CN102823726B - It is a kind of improve cottonseed meal content and can detoxification biofermentation method - Google Patents

Abstract

The invention discloses a bio-fermentation method capable of increasing the protein content of cottonseed meal and capable of detoxification, comprising: 1) respectively preparing fermentation liquids of Candida utilis, Aspergillus niger, Bacillus subtilis and Lactobacillus acidophilus as strains; 2) Prepare raw cotton meal; 3) Add water to the raw material, the weight ratio of raw material to water is 1:0.5-1.2; 4) Inoculate the strains prepared in step 1), respectively Candida utilis 0.5%-10 %, Aspergillus niger 0.5%-10%, Bacillus subtilis 0.5%-10%, Lactobacillus acidophilus 0.5%-10%; The product directly obtained by the method of the invention can effectively degrade free gossypol in the cottonseed meal, improve the protein quality of the cottonseed meal, improve the taste of the cottonseed meal, thereby effectively improving the utilization rate of the cottonseed meal in various livestock and poultry feeds.

Description

A bio-fermentation method that can increase the protein content of cottonseed meal and can detoxify

technical field

The invention belongs to the technical field of biological fermentation and relates to a biological fermentation method capable of increasing the protein content of cottonseed meal and capable of detoxification.

Background technique

Feed protein can be divided into animal protein and plant protein according to the source of raw materials. Among them, animal protein such as fish meal, meat and bone meal and blood meal has similar amino acid composition to the amino acid required for the growth of livestock and poultry, so this protein is not only The absorption rate is high, and it also has the advantages of fast absorption and less pollution. However, my country is a country that is extremely short of animal protein resources such as fishmeal. In particular, high-quality fishmeal mainly relies on imports, and its price is much higher than that of vegetable protein; More and more attention is being paid to the application of plant-based protein raw materials. Plant-based protein sources include soybean meal, rapeseed meal, cottonseed meal, peanut meal, etc., which are cheap and widely available. However, various anti-nutritional factors commonly found in these plant-based protein materials not only affect the palatability of animals, but more importantly The digestive enzyme activity of animals is greatly affected, thereby reducing the digestion and absorption rate of nutrients such as protein and amino acids. The level of digestibility will directly affect the utilization of plant protein sources by animals. In addition, some anti-nutritional factors can directly produce toxic effects on animals and disturb their normal physiological functions. At the same time, the imbalance of amino acid content is also an important factor restricting the proportion of vegetable protein used.

Cottonseed meal (CSM) is the by-product remaining after cottonseeds are husked, heated, flattened into thin flakes and extracted with solvent hexane. Cotton meal contains more than 40% protein, 1.59% lysine and 0.52% methionine. It is a very valuable raw material for livestock and poultry feed. Simultaneously my country is a big cotton-producing country, and my country produces more than 8,000,000 tons of cottonseeds annually, and after oil extraction, it is estimated that about 4,000,000 tons of cottonseed cakes (containing about 15% of the husk) can be produced. However, there are also many anti-nutritional factors in cotton meal, that is, a class of substances contained in cotton meal that have adverse effects on the digestion, absorption, and metabolism of nutrients and the health and production performance of animals. Due to the existence of these anti-nutritional factors, the application of cotton meal in livestock and poultry feed and aquatic feed is greatly restricted. The anti-nutritional factors in cottonseed meal mainly include free gossypol, cyclopropene fatty acid (CPFA), phytic acid and tannin.

Contents of the invention

In order to solve the above problems, the present invention provides a biological fermentation method that can increase the protein content of cottonseed meal and can detoxify.

The biological fermentation method provided by the invention comprises the steps of:

1) Prepare the fermentation broths of Candida utilis, Aspergillus niger, Bacillus subtilis and Lactobacillus acidophilus as strains respectively;

2) Prepare raw cotton meal;

3) Add water to the raw material, the weight ratio of raw material to water is 1:0.5～1.2;

4) The strains prepared in the inoculation step 1), the strains inoculated are based on the percentage of the volume of the culture medium containing the strains and the weight of the raw materials, respectively 0.5% to 10% of Candida utilis and 0.5% to 10% of Aspergillus niger , Bacillus subtilis 0.5% to 10%, Lactobacillus acidophilus 0.5% to 10%;

5) Temperature-controlled fermentation, the fermentation temperature is 25°C to 40°C.

