WO2020238151A1 - 一种咸味增强肽及其制备方法和用途 - Google Patents

一种咸味增强肽及其制备方法和用途 Download PDF

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WO2020238151A1
WO2020238151A1 PCT/CN2019/124743 CN2019124743W WO2020238151A1 WO 2020238151 A1 WO2020238151 A1 WO 2020238151A1 CN 2019124743 W CN2019124743 W CN 2019124743W WO 2020238151 A1 WO2020238151 A1 WO 2020238151A1
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salty taste
preparation
bream
protease
enhancing peptide
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PCT/CN2019/124743
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English (en)
French (fr)
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孙为正
李微
苏国万
赵谋明
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华南理工大学
广州现代产业技术研究院
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Publication of WO2020238151A1 publication Critical patent/WO2020238151A1/zh

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    • 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
    • A23L27/00—Spices; Flavouring agents or condiments; Artificial sweetening agents; Table salts; Dietetic salt substitutes; Preparation or treatment thereof
    • A23L27/88—Taste or flavour enhancing agents
    • 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
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/17—Amino acids, peptides or proteins
    • A23L33/18—Peptides; Protein hydrolysates
    • 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
    • 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 field of food additives, and specifically relates to a salty taste enhancing peptide and a preparation method and application thereof.
  • Salty enhancing peptides have received widespread attention due to their nutritional and functional properties. They have good development prospects and conform to the health concept of maintaining and improving physical health. The healthy China 2030 strategic plan proposed by the state.
  • the salty taste enhancing peptide is a proteolysis product obtained by using proteolysis technology. Its natural and wonderful characteristic flavor is combined with different taste components to give foods multi-level taste characteristics. The taste characteristics of peptides have a lot to do with the taste characteristics of amino acids. It has a buffering capacity due to the amphiphilic group containing amino and carboxyl groups, which has a subtle effect on the flavor of food, and can moderate the bitterness and astringency caused by other substitute ions such as K + .
  • salty taste enhancing peptide has pharmacological effects such as anti-oxidation, lowering blood sugar, lowering blood lipid and improving immune function. As a functional nutritious natural seasoning, the international high-end seasoning market has listed protein hydrolysate as an important research and development direction.
  • the primary purpose of the present invention is to provide a method for preparing salty taste enhancing peptides.
  • Another object of the present invention is to provide a salty taste enhancing peptide prepared by the above method.
  • the salty taste enhancing peptide has a fresh and salty taste, can enhance salty taste, and has nutritional and health care functions.
  • Another object of the present invention is to provide the use of the above salty taste enhancing peptide.
  • a method for preparing salty taste enhancing peptides includes the following steps:
  • Proteolysis After pretreatment of large yellow croaker, the minced fish and deionized water are evenly mixed, and then subjected to ultra-high pressure treatment, adding protease for enzymolysis for several hours, then inactivating the enzyme, freezing and centrifuging, and supernatant ultrafiltration to obtain the enzymatic hydrolysate;
  • Salty taste enhancement treatment the bream fish polypeptide powder is mixed with other non-sodium salts to obtain the solute, which is added to the large yellow croaker enzymatic hydrolysate, after the pulse electric field treatment, freeze-drying to obtain the salty taste enhancing peptide;
  • step (1) the pretreatment of large yellow croaker is to mince after removing the head and tail and viscera; the mincing is to use a meat grinder with an orifice plate diameter of 6 mm.
  • the protease is pancreatin.
  • step (1) the added amount of the protease is 0.5% of the protein content of large yellow croaker.
  • step (1) the pressure of the ultra-high pressure treatment is 100-150 MPa.
  • step (1) the enzymolysis time is 3 to 5 hours.
  • the ultrafiltration is to use a 5KDa ultrafiltration membrane to obtain a polypeptide mixture with a molecular weight below 5KDa.
  • step (2) the pretreatment of the bream is a process of removing the head and tail and guts, and the fish meat is crushed;
  • step (2) the feed-water ratio of bream meat to water is added at 1.0:1.5;
  • the proteases are papain and pancreatin, and papain is used first, and the hydrolysis time is 2 to 5 hours, and then pancreatin is used, and the hydrolysis time is 1 to 3 hours.
