Coproduction method of bovine-derived Muslim chondroitin sulfate and collagen peptide based on composite enzymolysis-membrane separation coupling technology

Abstract

The invention belongs to the technical field of biological product extraction, and particularly relates to a bovine-derived Muslim chondroitin sulfate and collagen peptide co-production method based on a composite enzymolysis-membrane separation coupling technology, which comprises the steps of pretreatment of bovine cartilage, enzymolysis, membrane analysis and filtration, extraction of chondroitin and collagen peptide and the like. According to the method, animal source enzyme is replaced by microbial enzyme, religious taboo is avoided, resources are efficiently utilized, two high-value products are synchronously extracted, and the raw material cost is reduced; a membrane separation technology replaces an organic solvent, and pollution is reduced.

Description

Technical Field

[0001] This invention belongs to the field of bioproduct extraction technology, and particularly relates to a method for the co-production of bovine halal chondroitin sulfate and collagen peptides based on a composite enzymatic hydrolysis-membrane separation coupling technology. Background Technology

[0002] Bovine cartilage is a highly differentiated connective tissue in cattle, providing support and protection. It is mainly composed of chondrocytes (10%-20%) and cartilage matrix (80%-90%). The core components of the matrix include collagen, chondroitin sulfate (CS), glucosamine (GlcN), and minerals. The collagen is primarily type II collagen, forming the fibrous framework of the cartilage and maintaining its structural stability. Chondroitin sulfate (CS), a glycosaminoglycan, combines with collagen to form proteoglycans, giving the cartilage elasticity and resistance to compression. Glucosamine (GlcN) is a precursor for the synthesis of the cartilage matrix and participates in cartilage repair. Minerals, such as calcium and phosphorus, are present in small amounts and help maintain cartilage stiffness.

[0003] Traditional chondroitin sulfate (CS) extraction often employs an alkaline extraction-trypsin method, which poses significant environmental pollution and risks to halal compliance. Furthermore, current technologies require multiple independent processes to separate CS from collagen (CPP), resulting in incomplete separation of collagen and chondroitin, a chondroitin yield of <15%, low efficiency, and the generation of large amounts of waste liquid. Additionally, imperfect application of membrane separation technology leads to an excessively wide molecular weight distribution, with a CV value >25%. Collagen peptide extraction often faces problems such as a wide molecular weight distribution, a strong bitter and fishy taste, and high salt content.

[0004] Chinese invention patent 201510007701.2 describes a method for the co-production of hyaluronic acid, chondroitin sulfate, collagen peptides, bone meal feed, and soap. The method involves soaking animal cartilage in water and pulverizing it to obtain cartilage powder. The cartilage powder is then enzymatically hydrolyzed with protease, adsorbed onto a resin, and the enzymatically hydrolysate is collected. The resin is then subjected to gradient elution with NaCl aqueous solution. Each eluent is desalted, concentrated, sterilized, and dried to obtain hyaluronic acid and chondroitin sulfate products, respectively. The adsorbed enzymatically hydrolysate is then decolorized, treated with a mixed column of anion and cation exchange resins, nanofiltration for impurity removal, concentrated, sterilized, and dried to obtain collagen peptides. This patent utilizes a resin adsorption-spray drying process to achieve high resource utilization of cartilage. However, the patent uses compound enzyme A, papain, trypsin, alkaline protease, and compound enzyme B, neutral protease, flavor protease, etc., which are enzymatically hydrolyzed in stages, but it does not clarify whether the source of the enzyme preparation is halal. For example, trypsin may come from pigs and does not meet halal requirements; and there are also technical problems such as insufficient separation purity and complex process. Summary of the Invention

[0005] To address the above technical issues, this invention provides a method for the co-production of bovine halal chondroitin sulfate and collagen peptides based on a composite enzymatic hydrolysis-membrane separation coupling technology. The raw materials are sourced under halal conditions, and microbial enzymes are used instead of animal-derived enzymes to avoid religious taboos, ensuring efficient resource utilization and simultaneous extraction of two high-value products, thus reducing raw material costs. Membrane separation technology replaces organic solvents, reducing pollution. The method offers high purity and a simple production process.

