CN104762234A - Complex bacterium for bean pulp fermentation and application of complex bacterium - Google Patents

Abstract

The invention provides a complex bacterium for bean pulp fermentation. The complex bacterium is composed of bacillus subtilis, bacillus coagulans, aspergillus niger, aspergillus oryzae, lactobacillus plantarum and saccharomyces cerevisiae. The complex bacterium provided by the invention is obtained through repeated scientific experiment and reasonable combination based on physiological metabolism feature of the microorganism, the complex bacterium is vigorous in growth metabolism, is capable of secreting a plurality of enzymes, acids and other metabolites, and is capable of efficiently decomposing macromolecule protein, anti-nutritional factors and sensitization factors in the bean pulp, obviously increasing content of soluble protein, small peptide, organic acid, protease and vitamin and the number of probiotics and antibacterial peptide, improving the digestion utilization rate, inhibiting the growth of pathogenic bacteria and reducing the use of antibiotic; and meanwhile, the complex bacterium has acid fragrance of fermentation and is good in food attractant effect. The bacteria strains forming the complex bacterium are included in a feed additive variety catalogue (2013) issued by the department of agriculture, the complex bacterium is safe and reliable, and can be directly used for the industrial production for the bean pulp fermentation.

Description

A kind of composite flora for soybean meal fermentation and its application

technical field

The invention belongs to the technical field of microorganisms, and in particular relates to a soybean meal fermentation technology, in particular to a complex flora used for soybean meal fermentation.

Background technique

With high crude protein content and balanced amino acid composition, soybean meal has always been one of the most important plant-source proteins for livestock and poultry. However, there are also anti-nutritional factors such as trypsin inhibitors, urease, soybean hemagglutinin and allergenic factors such as soybean antigen protein in soybean meal, which reduces the digestion and utilization rate of soybean meal and even adversely affects the growth and health of animals. The study found that the use of microbial fermentation to treat soybean meal can effectively reduce the content of anti-nutritional factors and allergenic factors in soybean meal, and at the same time improve the nutritional value and digestibility of soybean meal.

At present, there are many studies on soybean meal fermentation and production of fermented soybean meal enterprises, and the strains and processes used are also different. Generally speaking, the main problems are as follows: unreasonable selection of fermentation strains, incomplete fermentation of soybean meal, long fermentation period, The product quality is unstable and the nutritional value needs to be improved.

Contents of the invention

Aiming at the existing problems, the present invention is based on the physiological metabolism characteristics of microorganisms, a composite flora obtained through repeated scientific experiments and rational collocation, consisting of Bacillus subtilis, Bacillus coagulans, Aspergillus niger, Aspergillus oryzae, Lactobacillus plantarum and Saccharomyces cerevisiae. Among them, Bacillus subtilis is the preferred strain LM3403, and the strain LM3403 has been preserved on June 25, 2014 in the General Microbiology Center of China Committee for the Collection of Microbial Cultures (Institute of Microbiology, Chinese Academy of Sciences, No. 1, Beichen West Road, Chaoyang District, Beijing), The deposit number is CGMCC No.9377.

Beneficial effects of the present invention:

The composite bacterial flora used in the fermentation of soybean meal provided by the present invention is all included in the "Catalogue of Feed Additives (2013)" issued by the Ministry of Agriculture, is safe and reliable, and can be directly used in the industrial production of fermented soybean meal.

The composite flora provided by the present invention is obtained through repeated scientific experiments and reasonable collocation based on the physiological and metabolic characteristics of microorganisms. Through the synergistic effect of multiple strains, it grows rapidly and metabolizes vigorously. The soybean meal is fully fermented, the nutritional value is improved, and the production efficiency is improved. Production costs are reduced.

The composite flora provided by the invention has vigorous growth and metabolism, can secrete various enzymes, acids and other metabolites, can efficiently decompose macromolecular proteins, antinutritional factors and allergenic factors in soybean meal, and significantly increase soluble proteins, small peptides, The content of organic acids, proteases, vitamins, etc. and the number of beneficial bacteria and antimicrobial peptides can improve digestion and utilization, inhibit the growth of pathogenic bacteria, and reduce the use of antibiotics.

After complex bacterial fermentation, compared with soybean meal raw materials, the finished product has a fermented sour aroma, the content of crude protein increases by more than 17%, the content of small peptides increases by 11 times, the content of trypsin inhibitors decreases by more than 80%, and the content of soybean hemagglutinin decreases by 99%. above.

Detailed ways

In order to make the present invention easier to understand, the present invention will be further elaborated below through examples. It should be understood that these examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention.

The specific experimental methods not mentioned in the following examples are usually carried out according to conventional experimental methods.

