WO2019196513A1 - Preparation method for enhanced gelatinous myofibrillar protein - Google Patents
Preparation method for enhanced gelatinous myofibrillar protein Download PDFInfo
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- WO2019196513A1 WO2019196513A1 PCT/CN2018/125564 CN2018125564W WO2019196513A1 WO 2019196513 A1 WO2019196513 A1 WO 2019196513A1 CN 2018125564 W CN2018125564 W CN 2018125564W WO 2019196513 A1 WO2019196513 A1 WO 2019196513A1
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- 108090000623 proteins and genes Proteins 0.000 title claims abstract description 62
- 102000004169 proteins and genes Human genes 0.000 title claims abstract description 62
- 238000002360 preparation method Methods 0.000 title claims abstract description 12
- 239000012460 protein solution Substances 0.000 claims abstract description 15
- WQGWDDDVZFFDIG-UHFFFAOYSA-N pyrogallol Chemical compound OC1=CC=CC(O)=C1O WQGWDDDVZFFDIG-UHFFFAOYSA-N 0.000 claims abstract description 11
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- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 36
- 239000002244 precipitate Substances 0.000 claims description 26
- 238000000926 separation method Methods 0.000 claims description 24
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- DEFVIWRASFVYLL-UHFFFAOYSA-N ethylene glycol bis(2-aminoethyl)tetraacetic acid Chemical compound OC(=O)CN(CC(O)=O)CCOCCOCCN(CC(O)=O)CC(O)=O DEFVIWRASFVYLL-UHFFFAOYSA-N 0.000 claims description 9
- LNTHITQWFMADLM-UHFFFAOYSA-N gallic acid Chemical compound OC(=O)C1=CC(O)=C(O)C(O)=C1 LNTHITQWFMADLM-UHFFFAOYSA-N 0.000 claims description 8
- 210000004317 gizzard Anatomy 0.000 claims description 8
- 239000006228 supernatant Substances 0.000 claims description 8
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- 238000010438 heat treatment Methods 0.000 claims description 3
- 230000003064 anti-oxidating effect Effects 0.000 claims description 2
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- KDXKERNSBIXSRK-YFKPBYRVSA-N L-lysine Chemical compound NCCCC[C@H](N)C(O)=O KDXKERNSBIXSRK-YFKPBYRVSA-N 0.000 description 1
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- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 description 1
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- 235000020640 mackerel Nutrition 0.000 description 1
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Images
Classifications
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- 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
- A23J3/22—Working-up of proteins for foodstuffs by texturising
-
- 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
- A23L17/70—Comminuted, e.g. emulsified, fish products; Processed products therefrom such as pastes, reformed or compressed products
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
Definitions
- the invention belongs to the preparation of myofibrillar protein, in particular to a preparation method of strong gel myofibrillar protein.
- Myofibrillar protein is a kind of salt-soluble protein with important biological functions. It is also the main protein that constitutes muscle fiber, accounting for 50-55% of total protein content, including myosin, actin and tropomyosin. , troponin and so on. Myofibrillar proteins are involved in muscle contraction, affecting tenderness of meat and functional properties of meat products such as gelatinity, rheological properties, water retention, elasticity, texture, and the like. Among them, the gel properties of myofibrillar proteins are the most important functional properties in the processing of surimi products. A gel is an elastic semi-solid that is between a solid and a liquid, has a uniform appearance, and maintains a certain shape.
- Proteins can form gels because proteins are regulated by external conditions or dissociated by non-covalent bonds after heating, and the structure of the surface changes. In particular, the exposure of hydrophobic groups promotes the interaction between proteins, making protein molecules A uniform network-like gel is formed under polymerization.
- the myofibrillar protein obtained in the step 1) was added to a cold solution, homogenized, placed at 0 ° C for 30 min, centrifuged, and the supernatant was taken and concentrated by an ultrafiltration tube.
- the myofibrillar protein obtained in the step 2) is added to the pyrogallic acid, mixed uniformly, and left for 1 to 5 hours to obtain a mixed protein solution system.
- the mixed protein solution obtained in the step 3) is vacuum-packed and placed under an ultra-high pressure of 100 to 600 MPa for 5 to 30 minutes.
- the sample obtained in the step 4) was placed in a water bath at 20 ° C to be programmed to a temperature of 85 ° C, a heating rate of 1 ° C / min, held at 85 ° C for 30 min, and then quenched at 0 ° C for 12 h to obtain a myofibrillar protein gel. .
- the pH of the first separation buffer is 6.9 to 7.1, including 0.08 to 0.12 M KCl, 1.8 to 2.2 mM MgCl 2 , 0.8 to 1.2 mM EGTA, 0.4 to 0.6 mM DTT. And 8-11 mM K 2 HPO 4 ; the second buffer has a pH of 5.9-6.1, including 0.08-0.12 M NaCl and 0.8-1.2 mM. NaN 3 .
- step 1) the homogenization conditions are 7000 to 10000 rpm, 3 to 8 minutes.
- the centrifugation conditions are 7000 to 9000 rpm, 10 to 30 minutes, and 4 to 8 °C.
- the solution is 0.5 to 1.0 M NaCl, and the pH is 7.0 to 8.0.
- step 3 the concentration of pyrogallic acid added is from 6 ⁇ mol/g protein to 300 ⁇ mol/g protein.
- the vacuum packaging comprises a vacuuming time of 15 to 60 s, a heat sealing time of 1.0 to 3.0 s, an ultrahigh pressure treatment pressure of 100 to 600 MPa, and a dwell time of 5 to 30 minutes.
- the invention has the beneficial effects that the present invention utilizes the oxidation resistance of pyrogallic acid to weaken the degree of oxidation of the myofibrillar protein system, and combines with ultra-high pressure treatment to change the internal structure of myofibrillar protein to form a uniform Three-dimensional network gel structure.
- the invention is suitable for the field of food processing accessories and regulates the quality of fish products.
- Figure 1 is a process flow diagram of the present invention.
- FIG. 2 is a scanning electron micrograph of the strongly gelatinous myofibrillar protein obtained under different treatment conditions in Examples 1 to 4 of the present invention.
