Process method for synchronously extracting omphisa fuscidentalis hampson protein peptide and grease based on enzymolysis

Abstract

The invention provides a technological method for synchronously extracting omphisa fuscidentalis hampson protein peptide and grease based on enzymolysis, and relates to the technical field of omphisa fuscidentalis hampson extraction. The technological method for synchronously extracting omphisa fuscidentalis hampson protein peptide and grease based on enzymolysis specifically comprises the following steps: S1, preparing a living body; s2, metabolism activation; s3, minimally invasive intervention; s4, carrying out in-situ separation; s5, product refining; and S6, insect body recovery. According to the technical scheme, through innovative combination of a living body metabolism engine and an in-situ separation technology, efficient, green and sustainable utilization of omphisa fuscidentalis hampson resources is achieved, the comprehensive advantages of the omphisa fuscidentalis hampson resources in the aspects of technical performance, economic benefits, ecological sustainability, application prospects and the like are achieved, the limitation of a traditional technology is broken through, and the technological cost is reduced. And a brand new technical normal form is provided for the field of biological resource refining.

Description

Technical Field

[0001] This invention relates to the field of bamboo worm extraction technology, specifically a process for the simultaneous extraction of bamboo worm protein peptides and oils based on enzymatic hydrolysis. Background Technology

[0002] Bamboo worms, also known as bamboo bees or bamboo maggots, are the larvae of bamboo weevils. They parasitize inside bamboo tubes, feeding on the fleshy inner walls of tender bamboo shoots, eating their way down from the tip to the base. Bamboo worms are plump and white, about 3 cm long, and rich in nutrients such as protein, amino acids, fatty acids, minerals, and vitamins. Extracting bamboo worm protein peptides and oils is primarily due to their multifaceted value. Bamboo worm protein peptides are a high-quality protein source with broad application prospects in food, health products, and medicine. For example, they can be added to food as a nutritional fortifier to enhance its nutritional value, and in the pharmaceutical field, they can be used to develop drugs or health products with specific functions. Bamboo worm oils are rich in unsaturated fatty acids and other components, offering certain health benefits, such as lowering cholesterol and preventing cardiovascular disease. They also have applications in the cosmetics and biofuel industries. In cosmetics, they can be used as a natural oil raw material, and in biofuels, they can be processed into green energy sources such as biodiesel.

[0003] Current technologies typically employ a "deactivation followed by processing" extraction method. When extracting protein peptides and oils from bamboo worms, the worms are first killed, and then the worm bodies are broken down using mechanical cell-wall breaking equipment. Next, exogenous enzymes are often added for enzymatic hydrolysis to promote protein peptide formation. In the separation stage, centrifugation is primarily used to initially separate oils and protein peptides, followed by multiple separation and purification steps, using organic solvents to extract and remove impurities. However, this extraction method has many drawbacks. For example, mechanical cell-wall breaking can damage the product structure and affect its activity; adding exogenous enzymes can easily trigger side reactions and may cause activity loss; centrifugation can cause mechanical damage to the product structure; using organic solvents not only increases costs but may also lead to solvent residues requiring additional post-processing; the entire process requires complex equipment and extreme conditions such as high temperatures, strong acids, and strong alkalis, resulting in high energy consumption and significant product damage; moreover, the worm residue is usually treated as waste, leading to low resource utilization and the generation of harmful byproducts, causing environmental pollution. Summary of the Invention

[0004] To address the shortcomings of existing technologies, this invention provides a process for the simultaneous extraction of bamboo worm protein peptides and oils based on enzymatic hydrolysis, which solves the problems of easily damaged product activity, numerous side reactions, structural damage during separation, solvent residue, high energy consumption, and low utilization rate in existing technologies.

[0005] To achieve the above objectives, the present invention provides the following technical solution: a process for simultaneously extracting bamboo worm protein peptides and oils based on enzymatic hydrolysis, specifically comprising the following steps: S1. Live animal preparation Select healthy bamboo worm larvae, and feed them a metabolic trigger solution after fasting for 48 hours; S2. Metabolic activation Placing the insect body in a low-temperature environment of 12℃ for 24 hours activates the endogenous enzyme system; S3. Minimally Invasive Intervention A dual-channel responsive conduit system was implanted in the third abdominal segment of the worm. S4. In-situ separation The exudated oil was collected at 30°C using a temperature-responsive mechanism, and the protein peptide solution was eluted at pH 5.0 using a pH-responsive mechanism. S5. Product Refining The oils were decolorized by molecular sieves and purified by low-temperature crystallization; the protein peptide solution was concentrated by ultrafiltration and then freeze-dried. S6. Insect Body Recycling The surviving insects are transferred to the recovery and breeding system for recycling.