[0006] The present invention discloses a method for the co-production of bovine halal chondroitin sulfate and collagen peptides based on a composite enzymatic hydrolysis-membrane separation coupling technology, comprising the following steps:

[0007] Includes the following steps:

[0008] (1) Raw material pretreatment: Take beef cartilage from a halal slaughterhouse, wash and crush it;

[0009] (2) Enzymatic hydrolysis: The enzyme is a complex enzyme, which is composed of alkaline protease, neutral protease, lipase and mesophilic amylase in a weight ratio of 4.8-5.1:2.9-3.2:1.4-1.8:0.4-0.6, pH 6.5-8; material-to-liquid ratio 1:7.8-8.2;

[0010] (3) Membrane separation system and filtration: The membrane separation system consists of a 10kDa filter membrane to remove macromolecular impurities, a 30kDa ultrafiltration membrane to separate CS and CPP, and a 1kDa nanofiltration membrane to enrich collagen peptides.

[0011] (4) Extraction of chondroitin and collagen peptides: electrodialysis desalting and refining chondroitin sulfate.

[0012] The enzyme in step (2) also includes chondroitinase, which is 1% of the total mass of the complex enzymes. Chondroitinase is an enzyme of microbial origin.

[0013] In step (2), the complex enzyme is composed of alkaline protease, neutral protease, lipase and mesophilic amylase in a weight ratio of 5:3:1.5:0.5.

[0014] The optimized formulation of the compound enzyme includes the following components by weight: 50% alkaline protease, 30% neutral protease, 15% lipase, and 5% mesophilic amylase.

[0015] The enzymatic hydrolysis conditions are as follows: material-to-liquid ratio 1:8, temperature 45-55℃, time 6-7h, and compound enzyme addition amount 0.6%.

[0016] Key to Halal: Both the compound enzyme and chondroitin enzyme are derived from microbial fermentation, contain no animal ingredients, and avoid animal-derived enzymes.

[0017] This invention establishes a fully halal production system that complies with Islamic law, preventing cross-contamination with non-halal materials throughout the process. Animal-derived reagents are used, and the enzyme preparations meet ISO halal certification standards. Halal-certified bovine proteases are used, preferably alkaline proteases produced by Bacillus subtilis fermentation.

[0018] The step-by-step enzymatic hydrolysis technique used in step (2) is as follows:

[0019] First stage: Add alkaline protease and neutral protease at 45℃-50℃, hydrolyze for 4-5 hours, pH 6.5;

[0020] Second stage: Add lipase and medium-temperature amylase at 50℃, hydrolyze for 1-2 hours, pH 8.0; release collagen peptides, hydrolyze collagen into low molecular weight peptides.

[0021] The third stage: chondroitinase-directed depolymerization at 50-55℃.

[0022] The membrane separation system in step (3):

[0023] Specifically, Level 1: 100kDa ceramic microfiltration membrane, to remove suspended impurities and undissociated tissue;

[0024] Secondary stage: Ultrafiltration membrane 30kDa, retaining chondroitin sulfate molecular weight cutoff of 20-50kDa; separating chondroitin sulfate from collagen peptides.

[0025] Stage 3: 200 Da nanofiltration membrane, desalting and concentrating collagen peptides <5 kDa. The nanofiltration membrane enriches collagen peptides. Membrane separation precisely controls peptide molecular weight, eliminating bitterness and fishy taste; water solubility >98%.

[0026] The product purification and extraction in step (4) are as follows:

[0027] Chondroitin sulfate: desalted by electrodialysis, precipitated with ethanol and dried, purity ≥95%.

[0028] Collagen peptides: spray-dried from nanofiltration permeate, with moisture content controlled at ≤5%, and 90% of the molecular weight concentrated in the 300-1000 Da range.

[0029] In step (1), the particles are washed and then crushed to a particle size ≤1mm.

[0030] In step (1), the fat is removed by boiling in water at 85-95℃ with a water-to-material ratio of 2.5-3.5:1; preferably, the fat is removed by boiling in water at 90℃ with a water-to-material ratio of 3:1. Alternatively, supercritical CO2 can be used for degreasing before pulverization.

[0031] The products produced in this invention meet halal certification standards and have been tested by the Ministry of Islamic Affairs and found to be free of external contamination. The co-production yield is significantly improved, with chondroitin yield at 21.3% and collagen peptide yield at ≥35%. The products have a concentrated molecular weight distribution and a CV value <12%. Compared to the acid-base method, wastewater COD is reduced by 70%. Simultaneous production of both products improves efficiency by 40% compared to traditional stepwise processes. The membrane life is >12 months, and the utilization rate of bovine bone resources reaches 90%.