- a method for preparing a gelled myofibrillar protein comprises the following steps:
- the first separation buffer (1) adding the first separation buffer to the grounded fish gizzard, and sequentially homogenizing and centrifuging to remove the soluble protein to obtain a first precipitate, and then adding a second separation buffer to the first precipitate, in turn The mixture is homogenized, filtered by three layers of gauze and centrifuged to obtain a second precipitate; the pH of the first separation buffer is 7.0, including 0.1 M KCl, 2 mM MgCl 2 , 1 mM EGTA, 0.5 mM DTT and 10 mM K 2 HPO.
- the pH of the second buffer is 6.0, including 0.1 M NaCl and 1 mM NaN 3 ; in this step, the conditions of homogenization treatment are 8000 rpm, 5 min, and the conditions of centrifugation are 8000 rpm, 15 min, 4 ° C;
- the material obtained in the step (4) is placed in a water bath at 20 ° C, heated to 85 ° C at a rate of 1 ° C / min, kept for 30 min, and then immediately placed at 0 ° C for 12 h to obtain the strong gel property.
- Myofibrillar protein Myofibrillar protein.
- the first separation buffer (1) adding the first separation buffer to the grounded fish gizzard, and sequentially homogenizing and centrifuging to remove the soluble protein to obtain a first precipitate, and then adding a second separation buffer to the first precipitate, in turn The mixture is homogenized, filtered by three layers of gauze and centrifuged to obtain a second precipitate; the pH of the first separation buffer is 7.0, including 0.1 M KCl, 2 mM MgCl 2 , 1 mM EGTA, 0.5 mM DTT and 10 mM K 2 HPO.
- the pH of the second buffer is 6.0, including 0.1 M NaCl and 1 mM NaN 3 ; in this step, the conditions of homogenization treatment are 8000 rpm, 5 min, and the conditions of centrifugation are 8000 rpm, 15 min, 4 ° C;
- step (3) The material obtained in the step (3) is placed in a water bath at 20 ° C, heated to 85 ° C at a rate of 1 ° C / min, kept for 30 min, and then immediately placed at 0 ° C for 12 h, which is obtained.
- the first separation buffer (1) adding the first separation buffer to the grounded fish gizzard, and sequentially homogenizing and centrifuging to remove the soluble protein to obtain a first precipitate, and then adding a second separation buffer to the first precipitate, in turn The mixture is homogenized, filtered by three layers of gauze and centrifuged to obtain a second precipitate; the pH of the first separation buffer is 7.0, including 0.1 M KCl, 2 mM MgCl 2 , 1 mM EGTA, 0.5 mM DTT and 10 mM K 2 HPO.
- the pH of the second buffer is 6.0, including 0.1 M NaCl and 1 mM NaN 3 ; in this step, the conditions of homogenization treatment are 8000 rpm, 5 min, and the conditions of centrifugation are 8000 rpm, 15 min, 4 ° C;
- the first separation buffer (1) adding the first separation buffer to the grounded fish gizzard, and sequentially homogenizing and centrifuging to remove the soluble protein to obtain a first precipitate, and then adding a second separation buffer to the first precipitate, in turn The mixture is homogenized, filtered by three layers of gauze and centrifuged to obtain a second precipitate; the pH of the first separation buffer is 7.0, including 0.1 M KCl, 2 mM MgCl 2 , 1 mM EGTA, 0.5 mM DTT and 10 mM K 2 HPO.
- the pH of the second buffer is 6.0, including 0.1 M NaCl and 1 mM NaN 3 ; in this step, the conditions of homogenization treatment are 8000 rpm, 5 min, and the conditions of centrifugation are 8000 rpm, 15 min, 4 ° C;
- Example 1 that is, the addition of pyrogallic acid and ultra-high pressure treatment of myofibrillar protein, the protein gel has small pores, uniform and fine particles, and the gel network is even and orderly, so the gel property is superior to other implementations. example.
- a method for preparing a gelatinous myofibrillar protein comprises the following steps:
- the pH of the first separation buffer is 6.9-7.1, including 0.08-0.12 M KCl, 1.8-2.2 mM MgCl 2 , 0.8-1.2 mM EGTA, 0.4-0.6. mM DTT and 8-11 mM K 2 HPO 4 ;
- the second buffer has a pH of 5.9-6.1, including 0.08-0.12 M NaCl and 0.8-1.2 mM NaN 3 ;
- the above dissolution buffer Is a 0.5-0.7M NaCl solution, the pH is 7.9 ⁇ 8.1;
- the material obtained in the step (4) is placed in a water bath at 18-22 ° C, heated to 82-86 ° C at a rate of 0.8-1.2 ° C / min, kept for 25-40 min, and then immediately placed at 0 ° C for 10 ⁇ At 13h, the strongly gelatinous myofibrillar protein was obtained.
- the conditions of the homogenization treatment in the step (1) are 7000 to 10000 rpm, and 3 to 8 minutes.
- the conditions of centrifugation in the steps (1) and (2) are 7000 to 9000 rpm, 10 to 30 min, and 4 to 8 °C.
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- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Marine Sciences & Fisheries (AREA)
- Zoology (AREA)
- Health & Medical Sciences (AREA)
- Nutrition Science (AREA)
- Biochemistry (AREA)
- Peptides Or Proteins (AREA)
- Meat, Egg Or Seafood Products (AREA)
Abstract
A preparation method for an enhanced gelatinous myofibrillar protein, comprising the following steps: (1) preparing a myofibrillar protein; (2) dissolving the myofibrillar protein, and concentrating same to obtain a concentrate; (3) adding a pyrogallic acid to said concentrate, mixing uniformly, and placing the resultant mixture at normal temperature to obtain a mixed protein solution; (4) vacuum packaging said mixed protein solution and placing same under an ultra-high pressure; and (5) subjecting the material in step (4) to programmed warming, and then performing quenching, so as to obtain the enhanced gelatinous myofibrillar protein.
Description
本发明属于肌原纤维蛋白的制备，具体涉及一种强凝胶性肌原纤维蛋白的制备方法。The invention belongs to the preparation of myofibrillar protein, in particular to a preparation method of strong gel myofibrillar protein.
