CN111406827A - Method for extracting flavor peptide by using low-value marine products - Google Patents
Method for extracting flavor peptide by using low-value marine products Download PDFInfo
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- CN111406827A CN111406827A CN202010425726.5A CN202010425726A CN111406827A CN 111406827 A CN111406827 A CN 111406827A CN 202010425726 A CN202010425726 A CN 202010425726A CN 111406827 A CN111406827 A CN 111406827A
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- fermentation
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- lactobacillus plantarum
- fermented
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- Seasonings (AREA)
Abstract
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.
Description
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.
Example 1
A method for extracting flavor peptide from low-value marine product comprises the following steps:
(1) screening, cleaning and crushing krill sold in the market, filtering with 50 meshes, adding water according to the weight ratio of 1: 4 to mix into a raw material to be fermented, and pumping 400L raw material to be fermented into a fermentation tank;
(2) acid adjustment: measuring the pH value, adding citric acid to regulate the pH value to 7.0;
(3) adding fermentation regulator, namely mixing inulin, lactose and nisin according to parts by weight of 50, 35 and 20 to prepare the fermentation regulator, adding the fermentation regulator into a fermentation tank according to 0.1 percent (mass volume fraction, unit g/L) of the raw material to be fermented, and uniformly mixing the fermentation regulator with the raw material to be fermented;
(4) adding Lactobacillus plantarum HX1, pre-culturing Lactobacillus plantarum HX1 to strain concentration of 2.0 × 108CFU/m L, adding the bacterial liquid into the fermented mash in the step (3) according to the volume fraction of 10%;
(5) controlling the temperature to be 28 ℃, and naturally fermenting for 5d under an anaerobic condition to obtain a fermentation raw material; controlling the temperature and regularly monitoring the pH value and the fat content of the fermentation liquor in the fermentation process;
(6) after fermentation, readjusting pH of the fermentation product to 7.0, heating with high temperature steam, boiling for 10min, and cooling to 40 deg.C;
(7) adding neutral protease into the fermentation broth according to a ratio of 600U/g, performing enzymolysis reaction at 40 deg.C for 4h, heating with high temperature steam, boiling for 10min, inactivating enzyme, and cooling to room temperature;
(8) filtering the solution by a 100-mesh sieve, then performing ultrafiltration by using a 10kDa ultrafiltration membrane, collecting a filtrate with the molecular weight of less than 10kDa, and determining the contents of protein and amino acid;
(9) spray drying the filtrate in (8) into powder, packaging, and storing at 10 deg.C.
Example 2
A method for extracting flavor peptide from low-value marine product comprises the following steps:
(1) screening, cleaning and crushing krill sold in the market, filtering with 50 meshes, adding water according to the weight ratio of 1: 5 to mix into a raw material to be fermented, and pumping 400L raw material to be fermented into a fermentation tank;
(2) acid adjustment: measuring the pH value, adding citric acid to adjust the pH value to 6.0;
(3) adding fermentation regulator, mixing inulin, lactose and nisin 55, 30 and 30 by weight parts to prepare the fermentation regulator, adding the fermentation regulator into a fermentation tank according to 0.15% (mass volume fraction, unit g/L) of the raw material to be fermented, and uniformly mixing the fermentation regulator with the raw material to be fermented;
(4) adding Lactobacillus plantarum HX1, pre-culturing Lactobacillus plantarum HX1 to strain concentration of 9.0 × 108CFU/m L, adding the bacterial liquid into the fermented mash in the step (3) according to the volume fraction of 8%;
(5) controlling the temperature to be 30 ℃, and naturally fermenting for 3d under an anaerobic condition to obtain a fermentation raw material; controlling the temperature and regularly monitoring the pH value and the fat content of the fermentation liquor in the fermentation process;
(6) after fermentation, readjusting pH of the fermentation product to 6.5, heating with high temperature steam, boiling for 10min, and cooling to 35 deg.C;
(7) adding flavourzyme into the fermentation liquor according to the proportion of 600U/g, carrying out enzymolysis reaction for 5h at the temperature of 45 ℃, heating by using high-temperature steam, boiling for 10min, inactivating enzyme, and cooling to normal temperature;
(8) filtering the solution by a 100-mesh sieve, then performing ultrafiltration by using a 10kDa ultrafiltration membrane, collecting a filtrate with the molecular weight of less than 10kDa, and determining the contents of protein and amino acid;
(9) spray drying the filtrate in (8) into powder, packaging, and storing at 10 deg.C.