[0032] The bovine halal chondroitin sulfate and collagen peptides extracted in this invention are suitable for the production of raw materials for halal food, pharmaceuticals and cosmetics. Detailed Implementation

[0033] The present invention will be further described below with reference to specific embodiments. The following reagents were purchased on the market, and the instruments used are commonly used instruments on the market: bovine cartilage was obtained in accordance with halal slaughter standards; chondroitinase was purchased from Chifeng Pharmaceutical Co., Ltd., and was of microbial origin; neutral protease was purchased from Henan Jushuo Biotechnology Co., Ltd., and was extracted by fermentation of Bacillus subtilis, with an enzyme activity of 50,000-200,000 U / g; alkaline protease was purchased from Hubei Jikaixing Biotechnology Co., Ltd., and was produced by fermentation using genetically engineered strains (Bacillus subtilis); lipase was purchased from Shenzhen Lvweikang Biotechnology Co., Ltd., and was produced by microbial fermentation (LVK-F series), with an enzyme activity of 5,000-50,000 U / g; mesophilic α-amylase was purchased from Wuhan Jiyesheng Chemical Co., Ltd., and was produced by fermentation using Bacillus subtilis.

[0034] Example 1

[0035] A method for the co-production of bovine halal chondroitin sulfate and collagen peptides based on a composite enzymatic hydrolysis-membrane separation coupling technology, the specific steps of which are as follows:

[0036] (1) Raw material processing: Take the cartilage of cattle slaughtered in halal areas. The cartilage has a water content of 65%. After washing, crush it into 0.5mm particles. Boil it in water at 90℃ to remove fat. The water-to-material ratio is 3:1.

[0037] Degreasing: Supercritical CO2 degreasing is used at a pressure of 25 MPa and a temperature of 45℃, with residual fat content <0.5%.

[0038] Further pulverize the enzyme by freezing it with liquid nitrogen to 80 mesh, increasing the enzyme contact area.

[0039] (2) Enzymatic hydrolysis: Add pH 7.5 buffer at a material-to-liquid ratio of 1:8, and add 0.6% of complex enzyme, which consists of 50% alkaline protease, 30% neutral protease, 15% lipase and 5% medium-temperature amylase; Inactivate enzyme: maintain at 80℃ for 15 min, for a total of 7 h.

[0040] The step-by-step enzymatic hydrolysis-membrane separation coupling technology employed:

[0041] First stage: Add 50% alkaline protease and 30% neutral protease at 45℃, pH 6.5, and hydrolyze for 4 hours; release glycosaminoglycans containing chondroitin sulfate while preserving the structural integrity of collagen.

[0042] Second stage: Add 15% lipase and 5% medium-temperature amylase at 50℃, pH 8.0, and hydrolyze for 2 hours; release collagen peptides, and hydrolyze collagen into low molecular weight peptides.

[0043] The third stage involves targeted depolymerization of chondroitin sulfate by specific chondroitinase at 55℃ for 1 hour at pH 7.0, further hydrolyzing chondroitin sulfate into low molecular weight chondroitin sulfate.

[0044] (3) Membrane separation: a 10kDa filter membrane removes macromolecular impurities, a 30kDa ultrafiltration membrane separates CS and CPP, and a 1kDa nanofiltration membrane enriches collagen peptides.

[0045] Specifically, Level 1: 100kDa ceramic membrane, to remove suspended impurities and undissociated tissue;

[0046] Secondary stage: Ultrafiltration membrane (UF) 30kDa (molecular weight cutoff), retaining CS molecular weight 30-50kDa; separating CS and CPP.

[0047] The retention solution was a glycosaminoglycan complex containing chondroitin sulfate; the permeate solution contained collagen peptides and small molecule impurities.

[0048] Stage 3: The nanofiltration membrane (NF) is a 1 kDa polyamide composite membrane that desalinates and concentrates collagen peptides <5 kDa. The retainer is concentrated collagen peptides with a molecular weight of 300-1000 Da and a purity >95%; the permeate is a solution with inorganic salts and free amino acids discarded.

[0049] Grouped membrane separation enables the directional separation of chondroitin sulfate with a molecular weight of 40–80 kDa from collagen peptides with a molecular weight of 0.3–1 kDa.

