Solid anaerobic fermentation cottonseed meal as well as preparation method and application thereof
Through solid-state deep oxygen-sensitive fermentation technology and the use of composite bacteria, the problems of low protein content and high residual gossy phenol in the existing cotton meal fermentation technology are solved, and efficient and safe cotton meal fermentation effect is achieved.
Patent Information
- Application Number
- CN202411723665.5
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-11-28
- Publication Date
- 2025-02-11
AI Technical Summary
In the existing cotton meal fermentation technology, the acid-soluble protein/crude protein content is low, the residual gossy phenol cannot be fully reduced, and prolonging the fermentation time will lead to weight loss of materials and contamination of miscellaneous bacteria.
The solid-state deep oxygen-sensitive fermentation process is adopted, and the fermentation is carried out using complex bacteria such as Bacillus subtilis M-15 and Bacillus subtilis JQ-2 or Candidautilis QC-1. Control the anaerobic environment to inhibit the growth of aerobic bacteria.
The content of acid-soluble protein and crude protein in cotton meal is significantly improved, the residual gossipol is reduced, the weight loss of materials and contamination of miscellaneous bacteria is reduced, and the safety and efficiency of fermented cotton meal is improved.
Smart Images
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Abstract
Description
Technical Field
[0001] The invention belongs to the technical field of fermented feed preparation, and specifically relates to solid anaerobic fermented cottonseed meal and a preparation method and application thereof. Background Art
[0002] With the increasing global health awareness, the demand for high-protein foods has been steadily increasing. Animal products such as meat, eggs and dairy products are the main sources of dietary protein for humans. Therefore, sustainable protein feed is needed to maintain animal production levels. However, there is a serious shortage of protein feed in my country, so the use of agricultural product processing by-products such as rapeseed meal, cottonseed meal and peanut meal is a feasible measure to fill the gap in feed protein demand.
[0003] Cottonseed, commonly known as "white gold", is one of the main sources of edible oil. In recent years, the total cottonseed production in my country has been generally stable, maintaining at more than 10 million tons. It is reported that about 50% of cottonseed meal is produced after cottonseed oil is pressed. It is rich in crude protein (40%-45%), B vitamins, E vitamins, and has a balanced amino acid composition. It is regarded as the most promising protein feed resource. However, the free cottonpol in cottonseed meal has active hydroxyl groups and aldehyde groups that are toxic. Although ruminants are more tolerant to free cottonpol, monogastric animals are more susceptible to the toxicity of free cottonpol, and excessive intake of free cottonpol can cause animals to have a variety of clinical symptoms such as anorexia, metabolic disorders, growth inhibition, and abnormal visceral organ function, which may lead to animal death in severe cases. Therefore, detoxification of cottonseed meal to a safe level is a prerequisite for its use as an animal protein feed resource.
[0004] Microbial fermentation is widely used in cottonpol detoxification due to its high efficiency, low cost and environmental friendliness. At present, most production mainly adopts open aerobic fermentation process. The acid-soluble protein / crude protein content in the fermented cottonseed meal products still has room for improvement, and the residual cottonpol still needs to be further reduced. Prolonging the fermentation time can solve this problem, but it will also cause problems such as serious weight loss of materials and contamination by foreign bacteria. The solid-state deep anaerobic fermentation process can provide an anaerobic environment in the middle and late stages of fermentation, which is conducive to inhibiting the growth of aerobic bacteria, reducing material weight loss, and improving production efficiency. Combining facultative anaerobic bacteria as fermentation strains can effectively prolong the fermentation time of cottonseed meal, and control material weight loss and foreign bacterial contamination. Safe and efficient strains and suitable fermentation processes are the key to obtaining excellent fermented cottonseed meal. Summary of the invention
[0005] The purpose of the present invention is to provide a solid-state anaerobic fermentation cottonseed meal and a preparation method and application thereof.
[0006] The invention discloses solid anaerobic fermented cottonseed meal, which is prepared by fermenting composite bacteria.
[0007] Preferably, the composite bacterial species are Bacillus subtilis M-15 and Bacillus subtilis JQ-2; the preservation number of the Bacillus subtilis M-15 is CGMCC No.23483, and the preservation number of the Bacillus subtilis JQ-2 is CGMCC No.25660.