  • step (2) based on the mass of bream (meat) as a calculation standard, the added amount of the protease is 0.1% to 1.0%.
  • step (2) the temperature of the hydrolysis is 40-60°C, and the pH of the hydrolysis system is 5.4-7.0.
  • step (2) the inactivation is inactivation by boiling water, and the inactivation time is 10-30 min.
  • step (2) the centrifugal separation time is 20-60 minutes, the temperature is 4°C, and the rotation speed is 4500-8000 rpm.
  • the other non-sodium salts are sodium chloride, potassium chloride and calcium lactate.
  • step (3) is composed of the following components by weight percentage:
  • the pulse electric field parameters are 0.5-1.0 kV/cm, the number of pulses is 10-20, the pulse width is 10-15 ⁇ s, and the pulse frequency is 1 Hz.
  • the present invention uses large yellow croaker (Larimichthys crocea) as a raw material, prepares salty taste-enhancing proteolysis product through high-efficiency proteolysis, and adds it to the prepared low-sodium salty taste agent, which enhances the saltiness of the low-sodium salty taste agent Degree of taste.
  • the small-molecule mixed peptide obtained by enzymatic hydrolysis of large yellow croaker meat presents more salty and umami taste. Its addition makes it possible to reconcile and relieve the metallic bitterness caused by other substitute ions such as K + .
  • the invention not only enhances the saltiness value of the compound salt, reduces the added amount of sodium ions, but also provides a new salty agent with nutritional and health care functions.
  • the salty taste enhancing peptide prepared by the above method can be applied in the food field.
  • the present invention has the following advantages and effects:
  • the invention reduces the salt content under the same saltiness, and reduces the sodium ion content by 40%-50%.
  • small molecule peptides as protein hydrolysates, play an important role in the body's immune regulation, antibacterial, antiviral, and antioxidant properties. effect. Therefore, the salty taste enhancing peptide prepared by the method is a salty taste enhancer that has rich nutritional value and can also maintain the original flavor balance of the food, thereby effectively enhancing the salty taste.
  • a salty taste enhancing peptide prepared by the following steps:
  • the mass ratios were respectively: potassium chloride 0.45, sodium chloride 0.30, calcium lactate 0.15, and bream polypeptide powder 0.10 and mixed uniformly, and 1 g was dissolved in 100 g of water to make a salt solution.
  • the salty value at this time was 6.02.
  • the salty taste enhancing peptide with a mass concentration of 0.20% was added to the salty taste mixture, and after 20 times of pulse electric field pulses of 0.5kV/cm, 15 ⁇ s, 1Hz, freeze-dried to obtain the product, the salty value increased to 6.42.
  • the product is dissolved in water (with a concentration of 0.45%) and can be obtained by sensory evaluation.
  • the solution is clear and transparent in color, moderate in saltiness, fresh and salty taste, no unpleasant odor, and high nutritional value.
  • a salty taste enhancing peptide prepared by the following steps:
  • the mass ratios are respectively: potassium chloride 0.25, sodium chloride 0.30, calcium lactate 0.05, and bream polypeptide powder 0.40 and mixed uniformly, and 1 g is dissolved in 100 g of water to make a salt solution.
  • the salty value at this time is 6.31.
  • the salty taste enhancing peptide with a mass concentration of 0.20% was added to the salty taste mixture, and after 10 times of pulse electric field pulses of 1.0 kV/cm, 20 ⁇ s, 1 Hz, the product was freeze-dried to obtain a product with a salty taste value increased to 6.57.
  • the product is dissolved in water (with a concentration of 0.45%), and it can be obtained by sensory evaluation.
  • the solution is clear and transparent in color, pure in salty taste, slightly salty taste, no unpleasant odor, and rich in nutrients.
