JP7422236B2 - ハマグリ活性ペプチドの製造方法 - Google Patents
ハマグリ活性ペプチドの製造方法 Download PDFInfo
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- JP7422236B2 JP7422236B2 JP2022543690A JP2022543690A JP7422236B2 JP 7422236 B2 JP7422236 B2 JP 7422236B2 JP 2022543690 A JP2022543690 A JP 2022543690A JP 2022543690 A JP2022543690 A JP 2022543690A JP 7422236 B2 JP7422236 B2 JP 7422236B2
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- clam
- protease
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- 238000004519 manufacturing process Methods 0.000 title claims description 51
- 235000013372 meat Nutrition 0.000 claims description 84
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- 238000000034 method Methods 0.000 claims description 43
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- 238000006243 chemical reaction Methods 0.000 description 12
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 9
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 9
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- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 8
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- 241000237519 Bivalvia Species 0.000 description 4
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- XOAAWQZATWQOTB-UHFFFAOYSA-N taurine Chemical compound NCCS(O)(=O)=O XOAAWQZATWQOTB-UHFFFAOYSA-N 0.000 description 4
- 102000004157 Hydrolases Human genes 0.000 description 3
- 108090000604 Hydrolases Proteins 0.000 description 3
- BUGBHKTXTAQXES-UHFFFAOYSA-N Selenium Chemical compound [Se] BUGBHKTXTAQXES-UHFFFAOYSA-N 0.000 description 3
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- QIAFMBKCNZACKA-UHFFFAOYSA-N N-benzoylglycine Chemical compound OC(=O)CNC(=O)C1=CC=CC=C1 QIAFMBKCNZACKA-UHFFFAOYSA-N 0.000 description 2
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- NWOJMNXIJBZMPK-QJHJCNPRSA-N (2s)-2-[[(2s)-2-[(2-benzamidoacetyl)amino]-3-(1h-imidazol-5-yl)propanoyl]amino]-4-methylpentanoic acid;hydrate Chemical compound O.C([C@@H](C(=O)N[C@@H](CC(C)C)C(O)=O)NC(=O)CNC(=O)C=1C=CC=CC=1)C1=CN=CN1 NWOJMNXIJBZMPK-QJHJCNPRSA-N 0.000 description 1
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- 102000004142 Trypsin Human genes 0.000 description 1
- 108090000631 Trypsin Proteins 0.000 description 1
- HCHKCACWOHOZIP-UHFFFAOYSA-N Zinc Chemical compound [Zn] HCHKCACWOHOZIP-UHFFFAOYSA-N 0.000 description 1
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Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/14—Extraction; Separation; Purification
- C07K1/34—Extraction; Separation; Purification by filtration, ultrafiltration or reverse osmosis
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L17/00—Food-from-the-sea products; Fish products; Fish meal; Fish-egg substitutes; Preparation or treatment thereof
- A23L17/10—Fish meal or powder; Granules, agglomerates or flakes
-
- 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
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
- A23J1/00—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites
- A23J1/04—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites from fish or other sea animals
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
- A23J3/00—Working-up of proteins for foodstuffs
- A23J3/04—Animal proteins
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
- A23J3/00—Working-up of proteins for foodstuffs
- A23J3/30—Working-up of proteins for foodstuffs by hydrolysis
- A23J3/32—Working-up of proteins for foodstuffs by hydrolysis using chemical agents
- A23J3/34—Working-up of proteins for foodstuffs by hydrolysis using chemical agents using enzymes
- A23J3/341—Working-up of proteins for foodstuffs by hydrolysis using chemical agents using enzymes of animal proteins
