CN110403153A - A kind of preparation method of flavor squid nutrients biological product - Google Patents
A kind of preparation method of flavor squid nutrients biological product Download PDFInfo
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- CN110403153A CN110403153A CN201910801216.0A CN201910801216A CN110403153A CN 110403153 A CN110403153 A CN 110403153A CN 201910801216 A CN201910801216 A CN 201910801216A CN 110403153 A CN110403153 A CN 110403153A
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- 238000002360 preparation method Methods 0.000 title claims abstract description 32
- 239000000796 flavoring agent Substances 0.000 title claims abstract description 18
- 235000019634 flavors Nutrition 0.000 title claims abstract description 18
- 235000015097 nutrients Nutrition 0.000 title abstract description 5
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- 238000000855 fermentation Methods 0.000 claims abstract description 35
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- 235000016709 nutrition Nutrition 0.000 claims description 42
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- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 2
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- DLGJWSVWTWEWBJ-HGGSSLSASA-N chondroitin Chemical compound CC(O)=N[C@@H]1[C@H](O)O[C@H](CO)[C@H](O)[C@@H]1OC1[C@H](O)[C@H](O)C=C(C(O)=O)O1 DLGJWSVWTWEWBJ-HGGSSLSASA-N 0.000 description 1
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Classifications
-
- 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
- A23L3/00—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs
- A23L3/34—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals
- A23L3/3454—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals in the form of liquids or solids
- A23L3/3463—Organic compounds; Microorganisms; Enzymes
- A23L3/3472—Compounds of undetermined constitution obtained from animals or plants
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L17/00—Food-from-the-sea products; Fish products; Fish meal; Fish-egg substitutes; Preparation or treatment thereof
- A23L17/50—Molluscs
-
- 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
- A23L17/65—Addition of, or treatment with, microorganisms or enzymes
-
- 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
- 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
- A23L3/00—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs
- A23L3/34—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals
- A23L3/3454—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals in the form of liquids or solids
- A23L3/3463—Organic compounds; Microorganisms; Enzymes
- A23L3/3571—Microorganisms; Enzymes
-
- 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
- A23L33/105—Plant extracts, their artificial duplicates or their derivatives
-
- 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
-
- 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
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/21—Streptococcus, lactococcus
- A23V2400/249—Thermophilus
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- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Polymers & Plastics (AREA)
- Health & Medical Sciences (AREA)
- Nutrition Science (AREA)
- Engineering & Computer Science (AREA)
- Food Science & Technology (AREA)
- Microbiology (AREA)
- Zoology (AREA)
- Marine Sciences & Fisheries (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Botany (AREA)
- Mycology (AREA)
- Meat, Egg Or Seafood Products (AREA)
Abstract
The present invention provides a kind of preparation method of flavor squid nutrients biological product, belongs to sleeve-fish product technical field, includes the following steps: S1: Squid By-products are homogenized, and sterilize to obtain squid homogenate；S2: distilled water will first be added in the squid homogenate and flavor protease digests, enzyme deactivation；S3: it ferments thermophilus bacterium solution is added in the enzymatic hydrolysis squid homogenate, fermentation liquid is freeze-dried to get flavor squid nutrients biological product.The soluble protein high conversion rate of preparation method of the present invention, squid nutrients biological product contains higher unsaturated fatty acid in flavor obtained, has good sense organ flavor, can inhibit the growth of harmful bacteria, long fresh-keeping period.
Description
Technical Field
The invention belongs to the technical field of squid products, and particularly relates to a preparation method of a flavored squid nutritional biological product.
Background
Squid, also called as China squid, is a marine mollusk. The squid can be divided into three parts on the whole: the squid meat, the head and the feet, the squid skin and the internal organs respectively account for 50 percent, 25 percent and 25 percent of the weight of the squid, wherein the edible part accounts for 75 percent of the weight of the squid and is approximately 20 percent higher than most of other fishes, so that the overall utilization rate of the squid is still higher. The protein content of the squid is high, about 20 percent, the types of the contained amino acids are rich, and a plurality of essential amino acids required by a human body are included; the crude fat content of the squid is lower, wherein phospholipid is the main component of the squid, and accounts for about 40-50% of the total lipid content. The processing treatment of the squid has many byproducts, such as squid head, eyes, epidermis, cartilage and viscera, which are approximately 20% -25% of the total weight of the squid. The squid processing by-products are rich in nutrition, not only contain a large amount of protein and lipid, but also are rich in taurine, vitamins, chondroitin and the like, and people neglect the high-value utilization of the squid processing by-products. However, the water ratio of the squid is larger, in addition, the enzyme in the body of the squid is very active, so that the leftovers can not be stored for a long time, and the leftovers are accompanied with very strong fishy smell along with the prolonging of the storage time. For processing the squid leftovers, the processing difficulty is higher due to the lack of a mature processing technology, so that the conditions of high cost and low profit are caused, and thus the condition is forbidden for many people.
