阅读说明：本技术 一种利用低价值海产品提取风味肽的方法 (Method for extracting flavor peptide by using low-value marine products ) 是由 何熹 于海涛 臧汝瑛 邓基建 吉云倩 黄慧影 于 2020-05-19 设计创作，主要内容包括：本发明涉及一种利用低价值海产品提取风味肽的方法。包括如下步骤：(1)将低价值海产品经洗涤、筛选、破碎后,制得待发酵原料；(2)将待发酵原料按照质量比1：(3～5)的比例加水混合,调节pH值至6.0～7.0,制得发酵醪液,然后接种植物乳杆菌HX1,进行发酵,制得发酵原料；(3)将发酵原料经灭菌后,加入蛋白酶,经酶解,提纯,制得风味肽。本发明以低值海产品为原料,采用特定的植物乳杆菌HX1发酵后,再经蛋白酶酶解提取风味肽,通过实际生产,获得的风味肽可以显著去除海产品所特有的腥臭味,同时去除了风味肽中的苦味。(The invention relates to a method for extracting flavor peptide by using low-value marine products. The method comprises the following steps: (1) washing, screening and crushing low-value marine products to obtain raw materials to be fermented; (2) raw materials to be fermented are mixed according to a mass ratio of 1: (3-5) adding water to mix, adjusting the pH value to 6.0-7.0 to prepare fermented mash, inoculating lactobacillus plantarum HX1, and fermenting to prepare a fermentation raw material; (3) sterilizing the fermentation raw materials, adding protease, performing enzymolysis, and purifying to obtain flavor peptide. The invention takes low-value marine products as raw materials, and adopts specific lactobacillus plantarum HX1 for fermentation, and then flavor peptides are extracted by protease enzymolysis, and through actual production, the obtained flavor peptides can obviously remove the peculiar fishy smell of the marine products and simultaneously remove the bitter taste in the flavor peptides.)

1. A method for extracting flavor peptide by using low-value marine products is characterized by comprising the following steps:

(1) washing, screening and crushing low-value marine products to obtain raw materials to be fermented;

(2) mixing the raw materials to be fermented in the step (1) according to a mass ratio of 1: (3-5) adding water to mix, adjusting the pH value to 6.0-7.0 to prepare fermented mash, inoculating lactobacillus plantarum HX1, and fermenting to prepare a fermentation raw material;

the lactobacillus plantarum HX1 has a preservation number of: CCTCC No. M2017522;

(3) and (3) sterilizing the fermentation raw material prepared in the step (2), adding protease, performing enzymolysis, and purifying to obtain the flavor peptide.

2. The method of claim 1, wherein in step (1), the low value seafood is protein-rich tissue of fish, shrimp, shellfish or protein-rich offal left after processing of fish, shrimp, shellfish.

3. The method according to claim 1, wherein in the step (2), the inoculation amount of the lactobacillus plantarum HX1 is 8-15% of the volume of the fermented mash; inoculation ofThe lactobacillus plantarum HX1 is a bacterial liquid, and the concentration of the bacterial in the bacterial liquid is 1.0-9.0 × 10⁸CFU/mL。

4. The method according to claim 1, wherein the step (2) further comprises the step of adding a fermentation regulator after adjusting the pH value, wherein the fermentation regulator is added in a mass volume percentage of 0.1-0.2% in g/L;

further preferably, the fermentation regulator comprises the following components in parts by weight:

45-55 parts of inulin, 35-45 parts of lactose and 10-30 parts of nisin;

more preferably, the fermentation regulator comprises the following components in parts by weight:

50 parts of inulin, 40 parts of lactose and 20 parts of nisin (nisin).

5. The method of claim 1, wherein in step (2), the fermentation conditions are: fermenting for 3-5 days under the anaerobic condition at the temperature of 20-30 ℃.

6. The method according to claim 1, wherein in the step (2), the pH regulator for regulating the pH value is citric acid or sodium bicarbonate.

7. The method according to claim 1, wherein in the step (3), the sterilization condition is boiling for 8-15 min.

8. The method according to claim 1, wherein in the step (3), the protease is neutral protease or flavourzyme, and the addition amount is 500-1000U/g.