Example 3
A method for extracting flavor peptide from low-value marine product comprises the following steps:
(1) screening, cleaning and crushing krill sold in the market, filtering with 50 meshes, adding water according to the weight ratio of 1: 3 to mix into a raw material to be fermented, and pumping 400L raw material to be fermented into a fermentation tank;
(2) acid adjustment: measuring the pH value, adding citric acid to adjust the pH value to 6.5;
(3) adding fermentation regulator, namely mixing inulin, lactose and nisin according to parts by weight of 45, 40 and 20 to prepare the fermentation regulator, adding the fermentation regulator into a fermentation tank according to 0.2 percent (mass volume fraction, unit g/L) of the raw material to be fermented, and uniformly mixing the fermentation regulator with the raw material to be fermented;
(4) adding Lactobacillus plantarum HX1, pre-culturing Lactobacillus plantarum HX1 to strain concentration of 1.0 × 108CFU/m L, adding the bacterial liquid into the fermented mash in the step (3) according to the volume fraction of 15%;
(5) controlling the temperature at 25 ℃, and naturally fermenting for 5d under an anaerobic condition to obtain a fermentation raw material; controlling the temperature and regularly monitoring the pH value and the fat content of the fermentation liquor in the fermentation process;
(6) after fermentation, readjusting pH of the fermentation product to 6.5, heating with high temperature steam, boiling for 10min, and cooling to 35 deg.C;
(7) adding neutral protease into the fermentation broth according to a ratio of 600U/g, performing enzymolysis reaction at 35 deg.C for 5h, heating with high temperature steam, boiling for 10min, inactivating enzyme, and cooling to room temperature;
(8) filtering the solution by a 100-mesh sieve, then performing ultrafiltration by using a 10kDa ultrafiltration membrane, collecting a filtrate with the molecular weight of less than 10kDa, and determining the contents of protein and amino acid;
(9) spray drying the filtrate in (8) into powder, packaging, and storing at 10 deg.C.
Example 4
The method for extracting flavor peptides from krill by microbial fermentation and enzymolysis as described in example 1, except that no fermentation regulator is added in step (3).
Comparative example 1
The method for extracting flavor peptides by microbial fermentation and enzymolysis of krill as described in example 1, except that in step (3), inulin, lactose and nisin are mixed in parts by weight of 25, 20 and 5, prepared as fermentation regulators, and added to the grape pulp in a mass volume fraction of 0.1% (g/L).
Comparative example 2
The method for extracting flavor peptides from krill by microbial fermentation and enzymolysis as described in example 1, except that lactobacillus plantarum HX1 was replaced by lactobacillus plantarum WCSF1 in step (4).
Comparative example 3
The method for extracting flavor peptides from krill by microbial fermentation and enzymolysis as described in example 1, except that lactobacillus plantarum HX1 is not added in step (4).
Comparative example 4
The method for extracting flavor peptides from krill by microbial fermentation and enzymolysis as described in example 1, except that acetic acid is used to adjust the pH in step (2).
Examples of the experiments
Detection of flavor peptide-related index
pH value: measured according to the method specified in GB/T10468;
fat determination: measuring fat in food according to the method of GB/T5009.6-2003;
protein: measuring according to the method of GB 5009.5-2016 protein in food;
amino acids: measuring amino acids in food according to GB 5009.124-2016 method;
evaluation of fishy smell and evaluation of bitterness: a taste panel was composed of 10 sensory evaluators, and appropriate training was performed on the sensory evaluators before the sensory evaluation. With reference to "food sensory evaluation" (published by Zhang Yonghua, published by the university of south China publisher 2006) and "food sensory evaluation principle and practice" (published by Wang-dong-translation, etc., by the light industry, China publisher 2001), the freshness, fishy smell, bitter taste, etc. of the enzymatic hydrolysate were evaluated by 5-point system according to the principle of the comprehensive scoring method. Wherein, 1 point represents "slightly", 2 points represents "weak", 3 points represents "normal", 4 points represents "heavy", and 5 points represents "heavy".