The biotechnology is a hotspot of current research, the microbial fermentation is novel, has been applied in actual production and proved to be a safe and reliable technology, and the application of the microbial fermentation technology also obtains good economic benefit and application value, so the microbial fermentation technology has wide development space. Probiotics are a viable microbial dietary supplement that beneficially affects the host through its action in the gut. Some health-related effects associated with the ingestion of probiotics have been reported in human studies, including the relief of lactose intolerance and immune enhancement, the promotion of food digestion, the production of useful products to destroy bad microorganisms, the supplementation of missing digestive enzymes (due to missing or defective genes), and the maintenance of the pH of the digestive system, among others.
Disclosure of Invention
The invention aims to provide a preparation method of a flavor squid nutritional biological product with high soluble protein conversion rate, wherein the prepared flavor squid nutritional biological product contains higher unsaturated fatty acid, has good sensory flavor and has long fresh-keeping period.
The technical scheme adopted by the invention for realizing the purpose is as follows:
a preparation method of a flavored squid nutritional biological product comprises the following steps:
s1: homogenizing the squid byproduct, and sterilizing to obtain squid homogenate;
s2: adding distilled water and flavourzyme into the squid homogenate for enzymolysis, and inactivating enzyme;
s3: and adding thermophilic streptococcus liquid into the enzymolysis squid homogenate for fermentation, and freeze-drying the fermentation liquor to obtain the flavored squid nutritional biological product.
The squid processing by-product is rich in high-quality protein, in the storage process, the original spatial structure of the protein in the squid by-product is destroyed, the hydrophobic amino acid groups on the surface of the squid by-product are gradually exposed, so that the hydrophobicity of the surface of the squid is gradually increased, and the protein is condensed due to hydrophobic interaction, so that the solubility of the protein in the squid by-product is low, and the utilization of organisms is not facilitated. According to the preparation method, the combined action of streptococcus thermophilus and the bacterium enzyme of the flavourzyme can be used, so that the conversion rate of soluble protein can be improved, the content of small molecular protein in the obtained flavored squid nutritional biological product can be improved, the content of unsaturated fatty acid can be improved, and the problems of environmental pollution and resource waste of squid processing byproducts are solved; in addition, the transformation of the streptococcus thermophilus can inhibit the growth of harmful bacteria, and the streptococcus thermophilus and the flavor enzyme can endow the product with good sensory flavor together, so that the streptococcus thermophilus has a better fresh-keeping effect.
Preferably, the addition amount of the flavourzyme in S2 is 5000-; the enzymolysis temperature is 45-60 deg.C, pH is 6.5-7.5, and the time is 2-5 h.
Preferably, S3S Streptococcus thermophilus liquid is obtained by culturing Streptococcus thermophilus in MRS culture medium until the thallus reaches the end of logarithmic growth.
Preferably, the cell density of the Streptococcus thermophilus solution in S3 is not less than 1X 1014CFU/L。
Preferably, the addition amount of the streptococcus thermophilus in the S3 is 10-20%, the fermentation temperature is 30-40 ℃, the pH value is 6.5-7.5, the rotation speed is 100-150r/min, and the time is 30-40 h.
Preferably, the preparation process has a soluble protein conversion of > 25%.
Preferably, sage extract is added to distilled water in S2. The protein hydrolysate of oligopeptide and amino acid has a promoting effect on the proliferation of streptococcus thermophilus, is an ideal nitrogen source for culturing streptococcus thermophilus, the sage extract can also promote the flavourzyme to carry out enzymolysis on the protein in the squid byproduct homogenate to obtain the enzymolysis squid homogenate rich in oligopeptide and amino acid, particularly the content of glutamic acid in the enzymolysis squid homogenate is increased, the growth and proliferation of streptococcus thermophilus are facilitated, and thus the conversion rate of soluble protein is increased, and the conversion rate of the soluble protein is more than 30%. In addition, the content of unsaturated fatty acid in the biological product can be increased by the presence of the sage extract, and the content of unsaturated fatty acid in the biological product is more than 80g/100 g; the sage extract has stronger inoxidizability, can better inhibit the oxidative denaturation of unsaturated fatty acid, and effectively keeps the freshness of biological products; in addition, the growth of bacteria is inhibited, so that the preservation period of the biological product is prolonged.