  • a salty taste enhancing peptide prepared by the following steps:
  • the mass ratios were respectively: potassium chloride 0.37, sodium chloride 0.30, calcium lactate 0.15, and bream polypeptide powder 0.18 mixed uniformly, and 1 g was dissolved in 100 g of water to make a salt solution. At this time, the saltiness value was 6.14.
  • the salty taste enhancing peptide with a mass concentration of 0.20% was added to the salty taste mixture, and after 10 times of pulse electric field pulses of 1.0 kV/cm, 10 ⁇ s, 1 Hz, the product was freeze-dried and the salty taste value increased to 6.54.
  • the product is dissolved in water (with a concentration of 0.45%), and it can be obtained by sensory evaluation that the solution is transparent and slightly yellow, moderately salty, strong in flavor, and rich in nutrients.
  • Table 1 shows the salty scores of the bream enzymatic hydrolysate product BP and salts after compounding
  • Table 2 shows the change of the salty value after adding 0.20% of the large yellow croaker proteolytic product HP. It can be seen from the table that, corresponding to each group of samples in Table 1, the salty value of the complex salt increased after the addition of the large yellow croaker proteolysis product HP, which fully indicates that the large yellow croaker proteolysis product has a significant salty enhancement effect.

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  • Molecular Biology (AREA)
  • Polymers & Plastics (AREA)
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Abstract

一种咸味增强肽及其制备方法和用途，该咸味增强肽的制备方法包括以下步骤：大黄鱼预处理后，加入蛋白酶酶解数小时，冷冻离心，上清液超滤得到酶解液；鳊鱼预处理后，加入水和蛋白酶进行水解，冷冻离心，冷冻干燥，得到鳊鱼多肽粉；将鳊鱼多肽粉与其他非钠盐混合，得到溶质，加入到大黄鱼酶解液中，经脉冲电场处理后，冷冻干燥得到咸味增强肽。所述方法在同等咸度下降低了食盐含量，使钠离子含量降低了40％～50％。因此，所述方法制备的咸味增强肽是一种具有丰富的营养价值和也能够保持食品原本的风味平衡、进而有效地增强咸味的咸味增强剂。

