CN110607293A - Complex enzyme and application thereof in animal protein production - Google Patents
Complex enzyme and application thereof in animal protein production Download PDFInfo
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- CN110607293A CN110607293A CN201910988470.6A CN201910988470A CN110607293A CN 110607293 A CN110607293 A CN 110607293A CN 201910988470 A CN201910988470 A CN 201910988470A CN 110607293 A CN110607293 A CN 110607293A
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- C12N9/48—Hydrolases (3) acting on peptide bonds (3.4)
- C12N9/50—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25)
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P21/00—Preparation of peptides or proteins
- C12P21/06—Preparation of peptides or proteins produced by the hydrolysis of a peptide bond, e.g. hydrolysate products
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y304/00—Hydrolases acting on peptide bonds, i.e. peptidases (3.4)
- C12Y304/22—Cysteine endopeptidases (3.4.22)
- C12Y304/22002—Papain (3.4.22.2)
Abstract
The invention relates to the field of animal protein raw material processing, in particular to a complex enzyme and application thereof in animal protein production. The compound enzyme can efficiently carry out enzymolysis on animal protein raw materials through the synergistic effect of neutral protease, NSP enzyme, papain and disodium dihydrogen pyrophosphate, so that the content of easily digested and absorbed proteins such as small peptides, amino acids and the like is increased, the palatability of the animal protein is improved, and the protein utilization rate is increased.
Description
Technical Field
The invention relates to the field of animal protein raw material processing, in particular to a complex enzyme and application thereof in animal protein production.
Technical Field
The feed industry in China continues to develop at a high speed for thirty years, the yield and the sales volume jump the first place in the world, but the problem of non-negligible raw material shortage is also exposed, and the problem of contradiction between raw material supply and demand is increasingly prominent. Especially, protein raw materials mainly depend on import, and the continuous and healthy development of the feed industry and the breeding industry in China is severely restricted. Therefore, the search for the development of novel protein feed resources and the improvement of the protein utilization rate become important subjects for relieving the shortage of protein feed raw materials in China.
The existing protein degradation methods mainly comprise physical methods, chemical methods, biological methods and the like. The physical degradation method has large equipment investment and narrow modification range; the chemical degradation method has harsh reaction conditions, damaged amino acid and serious environmental pollution caused by residual chemical substances, so that the chemical degradation method and the amino acid are not emphasized by enterprises and industries in marketization competition. In recent years, with the continuous expansion of the application field of biotechnology and the continuous and deep research on enzyme preparation technology and immobilized enzyme technology, people gradually find that the protein hydrolysis by adopting the biological enzyme method has high efficiency, mild conditions, green and environmental protection, and the hydrolysate is superior to the physical and chemical hydrolysis method in the aspects of nutrition, flavor, technology and the like. The main products of the hydrolyzed protein by the enzyme method are various functional small peptides which can be combined with special receptors in the intestinal tract, promote the growth and development of the gastrointestinal tract of animals, improve the digestion and absorption functions of the gastrointestinal tract, ensure that part of the small peptides can be absorbed into a blood circulation system to adjust the immune function of the animals, regulate the growth of the animals through a growth axis and fully play the production potential of the animals. Therefore, the improvement of the conversion and utilization efficiency of protein feed resources by the enzymolysis digestion technology has become a hot spot of great concern in the industry.
Animal protein raw materials are mostly low-value and low-conversion and absorption materials, and a considerable part of the animal protein raw materials are not easily digested due to interaction (combination, coating and the like) of sugar, fat and other substances, and can also influence the degradation of the animal protein raw materials by pepsin. Therefore, how to improve the conversion and utilization efficiency of protein raw materials and increase the content of easily digested and absorbed proteins such as small peptides and amino acids by an enzymolysis and digestion technology has become a problem to be solved in the industry.
Disclosure of Invention
The invention provides a complex enzyme and application thereof in animal protein production for solving the problems of the prior art. The complex enzyme can efficiently carry out enzymolysis on animal protein raw materials through the synergistic effect of a plurality of enzymes, so that the content of easily digested and absorbed proteins such as small peptides, amino acids and the like is increased, the palatability of the animal protein is improved, and the protein utilization rate is increased.