More preferably, the weight concentration of the sage extract is 0.2-2%.
Compared with the prior art, the invention has the beneficial effects that: according to the invention, through the combined action of the streptococcus thermophilus and the flavourzyme after enzyme, the conversion rate of soluble protein can be improved, the content of small molecular protein in the obtained flavored squid nutritional biological product can be improved, the content of unsaturated fatty acid can be improved, and the problems of environmental pollution and resource waste of squid processing byproducts are solved; in addition, the transformation of the streptococcus thermophilus can inhibit the growth of harmful bacteria, so that the flavored squid nutritional biological product has a longer fresh-keeping period, and can endow the product with good sensory flavor together with the flavor enzyme, thereby having a better fresh-keeping effect.
The preparation method of the flavored squid nutritional biological product provided by the invention overcomes the defects of the prior art, and is reasonable in design and convenient to operate.
Drawings
FIG. 1 shows the glutamic acid content in the enzymatic squid homogenate of test example 1 of the present invention;
FIG. 2 is a result of measurement of the soluble protein conversion in test example 1 of the present invention;
FIG. 3 shows the content of unsaturated fatty acids in the nutritional biological product of flavor squid in test example 1 of the present invention;
FIG. 4 shows the results of measuring pH and titrated acidity of squid byproduct fermentation broth in test example 2 of the present invention.
Detailed Description
The invention is further illustrated by the following examples. It is to be understood that the examples are for illustrative purposes only and are not intended to limit the scope and spirit of the present invention.
Example 1:
a preparation method of a flavored squid nutritional biological product comprises the following steps:
s1: cleaning squid byproduct, crushing, homogenizing, sterilizing in a high pressure steam sterilization pot at 120 deg.C for 30min, and storing in a refrigerator at-20 deg.C;
s2: inoculating Streptococcus thermophilus into MRS broth conical flask, shake culturing at 37 deg.C and 120r/min for 24 hr to logarithmic growth end stage to obtain Streptococcus thermophilus liquid with thallus concentration of 1 × 10 or more14CFU/L；
S3: putting 2g of squid homogenate into a 50ml centrifugal tube, adding 10g of distilled water into flavourzyme according to the solid-to-liquid ratio of 1:5, adding the flavourzyme according to the ratio of 6000u/g, adjusting the pH value of the system to 7.0, then placing the centrifugal tube into a 50 ℃ incubator for enzymolysis for 3h, and then inactivating for 30min at 90 ℃;
s4: adding 20% (400ul) of streptococcus thermophilus liquid into the enzymolysis squid homogenate, fermenting for 36h at 40 ℃ at 120r/min, and freeze-drying the fermentation liquid to obtain the flavored squid nutritional biological product.
Example 2:
a preparation method of a flavored squid nutritional biological product comprises the following steps:
s1: cleaning squid byproduct, crushing, homogenizing, sterilizing in a high pressure steam sterilization pot at 120 deg.C for 30min, and storing in a refrigerator at-20 deg.C;
s2: streptococcus thermophilus was inoculated in MRS broth Erlenmeyer flasks at 37 deg.CCulturing in a shaking table at 120r/min for 24 hr to reach the final stage of logarithmic growth to obtain Streptococcus thermophilus liquid with thallus concentration not less than 1 × 1014CFU/L；
S3: putting 2g of squid homogenate into a 50ml centrifugal tube, adding 10g of distilled water into flavourzyme according to the solid-to-liquid ratio of 1:5, adding 0.3% of sage extract into the distilled water according to the mass concentration of 6000u/g, adjusting the pH value of the system to 7.0, then placing the centrifugal tube into a 50 ℃ incubator for enzymolysis for 3h, and then inactivating for 30min at 90 ℃;
s4: adding 20% (400ul) of streptococcus thermophilus liquid into the enzymolysis squid homogenate, fermenting for 36h at 40 ℃ at 120r/min, and freeze-drying the fermentation liquid to obtain the flavored squid nutritional biological product.