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L17/00—Food-from-the-sea products; Fish products; Fish meal; Fish-egg substitutes; Preparation or treatment thereof
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L17/00—Food-from-the-sea products; Fish products; Fish meal; Fish-egg substitutes; Preparation or treatment thereof
- A23L17/40—Shell-fish
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L17/00—Food-from-the-sea products; Fish products; Fish meal; Fish-egg substitutes; Preparation or treatment thereof
- A23L17/50—Molluscs
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Biochemistry (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Wood Science & Technology (AREA)
- Nutrition Science (AREA)
- Genetics & Genomics (AREA)
- Molecular Biology (AREA)
- General Health & Medical Sciences (AREA)
- Marine Sciences & Fisheries (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Microbiology (AREA)
- General Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biotechnology (AREA)
- Water Supply & Treatment (AREA)
- Analytical Chemistry (AREA)
- Biophysics (AREA)
- Medicinal Chemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Meat, Egg Or Seafood Products (AREA)
- Peptides Or Proteins (AREA)
Description
生肉:紅島ハマグリ肉
試験酵素:中性プロテアーゼ、アルカリ性プロテアーゼ、パパイン、複合プロテアーゼ
酵素分解のプロセスフロー:冷凍の紅島ハマグリ肉を解凍した後、ハマグリ肉:水=1:1の重量比で脱イオン水を加えて均質化した。中性プロテアーゼ、アルカリ性プロテアーゼ、パパイン、複合プロテアーゼ(処方は中性プロテアーゼ:アルカリ性プロテアーゼ:風味プロテアーゼ=2:1:1であった)の酵素分解温度を50℃として、酵素添加量をハマグリ肉スラリーの0.13%とし、pHを自然のpH値にした。一定温度で6時間酵素分解し、85℃で酵素を10分間失活し、4000r/minで30分間遠心分離し、上澄みを取った。
生肉:紅島ハマグリ肉
試験酵素:複合プロテアーゼ
酵素分解プロセスフロー:冷凍の紅島ハマグリ肉を解凍した後、ハマグリ肉:水=1:1の重量比で脱イオン水を加えて均質化し、最終的にハマグリ肉:水=1:2の重量比になるように、ハマグリ肉スラリーに一定量の脱イオン水を加えた。複合プロテアーゼ(処方は中性プロテアーゼ:アルカリ性プロテアーゼ:風味プロテアーゼ=2:1:1であった)の酵素分解温度を50℃とし、酵素添加量をハマグリ肉スラリーの0.13%とし、pHを自然のpH値にした。一定温度下で4時間酵素分解し、85℃で酵素を10分間失活し、16000r/minで遠心分離し、上澄みを膜ろ過し、噴霧乾燥させてハマグリ活性ペプチドを得た。
生肉:紅島ハマグリ肉
試験酵素:複合プロテアーゼ
酵素分解プロセスフロー:冷凍の紅島ハマグリ肉を解凍した後、ハマグリ肉:水=1:1の重量比で脱イオン水を加えて均質化し、最終的にハマグリ肉:水=1:3の重量比になるように、ハマグリ肉スラリーに一定量の脱イオン水を加えた。複合プロテアーゼ(処方は中性プロテアーゼ:アルカリ性プロテアーゼ:風味プロテアーゼ=2:1:1であった)の酵素分解温度を50℃とし、酵素添加量をハマグリ肉スラリーの0.13%とし、pHを自然のpH値にした。一定温度下で4時間酵素分解し、85℃で酵素を10分間失活し、16000r/minで遠心分離し、上澄みを膜ろ過し、噴霧乾燥させてハマグリ活性ペプチドを得た。
生肉:紅島ハマグリ肉
試験酵素:複合プロテアーゼ
酵素分解プロセスフロー:冷凍の紅島ハマグリ肉を解凍した後、ハマグリ肉:水=1:1の重量比で脱イオン水を加えて均質化し、最終的にハマグリ肉:水=1:2の重量比になるように、ハマグリ肉スラリーに一定量の脱イオン水を加えた。複合プロテアーゼ(処方は中性プロテアーゼ:アルカリ性プロテアーゼ:風味プロテアーゼ=2:1:1であった)の酵素分解温度を60℃とし、酵素添加量をハマグリ肉スラリーの0.2%とし、pHを自然のpH値にした。一定温度下で4時間酵素分解し、85℃で酵素を10分間失活し、16000r/minで遠心分離し、上澄みを膜ろ過し、噴霧乾燥させてハマグリ活性ペプチドを得た。
生肉:紅島ハマグリ肉
試験酵素:複合プロテアーゼ
酵素分解プロセスフロー:冷凍の紅島ハマグリ肉を解凍した後、ハマグリ肉:水=1:1の重量比で脱イオン水を加えて均質化し、最終的にハマグリ肉:水=1:2の重量比になるように、ハマグリ肉スラリーに一定量の脱イオン水を加えた。複合プロテアーゼ(処方は中性プロテアーゼ:アルカリ性プロテアーゼ:風味プロテアーゼ=2:1:1であった)の酵素分解温度を50℃とし、酵素添加量をハマグリ肉スラリーの0.3%とし、pHを自然のpH値にした;一定温度で別々に6時間酵素分解し、85℃で酵素を10分間失活し、16000r/minで遠心分離し、上澄みを膜ろ過し、噴霧乾燥させてハマグリ活性ペプチドを得た。
各実施例で得られたハマグリ活性ペプチドを検出した結果を表3に示した。
各実施例で得られたハマグリ活性ペプチドを液体クロマトグラフィーで分析し、実施例2で得られたハマグリ活性ペプチドの結果を図3に示した。
Claims (7)
- 新鮮なハマグリ肉を水で洗浄し、水を加えてコロイドミルにより均質化してハマグリ肉スラリーを製造する工程と、
水と複合プロテアーゼを加えてハマグリ肉スラリーを酵素分解させ、酵素分解後に酵素を失活するために加熱する工程と、
遠心分離して酵素加水分解液体を回収し、精密ろ過膜-限外ろ過膜-ナノろ過膜でろ過して分子量2KDa以下の酵素加水分解液体を採取し、乾燥させてハマグリ活性ペプチドを得る工程とを含み、
前記複合プロテアーゼが中性プロテアーゼと、アルカリ性プロテアーゼと、風味プロテアーゼとからなり、
各酵素の添加割合が中性プロテアーゼ:アルカリ性プロテアーゼ:風味プロテアーゼとして2:1:1であり、
前記複合プロテアーゼの添加量がハマグリ肉スラリーの質量に対する0.1~0.3%であり、
複合プロテアーゼを十分に利用し、酵素分解を完全にするように、前記酵素分解中に連続的に攪拌する、ハマグリ活性ペプチドの製造方法。 - 前記酵素分解に用いられるハマグリ肉と水との重量比が1:1~1:3である、請求項1に記載の製造方法。
- 前記ハマグリ肉が紅島ハマグリ肉であり、
前記洗浄が脱イオン水による洗浄であり、
前記均質化には、ハマグリ肉:水の重量比を1:1として、脱イオン水を加え、
前記コロイドミルによる均質化中には、コロイドミルの粒子の隙間が0~5に制御され、ハマグリ肉の粒子サイズが小さく均一になるようにする、請求項1に記載の製造方法。 - 前記酵素分解が、自然のpH条件下、50~60℃で4~6時間酵素分解することである、請求項1に記載の製造方法。
- 前記酵素を失活するための加熱が、酵素加水分解液体を85℃に加熱して10分間保持することである、請求項1に記載の製造方法。
- 前記遠心分離が、酵素加水分解液体を40℃以下に冷却し、200~300メッシュのふるいに通液してろ過した後、16000r/minで遠心分離することである、請求項1に記載の製造方法。
- 前記乾燥が、膜ろ過後のろ液を乾燥塔に移送して噴霧乾燥することである、請求項1に記載の製造方法。
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