Description

一种咸味增强肽及其制备方法和用途 技术领域
本发明属于食品添加剂领域，具体涉及一种咸味增强肽及其制备方法和用途。
背景技术
随着人们生活水平的提高，人们越来越重视饮食健康，咸味增强肽以其营养性和功能性受到广泛的关注，具有良好的发展前景，符合保持、提高身体健康素质的健康理念，符合国家提出的健康中国2030战略规划。
据研究表明，长期摄入高钠食物容易导致钠的滞留，从而增加高血压、肾脏病、心血管疾病以及骨质疏松症等疾病的发病率。随着经济发展以及消费水平的提升，消费者的饮食观念逐步趋于健康安全，因而开发一种低钠产品具有广阔的市场前景。
咸味增强肽是利用蛋白酶解技术得到的蛋白酶解产物，其天然、奇妙的特征风味与不同呈味组分相结合，给予食品多层次的呈味特征。多肽的呈味特性与其组成的氨基酸呈味特性有着很大关系。它由于含有氨基和羧基两性基团而具有缓冲能力，对食品的风味有微妙的影响，可以对其他替代离子如K +而导致的金属苦涩味起到调和缓解作用。同时，研究表明，咸味增强肽作为一种蛋白水解产物，它具有抗氧化、降血糖、降血脂以及提高免疫功能等药理功效作用。作为功能性营养型的天然调味料，国际高档调味料市场已将蛋白水解物列为研究和发展的一个重要方向。
发明内容
为了降低钠含量，保证咸味纯度，本发明的首要目的在于提供一种咸味增强肽的制备方法。
本发明的另一目的在于提供由上述方法制得的咸味增强肽，该咸味增强肽 口感鲜咸、能增强咸味、具有营养保健功能。
本发明的再一目的在于提供上述咸味增强肽的用途。
本发明的目的通过下述技术方案实现：
一种咸味增强肽的制备方法，包括如下步骤：
(1)蛋白酶解：大黄鱼预处理后，鱼肉糜与去离子水混合均匀后，进行超高压处理，加入蛋白酶酶解数小时，然后灭酶，冷冻离心，上清液超滤得到酶解液；
(2)鳊鱼酶解：鳊鱼预处理后，加入水和蛋白酶进行水解，然后灭活蛋白酶，冷冻离心，滤去残渣和浮油，得到鳊鱼酶解液，冷冻干燥，得到鳊鱼多肽粉；
(3)咸味增强处理：将鳊鱼多肽粉与其他非钠盐混合，得到溶质，加入到大黄鱼酶解液中，经脉冲电场处理后，冷冻干燥得到咸味增强肽；
进一步地，步骤(1)中，所述的大黄鱼预处理是去头去尾去内脏后，绞碎；所述绞碎是使用孔板直径为6mm的绞肉机。
进一步地，步骤(1)中，所述蛋白酶为胰酶。
更进一步地，步骤(1)中，所述蛋白酶添加量为大黄鱼蛋白质含量的0.5％。
进一步地，步骤(1)中，所述超高压处理，其压力为100-150MPa。
进一步地，步骤(1)中，所述酶解，其酶解时间为3～5h。
进一步地，步骤(1)中，所述超滤为使用5KDa超滤膜，得到组分为5KDa以下分子量段的多肽混合物。
进一步地，步骤(2)中，所述的鳊鱼预处理是经过去头去尾去内脏，鱼肉搅碎处理；
进一步地，步骤(2)中，鳊鱼肉与水的料水比按1.0:1.5添加；
进一步地，步骤(2)中，所述蛋白酶是木瓜蛋白酶和胰酶，且先用木瓜蛋白酶，水解时间为2～5h，后用胰酶，水解时间为1～3h。
更进一步地，步骤(2)中，以鳊鱼(肉)的质量为计算基准，所述蛋白酶的添加量为0.1％～1.0％。
进一步地，步骤(2)中，所述水解的温度为40～60℃，水解体系的pH值为5.4～7.0。
进一步地，步骤(2)中，所述灭活是采用沸水灭活，灭活时间为10～30min。
进一步地，步骤(2)中，所述离心分离时间为20～60min，温度4℃，转速为4500～8000rpm。
进一步地，步骤(3)中，所述其他非钠盐为氯化钠、氯化钾和乳酸钙。
进一步地，步骤(3)中所述的溶质由以下重量百分比的成分组成：
Figure PCTCN2019124743-appb-000001
更进一步地，所述脉冲电场参数为0.5～1.0kV/cm，脉冲次数10～20次，脉冲宽度10～15μs，脉冲频率1Hz。