Example 3:
a preparation method of a flavored squid nutritional biological product comprises the following steps:
s1: cleaning squid byproduct, crushing, homogenizing, sterilizing in a high pressure steam sterilization pot at 120 deg.C for 30min, and storing in a refrigerator at-20 deg.C;
s2: inoculating Streptococcus thermophilus into MRS broth conical flask, shake culturing at 37 deg.C and 120r/min for 24 hr to logarithmic growth end stage to obtain Streptococcus thermophilus liquid with thallus concentration of 1 × 10 or more14CFU/L；
S3: putting 2g of squid homogenate into a 50ml centrifugal tube, adding 10g of distilled water into flavourzyme according to the solid-to-liquid ratio of 1:5, adding 1.2% of sage extract into the distilled water according to the mass concentration of 6000u/g, adjusting the pH value of the system to 7.0, then placing the centrifugal tube into a 50 ℃ incubator for enzymolysis for 3h, and then inactivating for 30min at 90 ℃;
s4: adding 20% (400ul) of streptococcus thermophilus liquid into the enzymolysis squid homogenate, fermenting for 36h at 40 ℃ at 120r/min, and freeze-drying the fermentation liquid to obtain the flavored squid nutritional biological product.
Example 4:
the squid byproduct is rich in various nutrient substances for the growth of microorganisms, so that favorable conditions are provided for the propagation of the squid byproduct and exterior bacteria, and the product is easy to decay and deteriorate and has food safety problems. Therefore, the preservation of the flavored squid nutritional biological product mainly controls the growth and the propagation of microorganisms in the product. In order to further improve the harmful bacterium inhibition ability of the streptococcus thermophilus and the preservation period of the flavored squid nutritional biological product, glycocholic acid and berberine hydrochloride are added in the fermentation process, the acid production ability and the acid resistance of the streptococcus thermophilus can be improved, so that the accumulation amount of organic acid in fermentation liquor is increased, the bacteriostatic activity of squid byproduct fermentation liquor is improved, the synergistic effect of the glycocholic acid and the sage extract is realized, the growth and the propagation of harmful bacteria of putrefying bacteria are inhibited, and the preservation period of the biological product is prolonged.
The preparation method of the flavored squid nutritional biological product comprises the following steps:
s1: cleaning squid byproduct, crushing, homogenizing, sterilizing in a high pressure steam sterilization pot at 120 deg.C for 30min, and storing in a refrigerator at-20 deg.C;
s2: inoculating Streptococcus thermophilus into MRS broth conical flask, shake culturing at 37 deg.C and 120r/min for 24 hr to logarithmic growth end stage to obtain Streptococcus thermophilus liquid with thallus concentration of 1 × 10 or more14CFU/L；
S3: putting 2g of squid homogenate into a 50ml centrifugal tube, adding 10g of distilled water into flavourzyme according to the solid-to-liquid ratio of 1:5, adding 1.2% of sage extract into the distilled water according to the mass concentration of 6000u/g, adjusting the pH value of the system to 7.0, then placing the centrifugal tube into a 50 ℃ incubator for enzymolysis for 3h, and then inactivating for 30min at 90 ℃;
s4: adding 20 percent (400ul) of thermophilic streptococcus liquid into the enzymolysis squid homogenate, fermenting for 36 hours at 40 ℃ under the condition of 120r/min, adding glycocholic acid and berberine hydrochloride when fermenting for 12 hours to ensure that the contents of glycocholic acid and berberine hydrochloride in an anti-glue system are 0.02 per mill and 0.33 percent respectively, and freeze-drying the fermentation liquor to obtain the flavored squid nutritional biological product.