Embodiment 1: Preparation of liquid complex flora

The bacterial strains of the composite flora are respectively subjected to plate activation, culture in small shake flasks, culture in large shake flasks, and step-by-step amplified culture in fermentors using their corresponding culture medium to obtain bacterial fluids of various strains. The number of viable bacteria in Bacillus subtilis, Bacillus coagulans, Lactobacillus plantarum and Saccharomyces cerevisiae is not less than 1×10 ⁸ cfu/mL, and the number of viable bacteria in Aspergillus niger and Aspergillus oryzae is not less than 1×10 ⁷ cfu /mL. Mix the bacterial liquids of various strains in proportion to obtain the liquid composite bacterial group. The number of live bacteria in the liquid composite flora is not less than 5×10 ⁸ cfu/mL.

The liquid composite flora can also be obtained by mixing commercially available liquid strains of various strains in proportion.

Embodiment 2: Preparation of solid composite flora

The bacterial strains of the composite flora are respectively subjected to plate activation, culture in small shake flasks, culture in large shake flasks, and step-by-step amplified culture in fermentors using their corresponding culture medium to obtain bacterial fluids of various strains. The number of viable bacteria in Bacillus subtilis, Bacillus coagulans, Lactobacillus plantarum and Saccharomyces cerevisiae is not less than 5×10 ⁸ cfu/mL, and the number of viable bacteria in Aspergillus niger and Aspergillus oryzae is not less than 1×10 ⁸ cfu /mL. The solid bacterial agents are obtained by carrier adsorption, freeze-drying, spray-drying or solid matrix culture, etc., and the solid bacterial agents of each bacterial species are mixed in proportion to obtain a solid composite bacterial group. The number of live bacteria in solid composite flora is not less than 2×10 ⁸ cfu/g.

The solid composite bacterial group can also be obtained by mixing commercially available solid microbial agents of various bacterial species in proportion.

Embodiment 3: detection method

Moisture content is tested according to GB/T 6435-2006, crude protein content is tested according to GB/T 6432-1994, small peptide content is determined by TCA method, and trypsin inhibitor is detected by BAPA method with reference to literature (Li Sufen, Huo Guicheng, Yang Lijie , et al. Study on the content of trypsin inhibitor in the main soybean varieties in Heilongjiang Province[J]. Assay [J]. Biotechnology, 1996, 6(5): 44-46.). After testing, the moisture content of soybean meal raw materials is 12.1%, the crude protein content is 45.5%, the small peptide content is 1.1%, the trypsin inhibitor content is 1872TIU/mg, the soybean hemagglutinin potency is 10300, and the beany smell.

Example 4:

Mix 1kg of soybean meal, 0.1kg of bran, and 0.7L of water, keep the pH natural, sterilize at 121°C for 20 minutes, and cool down for later use. Add 0.1L of the liquid complex flora to the material that has dropped to room temperature, mix well, start the fermentation at 35°C, and ferment for 44 hours, turning the material every 5 hours for ventilation. After the fermentation is finished, it is dried at 50°C-60°C, crushed and sieved to obtain the finished product of fermented soybean meal. The number of viable bacteria in the liquid complex flora was 3×10 ⁹ cfu/mL, and the ratio of the number of each strain was 2:2:1:1:1:3. The finished fermented soybean meal has a fermented sour aroma, with a moisture content of 11.2%, a protein content of 54.8%, a small peptide content of 13.2%, a viable count of beneficial bacteria of 5×10 ⁸ cfu/g, a trypsin inhibitor content of 308TIU/mg, and soybean hemagglutinin Titer 34.

Example 5:

Mix 5kg of soybean meal, 0.3kg of bran, 0.3kg of rice bran, and 3.5L of water, keep the pH natural, sterilize at 121°C for 20 minutes, and cool down for later use. Add 0.8L of the liquid complex flora to the material that has dropped to room temperature, mix well, and start the fermentation at 35°C, the fermentation time is 52 hours, and the material is turned over every 4 hours for ventilation. After the fermentation is finished, it is dried at 50°C-60°C, crushed and sieved to obtain the finished product of fermented soybean meal. The number of live bacteria in the liquid complex flora is 5×10 ⁹ cfu/mL, and the ratio of the number of each strain is 3:1:1:1:3:1. The finished fermented soybean meal has a fermented sour aroma, with a moisture content of 11.8%, a protein content of 56.2%, a small peptide content of 14.1%, a viable count of beneficial bacteria of 2×10 ⁹ cfu/g, a trypsin inhibitor content of 321TIU/mg, and soybean hemagglutinin Titer 47.