[0008] Preferably, the composite bacterial species are Bacillus subtilis M-15 and Candidautilis QC-1; the preservation number of Bacillus subtilis M-15 is CGMCC No.23483, and the preservation number of Candidautilis QC-1 is CGMCC No.9165.
[0009] A method for preparing solid-state anaerobic fermentation cottonseed meal is carried out according to the following steps:
[0010] (1) crushing cottonseed meal into a powder with a particle size of 10-15 mesh, adding a carbon source and ammonium sulfate to mix evenly, adding water, stirring evenly, and obtaining a solid fermentation culture medium;
[0011] (2) Inoculate the composite bacterial agent into the solid fermentation culture medium prepared in step (1), and ferment at room temperature (25° C.) for 8-12 days.
[0012] The carbon source is corn flour, molasses, sucrose or starch, and the added amount is 5-15% of the weight of cottonseed meal.
[0013] The amount of ammonium sulfate added is 1-3% of the weight of cottonseed meal.
[0014] The amount of water added is 70-90% of the weight of the cottonseed meal.
[0015] The addition amount of the composite bacterial agent is 5.0×10 9 -2.5×10 10 CFU / kg.
[0016] The application of the solid-state anaerobic fermented cottonseed meal in preparing feed.
[0017] The beneficial effects of the present invention are as follows: the present invention compounds Bacillus subtilis M-15 with Bacillus subtilis JQ-2 or Candida albicans QC-1, and performs solid-state anaerobic fermentation on cottonseed meal, thereby successfully preparing a fermented cottonseed meal feed raw material with low gossypol, high acid-soluble protein and crude protein content, significantly improving the utilization rate of cottonseed meal as a protein source feed, and helping to alleviate the current shortage of protein resources in the feed industry. DETAILED DESCRIPTION
[0018] In order to facilitate the understanding of the present invention, the present invention will be described more fully below. However, the present invention can be implemented in many different forms and is not limited to the embodiments described herein. On the contrary, the purpose of providing these embodiments is to make the understanding of the disclosure of the present invention more thorough and comprehensive.
[0019] The Bacillus subtilis M-15 used in the following examples was preserved in this laboratory, Bacillus subtilis JQ-2 and Candidautilis QC-1 were purchased from the China General Microbiological Culture Collection Center (CGMCC); the cottonseed meal raw material was purchased from Baoding Dongguang Oil Pressing Factory, and the total cottonpol content in the cottonseed meal raw material was 2066 mg / kg, and the free cottonpol content was 980 mg / kg.
[0020] Example 1
[0021] A method for preparing solid-state anaerobic fermentation cottonseed meal is carried out according to the following steps:
[0022] (1) Grinding cottonseed meal into powder with a particle size of 12 mesh, adding corn flour and ammonium sulfate, mixing evenly, adding water, stirring evenly, and obtaining a solid fermentation culture medium; the amount of corn flour added is 10% of the weight of the cottonseed meal; the amount of ammonium sulfate added is 2% of the weight of the cottonseed meal; and the amount of water added is 80% of the weight of the cottonseed meal;
[0023] (2) adding a composite bacterial agent to the solid fermentation culture medium prepared in step (1), and fermenting at room temperature (25°C) for 10 days; the composite bacterial species are Bacillus subtilis M-15 and Bacillus subtilis JQ-2; the addition amount of Bacillus subtilis M-15 is 5.0×10 9 CFU / kg; the addition amount of Bacillus subtilis JQ-2 was 5.0×10 9 CFU / kg.
[0024] Example 2
[0025] A method for preparing solid-state anaerobic fermentation cottonseed meal is carried out according to the following steps:
[0026] (1) Grinding cottonseed meal into a powder with a particle size of 15 mesh, adding sucrose and ammonium sulfate and mixing evenly, adding water and stirring evenly to obtain a solid fermentation culture medium; the amount of sucrose added is 8% of the weight of the cottonseed meal; the amount of ammonium sulfate added is 3% of the weight of the cottonseed meal; and the amount of water added is 75% of the weight of the cottonseed meal;
[0027] (2) A composite bacterial agent was added to the solid fermentation culture medium prepared in step (1), and the mixture was fermented at room temperature (25°C) for 12 days; the composite bacterial species were Bacillus subtilis M-15 and Candida utilis QC-1, and the amount of Bacillus subtilis M-15 added was 5.0×10 9 CFU / kg; the addition amount of Candida utilis QC-1 was 5.0×10 9 CFU / kg.