Comparative example 1:
a preparation method of a flavored squid nutritional biological product comprises the following steps:
s1: cleaning squid byproduct, crushing, homogenizing, sterilizing in a high pressure steam sterilization pot at 120 deg.C for 30min, and storing in a refrigerator at-20 deg.C;
s2: inoculating Streptococcus thermophilus into MRS broth conical flask, shake culturing at 37 deg.C and 120r/min for 24 hr to logarithmic growth end stage to obtain Streptococcus thermophilus liquid with thallus concentration of 1 × 10 or more14CFU/L；
S3: taking 2g of squid homogenate to be put into a 50ml centrifuge tube, firstly adding 10g of distilled water into streptococcus thermophilus according to the solid-to-liquid ratio of 1:5, sucking 20% and 400ul of bacterial liquid into the centrifuge tube by a liquid transfer gun, placing the centrifuge tube under the conditions of 40 ℃ and 120r/min for fermentation for 36h, and inactivating the squid homogenate at 90 ℃ for 30 min;
s4: adding flavourzyme into the fermented squid homogenate according to 6000u/g, then placing the centrifuge tube into a 50 ℃ incubator for enzymolysis for 3h, and freeze-drying the zymotic fluid after enzymolysis to obtain the flavoured squid nutritional biological product.
Comparative example 2:
a preparation method of a flavored squid nutritional biological product comprises the following steps:
s1: cleaning squid byproduct, crushing, homogenizing, sterilizing in a high pressure steam sterilization pot at 120 deg.C for 30min, and storing in a refrigerator at-20 deg.C;
s2: inoculating Streptococcus thermophilus into MRS broth conical flask, shake culturing at 37 deg.C and 120r/min for 24 hr to logarithmic growth end stage to obtain Streptococcus thermophilus liquid with thallus concentration of 1 × 10 or more14CFU/L；
S3: taking 2g of squid homogenate to be put into a 50ml centrifugal tube, firstly adding 10g of distilled water into streptococcus thermophilus according to the solid-to-liquid ratio of 1:5, sucking 20% and 400ul of the bacterial liquid into the centrifugal tube by a liquid transfer gun, placing the centrifugal tube in the condition of 40 ℃ and 120r/min for fermentation for 36h, and freeze-drying the fermentation liquid to obtain the flavored squid nutritional biological product.
Comparative example 3:
a preparation method of a flavored squid nutritional biological product comprises the following steps:
s1: cleaning squid byproduct, crushing, homogenizing, sterilizing in a high pressure steam sterilization pot at 120 deg.C for 30min, and storing in a refrigerator at-20 deg.C;
s2: taking 2g of squid homogenate to be placed in a 50ml centrifugal tube, adding 10g of distilled water into streptococcus thermophilus according to the solid-to-liquid ratio of 1:5, adding flavourzyme according to 6000u/g, adjusting the pH of the system to 7.0, then placing the centrifugal tube in a 50 ℃ incubator for enzymolysis for 3h, then inactivating the mixture at 90 ℃ for 30min, and freeze-drying the enzymolysis liquid to obtain the flavoured squid nutritional biological product.
Test example 1:
1. determination of glutamic acid content in enzymolysis squid homogenate
Referring to the determination of amino acids in GB/T5009.124-2003 food, specifically, the enzymatic squid homogenate is freeze-dried, 2g is weighed, 15mL of 0.02mol/L hydrochloric acid solution is added, after sufficient homogenization, the volume is fixed to 50mL by 0.02mol/L hydrochloric acid solution, and centrifugation is carried out for 10min at 5000 r/min. Taking 2mL of the supernatant, adding 2mL of 8g/100mL sulfosalicylic acid solution, standing in a refrigerator at 4 ℃ for 30min, and centrifuging at 10000r/min for 10 min. The supernatant was filtered through a 0.22 μm filter and then subjected to sample injection at a volume of 20 μ L. As shown in fig. 1, it can be seen that the content of glutamic acid in the enzymatic squid homogenate obtained by the enzymatic hydrolysis in examples 2 and 3 is higher than that in example 1, which indicates that the sage extract can promote the flavourzyme to carry out the enzymatic hydrolysis on the protein in the squid byproduct homogenate, increase the content of glutamic acid in the enzymatic squid homogenate, and is beneficial to the growth and proliferation of streptococcus thermophilus.
2. Determination of soluble protein conversion
Soluble protein conversion was calculated according to the following formula:
soluble protein conversion (%) - (M-N)/P × 100%;
wherein,
m is the soluble protein content in the biological product (measured by the Folin phenol method);
n is the content of soluble protein in the squid by-product (measured by adopting a forskolin phenol method);
and P is the total protein content of the squid byproduct (measured by adopting a Kjeldahl method).