[0050] (4) Drying: Spray drying of the permeate (collagen peptides) and the retentate (chondroitin) separately to refine and extract.

[0051] Chondroitin sulfate: The ultrafiltration concentrate is desalted by electrodialysis, precipitated with ethanol and dried, with a purity of ≥90%.

[0052] Collagen peptides: Spray-dried from nanofiltration permeate, with moisture content controlled at ≤5%, and 80% of the molecular weight concentrated in the range of 500-3000 Da.

[0053] This invention uses no animal-derived reagents throughout the entire process and complies with Halal certification. Through staged enzymatic hydrolysis and multi-level membrane matching, it achieves efficient separation of chondroitin sulfate and collagen peptides in one step, with a cross-contamination rate of <5%. The purity of chondroitin sulfate is ≥90%, and 90% of the collagen peptides (CPP) have a molecular weight of 300-1000 Da.

[0054] Example 2

[0055] A method for the co-production of bovine halal chondroitin sulfate and collagen peptides based on a composite enzymatic hydrolysis-membrane separation coupling technology, comprising the following steps:

[0056] (1) Raw material pretreatment: Take beef cartilage from a halal slaughterhouse, wash and crush it, boil it in water at 95℃ to remove fat, with a water-to-material ratio of 3.5:1; after washing, crush it to a particle size ≤1mm.

[0057] (2) Enzymatic hydrolysis: The enzyme is a complex enzyme, composed of alkaline protease, neutral protease, lipase and mesophilic amylase in a weight ratio of 5.1:3.2:1.8:0.6. The enzyme also includes chondroitinase, which accounts for 1% of the total mass of the complex enzyme. The chondroitinase is an enzyme of microbial origin.

[0058] Enzymatic hydrolysis conditions: material-to-liquid ratio 1:8, pH 8.5, hydrolysis temperature 45-55℃, pH 6.5-8, time 7h, compound enzyme addition amount 0.6%.

[0059] Specifically, the step-by-step enzymatic hydrolysis technique:

[0060] First stage: Add alkaline protease and neutral protease at 45℃ and hydrolyze for 4-5 hours;

[0061] Second stage: Add lipase and medium-temperature amylase at 50℃, hydrolyze for 1-2 hours, pH 8.0; release collagen peptides, hydrolyze collagen into low molecular weight peptides.

[0062] Third stage: directional depolymerization of chondroitin at 55℃.

[0063] (3) Membrane separation system and filtration: The membrane separation system consists of a 10kDa filter membrane to remove macromolecular impurities, a 30kDa ultrafiltration membrane to separate CS and CPP, and a 1kDa nanofiltration membrane to enrich collagen peptides.

[0064] Specifically, Level 1: 100kDa ceramic microfiltration membrane, to remove suspended impurities and undissociated tissue;

[0065] Secondary stage: Ultrafiltration membrane 30kDa, retaining chondroitin sulfate molecular weight cutoff of 20-50kDa; separating chondroitin sulfate from collagen peptides.

[0066] Stage 3: 200 Da nanofiltration membrane, desalting and concentrating collagen peptides <5 kDa. The nanofiltration membrane enriches collagen peptides. Membrane separation precisely controls peptide molecular weight, eliminating bitterness and fishy taste; water solubility >98%.

[0067] (4) Extraction of chondroitin and collagen peptides: electrodialysis desalting and refining chondroitin sulfate.

[0068] Chondroitin sulfate: desalted by electrodialysis, precipitated with ethanol and dried, purity ≥95%.

[0069] Collagen peptides: Spray-dried from nanofiltration permeate, with moisture content controlled to ≤5%, a.

[0070] Based on 100 kg of bovine cartilage, the enzymatic hydrolysate volume is 800 L, and the CS content is 1.8 g / L; ultrafiltration concentrates CS, yielding 18.5 kg with a purity of 96.2%; nanofiltration collagen peptides yield 35.2 kg, with a molecular weight distribution of 90% concentrated in the 300-1000D range.

[0071] Example 3

[0072] A method for the co-production of bovine halal chondroitin sulfate and collagen peptides based on a composite enzymatic hydrolysis-membrane separation coupling technology, comprising the following steps:

[0073] (1) Raw material pretreatment: Take beef cartilage from a halal slaughterhouse, wash and crush it, boil it in water at 85℃ to remove fat, with a water-to-material ratio of 2.5:1; after washing, crush it to a particle size ≤1mm.