The invention provides a complex enzyme which comprises neutral protease, NSP enzyme and papain.
The complex enzyme also comprises disodium dihydrogen pyrophosphate.
The composite enzyme comprises the following components in parts by weight: 20-24 parts of neutral protease, 3-6 parts of NSP enzyme, 4-9 parts of papain and 1-3 parts of disodium dihydrogen pyrophosphate.
Further preferably, the complex enzyme comprises the following components in parts by weight: 24 parts of neutral protease, 5 parts of NSP enzyme, 9 parts of papain and 2 parts of disodium dihydrogen pyrophosphate.
Further preferably, the complex enzyme comprises the following components in parts by weight: 20 parts of neutral protease, 3 parts of NSP enzyme, 6 parts of papain and 1 part of disodium dihydrogen pyrophosphate.
Further preferably, the complex enzyme comprises the following components in parts by weight: 21 parts of neutral protease, 6 parts of NSP enzyme, 4 parts of papain and 3 parts of disodium dihydrogen pyrophosphate.
The invention also provides the application of the complex enzyme in the production of animal protein.
The invention also provides a production method of the animal protein powder, which comprises the following steps:
feeding animal protein raw materials into a cooking machine after iron removal and impurity removal, and adding water according to 40-50% of the mass of the raw materials; rapidly heating to 105 deg.C, steaming at high temperature and high pressure to break cells and solidify and denature protein, pulverizing, superfine grinding, and pulping to obtain pulp; adding a proper amount of phosphoric acid into the slurry while stirring, and adjusting the pH to 6.5-7.0; rapidly heating to 55 ℃, adding the complex enzyme provided by the invention according to 1.5-3 per mill of the mass of the slurry while stirring, preserving heat, and continuously stirring for enzymolysis for 4-6 h; and (4) carrying out spray drying on the slurry after enzymolysis to prepare the animal protein powder.
Advantageous effects
The complex enzyme provided by the invention can promote the efficient degradation of animal protein raw materials and improve the content of acid soluble proteins such as small peptides, amino acids and the like. Compared with a blank control group, the content of the acid-soluble protein in the fishbone meat powder prepared by each treatment group added with the complex enzyme is generally increased by 201-217%, the content of the acid-soluble protein in the fish paste is generally increased by 189.5-195.8%, the content of the acid-soluble protein in chicken liver pulp is generally increased by 475-485%, and the effect is extremely obvious.
When the neutral protease, the NSP enzyme and the papain enzyme in the compound enzyme are added simultaneously, a synergistic promotion effect is generated, the comprehensive enzymolysis effect is obviously higher than that of each single enzyme, the single enzyme with the highest enzymolysis efficiency is further improved by 23.6 percent, and an unexpected technical effect is obtained.
Although disodium dihydrogen pyrophosphate in the complex enzyme has no degradation effect on protein, the addition of disodium dihydrogen pyrophosphate can improve the comprehensive enzymolysis effect of neutral protease, NSP enzyme and papain by 18.1 percent, and has a synergistic promotion effect. And the disodium dihydrogen pyrophosphate can play a role in synergistically promoting the enzymolysis effects of neutral protease, NSP (non-toxic substance) enzyme and papain only when being added in a proper amount, and can play an antagonistic role when being added in an excessive amount, so that the effects of the three enzymes are inhibited to a certain extent, and unexpected technical effects are obtained.
In addition, the animal protein prepared by the complex enzyme has the advantages of increased delicate flavor, better flavor and obviously improved quality, and is beneficial to wide application in the fields of food and feed.
Detailed Description
The present invention will be described in detail with reference to examples. The reagents and raw materials selected in the embodiment of the invention can be purchased from any commercially available reagents and raw materials. Wherein the neutral protease can be purchased from Weifang kang dien biotechnology limited company, and the enzyme activity is 5 ten thousand U/g; the NSP enzyme can be purchased from Weifang kang dien biotechnology limited company, and the enzyme activity is 5 ten thousand U/g; papain can be purchased from Nanning Dong Henghuadao biotechnology, Inc., and Nanning Pombo bioengineering, Inc., and has enzyme activity of 20 ten thousand U/g; disodium dihydrogen pyrophosphate was purchased from Sichuan Jindiya scientific Co., Ltd.