肌原纤维蛋白是一类具有重要生物学功能特性的盐溶性蛋白，也是构成肌肉纤维的主要蛋白质，占总蛋白含量的50～55％,主要包括肌球蛋白、肌动蛋白、原肌球蛋白、肌钙蛋白等。肌原纤维蛋白参与肌肉的收缩、影响肉的嫩度和肉制品的功能特性，如凝胶性、流变学特性、保水性、弹性、质地等。其中，肌原纤维蛋白的凝胶特性是鱼糜制品加工中最重要的功能特性。凝胶是一种介于固体和液体之间、外观均匀、并保持一定形态的弹性半固体。蛋白能够形成凝胶是因为蛋白质受外界条件的调节或受热后使非共价键解离，表面的结构发生改变，尤其是，疏水基团的暴露促进了蛋白质之间的相互作用，使蛋白质分子在聚合作用下形成均匀的网络状凝胶体。Myofibrillar protein is a kind of salt-soluble protein with important biological functions. It is also the main protein that constitutes muscle fiber, accounting for 50-55% of total protein content, including myosin, actin and tropomyosin. , troponin and so on. Myofibrillar proteins are involved in muscle contraction, affecting tenderness of meat and functional properties of meat products such as gelatinity, rheological properties, water retention, elasticity, texture, and the like. Among them, the gel properties of myofibrillar proteins are the most important functional properties in the processing of surimi products. A gel is an elastic semi-solid that is between a solid and a liquid, has a uniform appearance, and maintains a certain shape. Proteins can form gels because proteins are regulated by external conditions or dissociated by non-covalent bonds after heating, and the structure of the surface changes. In particular, the exposure of hydrophobic groups promotes the interaction between proteins, making protein molecules A uniform network-like gel is formed under polymerization.
研究表明，在漂洗及贮藏过程中，马鲭鱼肉易被氧化，使鱼肉中羰基含量明显增多(Sylvie,Carolinep et al.2015)；虹鳟鱼肉中蛋白质氧化改变了化学作用力，如疏水作用力、二硫键、非共价键等，促使蛋白质分子构象及其聚集方式发生变化(Herrera and Mackie 2004)，鱼肉中蛋白质氧化也是由自由基链式反应所引起的，自由基首先进攻带有活性侧链的肽链骨架(Sante-Lhoutellier,Aubry et al.2007)，如半胱氨酸、色氨酸、赖氨酸，会产生羰基衍生物，削减巯基的数量等，导致氧化后的鱼肉蛋白凝胶能力变差、弹性降低、持水力下降。Studies have shown that during rinsing and storage, mackerel meat is easily oxidized, resulting in a significant increase in carbonyl content in fish meat (Sylvie, Carolinep et al. 2015); protein oxidation in rainbow trout meat changes chemical forces, such as hydrophobic forces, Disulfide bonds, non-covalent bonds, etc., promote the conformation of protein molecules and their aggregation patterns (Herrera and Mackie 2004). Protein oxidation in fish meat is also caused by free radical chain reaction. The peptide chain backbone of the chain (Sante-Lhoutellier, Aubry et al. 2007), such as cysteine, tryptophan, lysine, will produce carbonyl derivatives, reduce the number of sulfhydryl groups, etc., resulting in oxidized fish protein clotting The gel ability is deteriorated, the elasticity is lowered, and the water holding capacity is lowered.
发明内容Summary of the invention
本发明的目的在于克服现有技术缺陷，提供一种强凝胶性肌原纤维蛋白的制备方法。It is an object of the present invention to overcome the deficiencies of the prior art and to provide a method for preparing a strongly gelatinous myofibrillar protein.
本发明的技术方案如下：The technical solution of the present invention is as follows:
1)肌原纤维蛋白的制备1) Preparation of myofibrillar protein
向搅碎后的鱼糜中加入第一分离缓冲液，均质处理，离心，除去可溶性蛋白，得沉淀，加入第二缓冲液，均质，纱布过滤，溶液离心，得沉淀的肌原纤维蛋白。Adding the first separation buffer to the mashed fish gizzard, homogenizing, centrifuging, removing soluble protein, obtaining a precipitate, adding a second buffer, homogenizing, filtering with gauze, centrifuging the solution to obtain precipitated myofibrillar protein .
2)肌原纤维蛋白溶解及浓缩2) Myofibrillar protein solubilization and concentration
将步骤1)得到的肌原纤维蛋白加入到冷的溶解液中，均质，0℃放置30min，离心，取上清液，用超滤管浓缩。The myofibrillar protein obtained in the step 1) was added to a cold solution, homogenized, placed at 0 ° C for 30 min, centrifuged, and the supernatant was taken and concentrated by an ultrafiltration tube.
3)肌原纤维蛋白的抗氧化3) Anti-oxidation of myofibrillar protein
将步骤2)得到的肌原纤维蛋白加入焦性没食子酸，混合均匀，放置1～5h后，得到混合蛋白溶液体系。The myofibrillar protein obtained in the step 2) is added to the pyrogallic acid, mixed uniformly, and left for 1 to 5 hours to obtain a mixed protein solution system.
4)超高压处理4) Ultra high pressure treatment
将步骤3)得到的混合蛋白溶液进行真空包装，置于超高压100～600MPa下处理5～30min。The mixed protein solution obtained in the step 3) is vacuum-packed and placed under an ultra-high pressure of 100 to 600 MPa for 5 to 30 minutes.
5)热诱导凝胶5) Thermally induced gel
将步骤4)得到的样品置于水浴20℃中程序升温至85℃，升温速率1℃/min，在85℃下保温30min，然后骤冷置于0℃，12h，得肌原纤维蛋白凝胶。The sample obtained in the step 4) was placed in a water bath at 20 ° C to be programmed to a temperature of 85 ° C, a heating rate of 1 ° C / min, held at 85 ° C for 30 min, and then quenched at 0 ° C for 12 h to obtain a myofibrillar protein gel. .
在上述技术方法中，在步骤1)中，上述第一分离缓冲液的pH为6.9～7.1，包括0.08～0.12M KCl，1.8～2.2mM MgCl
2，0.8～1.2mM EGTA、0.4～0.6mM DTT和8～11mM K
2HPO
4；上述第二缓冲液的pH为5.9～6.1，包括0.08～0.12M NaCl和0.8～1.2mM.NaN
3。
In the above technical method, in the step 1), the pH of the first separation buffer is 6.9 to 7.1, including 0.08 to 0.12 M KCl, 1.8 to 2.2 mM MgCl 2 , 0.8 to 1.2 mM EGTA, 0.4 to 0.6 mM DTT. And 8-11 mM K 2 HPO 4 ; the second buffer has a pH of 5.9-6.1, including 0.08-0.12 M NaCl and 0.8-1.2 mM. NaN 3 .
