A method for preparing calcium-containing amino acids using fish bones and discarded feathers by bioconversion
By using probiotic Bacillus subtilis fermented fish bones and discarded feathers, the problem of its difficulty in degradation and resource utilization has been solved, efficient degradation and high-value utilization have been achieved, environmental pollution has been reduced, and its application areas have been broadened.
Patent Information
- Application Number
- CN202411880051.8
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-12-19
- Publication Date
- 2025-04-04
AI Technical Summary
Due to structural characteristics and odor problems, fish bones and waste feathers are difficult to effectively degrade and utilize them in resource-based ways, resulting in environmental pollution and waste of resources.
Bacillus subtilis, which has probiotic function, is used as a microbial species, and fish bones and waste feathers are used as fermentation substrates to convert them into soluble calcium-containing amino acid solutions.
It improves the degradation rate and soluble protein content of fish bones and waste feathers, realizes high-value utilization of resources, reduces environmental pollution, and broadens its application value in the fields of animal feed and plant fertilizers.
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Figure CN119753046A_ABST
Abstract
Description
Technical Field
[0001] The invention belongs to the field of waste resource utilization, and specifically relates to a method for preparing calcium-containing amino acids by utilizing fish bones and waste feathers through a biological transformation method. Background Art
[0002] my country is a country with a large scale of fish farming in the world, with an annual fish production of about 37 million tons. Fish bones account for about 50% of the total weight of the fish body, and more than 18 million tons of fish bone by-products are produced each year during the fish processing process. Fish bones emit a fishy smell and are not easy to use. Many fish bone scraps are directly discarded, causing environmental pollution. On the other hand, fish bones contain components such as protein, moisture, lipids, ash, high levels of mineral elements such as calcium and phosphorus, and various trace elements such as iron and selenium. The fish bone calcium, chondroitin sulfate, gelatin, fish bone peptides and fish bone oil extracted from them can be widely used in the fields of food, medicine and cosmetics. The calcium content in fish bones is high, and the calcium-phosphorus ratio is appropriate. Converting it into a soluble state is not only suitable for the calcium source needed by the human body, but also can be used as a nutrient for animals and plants.
[0003] At present, as an organic waste rich in protein content, the degradation and resource utilization of discarded feathers have attracted much attention. Waste feathers are one of the by-products of livestock and poultry farming, with an annual output of millions of tons in my country, which is huge. Waste feathers are cheap and easy to obtain, with a crude protein content of more than 85%. It is a protein resource with a variety of amino acids and rich content, and can also be used to prepare feed and fertilizer suitable for animals and plants. Because the feather structure contains a large number of disulfide bonds and hydrogen bonds, the feather structure is densely cross-linked and insoluble in water, which makes the degradation of feathers difficult, hindering the high-value utilization of waste feathers. Waste feathers also have a peculiar smell. If not handled properly, they can easily cause environmental pollution and disease transmission problems. Compared with conventional physical and chemical methods (such as strong alkali method) and enzyme methods, microbial degradation of waste feathers is the most promising method for its high-value application.
[0004] At present, there are many studies on the extraction of peptides, oils, collagen and high-calcium preparations from fish bones, and the screening and degradation methods of waste feather degradation strains. However, there are no reports on the bioconversion of high-calcium fish bones and high-protein waste feathers into soluble calcium-containing amino acid solutions through microbial metabolism. The combined resource utilization of fish bones and waste feathers can not only solve their pollution to the environment, but also microorganisms produce beneficial metabolites such as proteases through metabolic activities under the induction of them as substrates, making them have higher application value in the fields of animal feed and plant fertilizers. Summary of the invention
[0005] In view of the shortcomings of the above waste comprehensive utilization technology, the present invention provides a method for preparing calcium-containing amino acids from fish bones and waste feathers by a bioconversion method. The present invention adopts a probiotic bacillus strain that can be safely used in the field of feed and fertilizer as a microbial strain, uses fish bones and waste feathers as two organic wastes as fermentation substrates, inoculates bacillus seed liquid, obtains a calcium-containing amino acid mixed liquid through fermentation, and improves the degradation efficiency of fish bones and the soluble protein content of waste feathers.
[0006] To achieve the above object, the present invention adopts the following technical solutions:
[0007] Bacillus subtilis was inoculated into a culture medium containing fish bones and feathers to degrade the fish bones and feathers and obtain calcium-containing amino acids.
[0008] Preferably, the fish bone comprises 1-2% fish bone powder by mass.