Example 1
A compound enzyme comprises the following components in percentage by weight: 24 parts of neutral protease, 5 parts of NSP enzyme, 9 parts of papain and 2 parts of disodium dihydrogen pyrophosphate.
Weighing each component of the complex enzyme according to the weight, and uniformly mixing to obtain the complex enzyme, namely the complex enzyme 1 #.
Example 2
A compound enzyme comprises the following components in percentage by weight: 20 parts of neutral protease, 3 parts of NSP enzyme, 6 parts of papain and 1 part of disodium dihydrogen pyrophosphate.
The preparation method is the same as example 1, and the prepared complex enzyme is named complex enzyme # 2.
Example 3
A compound enzyme comprises the following components in percentage by weight: 21 parts of neutral protease, 6 parts of NSP enzyme, 4 parts of papain and 3 parts of disodium dihydrogen pyrophosphate.
The preparation method is the same as example 1, and the prepared complex enzyme is named complex enzyme # 3.
Example 4 application of the Complex enzyme of the present invention in production of fishbone meat powder
100kg of fishbone crushed meat is sent into a cooking machine after iron and impurity removal, and 40kg to 50kg of water is added; rapidly heating to 105 deg.C, steaming at high temperature and high pressure to break cells and solidify and denature protein, pulverizing, grinding, and pulping to obtain fish bone and meat slurry; adding a proper amount of phosphoric acid into the fish bone and meat paste while stirring, and adjusting the pH to 6.6; rapidly heating to 55 ℃, adding the complex enzyme provided by the embodiment of the invention according to 2 per mill of the mass of the fishbone meat paste while stirring, preserving heat, and continuously stirring for enzymolysis for 6 hours; and (4) carrying out spray drying on the fish bone and meat slurry subjected to enzymolysis to prepare the fish bone and meat powder.
Meanwhile, a blank control group is arranged, no enzyme is added, and other production processes are the same as above.
The content of acid soluble protein in the prepared fishbone meat powder is respectively detected, and specific results are shown in table 1.
High molecular weight proteins are easily precipitated under acidic conditions, while lower molecular weight proteins are soluble in acidic solutions, i.e., acid soluble proteins, whose hydrolysates include peptides and free amino acids. The content of the acid soluble protein is the percentage of the acid soluble protein in crude protein, and is an important index for judging the proteolysis level
The acid soluble protein content determination method comprises the following steps:
1. principle of
Trichloroacetic acid is used as protein precipitant to precipitate protein and peptide with longer peptide chain, short-chain small peptide is dissolved out by acid, and the protein content is expressed by percentage of coarse protein in sample after filtration, centrifugation, digestion and distillation.
2. Procedure for the preparation of the
Accurately weighing 6g of a sample in a 100mL beaker, accurately adding 50mL of 15% trichloroacetic acid solution, uniformly mixing, and standing for 5 min; dry-filtering with medium-speed qualitative filter paper, discarding a little initial filtrate, transferring the filtrate to a centrifuge tube, centrifuging 10 times at the rotating speed of 4000 rpm, accurately transferring 10mL of supernatant into a digestion tube, measuring the protein content by a Kjeldahl method, and simultaneously performing a blank test and a Kjeldahl method to measure the crude protein content.
3. Results and calculations
Formula for calculation
Acid soluble protein (%) [ (V1-V0) × C × 6.25 × 0.014 × 5]/(m × cp) × 100%.
V1-volume of hydrochloric acid standard solution consumed by distillate, mL;
v0-blank test consumption hydrochloric acid Standard volume, mL
C-molar concentration of hydrochloric acid, mol/L;
6.25X 0.014-protein conversion coefficient;
m-weighing the mass of the sample, g;
cp — crude protein content of sample,%.