本发明以大黄鱼(Larimichthys crocea)为原料，经高效蛋白酶解制备咸味增强蛋白酶解物，并将其加入制备的低钠咸味剂中，在原有基础上增强了低钠咸味剂的咸味程度。大黄鱼肉酶解得到的小分子混合肽更多地呈现咸味和鲜味，它的加入使得原有可以对其他替代离子如K +而导致的金属苦涩味起到调和缓解作用。本发明不仅增强了复配盐的咸味值，减少了钠离子的添加量，还提供了一种具有营养保健功能的新型咸味剂。
由上述方法制得的咸味增强肽可以应用在食品领域中。
本发明相对于现有技术具有如下的优点及效果：
本发明在同等咸度下降低了食盐含量，使钠离子含量降低了40％～50％，此外，小分子肽作为蛋白水解产物，对机体调节免疫、抗菌、抗病毒、抗氧化等起着重要作用。因此本方法制备的咸味增强肽是一种具有丰富的营养价值和也能够保持食品原本的风味平衡、进而有效地增强咸味的咸味增强剂。
具体实施方式
下面结合实施例对本发明作进一步详细的描述，但本发明的实施方式不限于此。
实施例1
一种咸味增强肽，通过以下步骤制得：
(1)蛋白酶解
取50g大黄鱼肉糜加入50g去离子水混合均匀后，经100MPa超高静压处理20min，加入蛋白含量0.5％的胰酶，55℃下酶解5h，然后进行沸水灭酶10min，4℃8000rpm冷冻离心30min，过滤得到酶解液后用5KDa的超滤膜过滤，滤过液即为咸味增强蛋白酶解物。
(2)鳊鱼酶解：
取鳊鱼与水质量比1:1.5混合，加入鳊鱼质量的1％木瓜蛋白酶，对其进行水解，水解温度为40℃，pH值为5.4，时间为2h，后加入胰酶，继续酶解3h。采用高温灭酶30min，4℃，7000rpm冷冻离心30min，滤去残渣和浮油，得鳊鱼酶解液，对滤液进行真空冷冻干燥，制得鳊鱼多肽粉。
(3)咸味混合物的制备
将质量比分别为：氯化钾0.45、氯化钠0.30、乳酸钙0.15、鳊鱼多肽粉0.10混合均匀，取1g溶于100g水中制成盐溶液，此时的咸味值为6.02。
(4)咸味增强作用
将质量浓度为0.20％的咸味增强肽加入咸味混合物中，经0.5kV/cm、15μs、1Hz的脉冲电场脉冲次数20次处理后，冷冻干燥得到产品，咸味值增加至6.42。
(5)感官评价
将产品溶于水(浓度为0.45％)，经感官评价可得，该溶液颜色澄清透明、咸度适中、口感鲜咸、无不良异味、营养价值高。
实施例2
一种咸味增强肽，通过以下步骤制得：
(1)蛋白酶解
取50g鳊鱼肉糜加入50g去离子水混合均匀后，经125MPa超高静压处理20min，加入鳊鱼蛋白0.5％的胰酶，50℃下酶解4h，水解完成后进行沸水灭酶20min，4℃8000rpm冷冻离心25min，过滤得到酶解液后用5KDa的超滤膜过滤，滤过液即为咸味增强蛋白酶解物。
(2)鳊鱼酶解：
取鳊鱼与水质量比1:1.5混合，加入鳊鱼质量的1.5％碱性蛋白酶，对其进行 水解，水解温度为50℃，pH值为6.0，时间，2.5h，后添加胰酶，继续酶解2h。采用高温沸水灭酶30min，4℃，7500rpm冷冻离心25min，滤去残渣和浮油，得鳊鱼酶解液，对滤液进行真空冷冻干燥，制得鳊鱼多肽粉。
(3)咸味混合物的制备
将质量比分别为：氯化钾0.25、氯化钠0.30、乳酸钙0.05、鳊鱼多肽粉0.40混合均匀，取1g溶于100g水中制成盐溶液，此时的咸味值为6.31。
(4)咸味增强作用
将质量浓度为0.20％的咸味增强肽加入咸味混合物中，经1.0kV/cm、20μs、1Hz的脉冲电场脉冲次数10次处理后，冷冻干燥得到产品，咸味值增至6.57。
(5)感官评价
将产品溶于水(浓度为0.45％)，经感官评价可得，该溶液颜色清亮透明、咸味纯正、口感略带咸味、无不良异味、富含营养。
实施例3
一种咸味增强肽，通过以下步骤制得：
(1)蛋白酶解
取50g鳊鱼肉糜加入50g去离子水混合均匀后，经150MPa超高静压处理20min，加入鳊鱼蛋白0.5％的胰酶，50℃下酶解3h，水解完成后进行沸水灭酶20min，4℃8000rpm冷冻离心20min，过滤得到酶解液后用5KDa的超滤膜过滤，滤过液即为咸味增强蛋白酶解物。