[0028] Example 3
[0029] A method for preparing solid-state anaerobic fermentation cottonseed meal is carried out according to the following steps:
[0030] (1) crushing cottonseed meal into a powder with a particle size of 10 mesh, adding molasses and ammonium sulfate to mix evenly, adding water, stirring evenly, and obtaining a solid fermentation culture medium; the amount of molasses added is 12% of the weight of the cottonseed meal, the amount of ammonium sulfate added is 1% of the weight of the cottonseed meal, and the amount of water added is 90% of the weight of the cottonseed meal;
[0031] (2) adding a composite bacterial agent to the solid fermentation culture medium prepared in step (1), and fermenting at room temperature (25°C) for 9 days; the composite bacterial species are Bacillus subtilis M-15 and Bacillus subtilis JQ-2; the addition amount of Bacillus subtilis M-15 is 8.0×10 9 CFU / kg; the addition amount of Bacillus subtilis JQ-2 was 8.0×10 9 CFU / kg.
[0032] Comparative Example 1
[0033] A method for preparing solid-state anaerobic fermentation cottonseed meal is carried out according to the following steps:
[0034] (1) Grinding cottonseed meal into powder with a particle size of 12 mesh, adding corn flour and ammonium sulfate, mixing evenly, adding water, stirring evenly, and obtaining a solid fermentation culture medium; the amount of corn flour added is 10% of the weight of the cottonseed meal; the amount of ammonium sulfate added is 2% of the weight of the cottonseed meal; and the amount of water added is 80% of the weight of the cottonseed meal;
[0035] (2) Inoculate the solid fermentation medium prepared in step (1) with the Bacillus subtilis M-15, and ferment at room temperature (25°C) for 10 days; the amount of the Bacillus subtilis M-15 added is 1.0×10 10 CFU / kg.
[0036] Comparative Example 2
[0037] A method for preparing solid-state anaerobic fermentation cottonseed meal is carried out according to the following steps:
[0038] (1) Grinding cottonseed meal into powder with a particle size of 12 mesh, adding corn flour and ammonium sulfate, mixing evenly, adding water, stirring evenly, and obtaining a solid fermentation culture medium; the amount of corn flour added is 10% of the weight of the cottonseed meal; the amount of ammonium sulfate added is 2% of the weight of the cottonseed meal; and the amount of water added is 80% of the weight of the cottonseed meal;
[0039] (2) Bacillus subtilis JQ-2 was inoculated into the solid fermentation medium prepared in step (1), and fermented at room temperature (25°C) for 10 days; the composite strains were Bacillus subtilis M-15 and; the amount of Bacillus subtilis JQ-2 added was 1.0×10 10 CFU / kg.
[0040] Comparative Example 3
[0041] A method for preparing solid-state anaerobic fermentation cottonseed meal is carried out according to the following steps:
[0042] (1) Grinding cottonseed meal into a powder with a particle size of 15 mesh, adding sucrose and ammonium sulfate and mixing evenly, adding water and stirring evenly to obtain a solid fermentation culture medium; the amount of sucrose added is 8% of the weight of the cottonseed meal; the amount of ammonium sulfate added is 3% of the weight of the cottonseed meal; and the amount of water added is 75% of the weight of the cottonseed meal;
[0043] (2) Bacillus subtilis M-15 was added to the solid fermentation medium prepared in step (1), and fermented at room temperature (25°C) for 12 days; the amount of Bacillus subtilis M-15 added was 1.0×1010 CFU / kg.
