CN113718006B - Oyster peptide extraction method - Google Patents

Oyster peptide extraction method Download PDF

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CN113718006B
CN113718006B CN202110906656.XA CN202110906656A CN113718006B CN 113718006 B CN113718006 B CN 113718006B CN 202110906656 A CN202110906656 A CN 202110906656A CN 113718006 B CN113718006 B CN 113718006B
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dispersion liquid
enzymolysis
microcrystalline cellulose
oyster
stirring
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CN113718006A (en
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林润暄
陈钰淇
林灼华
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Jamesons Brewing Fujian Co ltd
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    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12PFERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P21/00Preparation of peptides or proteins
    • C12P21/06Preparation of peptides or proteins produced by the hydrolysis of a peptide bond, e.g. hydrolysate products
    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07KPEPTIDES
    • C07K1/00General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
    • C07K1/14Extraction; Separation; Purification
    • C07K1/34Extraction; Separation; Purification by filtration, ultrafiltration or reverse osmosis

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  • Organic Chemistry (AREA)
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  • General Engineering & Computer Science (AREA)
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  • Biotechnology (AREA)
  • Water Supply & Treatment (AREA)
  • Analytical Chemistry (AREA)
  • Biophysics (AREA)
  • Medicinal Chemistry (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Coloring Foods And Improving Nutritive Qualities (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)

Abstract

The application discloses an oyster peptide extraction method, which comprises the following steps: (1) crushing: pulverizing cleaned and dried Concha Ostreae and Carnis Ostreae into Concha Ostreae fine powder; (2) dispersing: mixing oyster fine powder, calcium bicarbonate, sodium bicarbonate, microcrystalline cellulose and water according to a mass ratio of 1:0.02-0.04:0.04-0.06:0.01-0.03:23-28, stirring, and uniformly dispersing to obtain a dispersion liquid; (3) enzymolysis: adding a decomposing enzyme into the dispersion liquid to carry out enzymolysis on the dispersion liquid; (4) heavy metal removal: adding EDTA disodium into the dispersion liquid after enzymolysis, and stirring and reacting for 25-35 min; (5) filtering: filtering the dispersion liquid after metal removal, and taking filtrate; (6) concentrating, drying and packaging; has the advantages of removing heavy metals and improving the recovery rate of protein.