TABLE 1 Effect of Complex enzymes on the enzymolysis Effect of fishbone and meat
| Experimental group | Class of enzyme | Acid soluble protein content |
| Blank control group | - | 22.33％ |
| Treatment group 1 | The complex enzyme 1 #provided by the invention | 68.21％ |
| Treatment group 2 | The complex enzyme 2 #provided by the invention | 70.52％ |
| Treatment group 3 | The complex enzyme 3# provided by the invention | 67.31％ |
As can be seen from the data in Table 1, compared with the blank control group, the content of acid soluble protein in the fishbone meat powder prepared by each treatment group added with the complex enzyme is generally increased by 201-217%, and the effect is extremely obvious. Therefore, the complex enzyme provided by the invention can efficiently promote the degradation of protein in the fish skeleton minced meat, greatly improve the enzymolysis efficiency and obtain unexpected technical effects.
Example 5 synergistic Effect between Complex enzyme Components to promote proteolytic efficiency
The applicant adopts the production process of the fishbone meat powder described in example 4, and further analyzes the influence of each component in the complex enzyme on the proteolysis effect of the fishbone minced meat under the condition of the same addition amount (2 per thousand mass ratio), and the specific data is shown in table 2.
TABLE 2 influence of the Components of Complex enzyme on the enzymolysis Effect of fishbone and meat
As can be seen from the results in table 2, compared with the blank control group, the acid soluble protein content of the fishbone meat powder obtained by the treatment groups 4 and 6, in which the neutral protease and the papain are added separately, is increased by 116.9% and 110.1%, respectively, and the effect is very significant; the content of acid soluble protein in the fishbone meat powder obtained by the processing group 5 with the NSP enzyme added alone is not obviously improved; the content of acid soluble protein in the fishbone meat powder obtained by the treatment group 8 with the single addition of the disodium dihydrogen pyrophosphate is the same as that of the blank control group, and is not improved at all. Therefore, the neutral protease and the papain can carry out high-efficiency enzymolysis on the protein in the fish bone skeleton minced meat, the enzymolysis effect of the NSP enzyme is not obvious, and the disodium dihydrogen pyrophosphate has no degradation effect on the protein.
Compared with the treatment groups 4-6 added with single enzyme, the acid soluble protein content of the fishbone meat powder obtained by the treatment group 7 added with three enzymes of neutral protease, NSP enzyme and papain is greatly improved to 59.87 percent, and is further improved by 23.6 percent compared with the treatment group 4 with the highest acid soluble protein content in the treatment groups 4-6. Therefore, the three enzymes of neutral protease, NSP enzyme and papain generate a synergistic effect when being added simultaneously, the comprehensive enzymolysis effect is obviously higher than that of each single enzyme, and unexpected technical effects are obtained.
Compared with the treatment group 7, the content of acid soluble protein in the fishbone meat powder obtained by the treatment group 10 added with the complex enzyme No. 2 is improved by 18.1 percent, and the effect is obvious. The compound enzyme 2# further comprises disodium dihydrogen pyrophosphate besides three single enzymes of neutral protease, NSP enzyme and papain. Therefore, although the disodium dihydrogen pyrophosphate has no degradation effect on the protein, the addition of the disodium dihydrogen pyrophosphate can obviously improve the comprehensive enzymolysis effect of the neutral protease, the NSP enzyme and the papain on the protein, and has a synergistic promotion effect, thereby achieving unexpected technical effects.
Example 6 Effect of the amount of disodium dihydrogen pyrophosphate added on the enzymolysis Effect of Complex enzymes
The applicant adopts the production process of the fishbone meat powder described in example 4 to further analyze the influence of the addition amount of disodium dihydrogen pyrophosphate in the complex enzyme on the proteolysis effect of the fishbone minced meat under the condition of the same addition amount (2 per thousand mass ratio), and the specific data are shown in table 3.
TABLE 3 Effect of disodium dihydrogen pyrophosphate on Complex enzyme enzymolysis Effect
As can be seen from the data in table 3, the content of acid-soluble protein in the fishbone meat powder obtained in treatment group 11 was not increased when a small amount of disodium dihydrogen pyrophosphate was added as compared to the control group; when the content of the disodium dihydrogen pyrophosphate is further increased, the mass ratio of the three enzymes to the disodium dihydrogen pyrophosphate is up to 20: 3: 6: when 0.5 hour, the content of acid soluble protein in the fishbone meat powder obtained by the treatment group 12 is greatly improved by 12.6 percent compared with that of the control group; when the content of the disodium dihydrogen pyrophosphate is continuously increased, the mass ratio of the three enzymes to the disodium dihydrogen pyrophosphate is up to 20: 3: 6: 1 (complex enzyme # 2), the content of acid soluble protein in the fishbone meat powder obtained in the treatment group 13 is further increased by 18.1 percent compared with the control group; however, when the content of disodium dihydrogen pyrophosphate was further increased, the content of acid-soluble protein in the fishbone meal obtained in treatment groups 14 and 15 began to decrease significantly.
