Preparation process of low-fishy-smell marine fish skin collagen peptide

Abstract

The invention belongs to the technical field of collagen peptide extraction, and discloses a preparation process of low-fishy-smell marine fish skin collagen peptide, which is used for solving the problems of incomplete fishy smell removal and high protein loss rate in the traditional fishy smell removal process. Comprising the following steps: (1) immersing fish skin into water, adding lipase, cleaning the fish skin with pure water, and homogenizing to obtain homogenate; (2) adding saccharomyces cerevisiae and purple perilla juice into the homogenate, carrying out heat preservation and enzymolysis at 30 DEG C, then heating to 45 DEG C, adding compound enzyme for enzymolysis, and heating for enzyme deactivation to obtain collagen peptide enzymatic hydrolysate; (3) adding activated carbon and diatomite into the collagen peptide enzymatic hydrolysate, keeping the temperature at 70 DEG C, stirring, and carrying out filter pressing and refined filtration to obtain a collagen peptide decoloring solution; (4) sterilizing and concentrating the collagen peptide decoloring solution to obtain a collagen peptide concentrated solution; and (5) drying the collagen peptide concentrated solution to obtain collagen peptide powder. The collagen peptide prepared by the method is almost free of fishy smell and low in protein loss.

Description

Technical Field

[0001] The present invention belongs to the technical field of collagen peptide extraction, and particularly relates to a preparation process of low - fishy - smell marine fish skin collagen peptide. Background Art

[0002] Collagen, also known as collagen, is a helical fibrous protein formed by three peptide chains. It mainly exists in the skin, bones, ligaments and blood vessels of animals, and is an extremely important structural protein of connective tissue, playing the functions of supporting organs and protecting the body. Since collagen is rich in glycine, proline and hydroxyproline, its hydrolysis product - collagen peptide has good physiological activity functions and has great application prospects in the fields of medicine, health care and beauty. Marine fish skin is a high - quality raw material for preparing collagen peptide, without the residues of antibiotics and hormones, and also without the risk of immune diseases of terrestrial animals (such as pigs, cows, etc.), with high safety.

[0003] Commercially available marine fish skin collagen peptide has always had the disadvantage of a strong fishy smell. Traditional deodorization processes include: (1) Physical methods: such as activated carbon adsorption and β - cyclodextrin embedding. Activated carbon adsorption has the disadvantages of low adsorption rate and high protein loss; β - cyclodextrin embedding has the disadvantages of unsatisfactory deodorization effect and inability to completely embed macromolecular fishy - smell substances. (2) Chemical methods: such as acid - base treatment or Maillard reaction. Acid - base treatment will damage the protein structure, affect the activity of collagen peptide and produce a large amount of waste water, with high treatment cost; the Maillard reaction will introduce a burnt smell, resulting in poor sensory properties. (3) Biological methods: microbial fermentation, such as lactic acid bacteria and yeast metabolizing fishy - smell substances. This method has the disadvantages of high protein loss rate and long fermentation time.

[0004] The above - mentioned methods generally have problems such as incomplete deodorization, high protein loss rate, and complex processes, and it is difficult for a single means to take into account the deodorization effect, product quality and production cost. Summary of the Invention

[0005] The purpose of the present invention is to provide a preparation process of low - fishy - smell marine fish skin collagen peptide, effectively solving the problems of incomplete deodorization, high protein loss rate, complex process, etc. existing in traditional deodorization processes.

[0006] To solve the above - mentioned technical problems, the technical solution adopted by the present invention is:

[0007] A preparation process of low - fishy - smell marine fish skin collagen peptide, comprising the following steps: S1. Immerse the fish skin in water with a volume 3 - 8 times that of the fish skin, add lipase and stir for 1h - 3h, then wash the fish skin with pure water and homogenize it to obtain a homogenate. The addition amount of lipase is 0.2% - 2% of the mass of the fish skin.

[0008] S2. Add Saccharomyces cerevisiae and perilla juice to the homogenate, incubate at 30 °C for enzymatic hydrolysis for 1 h to 2 h, then raise the temperature to 45 °C to 55 °C, add a complex enzyme for enzymatic hydrolysis for 1 to 3 h, and heat to 100 °C for 10 min to inactivate the enzyme, obtaining a collagen peptide enzymatic hydrolysate.