The flavor peptides are respectively extracted from the same raw materials of the same batch by the methods of examples 1-4 and comparative examples 1-4, and the detection results of the enzymolysis solution products and the like are shown in Table 1:
TABLE 1 Effect of different treatments on the quality of the final flavor peptide product
The biological content of the fermentation products in the preparation process of the methods of examples 1-4 and comparative examples 1-4 was measured by plate counting method, and the results are shown in Table 2:
TABLE 2 different microorganism contents in the fermentation broth after 48 hours of different treatments (CFU/m L)
Note that the isolation culture medium of 3 microorganisms is Lactobacillus plantarum MRS culture medium, Bacillus subtilis L B culture medium, Vibrio maritime, Zobelli 2216E culture medium.
Analysis of results
Examples 1 andthe comparison of the method of example 4 shows that the growth of the engineering bacterium Lactobacillus plantarum HX1 of example 1 can reach 1.0 × 10 after 24h8CFU/m L, 72h can decompose 80% fat (the fat content in the raw material is about 8.5 g/L), and the time for the embodiment 4 to achieve the above effect is doubled, so the growth rate of the lactobacillus plantarum HX1 adopting the embodiment 4 is obviously reduced, the fat hydrolysis efficiency is greatly reduced, the fermentation time is prolonged, and the production period and the energy consumption are increased.
Comparing the example 1 with the comparative example 1, the method of the comparative example 1 is adopted, namely, inulin, lactose and nisin are mixed according to the parts by weight of 25, 20 and 5 to prepare a fermentation regulator, after 48h of fermentation, the appearance of mixed bacteria such as bacillus subtilis, vibrio marinus and the like in the fermentation liquid is found (table 2), and as can be seen from the table 1, due to the existence of the mixed bacteria, the growth of the engineering bacteria lactobacillus plantarum HX1 is inhibited, bad substances such as fishy smell and the like are generated, the quality of the product is adversely affected, and in addition, the mixed bacteria can utilize abundant proteins in the fermentation liquid for growth, so that the content of the protein and amino acid of the final enzymolysis product of the comparative example 1 is reduced, and the final yield of the flavor peptide product is affected. Therefore, the mixed fermentation regulator in the weight portion can not completely inhibit the growth of other mixed bacteria.
Comparing the results of example 1 and comparative example 2, it can be seen that the hydrolysis effect of lactobacillus plantarum HX1 on fat after 36h was more than 4 times that of control lactobacillus plantarum WCSF1 with the same 1% lactobacillus plantarum addition, as shown in fig. 1. It can also be seen from Table 1 that the yields of proteins and amino acids, which are closely related to the flavor peptide yield, are reduced, the umami sensory level is reduced, and the fishy components are increased in comparative example 2 compared to example 1 due to the presence of a portion of fat.
Comparing the data of example 1 and comparative example 3 in table 1, it can be seen that the fat content in example 1 is significantly lower than that in comparative example 3 after the same 5d fermentation, which indicates that lactobacillus plantarum HX1 exerts a good fat decomposition effect. Meanwhile, because the fat content in the solution is reduced, the hydrolysis inhibition effect of the fat on the protease is reduced, and the hydrolysis efficiency of small molecular proteins of the protease is favorably improved, so that the content of the proteins and amino acids meeting the requirements is obviously increased. In addition, due to the presence of fat, rancidity occurs after oxidation, and the product has fishy smell and bitter taste.
Comparing the data of example 1 and comparative example 4 in table 1, it can be seen that the adjustment of pH in comparative example 4 has a great influence on the flavor, especially umami taste of the product, since acetic acid has a stronger taste than citric acid. In addition, acetic acid has certain volatility and corrosivity, the use cost can be increased, the citric acid is good in stability and low in corrosion type, most importantly, the flavor is soft, the flavor of the final product cannot be adversely affected, and therefore the method is more suitable for adjusting the pH value of the flavor peptide product fermentation liquor compared with acetic acid.
Claims (10)
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.
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Date of cancellation: 20230615 Granted publication date: 20210625 Pledgee: Industrial and Commercial Bank of China Limited Weihai Wendeng sub branch Pledgor: SHANDONG WEIZHENGPINKANG FOOD TECHNOLOGY CO.,LTD. Registration number: Y2022980007404 |
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| PE01 | Entry into force of the registration of the contract for pledge of patent right |
Denomination of invention: A Method for Extracting Flavor Peptides from Low Value Seafood Effective date of registration: 20230620 Granted publication date: 20210625 Pledgee: Industrial and Commercial Bank of China Limited Weihai Wendeng sub branch Pledgor: SHANDONG WEIZHENGPINKANG FOOD TECHNOLOGY CO.,LTD. Registration number: Y2023980044418 |