在步骤1)中，均质条件为7000～10000rpm，3～8min。离心条件为7000～9000rpm，10～30min，4～8℃。In step 1), the homogenization conditions are 7000 to 10000 rpm, 3 to 8 minutes. The centrifugation conditions are 7000 to 9000 rpm, 10 to 30 minutes, and 4 to 8 °C.
在步骤2)中，溶解液为0.5～1.0M NaCl，pH 7.0～8.0。In the step 2), the solution is 0.5 to 1.0 M NaCl, and the pH is 7.0 to 8.0.
在步骤3)中，添加的焦性没食子酸浓度为6μmol/g蛋白～300μmol/g蛋白。In step 3), the concentration of pyrogallic acid added is from 6 μmol/g protein to 300 μmol/g protein.
在步骤4)中，真空包装包括抽真空时间为15～60s、热封时间1.0～3.0s，超高压处理的压力为100～600MPa，保压时间为5～30min。In step 4), the vacuum packaging comprises a vacuuming time of 15 to 60 s, a heat sealing time of 1.0 to 3.0 s, an ultrahigh pressure treatment pressure of 100 to 600 MPa, and a dwell time of 5 to 30 minutes.
本发明的有益效果是：本发明利用焦性没食子酸的抗氧化性将肌原纤维蛋白的体系被氧化的程度弱化，并结合超高压处理，从而改变肌原纤维蛋白的内部结构，形成均匀的三维网络凝胶结构。本发明适用于食品加工辅料领域，调节鱼肉制品的品质。The invention has the beneficial effects that the present invention utilizes the oxidation resistance of pyrogallic acid to weaken the degree of oxidation of the myofibrillar protein system, and combines with ultra-high pressure treatment to change the internal structure of myofibrillar protein to form a uniform Three-dimensional network gel structure. The invention is suitable for the field of food processing accessories and regulates the quality of fish products.
图1为本发明的工艺流程图。Figure 1 is a process flow diagram of the present invention.
图2为本发明实施例1至4中不同处理条件下所获得的强凝胶性肌原纤维蛋白的扫描电镜图。2 is a scanning electron micrograph of the strongly gelatinous myofibrillar protein obtained under different treatment conditions in Examples 1 to 4 of the present invention.
以下通过具体实施方式结合附图对本发明的技术方案进行进一步的说明和描述。The technical solutions of the present invention will be further described and described below with reference to the accompanying drawings.
实施例1Example 1
如图1所示，一种增强凝胶性肌原纤维蛋白的制备方法，包括如下步骤：As shown in FIG. 1, a method for preparing a gelled myofibrillar protein comprises the following steps:
(1)向搅碎后的鱼糜中加入第一分离缓冲液，依次经均质处理和离心，除去可溶性蛋白，得第一沉淀，再向该第一沉淀中加入第二分离缓冲液，依次经均质处理、三层纱布过滤得液体和离心，得第二沉淀；上述第一分离缓冲液的pH为7.0，包括0.1M KCl，2mM MgCl
2，1mM EGTA、0.5mM DTT和10mM K
2HPO
4；上述第二缓冲液的pH为6.0，包括0.1M NaCl和1mM NaN
3；该步骤中，均质处理的条件为8000rpm，5min，离心的条件为8000rpm，15min，4℃；
(1) adding the first separation buffer to the grounded fish gizzard, and sequentially homogenizing and centrifuging to remove the soluble protein to obtain a first precipitate, and then adding a second separation buffer to the first precipitate, in turn The mixture is homogenized, filtered by three layers of gauze and centrifuged to obtain a second precipitate; the pH of the first separation buffer is 7.0, including 0.1 M KCl, 2 mM MgCl 2 , 1 mM EGTA, 0.5 mM DTT and 10 mM K 2 HPO. 4 ; The pH of the second buffer is 6.0, including 0.1 M NaCl and 1 mM NaN 3 ; in this step, the conditions of homogenization treatment are 8000 rpm, 5 min, and the conditions of centrifugation are 8000 rpm, 15 min, 4 ° C;
(2)向上述第二沉淀中加入冷的溶解缓冲液，均质后于0℃放置30min，然后离心取上清，再用超滤管浓缩，得浓度为20mg/mL的浓缩液；上述溶解缓冲液为0.6M NaCl溶液，pH为8.0；该步骤中离心的条件为8000rpm，15min，4℃(2) adding a cold dissolving buffer to the second precipitate, homogenizing and then placing at 0 ° C for 30 min, then centrifuging to obtain a supernatant, and then concentrating with an ultrafiltration tube to obtain a concentrated solution having a concentration of 20 mg/mL; The buffer solution was 0.6 M NaCl solution and the pH was 8.0; the centrifugation conditions in this step were 8000 rpm, 15 min, 4 ° C.
(3)向上述浓缩液中加入焦性没食子酸，混合均匀，常温下放置2h后，得到混合蛋白溶液；上述焦性没食子酸在浓缩液中的浓度为300μmol/g蛋白；(3) adding coke gallic acid to the above concentrated solution, mixing uniformly, and placing it at room temperature for 2 hours to obtain a mixed protein solution; the concentration of the above-mentioned pyrogallic acid in the concentrated solution is 300 μmol/g protein;
(4)将上述混合蛋白溶液进行袋装抽真空30s、热封2.5s真空包装，置于超高压400MPa下处理15min；(4) The above mixed protein solution was vacuum-packed for 30 s, heat-sealed for 2.5 s, and placed under an ultra-high pressure of 400 MPa for 15 min;
(5)将步骤(4)所得的物料置于20℃的水浴中，以1℃/min的速率升温至85℃，保温30min，然后立即于0℃放置12h，即得所述强凝胶性的肌原纤维蛋白。(5) The material obtained in the step (4) is placed in a water bath at 20 ° C, heated to 85 ° C at a rate of 1 ° C / min, kept for 30 min, and then immediately placed at 0 ° C for 12 h to obtain the strong gel property. Myofibrillar protein.