[0009] S3. Add activated carbon and diatomaceous earth to the collagen peptide enzymatic hydrolysate, incubate and stir at 70 °C to 90 °C for 20 min to 40 min, and perform plate and frame pressure filtration and fine filtration through a titanium rod activated carbon filter to obtain a clarified collagen peptide decolorized solution.

[0010] S4. Subject the collagen peptide decolorized solution to ultra-high temperature instantaneous sterilization and then concentrate it by reverse osmosis technology to obtain a collagen peptide concentrate.

[0011] S5. Spray-dry the collagen peptide concentrate to obtain a white, low-odor collagen peptide powder.

[0012] Furthermore, in step S2, the complex enzyme is composed of two or more of alkaline protease, neutral protease, bromelain, papain, and ficin, and the total addition amount of the complex enzyme is 0.35% to 0.85% of the mass of fish skin.

[0013] Furthermore, the enzyme activity of alkaline protease is 150000 U / g to 400000 U / g, the enzyme activity of neutral protease is 100000 U / g to 200000 U / g, the enzyme activity of bromelain is 100000 U / g to 800000 U / g, the enzyme activity of papain is 100000 U / g to 600000 U / g, and the enzyme activity of ficin is 300000 U / g to 1200000 U / g.

[0014] Furthermore, in step S2, the perilla juice is prepared by homogenizing fresh perilla leaves after washing, and the addition amount of perilla leaves is 0.2% to 0.8% of the mass of fish skin.

[0015] Furthermore, in step S1, the lipase is a low-temperature lipase derived from Yarrowia lipolytica.

[0016] Furthermore, in step S2, the addition amount of Saccharomyces cerevisiae is 0.05% to 0.2% of the mass of fish skin.

[0017] Furthermore, in step S3, the addition amount of diatomaceous earth is 0.1% to 0.3% of the mass of fish skin.

[0018] Furthermore, in step S3, the addition amount of activated carbon is 0.02% to 1% of the mass of fish skin, and the specific surface area of the activated carbon is not less than 1000 m 2 / g.

[0019] Further, in step S1, the fish skin is washed twice with pure water and then homogenized.

[0020] Compared with the prior art, the beneficial technical effects of the present invention are as follows:

[0021] (1) By using perilla leaves in combination with Saccharomyces cerevisiae and activated carbon in the preparation of collagen peptides, the prepared collagen peptides have a low fishy smell approaching zero, low protein loss, and high extraction rate.

[0022] (2) The present invention uses perilla leaves to remove fishy smell, which is green, mild, safe and environmentally friendly. At the same time, it realizes multi-channel collaborative deodorization, achieves all-round reduction of fishy smell substances, is green and environmentally friendly, and has zero chemical reagent residues.

[0023] (3) The present invention uses perilla leaves in combination with Saccharomyces cerevisiae and activated carbon to remove fishy smell, reduces the usage amount of Saccharomyces cerevisiae and activated carbon while reducing protein loss, saves deodorization time, improves the deodorization effect, and maximally retains the activity of collagen peptides. It is conducive to realizing industrial production.

[0024] (4) The present invention uses low-temperature lipase for degreasing treatment, avoids the temperature limitation of conventional lipase, uses room temperature water for degreasing, reduces the complexity of the process and production energy consumption, and is suitable for large-scale industrial use. Specific Embodiments

[0025] Example 1: The preparation process of low-fishy-smell marine fish skin collagen peptides provided in this example includes the following steps: S1. Immerse the fish skin in 8 times the volume of water, add lipase and stir for 1.5 h, then wash the fish skin with pure water and homogenize it to obtain a homogenate. The addition amount of lipase is 1% of the fish skin mass.

[0026] The lipase used in this example is low-temperature lipase, and the source is Yarrowia lipolytica.

[0027] S2. Add Saccharomyces cerevisiae and perilla juice to the homogenate, incubate at 30 °C for enzymatic hydrolysis for 2 h, then raise the temperature to 50 °C, add a compound enzyme and enzymatically hydrolyze for 2 h, heat to 100 °C and incubate for 10 min to inactivate the enzyme, and obtain a collagen peptide enzymatic hydrolysate.

[0028] The compound enzyme used in this example is a compound of alkaline protease and neutral protease. The total addition amount of the compound enzyme is 0.65% of the fish skin mass, and the ratio of alkaline protease to neutral protease is 2:1. The enzyme activity of the alkaline protease used is 400000 U / g, and the enzyme activity of the neutral protease used is 200000 U / g.