[0044] Comparative Example 4
[0045] A method for preparing solid-state anaerobic fermentation cottonseed meal is carried out according to the following steps:
[0046] (1) Grinding cottonseed meal into a powder with a particle size of 15 mesh, adding sucrose and ammonium sulfate and mixing evenly, adding water and stirring evenly to obtain a solid fermentation culture medium; the amount of sucrose added is 8% of the weight of the cottonseed meal; the amount of ammonium sulfate added is 3% of the weight of the cottonseed meal; and the amount of water added is 75% of the weight of the cottonseed meal;
[0047] (2) Candida utilis QC-1 was inoculated into the solid fermentation medium prepared in step (1) and fermented at room temperature (25°C) for 12 days; the amount of Candida utilis QC-1 added was 1.0×10 10 CFU / kg.
[0048] Experimental example:
[0049] Determination of total gossypol (TGP) and free gossypol (FGP) content in fermented cottonseed meal: The free gossypol was determined by the national standard aniline method (GB / T 13086-2020), and the total gossypol was determined by the method recommended by the International Organization for Standardization ISO6866-1985. SPSS24.0 software was used for statistical analysis, and the quantitative data results were used (mean ± standard deviation) was expressed, and the Kolmogorov-Smirnov test was used for data normality test. For data that met the normal distribution, the mean difference between the two groups was compared using the t test, and the difference was considered statistically significant when P < 0.05. The test results are shown in Table 1-4:
[0050] Table 1
[0051]
[0052] Note: * represents P < 0.05 compared with the group in Example 1.
[0053] Table 2
[0054]
[0055] Note: * represents P < 0.05 compared with the group in Example 2.
[0056] Table 3
[0057]
[0058] Note: * represents P < 0.05 compared with the group in Example 1.
[0059] Table 4
[0060]
[0061] Note: * represents P < 0.05 compared with the group in Example 2.
[0062] The above-mentioned embodiments only express several implementation methods of the present invention, and the descriptions thereof are relatively specific and detailed, but they cannot be understood as limiting the scope of the invention patent. It should be pointed out that, for ordinary technicians in this field, several variations and improvements can be made without departing from the concept of the present invention, and these all belong to the protection scope of the present invention. Therefore, the protection scope of the patent of the present invention shall be subject to the attached claims.
Claims
1. A solid-state anaerobic fermentation cottonseed meal, characterized in that: The solid-state anaerobic fermented cottonseed meal is prepared by fermentation of composite bacteria.
2. The solid-state anaerobic fermentation of cottonseed meal according to claim 1, characterized in that: The composite bacterial species are Bacillus subtilis M-15 and Bacillus subtilis JQ-2; the preservation number of the Bacillus subtilis M-15 is CGMCC No.23483, and the preservation number of the Bacillus subtilis JQ-2 is CGMCC No.25660.
3. The solid-state anaerobic fermentation of cottonseed meal according to claim 1, characterized in that: The composite bacterial species are Bacillus subtilis M-15 and Candidautilis QC-1; the preservation number of the Bacillus subtilis M-15 is CGMCC No.23483, and the preservation number of the Candidautilis QC-1 is CGMCC No.9165.
4. A method for preparing solid-state anaerobic fermentation cottonseed meal, characterized in that: Follow these steps: (1) crushing cottonseed meal into a powder with a particle size of 10-15 mesh, adding a carbon source and ammonium sulfate to mix evenly, adding water, stirring evenly, and obtaining a solid fermentation culture medium; (2) The composite bacterial agent according to claim 1 is introduced into the solid fermentation culture medium prepared in step (1), and fermented at room temperature for 8-12 days.
5. The method for preparing solid-state anaerobic fermentation cottonseed meal according to claim 4, characterized in that: The carbon source is corn flour, molasses, sucrose or starch, and the added amount is 5-15% of the weight of cottonseed meal.
6. The method for preparing solid-state anaerobic fermentation cottonseed meal according to claim 4, characterized in that: The amount of ammonium sulfate added is 1-3% of the weight of cottonseed meal.
7. The method for preparing solid-state anaerobic fermentation cottonseed meal according to claim 4, characterized in that: The amount of water added is 70-90% of the weight of the cottonseed meal.
8. The method for preparing solid-state anaerobic fermentation cottonseed meal according to claim 4, characterized in that: The addition amount of the composite bacterial agent is 5.0×10 9 -2.5×10 10 CFU / kg.
9. Use of any solid-state anaerobic fermented cottonseed meal according to any one of claims 1 to 8 in preparing feed.