实施例2Example 2
(1)向搅碎后的鱼糜中加入第一分离缓冲液，依次经均质处理和离心，除去可溶性蛋白，得第一沉淀，再向该第一沉淀中加入第二分离缓冲液，依次经均质处理、三层纱布过滤得液体和离心，得第二沉淀；上述第一分离缓冲液的pH为7.0，包括0.1M KCl，2mM MgCl
2，1mM EGTA、0.5mM DTT和10mM K
2HPO
4；上述第二缓冲液的pH为6.0，包括0.1M NaCl和1mM NaN
3；该步骤中，均质处理的条件为8000rpm，5min，离心的条件为8000rpm，15min，4℃；
(1) adding the first separation buffer to the grounded fish gizzard, and sequentially homogenizing and centrifuging to remove the soluble protein to obtain a first precipitate, and then adding a second separation buffer to the first precipitate, in turn The mixture is homogenized, filtered by three layers of gauze and centrifuged to obtain a second precipitate; the pH of the first separation buffer is 7.0, including 0.1 M KCl, 2 mM MgCl 2 , 1 mM EGTA, 0.5 mM DTT and 10 mM K 2 HPO. 4 ; The pH of the second buffer is 6.0, including 0.1 M NaCl and 1 mM NaN 3 ; in this step, the conditions of homogenization treatment are 8000 rpm, 5 min, and the conditions of centrifugation are 8000 rpm, 15 min, 4 ° C;
(2)向上述第二沉淀中加入冷的溶解液，均质后于0℃放置30min，然后离心取上清，再用超滤管浓缩，得浓度为20mg/mL的浓缩液；上述溶解缓冲液为0.6M NaCl溶液，pH为8.0；该步骤中离心的条件为8000rpm，15min，4℃(2) adding a cold solution to the second precipitate, homogenizing and then placing at 0 ° C for 30 min, then centrifuging to obtain a supernatant, and then concentrating with an ultrafiltration tube to obtain a concentrated solution having a concentration of 20 mg/mL; The solution is 0.6M NaCl solution, the pH is 8.0; the centrifugation conditions in this step are 8000 rpm, 15 min, 4 ° C
(3)将上述浓缩液进行袋装抽真空30s、热封2.5s真空包装，置于超高压400MPa下处理15min；(3) The above concentrated liquid is bagged and vacuumed for 30s, heat-sealed for 2.5s vacuum packaging, and placed under ultra-high pressure 400MPa for 15min;
(4)将步骤(3)所得的物料置于20℃的水浴中，以1℃/min的速率升温至85℃，保温30min，然后立即于0℃放置12h，即得。(4) The material obtained in the step (3) is placed in a water bath at 20 ° C, heated to 85 ° C at a rate of 1 ° C / min, kept for 30 min, and then immediately placed at 0 ° C for 12 h, which is obtained.
实施例3Example 3
(1)向搅碎后的鱼糜中加入第一分离缓冲液，依次经均质处理和离心，除去可溶性蛋白，得第一沉淀，再向该第一沉淀中加入第二分离缓冲液，依次经均质处理、三层纱布过滤得液体和离心，得第二沉淀；上述第一分离缓冲液的pH为7.0，包括0.1M KCl，2mM MgCl
2，1mM EGTA、0.5mM DTT和10mM K
2HPO
4；上述第二缓冲液的pH为6.0，包括0.1M NaCl和1mM NaN
3；该步骤中，均质处理的条件为8000rpm，5min，离心的条件为8000rpm，15min，4℃；
(1) adding the first separation buffer to the grounded fish gizzard, and sequentially homogenizing and centrifuging to remove the soluble protein to obtain a first precipitate, and then adding a second separation buffer to the first precipitate, in turn The mixture is homogenized, filtered by three layers of gauze and centrifuged to obtain a second precipitate; the pH of the first separation buffer is 7.0, including 0.1 M KCl, 2 mM MgCl 2 , 1 mM EGTA, 0.5 mM DTT and 10 mM K 2 HPO. 4 ; The pH of the second buffer is 6.0, including 0.1 M NaCl and 1 mM NaN 3 ; in this step, the conditions of homogenization treatment are 8000 rpm, 5 min, and the conditions of centrifugation are 8000 rpm, 15 min, 4 ° C;
(2)向上述第二沉淀中加入冷的溶解液，均质后于0℃放置30min，然后离心取上清，再用超滤管浓缩，得浓度为20mg/mL的浓缩液；上述溶解缓冲液为0.6M NaCl溶液，pH为8.0；该步骤中离心的条件为8000rpm，15min，4℃(2) adding a cold solution to the second precipitate, homogenizing and then placing at 0 ° C for 30 min, then centrifuging to obtain a supernatant, and then concentrating with an ultrafiltration tube to obtain a concentrated solution having a concentration of 20 mg/mL; The solution is 0.6M NaCl solution, the pH is 8.0; the centrifugation conditions in this step are 8000 rpm, 15 min, 4 ° C
(3)向上述浓缩液中加入焦性没食子酸，混合均匀，常温下放置2h后，得到混合蛋白溶液；上述焦性没食子酸在浓缩液中的浓度为300μmol/g蛋白；(3) adding coke gallic acid to the above concentrated solution, mixing uniformly, and placing it at room temperature for 2 hours to obtain a mixed protein solution; the concentration of the above-mentioned pyrogallic acid in the concentrated solution is 300 μmol/g protein;
(4)将上述混合蛋白溶液置于20℃的水浴中，以1℃/min的速率升温至85℃，保温30min，然后立即于0℃放置12h，即得。(4) The above mixed protein solution was placed in a water bath at 20 ° C, heated to 85 ° C at a rate of 1 ° C / min, incubated for 30 min, and then immediately placed at 0 ° C for 12 h, which is obtained.
实施例4Example 4
(1)向搅碎后的鱼糜中加入第一分离缓冲液，依次经均质处理和离心，除去可溶性蛋白，得第一沉淀，再向该第一沉淀中加入第二分离缓冲液，依次经均质处理、三层纱布过滤得液体和离心，得第二沉淀；上述第一分离缓冲液的pH为7.0，包括0.1M KCl，2mM MgCl
2，1mM EGTA、0.5mM DTT和10mM K
2HPO
4；上述第二缓冲液的pH为6.0，包括0.1M NaCl和1mM NaN
3；该步骤中，均质处理的条件为8000rpm，5min，离心的条件为8000rpm，15min，4℃；
(1) adding the first separation buffer to the grounded fish gizzard, and sequentially homogenizing and centrifuging to remove the soluble protein to obtain a first precipitate, and then adding a second separation buffer to the first precipitate, in turn The mixture is homogenized, filtered by three layers of gauze and centrifuged to obtain a second precipitate; the pH of the first separation buffer is 7.0, including 0.1 M KCl, 2 mM MgCl 2 , 1 mM EGTA, 0.5 mM DTT and 10 mM K 2 HPO. 4 ; The pH of the second buffer is 6.0, including 0.1 M NaCl and 1 mM NaN 3 ; in this step, the conditions of homogenization treatment are 8000 rpm, 5 min, and the conditions of centrifugation are 8000 rpm, 15 min, 4 ° C;
(2)向上述第二沉淀中加入冷的溶解缓冲液，均质后于0℃放置30min，然后离心取上清，再用超滤管浓缩，得浓度为20mg/mL的浓缩液；上述溶解缓冲液为0.6M NaCl溶液，pH为8.0；该步骤中离心的条件为8000rpm，15min，4℃(2) adding a cold dissolving buffer to the second precipitate, homogenizing and then placing at 0 ° C for 30 min, then centrifuging to obtain a supernatant, and then concentrating with an ultrafiltration tube to obtain a concentrated solution having a concentration of 20 mg/mL; The buffer solution was 0.6 M NaCl solution and the pH was 8.0; the centrifugation conditions in this step were 8000 rpm, 15 min, 4 ° C.