As shown in FIG. 2, it can be seen that the soluble protein conversion rate in the preparation method of example 1 is as high as 25.15%, and the soluble protein conversion rates in the preparation methods of comparative examples 1-3 are 20.10%, 15.50% and 13.93%, respectively, so that the soluble protein conversion rate in the preparation method of example 1 is much higher than that in comparative examples 1-3, which shows that the preparation method of the present invention can greatly improve the soluble protein conversion rate by the combined action of Streptococcus thermophilus and flavourzyme (enzyme first and then bacterium); in addition, the conversion rate of soluble protein in the preparation method of example 1 is lower than that in examples 2-3, which shows that the sage extract can also promote the flavourzyme to enzymolyze protein in the squid by-product homogenate to obtain the enzymolyzed squid homogenate rich in oligopeptides and amino acids, particularly the content of glutamic acid in the enzymolyzed squid homogenate is increased, and the growth and proliferation of streptococcus thermophilus are facilitated, so that the conversion rate of soluble protein is finally increased.
3. Determination of content of unsaturated fatty acid in flavored squid nutritional biological product
Referring to the measurement of total fat, saturated fat (acid) and unsaturated fat (acid) in GB/T22223-2008 food, hydrolysis extraction-gas chromatography is adopted. The fatty acid composition of the flavor squid nutritional biological product is analyzed through gas chromatography detection, and the result is shown in fig. 3, so that the content of unsaturated fatty acid in the flavor squid nutritional biological product prepared in the example 1 is far higher than that in the comparative examples 1-3, which shows that the content of unsaturated fatty acid in the flavor squid nutritional biological product can be greatly improved through the combined action of streptococcus thermophilus and the bacterial enzymes of the flavourzyme (firstly enzyme and then bacteria); in addition, the content of unsaturated fatty acid in the flavored squid nutritional biological products prepared in the examples 2-3 is more than 80g/100g, which is far higher than that in the examples 2-3, and the content of unsaturated fatty acid in the biological products can be improved due to the sage extract.
Test example 2:
1. determination of pH value and titration acidity of squid byproduct fermentation liquor
The pH value of the squid byproduct fermentation liquor is measured by a pH meter, and the determination of the titration acidity is determined by referring to GB5413.34-2010NaOH titration method and is expressed by (T degree). The results are shown in fig. 4, and it can be seen that the pH value of the squid byproduct fermentation liquid obtained in example 4 is lower than that of the squid byproduct fermentation liquids obtained in examples 1 to 3 and comparative examples 1 to 2, and the acidity of the squid byproduct fermentation liquid obtained in example 4 is higher than that of the squid byproduct fermentation liquids obtained in examples 1 to 3 and comparative examples 1 to 2, which indicates that the acid production capacity and acid resistance of streptococcus thermophilus can be improved by adding glycocholic acid and berberine hydrochloride in the fermentation process, so that the accumulation amount of organic acids in the fermentation liquid can be increased.
2. Determination of bacteriostatic activity of squid byproduct fermentation liquor
In the test example, four common putrefying bacteria, namely escherichia coli, salmonella, staphylococcus aureus and bacillus cereus, are selected as indicator bacteria, and the antimicrobial activity of the squid byproduct fermentation liquor is measured by adopting an oxford cup method.
Preparing an indicator bacterium suspension: weighing MRS broth culture medium at a certain proportion, preparing 50ml, heating in water bath to dissolve completely, subpackaging 4 test tubes of 18ml with 6ml, sealing test tube port with broth culture medium with gauze and kraft paper, placing test tube and rubber plug in autoclave for sterilization at 121 deg.C for 20 min. After the sterilization is finished, the indicator bacteria cultured by the 4 test tube slants are respectively inoculated to the 4 sterilized broth culture medium test tubes by using an inoculating ring under the super clean bench sterile condition, a rubber plug is covered, and 4 different indicator bacteria are marked by using a marking pen. Shaking the 4 test tubes at 37 deg.C and 150r/min for 24h, at which time the concentration of indicator bacteria is 107-109cfu/ml。
An oxford cup method bacteriostasis test: placing the indicator bacterium in a culture dish, uniformly coating the indicator bacterium by using a sterile coater, then uniformly placing 3 sterilized oxford cups on a solid culture medium by using sterile tweezers, transferring 100 mu L of biological product solution (sterile water is dissolved) with the mass concentration of 1.5g/L into the oxford cups in the prepared detection plate, and placing the same liquid into the 3 oxford cups, namely, making 3 detection plates in parallel; after the fermentation liquid is added, the detection plate is placed in a refrigerator at 4 ℃ for 12 hours, then the detection plate is placed in a constant-temperature incubator at 37 ℃ for culturing for 16 hours, the culture dish is carefully taken out, and the diameter of the inhibition zone of the oxford cup is immediately measured by a caliper. As shown in Table 1, it can be seen that the solutions of the biologicals obtained in examples 1 to 4 and comparative examples 1 to 2 all have bacteriostatic effects on Escherichia coli, Salmonella, Staphylococcus aureus and Bacillus cereus, while the solution of the biologicals obtained in comparative example 3 has no bacteriostatic effects on Escherichia coli, Salmonella, Staphylococcus aureus and Bacillus cereus; meanwhile, the biological product solution obtained in the embodiment 4 has the best antibacterial effect on escherichia coli, salmonella, staphylococcus aureus and bacillus cereus, which shows that glycocholic acid and berberine hydrochloride are added in the fermentation process, so that the antibacterial activity of the squid byproduct fermentation liquid can be improved, the synergistic effect with the sage extract is realized, the growth and the propagation of harmful bacteria of putrefying bacteria are inhibited, and the preservation period of the biological product is prolonged.