Embodiment 6:

Mix 10kg of soybean meal, 1.2kg of bran, and 7L of water, keep the pH natural, sterilize at 121°C for 20 minutes, and cool down for later use. Add 1.7L of the liquid complex flora to the material that has dropped to room temperature, mix well, and ferment at an initial temperature of 35°C for a fermentation time of 56 hours, turning the material every 3 hours for ventilation. After the fermentation is finished, it is dried at 50°C-60°C, crushed and sieved to obtain the finished product of fermented soybean meal. The number of viable bacteria in the liquid composite flora was 8×10 ⁸ cfu/mL, and the ratio of the number of each strain was 1:3:2:1:2:1. The finished fermented soybean meal has a fermented sour aroma, with a moisture content of 11.2%, a protein content of 54.7%, a small peptide content of 12.8%, a viable count of beneficial bacteria of 5×10 ⁸ cfu/g, a trypsin inhibitor content of 345TIU/mg, and a soybean hemagglutinin effect. Price 50.

Embodiment 7:

5kg of soybean meal, 0.5kg of bran, and 5L of water are mixed evenly, the pH is natural, sterilized at 121°C for 20 minutes, and cooled for later use. Add 1.1 kg of solid composite bacteria to the material that has dropped to room temperature, mix well, start the fermentation at 35°C, and ferment for 60 hours, turning the material every 4 hours for ventilation. After the fermentation is finished, it is dried at 50°C-60°C, crushed and sieved to obtain the finished product of fermented soybean meal. The viable count of the solid composite flora was 5×10 ⁸ cfu/mL, and the ratio of the number of each strain was 1:1:1:2:3:2. The finished fermented soybean meal has a fermented sour aroma, with a moisture content of 11.6%, a protein content of 52.5%, a small peptide content of 12.2%, a viable count of beneficial bacteria of 2×10 ⁸ cfu/g, a trypsin inhibitor content of 364TIU/mg, and soybean hemagglutinin Titer 62.

Embodiment 8:

Cottonseed meal 2kg, rapeseed meal 1kg, peanut meal 1.5kg, bran 0.5kg, water 3.5L, mix evenly, pH is natural, sterilize at 121°C for 20 minutes, cool down for later use. Add 1L of the liquid complex flora to the material that has dropped to room temperature, mix well, and ferment the initial temperature at 35°C, the fermentation time is 60h, and turn the material every 4h for ventilation. After the fermentation is finished, it is dried at 50°C-60°C, crushed and sieved to obtain the finished product of fermented soybean meal. The number of live bacteria in the liquid complex flora is 2×10 ⁹ cfu/mL, and the ratio of the number of each strain is 2:1:1:2:2:2. The finished product has a fermented sour aroma, with a moisture content of 12.1%, a protein content of 43.5%, a small peptide content of 8.9%, and a viable count of beneficial bacteria of 5×10 ⁸ cfu/g.

For those of ordinary skill in the art, it is still possible to modify the technical solutions described in the above embodiments, or perform equivalent replacements for some of the technical features; and these modifications or replacements do not make the essence of the corresponding technical solutions deviate from this document. The spirit and scope of the technical solutions claimed by the invention.

Claims (7)

1. for a composite flora for bean pulp fermentation, it is characterized in that, composite flora is made up of subtilis, Bacillus coagulans, aspergillus niger, aspergillus oryzae, plant lactobacillus and yeast saccharomyces cerevisiae.

2. composite flora according to claim 1, is characterized in that, each bacterial classification number ratio is (1-3): (1-3): (1-2): (1-2): (1-3): (1-3).

3. composite flora according to claim 1, is characterized in that, the preferred preserving number of described subtilis is the bacterial strain of CGMCC No.9377.

4. composite flora according to claim 1, is characterized in that, can in liquid form or solid form be used for bean pulp fermentation.

5. described in claim 4, be characterised in that, the total viable count of liquid composite flora is not less than 2 × 10 ⁸cfu/mL, the total viable count of solid composite flora is not less than 2 × 10 ⁸cfu/g.

6. the application of composite flora according to claim 1 in bean pulp fermentation, is characterized in that, fermentation raw material weight ratio is dregs of beans 100 parts: wheat bran (or rice bran) 10-20 part; regulate fermentation raw material water content 45%-50%; pH nature, 121 DEG C of sterilizings 20 minutes, lower the temperature for subsequent use; The weight ratio of fermentation raw material and composite flora is fermentation raw material 100 parts: composite flora 10-20 part; Shallow tray fermentation after the fermentation raw material of room temperature is down in composite flora access, fermentation initial temperature 35 DEG C, every 3h-5h stirring ventilates, fermentation period 48h-60h, temperature of charge drops to less than 35 DEG C fermentation ends, and 50 DEG C-60 DEG C oven dry, pulverize and sieve and namely obtain fermented bean dregs finished product.

7. the application of composite flora according to claim 1 in the fermentations such as cottonseed meal, rapeseed meal, peanut meal and the mixing dregs of rice.