[0006] Preferably, the metabolic trigger is composed of disodium 5'-inosinate and quercetin in a mass ratio of 5:3.

[0007] Preferably, the metabolic trigger solution has a concentration of 10 g / L, is prepared using 0.1 mol / L phosphate buffer, and has a pH of 7.2.

[0008] Preferably, the dual-channel responsive catheter system includes: Oil phase channel: filled with poly(N-isopropylacrylamide-co-butyl methacrylate) temperature-responsive gel, accounting for 60% of the cross-sectional area, with a phase transition temperature of 28-32℃; Peptide phase channel: filled with chitosan / sodium alginate pH-responsive gel, accounting for 40% of the cross-sectional area, with a pH response threshold of 5.2.

[0009] Preferably, the in-situ separation step maintains the osmotic pressure of the parasite's hemolymph at 380-420 mOsm / kg, which is achieved by simultaneous perfusion of isotonic glucose solution.

[0010] Preferably, the ultrafiltration concentration uses an ultrafiltration membrane with a molecular weight cutoff of 5 kDa, and the proportion of components with a molecular weight ≤ 1 kDa in the obtained protein peptide product is ≥ 90%.

[0011] Preferably, the insect body recovery step specifically includes: Use a nutrient solution containing 2g / L sericin and 0.5g / L dragon's blood extract for wound repair; Repeat the operation after 7 days of rest, and it can be reused up to 3 times.

[0012] This invention provides a process for the simultaneous extraction of protein peptides and oils from bamboo worms based on enzymatic hydrolysis. It has the following beneficial effects: This invention provides a process for the simultaneous enzymatic extraction of protein peptides and oils from bamboo worms. This technology activates the bamboo worm's own metabolic system, achieving efficient conversion of protein peptides and oils in a living state. This avoids the activity loss caused by exogenous enzyme addition and mechanical cell disruption in traditional processes. The innovatively designed dual-channel responsive conduit system constructs a separation interface in situ within the worm, achieving spatiotemporal decoupling of the target product, significantly improving separation efficiency. Furthermore, it eliminates the need for organic solvents and chemical additives, fully complying with green manufacturing principles. The product quality is excellent, with concentrated molecular weight distribution of protein peptides, a high proportion of small peptides, and full preservation of the structural integrity of bioactive substances such as antimicrobial peptides. The oil exhibits strong oxidative stability and low risk of rancidity. The operation process is gentle and controllable. Through minimally invasive intervention and mild condition control, damage to the worm is minimized. Post-operatively, the worm can be recycled, significantly improving resource utilization. The technology is energy-efficient, has a short production cycle, requires no complex equipment, and reduces production costs. This technology not only overcomes the limitations of traditional processes but also provides a new technological paradigm for the field of biorefining, with broad application prospects. Attached Figure Description

[0013] Figure 1 This is a schematic diagram of the process of the present invention. Detailed Implementation

[0014] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0015] like Figure 1 As shown, this embodiment of the invention provides a process for simultaneously extracting bamboo worm protein peptides and oils based on enzymatic hydrolysis, specifically including the following steps: S1. Live animal preparation Healthy bamboo worm larvae were selected and fed a metabolic trigger solution after a 48-hour fast. The metabolic trigger solution consisted of disodium 5'-inosinate and quercetin in a mass ratio of 5:3. The concentration of the metabolic trigger solution was 10 g / L, prepared using 0.1 mol / L phosphate buffer at a pH of 7.2.

[0016] S2. Metabolic activation Placing the insect in a 12°C low-temperature environment for 24 hours activates the endogenous enzyme system.

[0017] S3. Minimally Invasive Intervention A dual-channel responsive conduit system was implanted in the third abdominal segment of the worm. The dual-channel responsive conduit system includes: Oil phase channel: filled with poly(N-isopropylacrylamide-co-butyl methacrylate) temperature-responsive gel, accounting for 60% of the cross-sectional area, with a phase transition temperature of 28-32℃; Peptide phase channel: filled with chitosan / sodium alginate pH-responsive gel, accounting for 40% of the cross-sectional area, with a pH response threshold of 5.2.

[0018] S4. In-situ separation The exudate oil was collected at 30℃ using a temperature response mechanism, and the protein peptide solution was eluted at pH 5.0 using a pH response mechanism. In the in-situ separation step, the osmotic pressure of the parasite's hemolymph was maintained at 380-420 mOsm / kg, which was achieved by simultaneous perfusion of isotonic glucose solution.