(3)将步骤(2)所得的浓缩液置于20℃的水浴中，以1℃/min的速率升温至85℃，保温30min，然后立即于0℃放置12h，即得。(3) The concentrated liquid obtained in the step (2) was placed in a water bath at 20 ° C, heated to 85 ° C at a rate of 1 ° C / min, kept for 30 min, and then immediately placed at 0 ° C for 12 h, which was obtained.
由图2可知，实施例1，即添加焦性没食子酸协同超高压处理的肌原纤维蛋白，蛋白凝胶孔洞小，颗粒均匀细小，凝胶网络均匀有序，所以凝胶特性优于其他实施例。As can be seen from Fig. 2, Example 1, that is, the addition of pyrogallic acid and ultra-high pressure treatment of myofibrillar protein, the protein gel has small pores, uniform and fine particles, and the gel network is even and orderly, so the gel property is superior to other implementations. example.
本领域普通技术人员可知，本发明的技术方案在下述范围内变化时，仍然能够得到与上述实施例相同或相近的技术效果，仍然属于本发明的保护范围：It will be apparent to those skilled in the art that when the technical solution of the present invention is changed within the following range, the same or similar technical effects as those of the above embodiment can still be obtained, and still belong to the protection scope of the present invention:
一种凝胶性肌原纤维蛋白的制备方法，包括如下步骤：A method for preparing a gelatinous myofibrillar protein comprises the following steps:
(1)向搅碎后的鱼糜中加入第一分离缓冲液，依次经均质处理和离心，除去可溶性蛋白，得第一沉淀，再向该第一沉淀中加入第二分离缓冲液，依次经均质处理、过滤和离心，得第二沉淀；上述第一分离缓冲液的pH为6.9～7.1，包括0.08～0.12M KCl，1.8～2.2mM MgCl
2，0.8～1.2mM EGTA、0.4～0.6mM DTT和8～11mM K
2HPO
4；上述第二缓冲液的pH为5.9～6.1，包括0.08～0.12M NaCl和0.8～1.2mM NaN
3；
(1) adding the first separation buffer to the grounded fish gizzard, and sequentially homogenizing and centrifuging to remove the soluble protein to obtain a first precipitate, and then adding a second separation buffer to the first precipitate, in turn After homogenization, filtration and centrifugation, a second precipitate is obtained; the pH of the first separation buffer is 6.9-7.1, including 0.08-0.12 M KCl, 1.8-2.2 mM MgCl 2 , 0.8-1.2 mM EGTA, 0.4-0.6. mM DTT and 8-11 mM K 2 HPO 4 ; the second buffer has a pH of 5.9-6.1, including 0.08-0.12 M NaCl and 0.8-1.2 mM NaN 3 ;
(2)向上述第二沉淀中加入冷的溶解液，均质后于～1～2℃放置25～40min，然后离心取上清，再用超滤管浓缩，得浓缩液；上述溶解缓冲液为0.5～0.7M NaCl溶液，pH为7.9～8.1；(2) adding a cold solution to the second precipitate, homogenizing and then placing at ~1 to 2 ° C for 25 to 40 minutes, then centrifuging to obtain a supernatant, and then concentrating with an ultrafiltration tube to obtain a concentrated solution; the above dissolution buffer Is a 0.5-0.7M NaCl solution, the pH is 7.9 ~ 8.1;
(3)向上述浓缩液中加入焦性没食子酸，混合均匀，常温下放置1～5h后，得到混合蛋白溶液；上述焦性没食子酸在浓缩液中的浓度为6μ～300μmol/g蛋白；(3) adding coke gallic acid to the above concentrated solution, uniformly mixing, and standing at normal temperature for 1 to 5 hours, to obtain a mixed protein solution; the concentration of the above-mentioned pyrogallic acid in the concentrated solution is 6 μ-300 μmol/g protein;
(4)将上述混合蛋白溶液进行真空包装，置于超高压100～600MPa下处理5～25min；(4) The above mixed protein solution is vacuum-packed and placed under an ultra-high pressure of 100-600 MPa for 5 to 25 minutes;
(5)将步骤(4)所得的物料置于18～22℃的水浴中，以0.8～1.2℃/min的速率升温至82～86℃，保温25～40min，然后立即于0℃放置10～13h，即得所述强凝胶性肌原纤维蛋白。(5) The material obtained in the step (4) is placed in a water bath at 18-22 ° C, heated to 82-86 ° C at a rate of 0.8-1.2 ° C / min, kept for 25-40 min, and then immediately placed at 0 ° C for 10 ~ At 13h, the strongly gelatinous myofibrillar protein was obtained.
所述步骤(1)中的均质处理的条件为7000～10000rpm，3～8min。所述步骤(1)和(2)中的离心的条件为7000～9000rpm，10～30min，4～8℃。The conditions of the homogenization treatment in the step (1) are 7000 to 10000 rpm, and 3 to 8 minutes. The conditions of centrifugation in the steps (1) and (2) are 7000 to 9000 rpm, 10 to 30 min, and 4 to 8 °C.
以上所述，仅为本发明的较佳实施例而已，故不能依此限定本发明实施的范围，即依本发明专利范围及说明书内容所作的等效变化与修饰，皆应仍属本发明涵盖的范围内。The above is only the preferred embodiment of the present invention, and thus the scope of the present invention is not limited thereto, and equivalent changes and modifications made in accordance with the scope of the present invention and the contents of the specification should still be covered by the present invention. In the range.