Description

Oyster peptide extraction method
Technical Field
The application relates to an oyster peptide extraction method.
Background
The sea food oyster has huge edible value and medicinal value, is the first large cultured shellfish in the world, and is cultured on a large scale in coastal areas such as Zhejiang province in China. Zhejiang Ninghaiqian is known as "oyster village", and there is 700 years of oyster cultivation history. The nutrition value of the oyster is extremely high, the protein content in the oyster is up to 45% -57%, and the amino acid composition is perfect. According to the assessment of the world grain and agriculture organization, the degree of completeness and mass proportion of essential amino acids in oyster meat are superior to those of human milk and cow milk. Ancient people called oyster the most noble of aquatic products, ancient romans had praised it as "sea delicacy-holy fish", western people as "goddess magic stone", "sea milk", and Japanese people as "root source".
Due to marine pollution, considerable heavy metals exist in modern marine organisms such as oysters, so that the technical problem needs to be considered when the oysters are treated.
Disclosure of Invention
The application aims to provide an oyster peptide extraction method which has the advantage of removing heavy metals.
The technical aim of the application is realized by the following technical scheme:
an oyster peptide extraction method comprises the following steps:
(1) Crushing: pulverizing cleaned and dried Concha Ostreae and Carnis Ostreae into Concha Ostreae fine powder;
(2) Dispersing: mixing oyster fine powder, calcium bicarbonate, sodium bicarbonate, microcrystalline cellulose and water, stirring, and dispersing uniformly to obtain a dispersion liquid; the input mass ratio of the oyster fine powder, the calcium bicarbonate, the sodium bicarbonate, the microcrystalline cellulose and the water is 1:0.02-0.04:0.04-0.06:0.01-0.03:23-28;
(3) Enzymolysis: adding a decomposing enzyme into the dispersion liquid to carry out enzymolysis on the dispersion liquid;
(4) Heavy metal removal: adding EDTA disodium into the dispersion liquid after enzymolysis, and stirring and reacting for 25-35 min;
(5) And (3) filtering: filtering the dispersion liquid after metal removal, and taking filtrate;
(6) Concentrating, drying, and packaging.
Further, in the step (3), the decomposing enzyme comprises neutral protease and lipase, and the enzymolysis step comprises the following two parts:
(1) regulating pH of the dispersion to 6.8-7.2, heating to 50-55deg.C, adding neutral protease, stirring for enzymolysis for 5-7 hr, heating to 75-85deg.C, and maintaining for 8-12 min to inactivate enzyme; the mass ratio of the dispersion liquid to the neutral protease is 100:0.2-0.4;
(2) regulating pH of the dispersion to 6.8-7.2, heating to 38-42 ℃, adding lipase, stirring for enzymolysis for 2-4 hr, heating to 85-95 ℃ after enzymolysis, and preserving heat for 4-5 min to inactivate enzyme; the mass ratio of the dispersion liquid to the lipase is 100:0.1-0.3.
Further, in the step (4), microcrystalline cellulose and ethyl acetate are added to the dispersion in addition to disodium EDTA; in the step (4), the mass ratio of the dispersion liquid after enzymolysis to the EDTA disodium, the microcrystalline cellulose and the ethyl acetate is 100:0.02-0.04:0.08-0.11:7-9.
Further, the method comprises the following steps:
(1) Crushing: pulverizing cleaned and dried Concha Ostreae and Carnis Ostreae into Concha Ostreae fine powder;
(2) Dispersing: mixing oyster fine powder, calcium bicarbonate, sodium bicarbonate, microcrystalline cellulose and water, stirring, and dispersing uniformly to obtain a dispersion liquid; the input mass ratio of the oyster fine powder, the calcium bicarbonate, the sodium bicarbonate, the microcrystalline cellulose and the water is 1:0.03:0.05:0.02:25;
(3) Enzymolysis:
(1) regulating pH of the dispersion to 7.0, heating to 52deg.C, adding neutral protease, stirring for enzymolysis for 6hr, heating to 80deg.C, and maintaining for 10min to deactivate enzyme; the mass ratio of the dispersion liquid to the neutral protease is 100:0.3;
(2) regulating pH of the dispersion to 7.0, heating to 40deg.C, adding lipase, stirring for enzymolysis for 3hr, heating to 90deg.C, and maintaining for 5min to deactivate enzyme; the mass ratio of the dispersion liquid to the lipase is 100:0.2;
(4) Heavy metal removal: adding EDTA disodium, microcrystalline cellulose and ethyl acetate into the dispersion liquid after enzymolysis, and stirring and reacting for 30min; in the step (4), the mass ratio of the dispersion liquid after enzymolysis to EDTA disodium, microcrystalline cellulose and ethyl acetate is 100:0.03:0.10:8;
(5) And (3) filtering: vacuum filtering the dispersion liquid after metal removal with a 0.5 μm filter membrane, and collecting filtrate;