The results show that the enzymolysis effects of the neutral protease, the NSP enzyme and the papain can be synergistically promoted only when the disodium dihydrogen pyrophosphate is added in a proper amount, so that the comprehensive effect of the complex enzyme is optimal; and when the disodium dihydrogen pyrophosphate is added in an excessive amount, antagonism can be realized, the action effect of three enzymes is inhibited to a certain extent, and unexpected technical effect is achieved.
Example 7 application of the Complex enzyme of the present invention in production of Fish paste
Feeding 100kg of iced fresh trash fish into a cooking machine after iron and impurity removal, rapidly heating to 100 ℃ for high-temperature cooking, so that cells are broken, and protein is coagulated and denatured; then, carrying out solid-liquid separation on the fish by using a screw press; the obtained liquid is further subjected to oil-water separation by an oil-water separator; mixing the separated protein water and the fish residue, and then carrying out superfine grinding and pulping to obtain slurry; then adding 10-30 kg of water and a proper amount of phosphoric acid while stirring, adjusting the viscosity of the slurry, and adjusting the pH to 6.8; quickly heating to 55 ℃, adding the complex enzyme provided by the invention according to 3.0 per mill of the mass of the slurry while stirring, preserving heat, and continuously stirring for enzymolysis for 5 hours. Placing the slurry after enzymolysis into a concentration tank, then quickly heating to 95 ℃, preserving heat for 0.5h, and killing viable bacteria and a small amount of residual enzyme; and concentrating the slurry until the water content is 45% to obtain the fish paste.
Meanwhile, a blank control group is arranged, no enzyme is added, and other production processes are the same as above.
The content of acid-soluble protein in the prepared fish paste is respectively detected, and the specific result is shown in table 4.
TABLE 4 Effect of Complex enzymes of the invention on production of Fish paste
| Experimental group | Class of enzyme | Acid soluble protein content |
| Blank control group | - | 26.37％ |
| Treatment group 16 | The complex enzyme 1 #provided by the invention | 78.01％ |
| Treatment group 17 | The complex enzyme 2 #provided by the invention | 76.35％ |
| Treatment group 18 | The complex enzyme 3# provided by the invention | 77.39％ |
As can be seen from the data in Table 4, compared with the blank control group, the content of acid soluble protein in the fish paste prepared by each treatment group added with the complex enzyme is generally increased by 189.5-195.8%, and the effect is extremely obvious. Therefore, the complex enzyme provided by the invention can efficiently promote the degradation of fish protein, greatly improve the enzymolysis efficiency and obtain unexpected technical effects.
Example 8 application of the Complex enzyme of the present invention in the production of Chicken liver pulp
100kg of iced fresh chicken liver is sent into a cooking machine after iron removal and impurity removal, 30 kg-50 kg of water is added, the temperature is rapidly heated to 100 ℃ for high-temperature cooking, cells are broken, and protein is solidified and denatured; then carrying out superfine grinding and pulping to obtain chicken liver pulp; adding a proper amount of phosphoric acid while stirring, and adjusting the pH value of the slurry to 7; quickly heating to 55 ℃, adding the complex enzyme according to 1.5 per mill of the mass of the slurry while stirring, preserving heat, and continuously stirring for enzymolysis for 4 hours. Placing the chicken liver slurry after enzymolysis into a concentration tank, rapidly heating to 95 ℃, preserving heat for 0.5h, and killing viable bacteria and a small amount of residual enzyme; then concentrating the chicken liver pulp until the water content is 65 percent to obtain the chicken liver pulp.
Meanwhile, a blank control group is arranged, no enzyme is added, and other production processes are the same as above.