[0019] S5. Product Refining The oil was decolorized by molecular sieve and purified by low-temperature crystallization. The protein peptide solution was concentrated by ultrafiltration and then freeze-dried. The ultrafiltration concentration was carried out using an ultrafiltration membrane with a molecular weight cutoff of 5 kDa. The proportion of components with a molecular weight ≤ 1 kDa in the obtained protein peptide product was ≥ 90%.

[0020] S6. Insect Body Recycling The surviving insects are transferred to the recovery and aquaculture system for recycling. The specific steps for insect recycling include: Use a nutrient solution containing 2g / L sericin and 0.5g / L dragon's blood extract for wound repair. Repeat the procedure after 7 days of rest, with a maximum of 3 cycles.

[0021] In protein enzymatic digestion (peptide formation), the general reaction formula is:

[0022] This allows for the activation of protease precursors (such as prozymes) at a low temperature of 12°C. Under pH 5.0 conditions (peptide phase channel environment), the activated proteases (such as cathepsins) hydrolyze proteins in hemolymph to generate small peptides.

[0023] The general reaction formula for lipid release reactions (partial hydrolysis) is:

[0024] The lipolysis pathway is activated by feeding a metabolic trigger (containing quercetin). When the temperature rises to 30°C, the HSL enzyme catalyzes the hydrolysis of triglycerides in adipocytes, releasing free fatty acids (lipid exudation). The temperature-responsive gel (hydrophobic phase transition) in the oil phase channel captures the free fatty acids.

[0025] Chemical effects of metabolic triggers: By feeding sodium 5'-inosine disodium (C 10 H 11 N4Na2O8P) and quercetin (C 15 H 10 O7 is absorbed into the blood and lymph via the intestines. Under low temperature conditions, it activates the AMPK signaling pathway.

[0026] The reaction equation for the protonation reaction of peptide phase channel gel is as follows:

[0027] Inject pH 5.0 citrate buffer, H + The gel contracts with chitosan amino groups to release protein peptides. Furthermore, during catheter implantation, the biocompatibility of the gel material (local inflammation rate <5%) and the nutrient (serin) inhibit the activity of apoptotic enzymes, thus ensuring an increased survival rate of the parasite.

[0028] Table 1: Separation Efficiency of Dual-Channel Catheter Systems

[0029] Table 2: Insect Recycling Performance Table (n=300 insects)

[0030] Table 3: Key Product Physicochemical Indicators

[0031] Table 4: Validation Table of Metabolic Pathway Regulation

[0032] The specific implementation is as follows: First, healthy bamboo worm larvae are selected and, after a 48-hour fast, fed a specially formulated metabolic trigger solution to activate the larvae's endogenous enzyme system. The larvae are then placed in a low-temperature environment for 24 hours to further enhance metabolic activity. Under aseptic conditions, a dual-channel responsive conduit system is implanted into the third abdominal segment of the larvae. This system includes a temperature-responsive oil phase channel and a pH-responsive peptide phase channel, each filled with a specific functional gel material. When the temperature is raised to 30 degrees Celsius, the oil phase channel undergoes a hydrophobic transition, continuously collecting exudated oil; simultaneously, a citrate buffer solution with a pH of 5.0 is injected into the peptide phase channel to elute the protein peptide solution. The collected oil is decolorized by molecular sieves and purified by low-temperature crystallization, while the protein peptide solution is concentrated through an ultrafiltration membrane and then freeze-dried. Post-operatively, the larvae survival rate remains high, allowing them to be transferred to a recovery and rearing system for recycling. The recovery and rearing system is equipped with wound repair nutrients to ensure the larvae recover to a reoperable state within seven days, and can be reused up to three times. Throughout the process, the osmotic pressure of the insect's hemolymph is maintained by synchronously injecting isotonic glucose solution, ensuring the stability of live handling. The final products, oils and protein peptides, both meet high-quality standards, with extremely low rancidity and peroxide values ​​in the oils, a high proportion of small-molecule peptides in the protein peptides, and excellent retention of antimicrobial peptide activity. This technology, through the innovative combination of a live metabolic engine and in-situ separation technology, achieves efficient, green, and sustainable utilization of bamboo worm resources, breaking through the limitations of traditional processes and providing a new technological paradigm for the field of biorefining.