(6) Concentrating under vacuum until the solid content is 50wt%, spray drying, and packaging.
The technical effects of the application are mainly as follows:
in the oyster processing process, EDTA disodium is used for heavy metal processing, so that the heavy metal content in the product is reduced, and the food safety is improved; in the treatment process, because of the existence of heavy metals, the possibility of chelation exists between the polypeptides and the heavy metals in the process, so that the chelated heavy metals exist in the product, the calcium bicarbonate is utilized to push and combine microcrystalline cellulose to disperse and block chelation under alkaline conditions and the action of the heavy metals, so that the chelation of the heavy metals and the polypeptides is reduced, the heavy metals are in a free state, so that the heavy metals act with EDTA disodium, the heavy metal removal effect is improved, and the loss of the polypeptides is also reduced;
the method has the advantages that the emulsion stability in the enzymolysis liquid obtained after oyster enzymolysis is good, the separation is not ideal, the separation time and the product quality are influenced, the interaction between protein and fat is destroyed by lipase enzymolysis, and the stability of emulsion is reduced, so that the oil-water separation is realized, the separation effect is improved, the process progress is accelerated, and the protein recovery rate is improved by the action of ethyl acetate and microcrystalline cellulose.
Detailed Description
Example 1: an oyster peptide extraction method comprises the following steps:
(1) Crushing: pulverizing cleaned and dried Concha Ostreae and Carnis Ostreae into Concha Ostreae fine powder;
(2) Dispersing: mixing oyster fine powder, calcium bicarbonate, sodium bicarbonate, microcrystalline cellulose and water, stirring, and dispersing uniformly to obtain a dispersion liquid; the input mass ratio of the oyster fine powder, the calcium bicarbonate, the sodium bicarbonate, the microcrystalline cellulose and the water is 1:0.03:0.05:0.02:25;
(3) Enzymolysis:
(1) regulating pH of the dispersion to 7.0, heating to 52deg.C, adding neutral protease, stirring for enzymolysis for 6hr, heating to 80deg.C, and maintaining for 10min to deactivate enzyme; the mass ratio of the dispersion liquid to the neutral protease is 100:0.3;
(2) regulating pH of the dispersion to 7.0, heating to 40deg.C, adding lipase, stirring for enzymolysis for 3hr, heating to 90deg.C, and maintaining for 5min to deactivate enzyme; the mass ratio of the dispersion liquid to the lipase is 100:0.2;
(4) Heavy metal removal: adding EDTA disodium, microcrystalline cellulose and ethyl acetate into the dispersion liquid after enzymolysis, and stirring and reacting for 30min; in the step (4), the mass ratio of the dispersion liquid after enzymolysis to EDTA disodium, microcrystalline cellulose and ethyl acetate is 100:0.03:0.10:8;
(5) And (3) filtering: vacuum filtering the dispersion liquid after metal removal with a 0.5 μm filter membrane, and collecting filtrate;
(6) Concentrating under vacuum until the solid content is 50wt%, spray drying, and packaging.
Example 1 oyster peptides have a molecular weight distribution as shown in table 1.
TABLE 1
Molecular weight range (Dalton) Percent peak area (%,. Lambda.220 nm)
M>3000 0.15
2000<M≤3000 0.44
1000<M≤2000 2.03
180<M≤1000 17.25
M≤180 80.13
Example 2: influence of step (2) on oyster peptide extraction method
Test object: example 1, control 1-5.
Control group 1: referring to example 1, the difference from example 1 is that the input mass ratio of oyster fine powder, monocalcium carbonate, sodium bicarbonate, microcrystalline cellulose and water in step (2) is 1:0.02:0.04:0.01:25.
Control group 2: referring to example 1, the difference from example 1 is that the input mass ratio of oyster fine powder, monocalcium carbonate, sodium bicarbonate, microcrystalline cellulose and water in step (2) is 1:0.04:0.06:0.03:25.
Control group 3: referring to example 1, the difference from example 1 is that no monocalcium carbonate, sodium bicarbonate or microcrystalline cellulose was added to the oyster powder in step (2), only water was added and the input mass ratio of the oyster powder to the water was 1:25.
Control group 4: referring to example 1, the difference from example 1 is that microcrystalline cellulose was not added to the oyster shell fine powder in step (2), and the input mass ratio of the oyster shell fine powder, the monocalcium carbonate, the sodium bicarbonate and the water was 1:0.03:0.05:25.
Control group 5: referring to example 1, the difference from example 1 is that the input mass ratio of oyster fine powder, monocalcium carbonate, sodium bicarbonate, microcrystalline cellulose and water in step (2) is 1:0.06:0.08:0.05:25.