Claims (13)
- 一种增强凝胶性肌原纤维蛋白的制备方法，其特征在于包括如下步骤：A method for preparing a gelled myofibrillar protein, comprising the steps of:(1)向搅碎后的鱼糜中加入第一分离缓冲液，依次经均质处理和离心，除去可溶性蛋白，得第一沉淀，再向该第一沉淀中加入第二分离缓冲液，依次经均质处理、过滤和离心，得第二沉淀；上述第一分离缓冲液的pH为6.9～7.1，包括0.08～0.12M KCl，1.8～2.2mM MgCl 2，0.8～1.2mM EGTA、0.4～0.6mM DTT和8～11mM K 2HPO 4；上述第二缓冲液的pH为5.9～6.1，包括0.08～0.12M NaCl和0.8～1.2mM NaN 3； (1) adding the first separation buffer to the grounded fish gizzard, and sequentially homogenizing and centrifuging to remove the soluble protein to obtain a first precipitate, and then adding a second separation buffer to the first precipitate, in turn After homogenization, filtration and centrifugation, a second precipitate is obtained; the pH of the first separation buffer is 6.9-7.1, including 0.08-0.12 M KCl, 1.8-2.2 mM MgCl 2 , 0.8-1.2 mM EGTA, 0.4-0.6. mM DTT and 8-11 mM K 2 HPO 4 ; the second buffer has a pH of 5.9-6.1, including 0.08-0.12 M NaCl and 0.8-1.2 mM NaN 3 ;(2)向上述第二沉淀中加入冷的溶解液，均质后于～1～2℃放置25～40min，然后离心取上清，再用超滤管浓缩，得浓缩液；上述溶解液为0.5～0.7M NaCl溶液，pH为7.9～8.1；(2) adding a cold solution to the second precipitate, homogenizing and placing at ~1 to 2 ° C for 25 to 40 minutes, then centrifuging to obtain a supernatant, and then concentrating with an ultrafiltration tube to obtain a concentrated solution; the above solution is 0.5 ~ 0.7M NaCl solution, pH is 7.9 ~ 8.1;(3)向上述浓缩液中加入焦性没食子酸，混合均匀，常温下放置1～5h后，得到混合蛋白溶液；上述焦性没食子酸在浓缩液中的浓度为6μ～300μmol/g蛋白；(3) adding coke gallic acid to the above concentrated solution, uniformly mixing, and standing at normal temperature for 1 to 5 hours, to obtain a mixed protein solution; the concentration of the above-mentioned pyrogallic acid in the concentrated solution is 6 μ-300 μmol/g protein;(4)将上述混合蛋白溶液进行真空包装，置于超高压100～600MPa下处理5～25min；(4) The above mixed protein solution is vacuum-packed and placed under an ultra-high pressure of 100-600 MPa for 5 to 25 minutes;(5)将步骤(4)所得的物料置于18～22℃的水浴中，以0.8～1.2℃/min的速率升温至82～86℃，保温25～40min，然后立即于0℃放置10～13h，即得所述强凝胶性的肌原纤维蛋白凝胶。(5) The material obtained in the step (4) is placed in a water bath at 18-22 ° C, heated to 82-86 ° C at a rate of 0.8-1.2 ° C / min, kept for 25-40 min, and then immediately placed at 0 ° C for 10 ~ At 13 h, the strongly gelatinous myofibrillar protein gel was obtained.
- 如权利要求1所述的制备方法，其特征在于：所述第一分离缓冲液的pH为7.0，包括0.1M KCl，2mM MgCl 2，1mM EGTA、0.5mM DTT和10mM K 2HPO 4。 The preparation method according to claim 1, wherein the first separation buffer has a pH of 7.0, and comprises 0.1 M KCl, 2 mM MgCl 2 , 1 mM EGTA, 0.5 mM DTT, and 10 mM K 2 HPO 4 .
- 如权利要求1所述的制备方法，其特征在于：所述第二缓冲液的pH为6.0，包括0.1M NaCl和1mM NaN 3。 The preparation method according to claim 1, wherein the second buffer has a pH of 6.0 and comprises 0.1 M NaCl and 1 mM NaN 3 .
- 如权利要求1所述的制备方法，其特征在于：所述溶解液为0.6M NaCl溶液，pH为8.0。The preparation method according to claim 1, wherein the solution is a 0.6 M NaCl solution and has a pH of 8.0.
- 如权利要求1所述的制备方法，其特征在于：所述步骤(1)中的均质处理的条件为7000～10000rpm，3～8min。The preparation method according to claim 1, wherein the conditions of the homogenization treatment in the step (1) are 7000 to 10000 rpm, 3 to 8 minutes.
- 如权利要求1所述的制备方法，其特征在于：所述步骤(1)和(2)中的离心的条件为7000～9000rpm，10～30min，4～8℃。The production method according to claim 1, wherein the conditions of the centrifugation in the steps (1) and (2) are 7,000 to 9000 rpm, 10 to 30 minutes, and 4 to 8 °C.
- 如权利要求1所述的制备方法，其特征在于：所述步骤(4)为：将上述 混合蛋白溶液进行真空包装，置于超高压400MPa下处理15min。The preparation method according to claim 1, wherein the step (4) is: vacuum-packing the mixed protein solution and placing it under an ultra-high pressure of 400 MPa for 15 minutes.
- 如权利要求1所述的制备方法，其特征在于：所述步骤(5)为：将步骤(4)所得的物料置于20℃的水浴中，以1℃/min的速率升温至85℃，保温30min，然后立即于0℃放置12h，即得所述强凝胶性的肌原纤维蛋白凝胶。The preparation method according to claim 1, wherein the step (5) is: the material obtained in the step (4) is placed in a water bath at 20 ° C, and the temperature is raised to 85 ° C at a rate of 1 ° C / min. The strong gelatinous myofibrillar protein gel was obtained by incubating for 30 min and then immediately at 0 ° C for 12 h.