9. The method according to claim 1, wherein in the step (3), the enzymolysis reaction is carried out at 35-45 ℃ for 3-5 h.

10. The method according to claim 1, wherein the step (3) further comprises a step of enzyme deactivation after enzymolysis, and the treatment is carried out for 5-10 min at a temperature of more than 100 ℃;

preferably, in the step (3), the purification is ultrafiltration purification, and the pore diameter of the membrane is less than 10 kDa.

Preferably, the step (3) further comprises a step of spray drying after purification.

Technical Field

The invention relates to a method for extracting flavor peptide by using low-value marine products, belonging to the technical field of fermentation and extraction of marine products.

Background

The term "food sensory peptide" refers to a general term for peptides added to foods to regulate the quality, appearance and taste of foods, and among them, peptides capable of improving the flavor and palatability of foods are called flavor peptides. The production process of the flavor peptide at present is most commonly biosynthetic and enzymolysis, and the method adopting enzymolysis is the most common production method. The technological process includes the main steps of using vegetable protein, such as soybean, etc. and animal protein, such as poultry, livestock, etc. as material, deoiling, pre-treatment, adding different kinds of proteinase to degrade the macro molecular protein into micro molecular peptide, and subsequent grading and purification to obtain the flavor peptide.

Chinese patent document CN110169563A (application number 201910450032.4) discloses Maillard flavor peptide with antioxidant function and a preparation method thereof, belonging to the field of food additives. The Maillard flavor peptide with potato flavor enhancement and oxidation resistance functions is prepared by taking corn protein peptide, methionine, cysteine and xylose as raw materials and based on the Maillard reaction technology, the prepared Maillard flavor peptide has different flavors by regulating and controlling the proportion of amino acids, can partially replace the use of a chemically synthesized antioxidant in food while enhancing the flavor characteristics of the food, is a safe, efficient and multifunctional food additive, increases the additional function of Maillard reaction products in the traditional sense, and widens the research and application range of the Maillard flavor peptide in the field of food additives. However, the flavor peptides derived from plants are significantly different in quality and taste from flavor peptides prepared from animal proteins, particularly marine products, due to the difference in the kinds of amino acids contained in the raw material components.

Chinese patent document CN109699988A (application No. 201910089665.7) discloses a preparation method of natural shrimp sauce by using flavor peptides of shrimps as main materials to guide generation, which solves the problems of long period and unstable batch of the fermentation of the traditional fish sauce, can obtain the shrimp sauce which has strong and mellow flavor and delicious shrimp taste in a short time, and can be used for synergistic action with other seasonings such as monosodium glutamate, chicken essence and the like to enhance the seasoning effect and the taste development force and make food more delicious. In the process of producing flavor peptide by enzymolysis, in order to improve the enzymolysis efficiency, the raw material is pretreated by desalting and deoiling, especially when animal protein such as fish and shrimp is used as the raw material, although the technology adopts edible oil for extraction, the effect is not ideal, and when industrial production is realized, the fat is often removed by adopting organic solvent such as petroleum ether or a method of adding acid and alkali. However, these methods have adverse effects such as solvent residue and deterioration of protein activity, which adversely affect the quality of the final flavor peptide.

In addition, the flavor peptide prepared by using marine products as raw materials has very strong fishy smell, which is more prominent in the preparation process, and the quality and market acceptance of the prepared flavor peptide are seriously influenced.

Disclosure of Invention

Aiming at the defects of the prior art, the invention provides a method for extracting flavor peptide by using low-value marine products.

Description of the terms

The marine product is a substance rich in animal protein and available for eating or using in the sea.

The technical scheme of the invention is as follows:

a method for extracting flavor peptide from low-value marine product comprises the following steps:

(1) washing, screening and crushing low-value marine products to obtain raw materials to be fermented;

(2) mixing the raw materials to be fermented in the step (1) according to a mass ratio of 1: (3-5) adding water to mix, adjusting the pH value to 6.0-7.0 to prepare fermented mash, inoculating lactobacillus plantarum HX1, and fermenting to prepare a fermentation raw material;

the lactobacillus plantarum HX1 has a preservation number of: CCTCC No. M2017522;

(3) and (3) sterilizing the fermentation raw material prepared in the step (2), adding protease, performing enzymolysis, and purifying to obtain the flavor peptide.