[0029] The perilla juice added in this example is prepared by homogenizing fresh perilla leaves after washing. The addition amount of perilla leaves is 0.8% of the fish skin mass; the addition amount of Saccharomyces cerevisiae is 0.05% of the fish skin mass.

[0030] S3. Add activated carbon and diatomaceous earth into the collagen peptide hydrolysate, keep stirring at 75 °C for 20 min, and then perform plate and frame pressure filtration and fine filtration through a titanium rod activated carbon filter to obtain a clarified decolorized collagen peptide solution.

[0031] In this example, the addition amount of diatomaceous earth is 0.1% of the fish skin mass, and the addition amount of activated carbon is 0.02% of the fish skin mass. The specific surface area of the activated carbon is not less than 1000 m 2 / g.

[0032] S4. Ultra-high temperature instantaneous sterilize the decolorized collagen peptide solution and then concentrate it by reverse osmosis technology to obtain a concentrated collagen peptide solution.

[0033] S5. Spray dry the concentrated collagen peptide solution to obtain a white collagen peptide powder with low fishy smell.

[0034] Example 2: The preparation process of low-fishy-smell marine fish skin collagen peptide provided in this example includes the following steps: S1. Immerse the fish skin in water with a volume 5 times that of the fish skin, add lipase and stir for 2 h, then wash the fish skin with pure water and homogenize it to obtain a homogenate. The addition amount of lipase is 0.2% of the fish skin mass.

[0035] The lipase used in this example is a low-temperature lipase, and its source is Yarrowia lipolytica.

[0036] S2. Add Saccharomyces cerevisiae and perilla juice into the homogenate, keep it at 30 °C for enzymatic hydrolysis for 1 h, then raise the temperature to 45 °C, add a compound enzyme and hydrolyze for 3 h, heat to 100 °C and keep it for 10 min to inactivate the enzyme, and obtain a collagen peptide hydrolysate.

[0037] The compound enzyme used in this example is compounded by alkaline protease, papain and ficin. The total addition amount of the compound enzyme is 0.35% of the fish skin mass, and the ratio of alkaline protease, papain and ficin is 2.5:1.5:1. The enzyme activity of the used alkaline protease is 400000 U / g, the enzyme activity of the used papain is 100000 U / g, and the enzyme activity of the used ficin is 1200000 U / g.

[0038] The perilla juice added in this example is prepared by homogenizing fresh perilla leaves after washing. The addition amount of perilla leaves is 0.2% of the fish skin mass; the addition amount of Saccharomyces cerevisiae is 0.2% of the fish skin mass.

[0039] S3. Add activated carbon and diatomaceous earth into the collagen peptide hydrolysate, keep stirring at 80 °C for 40 min, and then perform plate and frame pressure filtration and fine filtration through a titanium rod activated carbon filter to obtain a clarified decolorized collagen peptide solution.

[0040] In this example, the addition amount of diatomaceous earth is 0.3% of the mass of fish skin, and the addition amount of activated carbon is 1% of the mass of fish skin. The specific surface area of the activated carbon is not less than 1000m 2 / g.

[0041] S4. After subjecting the collagen peptide decolorized liquid to ultra-high temperature instantaneous sterilization, concentrate it by reverse osmosis technology to obtain a collagen peptide concentrate.

[0042] S5. Spray-dry the collagen peptide concentrate to obtain a white and low-odor collagen peptide powder.

[0043] Example 3: The preparation process of low-odor marine fish skin collagen peptide provided in this example includes the following steps: S1. Immerse the fish skin in water with a volume three times that of the fish skin, add lipase and stir for 2 h, then wash the fish skin with pure water and homogenize it to obtain a homogenate. The addition amount of lipase is 0.5% of the mass of fish skin.

[0044] The lipase used in this example is a low-temperature lipase, and its source is Yarrowia lipolytica.

[0045] S2. Add Saccharomyces cerevisiae and perilla juice to the homogenate, incubate at 30 °C for enzymatic hydrolysis for 2 h, then raise the temperature to 55 °C, add a complex enzyme for enzymatic hydrolysis for 1 h, and heat to 100 °C for 10 min to inactivate the enzyme to obtain a collagen peptide enzymatic hydrolysate.

[0046] The complex enzyme used in this example is composed of a neutral protease, bromelain, and ficin. The total addition amount of the complex enzyme is 0.85% of the mass of fish skin. The ratio of the neutral protease, bromelain, and ficin is 3.5:1.5:1. The enzyme activity of the used neutral protease is 200000 U / g, the enzyme activity of the used bromelain is 800000 U / g, and the enzyme activity of the used ficin is 300000 U / g.