The test content is as follows: and (3) respectively measuring crude proteins of the test objects and measuring total amounts of three heavy metals including lead, cadmium and chromium.
Measurement of crude protein: the Kjeldahl nitrogen determination method is adopted, and the standard is GB5009.5-2010. Protein recovery (%) = product protein content x 100%/raw material protein content, product protein content is the protein content in the product obtained after spray drying in step (6), and raw material protein content is the protein content of the oyster fine powder obtained in step (1).
Heavy metal determination: referring to GB15618-1995, the test object is the product obtained after spray drying in step (6).
The test results are shown in Table 2. Table 2 shows:
(1) Referring to the data of protein recovery rate, as the contents of the oyster fine powder, the calcium bicarbonate, the sodium bicarbonate and the microcrystalline cellulose in the step (2) are increased, the protein recovery rate tends to be increased firstly, then gradually decreased; oyster fine powder, calcium bicarbonate, sodium bicarbonate and microcrystalline cellulose=1:0.03:0.05:0.02 and 1:0.04:0.06:0.03, and the protein recovery rate is more than 85%;
(2) Referring to the total amount data of lead, cadmium and chromium, along with the increase of the contents of the calcium carbonate, the sodium bicarbonate and the microcrystalline cellulose in the oyster fine powder, the calcium bicarbonate, the sodium bicarbonate and the microcrystalline cellulose in the step (2), the total amount of the lead, the cadmium and the chromium shows a trend of decreasing firstly and then increasing; oyster fine powder, calcium bicarbonate, sodium bicarbonate and microcrystalline cellulose=1:0.03:0.05:0.02, and the total amount of lead, cadmium and chromium is 0.
TABLE 2
Example 3: influence of step (3) and step (4) on oyster peptide extraction method
Test object: example 1, control 6-9.
Control group 6: referring to example 1, the difference from example 1 is that the enzymatic hydrolysis step (3) includes only one of the following: regulating pH of the dispersion to 7.0, heating to 52deg.C, adding neutral protease, stirring for enzymolysis for 6hr, heating to 80deg.C, and maintaining for 10min to deactivate enzyme; the mass ratio of the dispersion liquid and the neutral protease is 100:0.3.
Control group 7: referring to example 1, the difference from example 1 is that only disodium EDTA was added in step (4), and microcrystalline cellulose or ethyl acetate was not added.
Control group 8: referring to example 1, the difference from example 1 is that only disodium EDTA and microcrystalline cellulose were added in step (4), and ethyl acetate was not added.
Control group 9: referring to example 1, the difference from example 1 is that only disodium EDTA and ethyl acetate were added in step (4), and microcrystalline cellulose was not added.
The test content is as follows: and (3) respectively measuring the time consumption of vacuum filtration of the test object, namely taking 1kg of the dispersion liquid treated in the step (4), performing vacuum filtration by using a 0.5 mu m filter membrane, wherein the suction filtration pressure is 0.1MPa, the suction filtration temperature is 25 ℃, counting time from the beginning of the suction filtration, and recording the total time consumption until the filtrate is completely filtered.
Measurement of crude protein: the Kjeldahl nitrogen determination method is adopted, and the standard is GB5009.5-2010. Protein recovery (%) = product protein content x 100%/raw material protein content, product protein content is the protein content in the product obtained after spray drying in step (6), and raw material protein content is the protein content of the oyster fine powder obtained in step (1).
The test results are shown in Table 3. Table 3 shows: compared with the control group 6 which is not subjected to the two-step treatment in the step (3), the vacuumized filtration in the embodiment 1 of the application consumes more time and has higher protein recovery rate; compared with the control group 7-9 without ethyl acetate and/or microcrystalline cellulose added in step (4), the vacuuming filtration of example 1 of the present application takes more time and the protein recovery rate is higher.
TABLE 3 Table 3
Vacuum filtration time-consuming (min) Protein recovery (%)
Example 1 10 85.2
Control group 6 92 63.0
Control group 7 53 70.2
Control group 8 88 68.1
Control group 9 82 66.4
Of course, the above is only a typical example of the application, and other embodiments of the application are also possible, and all technical solutions formed by equivalent substitution or equivalent transformation fall within the scope of the application claimed.