Preferably, in step (1), the low value marine product is protein-rich tissue of fish, shrimp, shellfish or protein-rich leftovers of fish, shrimp, shellfish after processing.

According to the invention, in the step (2), the inoculation amount of the lactobacillus plantarum HX1 is 8-15% of the volume of the fermented mash, the inoculated lactobacillus plantarum HX1 is a bacterial liquid, and the bacterial concentration in the bacterial liquid is 1.0-9.0 × 10⁸CFU/mL。

According to the invention, the preferable step (2) further comprises the step of adding a fermentation regulator after the pH value is adjusted, wherein the fermentation regulator is added in a mass volume percentage of 0.1-0.2% and a unit of g/L.

Further preferably, the fermentation regulator comprises the following components in parts by weight:

45-55 parts of inulin, 35-45 parts of lactose and 10-30 parts of nisin.

More preferably, the fermentation regulator comprises the following components in parts by weight:

50 parts of inulin, 40 parts of lactose and 20 parts of nisin (nisin).

Preferably, in step (2), the fermentation conditions are: fermenting for 3-5 days under the anaerobic condition at the temperature of 20-30 ℃.

According to the present invention, in the step (2), the pH adjusting agent for adjusting the pH is citric acid or sodium bicarbonate.

According to the invention, in the step (3), the sterilization condition is boiling for 8-15 min.

According to the invention, in the step (3), the protease is neutral protease or flavourzyme, and the addition amount is 500-1000U/g.

According to the invention, in the step (3), the enzymolysis is preferably carried out at 35-45 ℃ for 3-5 hours.

According to the invention, the step (3) preferably further comprises an enzyme deactivation step after enzymolysis, and the treatment is carried out for 5-10 min at the temperature of more than 100 ℃.

According to the present invention, in the step (3), the purification is ultrafiltration purification, and the membrane pore size is less than 10 kDa.

According to the present invention, preferably, the step (3) further comprises a step of spray drying after purification.

Advantageous effects

1. According to the invention, low-value marine products are used as raw materials, specific lactobacillus plantarum HX1 is adopted for fermentation, and then flavor peptides are extracted through protease enzymolysis, and through actual production, the obtained flavor peptides can obviously remove the peculiar fishy smell of the marine products and simultaneously remove the bitter taste in the flavor peptides;

2. according to the invention, the fermentation regulator is added before fermentation, so that the growth of lactobacillus plantarum HX1 is facilitated, the specific lipase activity of lactobacillus plantarum HX1 is utilized to accelerate the hydrolysis of fat in the raw material into glycerol and fatty acid, and compared with the traditional fat removal method, the method has mild reaction conditions, is low-carbon and is environment-friendly;

3. the pH value is adjusted by adopting citric acid or sodium bicarbonate, so that the fishy smell in the product can be removed, the prepared product meets the relevant regulations of food production and sanitation safety, the quality is good, the value is high, and a new way can be opened up for the comprehensive utilization of low-value marine products.

Drawings

FIG. 1 is a graph showing the degradation effect of two kinds of Lactobacillus plantarum used in example 1 and comparative example 3 on krill pulp fat;

Detailed Description

The following examples are intended to better illustrate the present invention and to enable those skilled in the art to better understand and understand the present invention with the aid of the examples. However, the scope of the claims of the present invention is not limited to the examples provided.

Materials and apparatus

Yellow croaker, scallop skirt and krill are all products sold in common markets;

neutral protease (5 × 10)⁴U/g) from Dada enzyme preparations, Inc., Shijiazhuang;

flavourzyme was purchased from Novoxin (China) Biotech Ltd

Inulin was purchased from Beneo Orafti s.a, belgium;

lactose was purchased from the oceanic biotechnology limited of zhhai city;

nisin was purchased from Shandongtongtai bioengineering, Inc.;

lactobacillus plantarum HX1 (L actinobacillus plantarum HX1), known strain, is from China center for type culture Collection, and has a collection number of CCTCC No. M2017522, and patent documents CN110846331A and CN108077826A are both described.

Fermentation tank 500L stainless steel anaerobic pilot fermentation tank.