[0009] Preferably, the feathers contain 1% feathers by mass.
[0010] Preferably, the feathers are discarded feathers.
[0011] Preferably, (1) bacterial cell activation: streaking the target Bacillus strain on a conventional LB solid medium, culturing at 37° C. for 20 to 30 hours, and growing a single colony;
[0012] (2) Seed liquid preparation: transplant a single colony into LB liquid medium and culture at 37°C and 160-220 rpm for 18-22 h;
[0013] (3) Degradation of fish bones and discarded feathers: The fish bones were boiled and heated, air-dried, and crushed to obtain fish bone powder; the discarded feathers were washed and air-dried, and the seed liquid was inoculated into the culture medium containing fish bones and discarded feathers at a volume ratio of 5-12%, respectively, and cultured at about 37°C and 160-220°C for 24-45 hours. Similarly, the same seed liquid was inoculated into the culture medium containing discarded feathers and fish bones (hair-bone culture medium), and cultured under the same conditions for 24-45 hours.
[0014] The waste feather culture medium comprises, by mass fraction, 0.01-0.07% KH2PO4, 0.02-0.08% K2HPO4, 1% waste feathers, a pH value of 8-9, and the balance being water.
[0015] The hair-bone culture medium comprises, by mass fraction, 1-2% fish bone powder, 1% discarded feathers, 0.01-0.07% KH2PO4, 0.02-0.08% K2HPO4, pH value is 8-9, and the balance is water.
[0016] Beneficial effects of the present invention:
[0017] (1) The combined degradation of fish bones and discarded feathers improves the degradation rate of fish bones.
[0018] (2) The present invention effectively achieves the dual reduction and value-added efficiency of fish bones and waste feathers, avoids environmental pollution, and broadens the application fields of the two.
[0019] (3) Use probiotic microbial strains commonly used in feed and fertilizer to degrade fish bones and discarded feathers to produce soluble calcium-containing amino acids, which can be used to prepare feed and fertilizer, thereby promoting the high-value utilization of waste.
[0020] (4) This microorganism, which is commonly used in feed and fertilizer, has probiotic functions for both animals and plants. In the process of degrading fish bones and discarded feathers to form a mixed liquid, it may also produce metabolites with probiotic and immune-enhancing effects, making it a promising alternative to antibiotics. BRIEF DESCRIPTION OF THE DRAWINGS
[0021] Figure 1 It is the crushing process of fish bones.
[0022] Figure 2 It is an apparent comparison of the degradation of fish bones and discarded feathers.
[0023] Figure 3 This is a case of combined degradation of fish bones and discarded feathers.
[0024] Figure 4 It is a comparison of the degradation effects of fish bones and discarded feathers, either individually or in combination. Fish bones represent the degradation rate of fish bones alone, feathers represent the degradation rate of feathers alone, (hair + bones)-hair represents the degradation rate of feathers in the simultaneous degradation of feathers and fish bones, and (hair + bones)-bones represents the degradation rate of fish bones in the simultaneous degradation of feathers and fish bones. Specific embodiments
[0025] The present invention is further explained below in conjunction with the accompanying drawings and specific embodiments, but does not constitute any limitation to the present invention.
[0026] This embodiment is described by taking Bacillus subtilis CICC 21095 suitable for biological feed and fertilizer as an example. The strain was purchased from the China Industrial Microbiological Culture Collection Management Center.
[0027] Example 1 Degradation of fish bones and discarded feathers by Bacillus subtilis CICC 21095
[0028] Bacillus subtilis CICC 21095 was streaked on LB solid medium and placed at 37°C for 24 hours to obtain a single colony; the single colony was transferred to LB liquid medium through an inoculation loop and cultured at 37°C and 160rpm for 20 hours to obtain a seed solution. Then, the seed solution was inoculated into fish bone culture medium (the mass content of fish bones was 1% and 2%, respectively) and discarded chicken feather culture medium (the mass content of chicken feathers was 1%) at a volume ratio of 7%, and degraded at 37°C and 200rpm for 40 hours to obtain a degradation solution. Figure 1 As shown, fish bones are boiled, air-dried and crushed to obtain fish bone powder that is easy to use. Through the degradation of Bacillus subtilis CICC 21095, it was found that the CICC 21095 strain grew well in the culture medium with fish bones as the substrate; at the same time, compared with 1% fish bones, the solution formed by it using 2% fish bones has a larger turbidity ( Figure 2 a, b). This indicates that the CICC 21095 strain can degrade fish bones and use them for growth. On the other hand, the CICC 21095 strain can convert discarded chicken feathers into a soluble state ( Figure 2 c), indicating that the strain is capable of degrading discarded feathers.
