阅读说明：本技术 一种含抗菌肽的无抗生素饲料添加剂及其制备方法 (Antibiotic-free feed additive containing antibacterial peptide and preparation method thereof ) 是由 张志东 刘培冉 赵学峰 冯艳忠 刘刚 于 2019-03-14 设计创作，主要内容包括：本发明涉及无抗饲料添加剂技术领域,特别涉及一种含抗菌肽的无抗生素饲料添加剂,包括麦芽、玉米秸秆、乳酸杆菌、枯草芽孢杆菌、硫色曲霉菌、吐温-80、梓叶、谷精草。本发明的有益效果：不含抗生素,在起到促进动物生长,提高动物抗病能力,提高日增重,降低料肉比,提高动物饲料利用效率等作用；采用3种复合菌种制备抗菌肽,克服了发酵过程PH改变对产肽的影响；秸秆壳与秸秆芯分离后使用,秸秆壳与秸秆芯分别发挥各自特点,物尽其用,提高了利用率。将梓叶与谷精草的有用物质提取出来,增强了使用效果与便利性。(The invention relates to the technical field of antibiotic-free feed additives, and particularly relates to an antibiotic-free feed additive containing antibacterial peptide, which comprises malt, corn straw, lactobacillus, bacillus subtilis, aspergillus thiochrus, tween-80, catalpa ovata and pipewort. The invention has the beneficial effects that: the feed additive does not contain antibiotics, and has the effects of promoting the growth of animals, improving the disease resistance of the animals, increasing the daily gain, reducing the feed conversion ratio, improving the utilization efficiency of the animal feed and the like; 3 kinds of composite strains are adopted to prepare the antibacterial peptide, so that the influence of pH change in the fermentation process on peptide production is overcome; the straw shells and the straw cores are used after being separated, and the straw shells and the straw cores respectively play their own characteristics, so that the straw shells and the straw cores are fully used, and the utilization rate is improved. The useful substances of catalpa leaves and pipewort are extracted, so that the use effect and convenience are enhanced.)

1. An antibiotic-free feed additive containing antibacterial peptide is characterized by comprising malt, corn straw, lactobacillus, bacillus subtilis, aspergillus thiochrans, tween-80, catalpa ovata and pipewort.

2. A method of preparing an antibiotic-free feed additive containing antimicrobial peptides according to claim 1, comprising the steps of:

(1) corn stalk shell and core separation

Separating and respectively crushing the corn straws by using a mechanical technology, crushing the straw shells into coarse powder, and crushing the straw cores into fine powder;

(2) wort saccharification

Malt was added with water at a ratio of 1: 4-1: 6, steaming the straw shell coarse powder in the step (1), cooling to 45-55 ℃, uniformly mixing with the malt homogenate, mixing in a saccharifying pot, keeping the temperature at 45-55 ℃ for 60-90min, decomposing protein, saccharifying for 2-4h at 55-60 ℃, heating to 75-78 ℃, saccharifying for 2-4h, wherein the mass ratio of the straw shell powder to the malt is 5: 1-15: 1;

(3) adding strain for fermentation

Adding more than two of lactobacillus, bacillus subtilis and aspergillus sulfureus into warm water containing 0.01% (m/m) glucose at 35-37 deg.C according to a certain proportion, activating for 1 hr, uniformly mixing the activated bacteria liquid with saccharified straw coarse powder, putting the mixture into a fermentation barrel, keeping the temperature at 35-37 ℃, continuously fermenting for 10 hours, adding 0.01% (m/m) of Tween-80 after fermentation is completed, uniformly stirring, carrying out filter pressing, centrifuging the filtrate at 10000-12000r/min for 8-10 minutes to obtain a supernatant of the fermentation liquid, carrying out ultrafiltration on the supernatant through an ultrafiltration membrane with the molecular weight cutoff of 20kDa, carrying out ultrafiltration on the collected filtrate through an ultrafiltration membrane with the molecular weight cutoff of 1000Da, discarding the filtrate, and obtaining the fermentation liquid containing the antibacterial peptide, wherein the activity of the antibacterial peptide reaches 6000U/ml, and the total weight of the bacteria powder is at least 1/10 of the weight of malt;

(4) mixing the fermentation liquid containing antibacterial peptide with corn cob powder

And (3) mixing the fermentation liquor containing the antibacterial peptide prepared in the step (3) with the straw core powder prepared in the step (1) to obtain the corn core powder carrying the antibacterial peptide.

(5) Preparation of plant extract

One or two of catalpa leaves and pipewort are put into 75 percent ethanol with the mass volume ratio of 20 times for reflux extraction, the extracting solution is decompressed and concentrated to be dry, crushed and sieved to obtain the plant extract;

(6) the corn cob powder carrying the antibacterial peptide and the plant extract are mixed according to the mass ratio of 1: 1, mixing uniformly to obtain the antibiotic-free feed additive.

3. The method for preparing antibiotic-free feed additive containing antibacterial peptide according to claim 2, wherein the malt in the step (2) is added with water in a ratio of 1: 4, preserving heat of the prepared malt homogenate by 50 ℃, steaming the straw shell coarse powder in the step (1), cooling by 50 ℃, uniformly mixing with the malt homogenate, mixing in a saccharifying pot, preserving heat for 60min at 50 ℃, performing protein decomposition, saccharifying for 3h at 58 ℃, heating to 78 ℃, and saccharifying for 3h, wherein the mass ratio of the straw shell powder to the malt is 9: 1.

4. the method for preparing the antibiotic-free feed additive containing the antibacterial peptide according to claim 2, wherein in the step (4), the mass ratio of the fermentation liquid to the straw core powder is 1: 3.