[0074] (2) Enzymatic hydrolysis: The enzyme is a complex enzyme, which is composed of alkaline protease, neutral protease, lipase and mesophilic amylase in a weight ratio of 4.8:2.9:1.4:0.4, with a pH of 7.0-8;

[0075] Enzymatic hydrolysis conditions: material-to-liquid ratio 1:8.2, temperature 45-50℃, time 6h, compound enzyme addition 0.6%.

[0076] Specifically, the step-by-step enzymatic hydrolysis technique:

[0077] First stage: Add alkaline protease and neutral protease at 45℃, hydrolyze for 5 hours, pH 7;

[0078] Second stage: Add lipase and medium-temperature amylase at 50℃, hydrolyze for 1 hour, pH 8.0; release collagen peptides, hydrolyze collagen into low molecular weight peptides.

[0079] (3) Membrane separation system and filtration: The membrane separation system consists of a 10kDa filter membrane to remove macromolecular impurities, a 30kDa ultrafiltration membrane to separate CS and CPP, and a 1kDa nanofiltration membrane to enrich collagen peptides.

[0080] Specifically, Level 1: 100kDa ceramic microfiltration membrane, to remove suspended impurities and undissociated tissue;

[0081] Secondary stage: Ultrafiltration membrane 30kDa, retaining chondroitin sulfate molecular weight cutoff of 20-50kDa; separating chondroitin sulfate from collagen peptides.

[0082] Stage 3: 200 Da nanofiltration membrane, desalting and concentrating collagen peptides <5 kDa. The nanofiltration membrane enriches collagen peptides. Membrane separation precisely controls peptide molecular weight, eliminating bitterness and fishy taste; water solubility >98%.

[0083] (4) Extraction of chondroitin and collagen peptides: electrodialysis desalting and refining chondroitin sulfate.

[0084] Chondroitin sulfate: desalted by electrodialysis, precipitated with ethanol and dried, purity ≥95%.

[0085] Collagen peptides: spray-dried from nanofiltration permeate, with moisture content controlled at ≤5%, and 90% of the molecular weight concentrated in the 300-1000 Da range.

[0086] Example 4

[0087] A method for the co-production of bovine halal chondroitin sulfate and collagen peptides based on a composite enzymatic hydrolysis-membrane separation coupling technology, comprising the following steps:

[0088] (1) Raw material pretreatment: Take 1 kg of bovine cartilage, defatt it with supercritical CO2 and then crush it with liquid nitrogen.

[0089] (2) Enzymatic hydrolysis: The enzyme is a complex enzyme, and the complex enzyme formula consists of the following components by weight: 50% alkaline protease, 30% neutral protease, 15% lipase and 5% mesophilic amylase.

[0090] Enzymatic hydrolysis conditions: material-to-liquid ratio 1:7.8, time 7h, compound enzyme addition 0.6%.

[0091] Specifically, the step-by-step enzymatic hydrolysis technique:

[0092] First stage: Add alkaline protease and neutral protease at 48℃, hydrolyze for 4.5 hours, pH 6.5;

[0093] Second stage: Lipase and medium-temperature amylase are added at 50℃ and hydrolyzed for 1.5 hours at pH 8.0; collagen peptides are released, and the hydrolyzed collagen is converted into low molecular weight peptides.

[0094] Third stage: Chondroitinase-directed depolymerization at 55℃ for 1 hour, pH 7.0.

[0095] (3) Membrane separation system and filtration: The membrane separation system consists of a 10kDa filter membrane to remove macromolecular impurities, a 30kDa ultrafiltration membrane to separate CS and CPP, and a 1kDa nanofiltration membrane to enrich collagen peptides.

[0096] Specifically, Level 1: 100kDa ceramic microfiltration membrane, to remove suspended impurities and undissociated tissue;

[0097] Second stage: Ultrafiltration membrane 30kDa, retaining chondroitin sulfate molecular weight of 20-50kDa; separating chondroitin sulfate from collagen peptides. Third stage: Nanofiltration membrane 200Da, desalting and concentrating collagen peptides to <5kDa. Nanofiltration membrane enriches collagen peptides. Membrane separation precisely controls peptide molecular weight, eliminating bitterness and fishy taste; water solubility >98%.

[0098] (4) Extraction of chondroitin and collagen peptides: electrodialysis desalting and refining chondroitin sulfate.