[0029] The fish bone culture medium comprises fish bone powder (1% and 2%) by mass fraction, and the balance is water, and is sterilized at high temperature for standby use.
[0030] The discarded chicken feather culture medium comprises, by mass fraction, 0.03% KH2PO4, 0.05% K2HPO4, 1% discarded chicken feathers, with a pH of 8.5 and the remainder being water, and is sterilized at high temperature for standby use.
[0031] Example 2 Degradation of fish bones and discarded feathers
[0032] Based on the ability of CICC 21095 strain to degrade fish bones and discarded feathers, in order to further improve the resource utilization value of the two, the two were combined for degradation. Similar to the method in Example 1, with 2% fish bones and 1% discarded chicken feathers as substrates, the CICC 21095 strain seed liquid was inoculated into the hair-bone culture medium at a volume ratio of 7%, and the degradation was carried out at 37°C and 200rpm for 40h. Figure 3 The results showed that the strain also significantly converted discarded chicken feathers into a soluble state, obtaining a calcium-containing amino acid mixture. Through testing, it was found that the degradation rate of the CICC 21095 strain for 2% fish bones was 68.1%, and its degradation rate for fish bones in the combined degradation process of fish bones and discarded feathers reached 83.5%, which was 22.6% higher than that of fish bones alone. At the same time, the degradation rate of discarded chicken feathers by the strain alone and in the combined degradation process of fish bones reached 100% ( Figure 4a). In addition, the soluble protein content produced by the CICC 21095 strain when degrading fish bones and discarded chicken feathers was 0.9 g / L and 3.4 g / L respectively, while the soluble protein content formed in the hair-bone medium reached 4.1 g / L, which was significantly higher than that of the two alone ( Figure 4 b).
[0033] The hair-bone culture medium comprises, by mass fraction, 2% fish bone powder, 1% discarded chicken feathers, 0.03% KH2PO4, 0.06% K2HPO4, pH value is 8.5, and the balance is water.
[0034] The above results show that when the CICC 21095 strain degrades fish bones and discarded feathers at the same time, it can not only completely degrade the discarded feathers, but also significantly enhance the ability to degrade fish bones, and can increase the soluble protein content, making it a calcium-containing amino acid mixture. At the same time, as a probiotic commonly used in feed and fertilizer, the CICC 21095 strain will produce beneficial metabolites through fermentation under the induction of both as substrates. This makes fish bones and discarded feathers have higher application value in the fields of animal feed and plant fertilizers.
[0035] The above are only specific embodiments of the present invention, but the technical features of the present invention are not limited thereto, and any simple modification, change or equivalent substitution made within the concept and principle of the present invention should be considered to still fall within the protection scope of the present invention.
Claims
1. A method for preparing calcium-containing amino acids using fish bones and discarded feathers by bioconversion, characterized in that: Bacillus subtilis was inoculated into a culture medium containing fish bones and feathers to degrade the fish bones and feathers and obtain calcium-containing amino acids.
2. The method according to claim 1, characterized in that The fish bone comprises 1-2% fish bone powder by mass fraction.
3. The method according to claim 1, characterized in that The feathers contain 1% feathers by mass.
4. The method according to claim 1, characterized in that: The feathers are discarded chicken feathers.
5. The method according to claim 1, characterized in that The specific steps are: Bacterial activation: Streak the target Bacillus strain on a conventional LB solid medium, incubate at 37°C for 20-30 hours, and grow a single colony; Seed solution preparation: transplant a single colony into LB liquid medium and culture at 37°C and 160-220 rpm for 18-22 hours; Degrading fish bones and waste feathers: The fish bones are boiled and heated, air-dried, and crushed to obtain fish bone powder; the waste feathers are washed and air-dried, and the seed liquid is inoculated into the culture medium containing the two wastes, fish bones and waste feathers, at a volume ratio of 5-12%, respectively, and cultured at about 37°C and 160-220°C for 24-45 hours.
6. The method according to claim 5, characterized in that The hair-bone culture medium comprises, by mass fraction, 1-2% fish bone powder, 1% discarded feathers, 0.01-0.07% KH2PO4, 0.02-0.08% K2HPO4, pH value is 8-9, and the balance is water.