The acid-soluble protein content of the chicken liver pulp obtained by the preparation method is respectively detected, and the specific results are shown in table 5.
TABLE 5 Effect of Complex enzymes of the present invention on Chicken liver pulp production
As can be seen from the data in Table 5, compared with the blank control group, the content of acid soluble protein in chicken liver pulp prepared by each treatment group added with the complex enzyme disclosed by the invention is generally improved by 475-485%, and the effect is extremely obvious. Therefore, the complex enzyme provided by the invention can efficiently promote the degradation of chicken liver protein, greatly improve the enzymolysis efficiency and obtain unexpected technical effects.
In addition, the chicken liver paste produced by the complex enzyme provided by the invention has better fluidity, less connective tissue fragments, increased delicate flavor and better flavor, and is more suitable for application in the fields of food and feed.
Claims (8)
1. A compound enzyme is characterized by comprising neutral protease, NSP enzyme and papain.
2. The complex enzyme of claim 1, wherein the complex enzyme further comprises disodium dihydrogen pyrophosphate.
3. The complex enzyme as claimed in claim 2, wherein the complex enzyme comprises the following components in parts by weight: 20-24 parts of neutral protease, 3-6 parts of NSP enzyme, 4-9 parts of papain and 1-3 parts of disodium dihydrogen pyrophosphate.
4. The complex enzyme as claimed in claim 3, wherein the complex enzyme comprises the following components in parts by weight: 24 parts of neutral protease, 5 parts of NSP enzyme, 9 parts of papain and 2 parts of disodium dihydrogen pyrophosphate.
5. The complex enzyme as claimed in claim 3, wherein the complex enzyme comprises the following components in parts by weight: 20 parts of neutral protease, 3 parts of NSP enzyme, 6 parts of papain and 1 part of disodium dihydrogen pyrophosphate.
6. The complex enzyme as claimed in claim 3, wherein the complex enzyme comprises the following components in parts by weight: 21 parts of neutral protease, 6 parts of NSP enzyme, 4 parts of papain and 3 parts of disodium dihydrogen pyrophosphate.
7. Use of the complex enzyme of any of claims 1-6 in the production of animal proteins.
8. A production method of animal protein powder comprises the following steps:
feeding animal protein raw materials into a cooking machine after iron removal and impurity removal, and adding water according to 40-50% of the mass of the raw materials; rapidly heating to 105 deg.C, steaming at high temperature and high pressure to break cells and solidify and denature protein, pulverizing, superfine grinding, and pulping to obtain pulp; adding a proper amount of phosphoric acid into the slurry while stirring, and adjusting the pH to 6.5-7.0; rapidly heating to 55 ℃, adding the complex enzyme according to 1.5-3 per mill of the mass of the slurry while stirring, preserving heat, and continuously stirring for enzymolysis for 4-6 hours; and (4) carrying out spray drying on the slurry after enzymolysis to prepare the animal protein powder.
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| WO1998018345A1 (en) * | 1996-10-29 | 1998-05-07 | Finnfeeds International Limited | Use of an enzyme for promoting pigment uptake from a feed |
| CN103621765A (en) * | 2013-11-29 | 2014-03-12 | 中国水产科学研究院长江水产研究所 | Method for preparing oligopeptide through mixed enzymolysis and fermentation of fish meal processing waste liquid and vegetable proteins |
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| CN108753892A (en) * | 2018-06-21 | 2018-11-06 | 吴忠市王国旗生物科技有限公司 | A kind of preparation process of peptone |
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| WO1998018345A1 (en) * | 1996-10-29 | 1998-05-07 | Finnfeeds International Limited | Use of an enzyme for promoting pigment uptake from a feed |
| CN103621765A (en) * | 2013-11-29 | 2014-03-12 | 中国水产科学研究院长江水产研究所 | Method for preparing oligopeptide through mixed enzymolysis and fermentation of fish meal processing waste liquid and vegetable proteins |
| CN105368876A (en) * | 2015-12-19 | 2016-03-02 | 青岛伟源达技术服务有限公司 | Process for preparing leftover fertilizer processed through globefish by adopting enzymolysis and fermentation method |
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