[0033] The present invention has the following advantages: This technology eliminates the need for complex mechanical cell-wall breaking equipment and expensive exogenous enzyme preparations, significantly reducing energy consumption. Simultaneously, the live-cell metabolic process reduces the use of organic solvents, lowering raw material costs and avoiding post-processing expenses associated with solvent residues. In traditional processes, insect residue is typically treated as waste, while this technology maximizes the utilization of insect resources through a recycling mechanism, significantly improving overall economic efficiency. Furthermore, the increased yield and activity of high-value-added products (such as antimicrobial peptides) further enhance market competitiveness. Although live-cell operations require a certain recovery time, the single-extraction efficiency is significantly improved, and the elimination of multi-stage separation and purification steps results in an overall production cycle superior to traditional processes. This technology eliminates the need for organic solvents and chemical additives throughout the entire process, avoiding environmental pollution. Moreover, the live-cell metabolic process occurs under natural physiological conditions, generating no harmful byproducts, aligning with green manufacturing principles. By establishing an insect recovery and breeding system, multi-batch recycling of resources is achieved, reducing raw material consumption and embodying the principles of a circular economy. Furthermore, the insects can still be used as breeding stock or feed after the procedure, further improving resource utilization. This technology has low energy consumption and does not require energy-intensive operations such as high temperature and high pressure, resulting in significantly lower carbon emissions than traditional processes.

[0034] Although embodiments of the invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made to these embodiments without departing from the principles and spirit of the invention, the scope of which is defined by the appended claims and their equivalents.

Claims

1. A process for simultaneously extracting protein peptides and oils from bamboo worms based on enzymatic hydrolysis, specifically comprising the following steps, characterized in that: S1. Live animal preparation Select healthy bamboo worm larvae, and feed them a metabolic trigger solution after fasting for 48 hours; S2. Metabolic activation Placing the insect body in a low-temperature environment of 12℃ for 24 hours activates the endogenous enzyme system; S3. Minimally Invasive Intervention A dual-channel responsive conduit system was implanted in the third abdominal segment of the worm. S4. In-situ separation The exudated oil was collected at 30°C using a temperature-responsive mechanism, and the protein peptide solution was eluted at pH 5.0 using a pH-responsive mechanism. S5. Product Refining The oils were decolorized by molecular sieves and purified by low-temperature crystallization; the protein peptide solution was concentrated by ultrafiltration and then freeze-dried. S6. Insect Body Recycling The surviving insects are transferred to the recovery and breeding system for recycling.

2. The process for simultaneous extraction of bamboo worm protein peptides and oils based on enzymatic hydrolysis according to claim 1, characterized in that: The metabolic trigger is composed of disodium 5'-inosinate and quercetin in a mass ratio of 5:

3.

3. The process for simultaneous extraction of bamboo worm protein peptides and oils based on enzymatic hydrolysis according to claim 1, characterized in that: The metabolic trigger solution was prepared with a concentration of 10 g / L using 0.1 mol / L phosphate buffer at a pH of 7.

2.

4. The process for simultaneous extraction of bamboo worm protein peptides and oils based on enzymatic hydrolysis according to claim 1, characterized in that: The dual-channel responsive catheter system includes: Oil phase channel: filled with poly(N-isopropylacrylamide-co-butyl methacrylate) temperature-responsive gel, accounting for 60% of the cross-sectional area, with a phase transition temperature of 28-32℃; Peptide phase channel: filled with chitosan / sodium alginate pH-responsive gel, accounting for 40% of the cross-sectional area, with a pH response threshold of 5.

2.

5. The process for simultaneous extraction of bamboo worm protein peptides and oils based on enzymatic hydrolysis according to claim 1, characterized in that: In the in-situ separation step, the osmotic pressure of the parasite's hemolymph is maintained at 380-420 mOsm / kg, which is achieved by simultaneous perfusion of isotonic glucose solution.

6. The process for simultaneous extraction of bamboo worm protein peptides and oils based on enzymatic hydrolysis according to claim 1, characterized in that: The ultrafiltration concentration uses an ultrafiltration membrane with a molecular weight cutoff of 5 kDa, and the proportion of components with a molecular weight ≤ 1 kDa in the obtained protein peptide product is ≥ 90%.

7. The process for simultaneous extraction of bamboo worm protein peptides and oils based on enzymatic hydrolysis according to claim 1, characterized in that: The insect body recovery step specifically includes: Use a nutrient solution containing 2g / L sericin and 0.5g / L dragon's blood extract for wound repair; Repeat the operation after 7 days of rest, and it can be reused up to 3 times.

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