(2)鳊鱼酶解：
取鳊鱼与水质量比1:1.5混合，加入鳊鱼质量的0.5％木瓜蛋白酶，对其进行酶解，水解温度为55℃，pH值为7.0，时间为4h，后添加胰酶，继续酶解2h。采用高温沸水灭酶30min，4℃，7500rpm冷冻离心25min，滤去残渣和浮油，得鳊鱼酶解液，对滤液进行真空冷冻干燥，制得鳊鱼多肽粉。
(3)咸味混合物的制备
将质量比分别为：氯化钾0.37、氯化钠0.30、乳酸钙0.15、鳊鱼多肽粉0.18混合均匀，取1g溶于100g水中制成盐溶液，此时咸味值为6.14。
(4)咸味增强作用
将质量浓度为0.20％的咸味增强肽加入咸味混合物中，经1.0kV/cm、10μs、1Hz的脉冲电场脉冲次数10次处理后，冷冻干燥得到产品，咸味值增至6.54。
(5)感官评价
将产品溶于水(浓度为0.45％)，经感官评价可得，该溶液颜色透明略带黄色、咸度适中、鲜味较浓、且营养丰富。
表1所示为鳊鱼酶解产物BP与盐类复配后的咸味得分，而表2所示为加入0.20％的大黄鱼蛋白酶解物HP后咸味值的变化情况。由表可知，对应表1中的各组样品，大黄鱼蛋白酶解物HP加入后复合盐的咸味值增强，这充分表明大黄鱼蛋白酶解产物具有明显的咸味增强作用。
表1.鳊鱼酶解物(BP)与盐类复配后的咸味值得分
Figure PCTCN2019124743-appb-000002
表2.大黄鱼蛋白酶解物(HP)混合加入后的咸味值得分
Figure PCTCN2019124743-appb-000003
上述实施例为本发明较佳的实施方式，但本发明的实施方式并不受上述实施例的限制，其他的任何未背离本发明的精神实质与原理下所作的改变、修饰、替代、组合、简化，均应为等效的置换方式，都包含在本发明的保护范围之内。

Claims (10)

  1. 一种咸味增强肽的制备方法，其特征在于包括如下步骤：
    (1)蛋白酶解：大黄鱼预处理后，鱼肉糜与去离子水混合均匀后，进行超高压处理，加入蛋白酶酶解数小时，然后灭酶，冷冻离心，上清液超滤得到酶解液；
    (2)鳊鱼酶解：鳊鱼预处理后，加入水和蛋白酶进行水解，然后灭活蛋白酶，冷冻离心，滤去残渣和浮油，得到鳊鱼酶解液，冷冻干燥，得到鳊鱼多肽粉；
    (3)咸味增强处理：将鳊鱼多肽粉与其他非钠盐混合，得到溶质，加入到大黄鱼酶解液，经脉冲电场处理后，冷冻干燥得到咸味增强肽。
  2. 根据权利要求1所述的制备方法，其特征在于：步骤(1)中，所述蛋白酶为胰酶，所述蛋白酶添加量为大黄鱼蛋白质含量的0.5％。
  3. 根据权利要求1所述的制备方法，其特征在于：步骤(1)中，所述超滤为使用5KDa超滤膜，得到组分为5KDa以下分子量段的多肽混合物。
  4. 根据权利要求1所述的制备方法，其特征在于：步骤(2)中，所述蛋白酶是木瓜蛋白酶和胰酶，且先用木瓜蛋白酶，水解时间为2～5h，后用胰酶，水解时间为1～3h。
  5. 根据权利要求1所述的制备方法，其特征在于：步骤(2)中，以鳊鱼(肉)的质量为计算基准，所述蛋白酶的添加量为0.1％～1.0％。
  6. 根据权利要求1所述的制备方法，其特征在于：步骤(3)中，所述其他非钠盐为氯化钠、氯化钾和乳酸钙。
  7. 根据权利要求1所述的制备方法，其特征在于：步骤(3)中所述的溶质由以下重量百分比的成分组成：
    Figure PCTCN2019124743-appb-100001
  8. 根据权利要求1所述的制备方法，其特征在于：步骤(3)所述脉冲电场处理，脉冲电场参数为0.5～1.0kV/cm，脉冲次数10～20次，脉冲宽度10～15μs， 脉冲频率1Hz。
  9. 一种咸味增强肽，其特征在于：是由权利要求1-8任一项所述的方法制得。
  10. 权利要求9所述的咸味增强肽在食品中的应用。
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