[0047] The perilla juice added in this example is prepared by homogenizing fresh perilla leaves after washing. The addition amount of perilla leaves is 0.6% of the mass of fish skin; the addition amount of Saccharomyces cerevisiae is 0.1% of the mass of fish skin.

[0048] S3. Add activated carbon and diatomaceous earth to the collagen peptide enzymatic hydrolysate, incubate at 90 °C with stirring for 30 min, and perform fine filtration through a plate and frame filter press and a titanium rod activated carbon filter to obtain a clarified collagen peptide decolorized liquid.

[0049] In this example, the addition amount of diatomaceous earth is 0.2% of the mass of fish skin, and the addition amount of activated carbon is 0.05% of the mass of fish skin. The specific surface area of the activated carbon is not less than 1000m 2 / g.

[0050] S4. After subjecting the collagen peptide decolorized solution to ultra-high temperature instantaneous sterilization, it is concentrated by reverse osmosis technology to obtain a collagen peptide concentrated solution.

[0051] S5. The collagen peptide concentrated solution is spray-dried to obtain a white collagen peptide powder with low fishy smell.

[0052] The molecular weights of the marine fish skin collagen peptides obtained in Example 1, Example 2, and Example 3 were measured according to the method shown in GB / T 22729 (Marine fish oligopeptide powder), and the measurement results are shown in Table 1.

[0053] Table 1 Molecular weights of marine fish skin collagen peptides extracted by different processes

[0054]

[0055]

[0056] It can be seen from Table 1 that the proportion of protein hydrolysates with a relative molecular weight of less than 1000 u in the low-fishy-smell marine fish skin collagen peptides obtained in each example is greater than 85%, which is more conducive to human digestion and absorption.

[0057] Comparative Example 1: The difference from Example 1 is that in this comparative example, only perilla juice is used for deodorization alone, and no Saccharomyces cerevisiae and activated carbon are added.

[0058] Comparative Example 2: The difference from Example 1 is that in this comparative example, only Saccharomyces cerevisiae is used for deodorization alone, and no perilla juice and activated carbon are added.

[0059] Comparative Example 3: The difference from Example 1 is that in this comparative example, only activated carbon is used for deodorization alone, and no perilla juice and Saccharomyces cerevisiae are added.

[0060] The fishy smell degrees of the marine fish skin collagen peptides obtained in Examples 1-3 and Comparative Examples 1-3 were evaluated by sensory evaluation. The sensory evaluation panel consisted of 20 sensory evaluators (10 males and 10 females), who respectively evaluated the fishy smell degrees of the marine fish skin collagen peptides prepared in Examples 1-3 and Comparative Examples 1-3.

[0061] The sensory scoring standard is: the score for fishy smell from light to heavy is from 0 to 5 points, 0 point means no fishy smell, 5 points means extremely strong fishy smell, the score can be accurate to one decimal place, and the final score is the average score, reserved to two decimal places.

[0062] The evaluation results of the fishy smell degree are shown in Table 2. The effect of using perilla leaves combined with Saccharomyces cerevisiae and activated carbon to remove fishy smell in Examples 1-3 is significantly improved compared with the effect of the three deodorizing alone in each comparative example, and the fishy smell of the marine fish skin collagen peptides prepared in Examples 1-3 is significantly reduced. And it can be found from the comparative examples that perilla juice is superior to yeast fermentation and activated carbon treatment in deodorization.

[0063] Table 2 shows the comparison of fishy smells of marine fish skin collagen peptides extracted by different processes

[0064] Degree of fishy smell Example 1 0.55 Example 2 1.12 Example 3 0.88 Comparative Example 1 1.71 Comparative Example 2 2.63 Comparative Example 3 2.19