In one embodiment of the present invention, step 1) the process of preparing the fermentation broth of Candida utilis, Aspergillus niger, Bacillus subtilis and Lactobacillus acidophilus is: the aerobic bacteria Candida utilis, Aspergillus niger, subtilis Bacillus was cultured by aerobic fermentation, and Lactobacillus acidophilus was cultured by anaerobic fermentation.

In another embodiment of the present invention, the weight ratio of raw materials to water in step 3) is preferably 1:0.8-1.0.

In another embodiment of the present invention, the strains inoculated in step 4) are respectively 2% to 8% of Candida utilis and 2% to 8%, Bacillus subtilis 2% to 8%, Lactobacillus acidophilus 2% to 8%.

In a preferred embodiment of the present invention, the bacterial species inoculated in step 4) are respectively Candida utilis 5%, Aspergillus niger 5%, Bacillus subtilis 5%, Lactobacillus acidophilus 5%.

In the present invention, the specific strains of Candida utilis, Aspergillus niger, Bacillus subtilis and Lactobacillus acidophilus are not particularly limited, and conventional strains of these bacteria can be used in the present invention.

The beneficial effects of the present invention are as follows: 1. Enzymolysis treatment is carried out on the raw material cotton meal during microbial fermentation, the nutritional value of cotton meal is significantly improved, and the crude protein content is increased by more than 15% compared with that before fermentation; 2. The content of free gossypol Significant reduction: after the cotton meal is fermented and produced by the method of the present invention, the free gossypol in the product can be reduced by more than 80% relative to the content in the raw cotton meal; 3. After the lactic acid bacteria utilize the nutrients in the cotton meal matrix, , along with the growth, reproduction and metabolism of the bacteria, certain organic acids such as lactic acid, acetic acid, propionic acid, butyric acid, etc. are produced, which reduce the pH value of the substrate and improve the taste of cottonseed meal.

detailed description

The following examples are used to illustrate the present invention, but are not intended to limit the scope of the present invention.

Example 1 Improve the crude protein content of cottonseed meal and the screening of strains that degrade gossypol

Raw cottonseed meal is derived from the by-products of cottonseed after oil extraction; the weight ratio of raw material to water is 1:0.8, and the prepared Aspergillus niger CICC41057 (purchased from China Institute of Food and Fermentation Industry), Candida utilis CICC 31188 (purchased from China Food Fermentation Industry Research Institute), Saccharomyces cerevisiae (CGMCC No.2388), Bacillus licheniformis (CGMCC No.5094), Lactobacillus salivarius CICC 21807 (purchased from China Food Fermentation Industry Research Institute) hospital); the fermentation temperature is 30°C, and the fermentation time is 48 hours. The test results are shown in Table 1.

Table 1 Effects of different bacterial strains on the crude protein content and gossypol of cottonseed meal

According to the results, Aspergillus niger and Candida utilis were selected as the strains to increase the crude protein content of cotton meal and degrade gossypol.

Example 2 Screening of bacterial strains that improve the mouthfeel of cotton meal

Raw cotton meal is derived from the by-products of cottonseed after oil extraction; the weight ratio of raw material to feed water is 1:0.8, and the prepared Bacillus subtilis (CGMCC No.4628), acidophilus milk bacillus (CGMCC No.5093), Enterococcus faecalis (CGMCC No.5092), Bacillus pumilus (CGMCC No.4756), Bacillus licheniformis (CGMCC No.5094), Lactobacillus salivarius CICC 21807; fermentation temperature is 30 ℃, fermentation The time is 48 hours. The test results are shown in Table 2.

Table 2 Effects of different strains on the pH of cottonseed meal

It can be seen from Table 2 that the pH of cotton meal fermented by Bacillus subtilis and Lactobacillus acidophilus is low, that is, more organic acids and other flavor substances are produced, so it is selected as the strain to improve the taste of cotton meal. Embodiment 3 Cotton Meal Biological Fermentation

Preparation of Aspergillus niger fermentation liquid: Inoculate Aspergillus niger into PDA slant medium, culture at 30°C for 72h, pick the mycelium on the slope, insert it into PDA liquid medium, and culture at 30°C for 16h with shaking at 150r/min.

Preparation of Candida utilis fermentation broth: insert Candida utilis into YPD slant medium, culture at 30°C for 48 hours, pick the lawn on the slant, insert it into YPD liquid medium, shake at 30°C, 150r/min Cultivate for 48h.

Preparation of Bacillus subtilis fermentation broth: Introduce Bacillus subtilis into LB slant medium, culture at 37°C for 24 hours, pick the bacterial lawn on the slope, insert it into LB liquid medium, and culture at 37°C, shaking at 170r/min for 24h.