- 一种增强凝胶性肌原纤维蛋白的制备方法，包括：A method for preparing a gelatinous myofibrillar protein, comprising:1)肌原纤维蛋白的制备：1) Preparation of myofibrillar protein:向搅碎后的鱼糜中加入第一分离缓冲液，均质处理，离心，除去可溶性蛋白，得沉淀；加入第二缓冲液，均质，纱布过滤，溶液离心，沉淀为肌原纤维蛋白；Adding the first separation buffer to the grounded fish gizzard, homogenizing, centrifuging, removing the soluble protein to obtain a precipitate; adding a second buffer, homogenizing, filtering with gauze, centrifuging the solution, and precipitating into myofibrillar protein;2)肌原纤维蛋白溶解及浓缩：2) Myofibrillar protein solubilization and concentration:将步骤1)得到的肌原纤维蛋白加入到冷的溶解液中，均质，0℃放置30min，离心，取上清液，用超滤管浓缩；The myofibrillar protein obtained in the step 1) is added to the cold solution, homogenized, placed at 0 ° C for 30 min, centrifuged, and the supernatant is taken and concentrated with an ultrafiltration tube;3)肌原纤维蛋白的抗氧化：3) Anti-oxidation of myofibrillar protein:将步骤2)得到的肌原纤维蛋白加入焦性没食子酸，混合均匀，放置1～5h后，得到混合蛋白溶液体系；The myofibrillar protein obtained in the step 2) is added to the pyrogallic acid, mixed uniformly, and left for 1 to 5 hours to obtain a mixed protein solution system;4)超高压处理4) Ultra high pressure treatment将步骤3)得到的混合蛋白溶液进行真空包装，置于超高压100～600MPa下处理5～30min；The mixed protein solution obtained in the step 3) is vacuum-packed, and placed under an ultra-high pressure of 100-600 MPa for 5 to 30 minutes;在上述技术方法中，在步骤1)中，第一分离缓冲液为0.08～0.12MKCl，1.8～2.2mM MgCl 2，0.8～1.2mMEGTA,0.4～0.6mM DTT和8～11mMK 2HPO 4，pH6.9-7.1，第二分离缓冲液为0.08～0.12M NaCl和0.8～1.2mMNaN 3，pH5.9-6.1； In the above technical method, in the step 1), the first separation buffer is 0.08 to 0.12 MKCl, 1.8 to 2.2 mM MgCl 2 , 0.8 to 1.2 mM EGTA, 0.4 to 0.6 mM DTT and 8 to 11 mM K 2 HPO 4 , pH 6. 9-7.1, the second separation buffer is 0.08-0.12 M NaCl and 0.8-1.2 mM NaN 3 , pH 5.9-6.1;在步骤2)中，溶解液为0.5～1.0M NaCl，pH 7.0～8.0。In the step 2), the solution is 0.5 to 1.0 M NaCl, and the pH is 7.0 to 8.0.
- 根据权利要求9所述的一种强凝胶性肌原纤维蛋白的制备方法，其特征还包括：The method for preparing a strong gel myofibrillar protein according to claim 9, further comprising:5)热诱导凝胶：5) Thermally induced gel:将步骤4)得到的样品置于水浴20℃中程序升温至85℃，升温速率1℃/min，在85℃下保温30min，然后骤冷置于0℃，12h，得肌原纤维蛋白凝胶。The sample obtained in the step 4) was placed in a water bath at 20 ° C to be programmed to a temperature of 85 ° C, a heating rate of 1 ° C / min, held at 85 ° C for 30 min, and then quenched at 0 ° C for 12 h to obtain a myofibrillar protein gel. .
- 根据权利要求9或10所述的一种增强凝胶性肌原纤维蛋白的制备方法，其特征在步骤1)中，均质条件为7000～10000rpm，3～8min，离心条件为7000～9000rpm，10～30min，4～8℃。The method for preparing a gelled myofibrillar protein according to claim 9 or 10, wherein in the step 1), the homogenization condition is 7000 to 10000 rpm, 3 to 8 minutes, and the centrifugation condition is 7000 to 9000 rpm. 10 to 30 min, 4 to 8 ° C.
- 根据权利要求9或10所述的一种强凝胶性肌原纤维蛋白的制备方法，其特征在步骤3)中，添加的焦性没食子酸浓度为6μmol/g蛋白～300μmol/g蛋白。The method for producing a strongly gelatinous myofibrillar protein according to claim 9 or 10, wherein in the step 3), the concentration of the pyrogallic acid added is from 6 μmol/g protein to 300 μmol/g protein.
- 根据权利要求9或10所述的一种增强凝胶性肌原纤维蛋白的制备方法，其特征在步骤4)中，真空包装包括抽真空时间为15～60s、热封时间1.0-3.0s，超高压处理的压力为100～600MPa，保压时间为5～30min。The method for preparing a gelled myofibrillar protein according to claim 9 or 10, wherein in the step 4), the vacuum packaging comprises a vacuuming time of 15 to 60 s and a heat sealing time of 1.0 to 3.0 s. The pressure of the ultrahigh pressure treatment is 100 to 600 MPa, and the pressure holding time is 5 to 30 minutes.
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| CN108850420A (en) * | 2018-06-08 | 2018-11-23 | 天津商业大学 | A method of improving chicken myofibril protein antioxidant and functional characteristic |
| CN109553675B (en) * | 2018-11-02 | 2022-05-20 | 广东海洋大学 | Preparation method of tilapia myosin heat-induced gel |
| CN109924339B (en) * | 2019-03-05 | 2022-08-26 | 大连工业大学 | Method for enhancing fish myofibrillar protein gel characteristics based on combination of catechin and polyphenol oxidase |
| CN109956999A (en) * | 2019-03-05 | 2019-07-02 | 大连工业大学 | A method of fish myofibrillar protein gel characteristic is improved in conjunction with polyphenol oxidase based on kelp polyphenol |
| CN109929028B (en) * | 2019-03-28 | 2021-02-09 | 浙江省农业科学院 | Method for improving properties of myofibrillar proteins in meat paste |
| CN110214851A (en) * | 2019-05-08 | 2019-09-10 | 大连工业大学 | A method of improving fish myofibrillar protein gel characteristic |
| CN110100942B (en) * | 2019-05-14 | 2022-04-12 | 浙江万里学院 | Method for regulating and controlling adsorption capacity of marine fish myofibrillar protein on flavor compounds |
| CN110845593B (en) * | 2019-11-27 | 2021-05-28 | 中国水产科学研究院黄海水产研究所 | Separation and identification method of patinopecten yessoensis myofibrils |
| CN114947077A (en) * | 2022-07-04 | 2022-08-30 | 吉林农业大学 | Method for improving myofibrillar protein gel structure and water retention of penaeus vannamei boone |
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