[0065] The present invention creatively uses perilla leaves in combination with Saccharomyces cerevisiae and activated carbon in the preparation of collagen peptides, resulting in collagen peptides with low fishy smell, almost no fishy smell, low protein loss, and high extraction rate. The main active ingredient perillyl alcohol in perilla leaves can convert key fishy substances (such as amines like trimethylamine and dimethylamine) into odorless carboxylic acid substances through redox reactions. Volatile terpene substances such as perillene and limonene in perilla leaves, with their strong aroma (a mixture of herbal fragrance and fruit fragrance), inhibit the brain's reception of fishy smell signals through the olfactory competition mechanism and reduce the fishy smell perception threshold. At the same time, polyphenolic substances in perilla juice can inhibit aldehyde and ketone fishy smell substances (such as hexanal and nonenal) produced by lipid oxidation. Perilla leaves can achieve multi-path collaborative deodorization, comprehensively reducing fishy smell substances, being green and environmentally friendly with zero chemical reagent residues. At the same time, in combination with Saccharomyces cerevisiae and activated carbon, it reduces the usage amounts of the two while reducing protein loss, saves deodorization time, improves the deodorization effect, and is conducive to industrial production.

[0066] Certainly, the above description is not a limitation of the present invention, and the present invention is not limited to the above examples. Changes, modifications, additions, or substitutions made by those skilled in the art within the scope of the essence of the present invention should also fall within the protection scope of the present invention.

Claims

1. A preparation process of low - fishy - smell marine fish skin collagen peptide, characterized in that, It includes the following steps: S1. Immerse fish skin in water with a volume 3 - 8 times that of the fish skin, add lipase and stir for 1 h - 3 h, then wash the fish skin with pure water and homogenize it to obtain a homogenate. The addition amount of lipase is 0.2% - 2% of the mass of the fish skin; S2. Add Saccharomyces cerevisiae and perilla juice to the homogenate, incubate at 30 °C for enzymatic hydrolysis for 1 h - 2 h, then raise the temperature to 45 °C - 55 °C, add a complex enzyme for enzymatic hydrolysis for 1 h - 3 h, heat to 100 °C and keep warm for 10 min to inactivate the enzyme, obtaining a collagen peptide enzymatic hydrolysate; S3. Add activated carbon and diatomaceous earth to the collagen peptide enzymatic hydrolysate, keep warm and stir at 70 °C - 90 °C for 20 min - 40 min, and filter through a plate and frame filter press and a titanium rod activated carbon filter to obtain a clarified collagen peptide decolorized solution; S4. Subject the collagen peptide decolorized solution to ultra-high temperature instantaneous sterilization and then concentrate it by reverse osmosis technology to obtain a collagen peptide concentrate; S5. Spray-dry the collagen peptide concentrate to obtain collagen peptide powder.

2. The preparation process of the low-fishy-smell marine fish skin collagen peptide according to claim 1, characterized in that, In step S2, the complex enzyme is composed of two or more of alkaline protease, neutral protease, bromelain, papain and ficin, and the total addition amount of the complex enzyme is 0.35% - 0.85% of the mass of the fish skin.

3. The preparation process of the low-odor marine fish skin collagen peptide according to claim 2, wherein, The enzyme activity of alkaline protease is 150000 U / g - 400000 U / g, the enzyme activity of neutral protease is 100000 U / g - 200000 U / g, the enzyme activity of bromelain is 100000 U / g - 800000 U / g, the enzyme activity of papain is 100000 U / g - 600000 U / g, and the enzyme activity of ficin is 300000 U / g - 1200000 U / g.

4. The preparation process of the low-odor marine fish skin collagen peptide according to claim 3, characterized in that, In step S2, the perilla juice is prepared by washing fresh perilla leaves and then homogenizing them. The addition amount of perilla leaves is 0.2% - 0.8% of the mass of the fish skin.

5. The preparation process of the low-fishy-smell marine fish skin collagen peptide according to claim 4, characterized in that, In step S1, the lipase is a low-temperature lipase derived from Yarrowia lipolytica.

6. The preparation process of the low-fishy-taste marine fish skin collagen peptide according to claim 5, wherein In step S2, the addition amount of Saccharomyces cerevisiae is 0.05% - 0.2% of the mass of the fish skin.

7. The preparation process of the low-odor marine fish skin collagen peptide according to claim 6, characterized in that, In step S3, the addition amount of diatomaceous earth is 0.1% - 0.3% of the mass of the fish skin.

8. The preparation process of the low-odor marine fish skin collagen peptide according to claim 7, characterized in that, In step S3, the addition amount of activated carbon is 0.02% to 1% of the mass of fish skin, and the specific surface area of the activated carbon is not less than 1000 m 2 / g.

9. The preparation process of the low-odor marine fish skin collagen peptide according to claim 8, characterized in that, In step S1, wash the fish skin with pure water twice and then homogenize it.

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