Preparation of Lactobacillus acidophilus fermentation broth: Insert Lactobacillus acidophilus into MRS slant medium, culture at 37°C for 24 hours, pick the lawn on the slope, insert it into MRS liquid medium, and place Lactobacillus acidophilus at 37°C Incubate for 24 hours.

Raw cotton meal comes from the by-products of cottonseed after oil extraction; the weight ratio of raw material to feed water is 1:0.8; 8% silk yeast, 8% Aspergillus niger, 8% Bacillus subtilis, 8% Lactobacillus acidophilus; the fermentation temperature is 25°C, and the fermentation time is 60 hours.

Embodiment 4 biological fermentation of cottonseed meal

Raw cotton meal comes from the by-products of cottonseed after oil extraction; the weight ratio of raw material to feed water is 1:1; Trichotrichum 3%, Aspergillus niger 3%, Bacillus subtilis 3%, Lactobacillus acidophilus 3%; fermentation temperature is 28°C, fermentation time is 72 hours.

Embodiment 5 biological fermentation of cottonseed meal

Raw cotton meal comes from the by-products of cottonseed after oil extraction; the weight ratio of raw material to feed water is 1:0.8; 6% Trichotrichum, 6% Aspergillus niger, 6% Bacillus subtilis, 6% Lactobacillus acidophilus; the fermentation temperature is 30°C, and the fermentation time is 48 hours.

Embodiment 6 Cotton Meal Biological Fermentation

Raw cotton meal comes from the by-products of cottonseed after oil extraction; the weight ratio of raw material to feed water is 1:0.5; Trichotrichum 5%, Aspergillus niger 5%, Bacillus subtilis 5%, Lactobacillus acidophilus 5%; the fermentation temperature is 37°C, and the fermentation time is 48 hours.

Example 7 Cotton Meal Biological Fermentation

Raw cotton meal comes from the by-products of cottonseed after oil extraction; the weight ratio of raw material to feed water is 1:1.2; Trichotrichum 2%, Aspergillus niger 2%, Bacillus subtilis 2%, Lactobacillus acidophilus 2%; fermentation temperature is 40°C, fermentation time is 48 hours.

Test example 1

Taking cotton meal without fermentation treatment as a control, the crude protein content and free gossypol content in the cotton meal treated in Examples 3-7 and the control were measured, and the results are shown in Table 3.

Table 3 Effects of Different Tests on Crude Protein Content and Free Gossypol

It can be seen that the crude protein content of cottonseed meal after microbial fermentation of the present invention has increased by more than 15% compared with that before fermentation, and the content of free gossypol can be reduced by more than 80% relative to the content of raw cottonseed meal, while fermentation also produces some organic acids Flavor substances reduce the pH of cottonseed meal and improve its palatability.

The above is only a preferred embodiment of the present invention, it should be pointed out that for those of ordinary skill in the art, without departing from the technical principle of the present invention, some improvements and modifications can also be made. These improvements and modifications It should also be regarded as the protection scope of the present invention.

Claims (3)

1. A biofermentation method that improves cottonseed meal protein content and can be detoxified, is characterized in that, comprises the steps: 1) Prepare the fermentation broths of Candida utilis, Aspergillus niger, Bacillus subtilis and Lactobacillus acidophilus as strains respectively; 2) Prepare raw cotton meal; 3) Add water to the raw material, the weight ratio of raw material to water is 1:0.8～1.0; 4) The strains prepared in the inoculation step 1), the strains inoculated are based on the percentage of the volume of the culture medium containing the strains and the weight of the raw materials, respectively 2% to 8% of Candida utilis and 2% to 8% of Aspergillus niger , Bacillus subtilis 2% to 8%, Lactobacillus acidophilus 2% to 8%; 5) Temperature-controlled fermentation, the fermentation temperature is 25°C to 40°C. 2. The biological fermentation method as claimed in claim 1, characterized in that, step 1) the process of preparing Candida utilis, Aspergillus niger, Bacillus subtilis and Lactobacillus acidophilus fermentation liquid is: making the aerobic bacteria Candida, Aspergillus niger, and Bacillus subtilis were cultured by aerobic fermentation, and Lactobacillus acidophilus was cultured by anaerobic fermentation. 3. The biological fermentation method as claimed in claim 1, characterized in that, in step 4), the strains inoculated are respectively 5% of Candida utilis, black Aspergillus 5%, Bacillus subtilis 5%, Lactobacillus acidophilus 5%.