Claims (1)

1. The oyster peptide extracting process includes the following steps:
(1) Crushing: pulverizing cleaned and dried Concha Ostreae and Carnis Ostreae into Concha Ostreae fine powder;
(2) Dispersing: mixing oyster fine powder, calcium bicarbonate, sodium bicarbonate, microcrystalline cellulose and water, stirring, and dispersing uniformly to obtain a dispersion liquid; the input mass ratio of the oyster fine powder, the calcium bicarbonate, the sodium bicarbonate, the microcrystalline cellulose and the water is 1:0.03:0.05:0.02:25;
(3) Enzymolysis:
(1) regulating pH of the dispersion to 7.0, heating to 52deg.C, adding neutral protease, stirring for enzymolysis for 6hr, heating to 80deg.C, and maintaining for 10min to deactivate enzyme; the mass ratio of the dispersion liquid to the neutral protease is 100:0.3;
(2) regulating pH of the dispersion to 7.0, heating to 40deg.C, adding lipase, stirring for enzymolysis for 3hr, heating to 90deg.C, and maintaining for 5min to deactivate enzyme; the mass ratio of the dispersion liquid to the lipase is 100:0.2;
(4) Heavy metal removal: adding EDTA disodium, microcrystalline cellulose and ethyl acetate into the dispersion liquid after enzymolysis, and stirring and reacting for 30min; in the step (4), the mass ratio of the dispersion liquid after enzymolysis to EDTA disodium, microcrystalline cellulose and ethyl acetate is 100:0.03:0.10:8;
(5) And (3) filtering: vacuum filtering the dispersion liquid after metal removal with a 0.5 μm filter membrane, and collecting filtrate;
(6) Concentrating under vacuum until the solid content is 50wt%, spray drying, and packaging.
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Citations (6)

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CN105028283A (en) * 2015-08-26 2015-11-11 浙江海洋学院 Preparation method and application of heavy-metal removing agent for temporary shellfish cultivation
CN106306326A (en) * 2016-08-17 2017-01-11 任浩楠 Preparation method of food-grade oyster polypeptide
CN108998041A (en) * 2018-06-01 2018-12-14 王玲 Treatment agent for heavy metal contaminated soil and preparation method thereof
CN110467669A (en) * 2019-07-22 2019-11-19 福建福瑞康信息技术有限公司 The extracting method of Zinc metal sulfur protein zinc in one seed oyster
CN111498972A (en) * 2020-04-27 2020-08-07 李洪 Heavy metal polluted wastewater treatment agent and preparation method and application thereof
KR20210017492A (en) * 2019-08-08 2021-02-17 주식회사 셀트리온제약 Animal-derived liver hydrolysate

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Publication number Priority date Publication date Assignee Title
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Publication number Priority date Publication date Assignee Title
CN105028283A (en) * 2015-08-26 2015-11-11 浙江海洋学院 Preparation method and application of heavy-metal removing agent for temporary shellfish cultivation
CN106306326A (en) * 2016-08-17 2017-01-11 任浩楠 Preparation method of food-grade oyster polypeptide
CN108998041A (en) * 2018-06-01 2018-12-14 王玲 Treatment agent for heavy metal contaminated soil and preparation method thereof
CN110467669A (en) * 2019-07-22 2019-11-19 福建福瑞康信息技术有限公司 The extracting method of Zinc metal sulfur protein zinc in one seed oyster
KR20210017492A (en) * 2019-08-08 2021-02-17 주식회사 셀트리온제약 Animal-derived liver hydrolysate
CN111498972A (en) * 2020-04-27 2020-08-07 李洪 Heavy metal polluted wastewater treatment agent and preparation method and application thereof

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