps militaris and then second-stage fermentation of the fermented corncobs by using phaffia rhodozyma; wherein the additive contains Cordyceps militaris essence not less than 100mg/kg, and astaxanthin not less than 300 mg/kg.

According to the second direction of the invention, the functional mixed feed additive is applied to livestock and poultry breeding

According to a third aspect of the invention, there is provided a process for preparing an additive comprising the steps of:

(1) mixing cordyceps militaris bacterium liquid and corncobs according to the ratio of 1: 1.3-1.5, and performing solid fermentation at 25-28 ℃;

(2) ventilating every 2-4 h; stirring and turning materials every 1-2 h; completing the fermentation in the first stage after 120-144 h;

(3) mixing the phaffia rhodozyma bacterial liquid and the fermented corncobs in the step (2) according to the ratio of 1: mixing at a mass ratio of 0.4-0.6, and performing solid fermentation at 26-30 deg.C;

(4) ventilating every 4-6 h; stirring and turning materials every 30-40 min; completing the second stage fermentation after 48-60 h;

(5) and (4) sterilizing the fermentation product obtained in the step (4) to obtain an additive finished product.

In the invention, the cordyceps militaris and the phaffia rhodozyma are adopted for step-by-step fermentation, and the corncobs are thoroughly fermented by utilizing the synergistic action of microorganisms. Wherein the corn cob contains sugar 54.5%, crude protein 2.2%, crude fat 0.4%, crude fiber 29.7%, and mineral 1.2%. Wherein, the sugar mainly exists in the form of sugar alcohol, the cordyceps sinensis can ferment the sugar alcohol into macromolecular polysaccharide, and then the phaffia rhodozyma is fermented to further ferment into micromolecular oligosaccharide, so that the synergistic effect of the strains is fully exerted.

Further, the preparation method of the cordyceps militaris bacterial liquid comprises the steps of inoculating cordyceps militaris to a seed culture medium, and carrying out shake culture at 25-27 ℃ and 150-180 rpm in a dark environment for 3-5 d; inoculating the seed liquid into a fermentation culture medium according to the volume percentage of 10-15%, wherein the seed liquid is inoculated for 96-120 h at the rotating speed of 160-220 rpm; when the pH value is 3.5, the fermentation is stopped.

Further, the cordyceps militaris seed culture medium comprises 130g of potato 110-.

Preferably, the cordyceps militaris seed culture medium is 120g of potato, 5g of silkworm chrysalis powder, 5g of malt, 10g of glucose, 7.5g of peptone, 6g of yeast extract, 1.2g of monopotassium phosphate, 1.2g of magnesium sulfate and 10.03g of vitamin B.

Boiling potato, silkworm chrysalis powder and malt for 30min, filtering residues to obtain supernatant, adding glucose, peptone, yeast extract, potassium dihydrogen phosphate, magnesium sulfate and vitamin B1, adding water to a constant volume of 1000ml, mixing, adjusting pH to 7.2, and sterilizing at 121 ℃ for 30min to obtain the cordyceps militaris strain seed culture medium.

Furthermore, the cordyceps militaris fermentation medium is 5-10% of soybean cake powder, 3-6% of cane sugar, 4-8% of silkworm chrysalis defatted powder, 0.2-0.5% of ammonium sulfate, 0.1-0.3% of sodium carbonate and 0.005% of vitamin B10,003.

Further, the preparation method of the phaffia rhodozyma bacterial liquid comprises the steps of inoculating the phaffia rhodozyma in a seed culture medium, and carrying out shake cultivation for 1d-2d at 26-28 ℃ and 130-160 rpm; inoculating the seed liquid into a fermentation culture medium according to the volume percentage of 8-18%, wherein the seed liquid is inoculated for 36-48 h, and the rotating speed is 130-180 rpm; when the pH value is 4.5, the fermentation is stopped.

Furthermore, the Phaffia rhodozyma seed culture medium is 10-12g of malt extract powder, 10-15g of glucose, 8-12g of peptone, 10-20g of sucrose, 1.1-1.7g of magnesium sulfate, 1.1-1.8g of sodium chloride and 7.0-7.5 of pH value.

Preferably, the Phaffia rhodozyma seed culture medium is 10g of malt extract powder, 10g of glucose, 8.5g of peptone, 15g of sucrose, 1.3g of magnesium sulfate and 1.5g of sodium chloride, and water is added to the culture medium until the volume is 1000ml, and the mixture is uniformly mixed, and the pH value is 7.2.

Malt extract powder, glucose, peptone, sucrose, magnesium sulfate and sodium chloride, adding water to a constant volume of 1000ml, mixing uniformly, sterilizing at the temperature of 121 ℃ for 30min and at the pH of 6.5 and 7.2 to obtain the Phaffia rhodozyma seed culture medium.

Furthermore, the fermentation medium of the phaffia rhodozyma comprises 6 to 10 percent of yeast extract, 4 to 7 percent of fructose, 4 to 8 percent of apple raw vinegar, 0.1 to 0.3 percent of ammonium sulfate and 0.2 to 0.4 percent of monopotassium phosphate.

Through the technical scheme, the invention provides a functional mixed feed additive and a preparation method thereof, and the functional mixed feed additive has the following advantages:

(1) the method firstly combines the fermentation technology of bioengineering with the physiological metabolism characteristics of different strains, adopts a staged fermentation mode, furthest exerts the characteristics of each strain, produces the novel feed additive taking corncobs as the main raw material, contains rich cordycepin, cordyceps polysaccharide, astaxanthin and other active substances, can be well used in the livestock breeding industry, has good experimental effect after multiple experiments, has simple process flow, is safe, environment-friendly, has no residue, and has low cost and very high market prospect and economic benefit.

(2) The invention optimizes the solid fermentation of two stages of corncobs and the like, can exert the metabolic characteristics of strains to the maximum extent, greatly improves the synergistic effect of cordyceps militaris and phaffia rhodozyma in the corncob fermentation process, produces various substances such as cordycepin, cordyceps polysaccharide, cordycepic acid, astaxanthin and the like, wherein the cordycepin is more than or equal to 100mg/kg, and the astaxanthin is more than or equal to 300mg/kg, and the obtained product is environment-friendly and accords with the national industrial policy.

Drawings

FIG. 1 shows a micrograph of the present invention of Cordyceps militaris.

FIG. 2 shows a micrograph of Phaffia rhodozyma of the present invention.

Fig. 3 shows a cordycepin standard curve HPLC picture.

FIG. 4 shows HPLC pictures of cordycepin produced by fermentation of Cordyceps in example 7 of the present invention.

Detailed Description

In order that the above objects, features and advantages of the present invention can be more clearly understood, the present invention will be described in further detail with reference to specific embodiments. It should be noted that the embodiments of the present invention and features of the embodiments may be combined with each other without conflict.

In the following description, numerous specific details are set forth in order to provide a thorough understanding of the present invention, however, the present invention may be practiced in other ways than those specifically described herein, and therefore the scope of the present invention is not limited by the specific embodiments disclosed below.

Cordyceps militaris and Phaffia rhodozyma are commercially available strains, the microstructure of the Cordyceps militaris and Phaffia rhodozyma is shown in figure 1-2, the Cordyceps militaris is purchased from Guangdong province microorganism strain preservation center, the purchase number of the Cordyceps militaris is GIM5.269, and the purchase number of the Phaffia rhodozyma is 2.177. The YEAST EXTRACT is YEAST powder YEAST EXTRACT purchased from the market, and the production area: OXOID.