Technical Field

The invention relates to the technical field of antibiotic-free feed additives, in particular to an antibiotic-free feed additive containing antibacterial peptide and a preparation method thereof.

Background

The abuse of antibiotics or long-term use of antibiotics can cause great potential safety hazard to the whole ecological environment, so that the animal body generates dependence, the function exertion of immune cells in vivo is limited, and the immunity and disease resistance of the animal are reduced; the antibiotic inhibits or kills harmful bacteria in animal intestinal tract, and beneficial microorganism, so that normal microorganism system is unbalanced; can cause antibiotic residue in animal bodies, threatens food safety and is not good for human health.

Antimicrobial peptides (AMPs) are a bioactive small molecule induced in vivo

The sub-polypeptide has a molecular weight of about 2000-7000 and consists of 20-60 amino acid residues. Most of the active polypeptides have the characteristic of broad-spectrum antibiosis. Compared with antibiotics, the antibacterial peptide has high safety, is harmless to normal cells of human and animals, belongs to a biological preparation without toxic and side effects, residues and bacterial drug resistance, and has wide application prospect in non-antibiotic feed.

Disclosure of Invention

In order to solve the problems of the prior art, the invention provides an antibiotic-free feed additive containing antibacterial peptide and a preparation method thereof, wherein the antibiotic-free feed additive is prepared from 3 composite strains, can promote the growth of animals and improve the disease resistance of the animals.

The technical scheme of the invention is as follows:

an antibiotic-free feed additive containing antibacterial peptide comprises fructus Hordei Germinatus, corn stalk, lactobacillus, Bacillus subtilis, Aspergillus thiochromus, Tween-80, catalpa ovata leaf, and flos Eriocauli.

A preparation method of antibiotic-free feed additive containing antibacterial peptide comprises the following specific steps:

(1) corn stalk shell and core separation

Separating and respectively crushing the corn straws by using a mechanical technology, crushing the straw shells into coarse powder, and crushing the straw cores into fine powder;

(2) wort saccharification

Malt was added with water at a ratio of 1: 4-1: 6, steaming the straw shell coarse powder in the step (1), cooling to 45-55 ℃, uniformly mixing with the malt homogenate, mixing in a saccharifying pot, keeping the temperature at 45-55 ℃ for 60-90min, decomposing protein, saccharifying for 2-4h at 55-60 ℃, heating to 75-78 ℃, saccharifying for 2-4h, wherein the mass ratio of the straw shell powder to the malt is 5: 1-15: 1;

(3) adding strain for fermentation

Adding more than two of lactobacillus, bacillus subtilis and aspergillus sulfureus into warm water containing 0.01% (m/m) glucose at 35-37 deg.C according to a certain proportion, activating for 1 hr, uniformly mixing the activated bacteria liquid with saccharified straw coarse powder, putting the mixture into a fermentation barrel, keeping the temperature at 35-37 ℃, continuously fermenting for 10 hours, adding 0.01% (m/m) of Tween-80 after fermentation is completed, uniformly stirring, carrying out filter pressing, centrifuging the filtrate at 10000-12000r/min for 8-10 minutes to obtain a supernatant of the fermentation liquid, carrying out ultrafiltration on the supernatant through an ultrafiltration membrane with the molecular weight cutoff of 20kDa, carrying out ultrafiltration on the collected filtrate through an ultrafiltration membrane with the molecular weight cutoff of 1000Da, discarding the filtrate, and obtaining the fermentation liquid containing the antibacterial peptide, wherein the activity of the antibacterial peptide reaches 6000U/ml, and the total weight of the bacteria powder is at least 1/10 of the weight of malt;

(4) mixing the fermentation liquid containing antibacterial peptide with corn cob powder

And (3) mixing the fermentation liquor containing the antibacterial peptide prepared in the step (3) with the straw core powder prepared in the step (1) to obtain the corn core powder carrying the antibacterial peptide.

(5) Preparation of plant extract

One or two of catalpa leaves and pipewort are put into 75 percent ethanol with the mass volume ratio of 20 times for reflux extraction, the extracting solution is decompressed and concentrated to be dry, crushed and sieved to obtain the plant extract;

(6) the corn cob powder carrying the antibacterial peptide and the plant extract are mixed according to the mass ratio of 1: 1, mixing uniformly to obtain the antibiotic-free feed additive.

Preferably, in step (2), malt is added with water 1: 4, preserving heat of the prepared malt homogenate by 50 ℃, steaming the straw shell coarse powder in the step (1), cooling by 50 ℃, uniformly mixing with the malt homogenate, mixing in a saccharifying pot, preserving heat for 60min at 50 ℃, performing protein decomposition, saccharifying for 3h at 58 ℃, heating to 78 ℃, and saccharifying for 3h, wherein the mass ratio of the straw shell powder to the malt is 9: 1.

in the step (4), the mass ratio of the fermentation liquor to the straw core powder is 1: 3.

the invention has the beneficial effects that:

(1) the feed additive does not contain antibiotics, has the effects of promoting the growth of animals, improving the disease resistance of the animals, increasing the daily gain, reducing the feed conversion ratio, improving the utilization efficiency of the animal feed and the like, and does not have the harm of the antibiotics.

(2) 3 kinds of composite bacteria are adopted to prepare the antibacterial peptide, so that the influence of pH change in the fermentation process on peptide production is overcome.

(3) The straw shells and the straw cores are used after being separated, and the straw shells and the straw cores respectively play their own characteristics, so that the straw shells and the straw cores are fully used, and the utilization rate is improved.

(4) The useful substances of catalpa leaves and pipewort are extracted, so that the use effect and convenience are enhanced.

Detailed Description

In order to make the objects, technical solutions and advantages of the present invention more apparent, the following detailed description is further provided in conjunction with the embodiments.