[0099] Chondroitin sulfate: desalted by electrodialysis, precipitated with ethanol and dried to obtain 92.3 g of CS with a purity of 91.5%.

[0100] Collagen peptides: spray-dried from nanofiltration permeate, moisture content controlled to ≤5%, CPP 68.4g, molecular weight distribution: 88% of the product is 500Da.

[0101] The above embodiments / experimental examples are merely illustrative and not intended to limit the implementation methods. Those skilled in the art will recognize that various variations or modifications can be made based on the above description. It is neither necessary nor possible to exhaustively list all possible implementation methods. However, obvious variations or modifications derived therefrom remain within the scope of this invention.

Claims

1. A method for the co-production of bovine halal chondroitin sulfate and collagen peptides based on a composite enzymatic hydrolysis-membrane separation coupling technology, characterized in that: Includes the following steps: (1) Raw material pretreatment: Take beef cartilage from a halal slaughterhouse, wash and crush it; (2) Enzymatic hydrolysis: The enzyme is a complex enzyme, which is composed of alkaline protease, neutral protease, lipase and mesophilic amylase in a weight ratio of 4.8-5.1:2.9-3.2:1.4-1.8:0.4-0.

6. (3) Three-stage membrane separation system and filtration: The three-stage membranes are: a 100kDa ultrafiltration membrane to remove macromolecular impurities, a 10kDa nanofiltration membrane to enrich collagen peptides and a 1kDa nanofiltration membrane to enrich collagen peptides. (4) Extraction of chondroitin and collagen peptides: electrodialysis desalting and refining of chondroitin.

2. The method for co-producing bovine halal chondroitin sulfate and collagen peptides based on composite enzymatic hydrolysis-membrane separation coupling technology according to claim 1, characterized in that: The enzyme in step (2) also includes chondroitinase, which is 1% by total mass of the complex enzyme.

3. The method for co-producing bovine halal chondroitin sulfate and collagen peptides based on composite enzymatic hydrolysis-membrane separation coupling technology according to claim 1 or 2, characterized in that: The compound enzyme formulation comprises the following components by weight: 50% alkaline protease, 30% neutral protease, 15% lipase, and 5% mesophilic amylase; the enzymatic hydrolysis temperature is 45-55℃, the pH is 6.5-8, the material-to-liquid ratio is 1:7.8-8.2, and the hydrolysis time is 6-7 hours.

4. The method for co-producing bovine halal chondroitin sulfate and collagen peptides based on composite enzymatic hydrolysis-membrane separation coupling technology according to claim 1 or 3, characterized in that: The enzymatic hydrolysis conditions are as follows: material-to-liquid ratio 1:7.8-8.2, temperature 45-55℃, time 6-7h, and compound enzyme addition amount 0.6%.

5. The method for co-producing bovine halal chondroitin sulfate and collagen peptides based on composite enzymatic hydrolysis-membrane separation coupling technology according to claim 3, characterized in that: The step-by-step enzymatic hydrolysis used in step (2) is as follows: First stage: Add alkaline protease and neutral protease at 45℃-50℃ and hydrolyze for 4-5 hours; Second stage: Add lipase and medium-temperature amylase at 50℃ and hydrolyze for 1-2 hours; Second stage: directional depolymerization of chondroitin at 50-55℃.

6. The method for co-producing bovine halal chondroitin sulfate and collagen peptides based on composite enzymatic hydrolysis-membrane separation coupling technology according to claim 1, characterized in that: The product purification and extraction in step (4) are as follows: Chondroitin sulfate: desalted by electrodialysis, precipitated with ethanol and dried; Collagen peptides: Spray-dried from nanofiltration permeate, with moisture content controlled to ≤5%.

7. The method for co-producing bovine halal chondroitin sulfate and collagen peptides based on composite enzymatic hydrolysis-membrane separation coupling technology according to claim 1, characterized in that: In step (1), the particles are washed and then crushed to a particle size ≤1mm.

8. The method for co-producing bovine halal chondroitin sulfate and collagen peptides based on composite enzymatic hydrolysis-membrane separation coupling technology according to claim 1, characterized in that: In step (1), the fat is removed by boiling in water at 85-95℃ with a water-to-material ratio of 2.5-3.5:1; preferably, the fat is removed by boiling in water at 90℃ with a water-to-material ratio of 3:1.