TABLE 1 inhibition zone (mm) of squid by-product fermentation broth for four pathogenic bacteria
| Escherichia coli | Salmonella | Staphylococcus aureus | Bacillus cereus | |
| Example 1 | 13.8±0.23 | 12.1±0.23 | 14.9±0.37 | 14.6±0.34 |
| Example 2 | 15.8±0.42 | 14.6±0.54 | 16.5±0.74 | 15.3±0.44 |
| Example 3 | 16.3±0.51 | 15.1±0.37 | 17.2±0.83 | 16.0±0.48 |
| Example 4 | 19.8±0.64 | 17.7±0.45 | 21.4±1.01 | 20.8±0.87 |
| Comparative example 1 | 10.5±0.22 | 9.8±0.27 | 12.4±0.32 | 9.4±0.24 |
| Comparative example 2 | 11.6±0.16 | 11.0±0.31 | 12.8±0.25 | 10.7±0.34 |
| Comparative example 3 | - | - | - | - |
Note: "-" indicates that no zone of inhibition was observed
Conventional techniques in the above embodiments are known to those skilled in the art, and therefore, will not be described in detail herein.
The above embodiments are merely illustrative, and not restrictive, and those skilled in the art can make various changes and modifications without departing from the spirit and scope of the invention. Therefore, all equivalent technical solutions also belong to the scope of the present invention, and the protection scope of the present invention should be defined by the claims.
Claims (8)
1. A preparation method of a flavored squid nutritional biological product is characterized by comprising the following steps: the method comprises the following steps:
s1: homogenizing the squid byproduct, and sterilizing to obtain squid homogenate;
s2: adding distilled water and flavourzyme into the squid homogenate for enzymolysis, and inactivating enzyme;
s3: and adding thermophilic streptococcus liquid into the enzymolysis squid homogenate for fermentation, and freeze-drying the fermentation liquor to obtain the flavored squid nutritional biological product.
2. The preparation method of the flavored squid nutritional biological product according to claim 1, characterized in that: the addition amount of the flavor protease in the S2 is 5000-; the enzymolysis temperature is 45-60 deg.C, pH is 6.5-7.5, and the time is 2-5 h.
3. The preparation method of the flavored squid nutritional biological product according to claim 1, characterized in that: the streptococcus thermophilus liquid in the S3 is obtained by culturing streptococcus thermophilus in an MRS culture medium until the end of logarithmic growth of thalli is reached.
4. The preparation method of the flavored squid nutritional biological product according to the claim 1 or 3, is characterized in that: the thallus concentration of the streptococcus thermophilus liquid in the S3 is more than or equal to 1 multiplied by 1014CFU/L。
5. The preparation method of the flavored squid nutritional biological product according to the claim 1 or 3, is characterized in that: the addition amount of the streptococcus thermophilus in the S3 is 10-20%, the fermentation temperature is 30-40 ℃, the pH value is 6.5-7.5, the rotation speed is 100-150r/min, and the time is 30-40 h.
6. The preparation method of the flavored squid nutritional biological product according to claim 1, characterized in that: the conversion rate of soluble protein in the preparation method is more than 25%.
7. The preparation method of the flavored squid nutritional biological product according to claim 1, characterized in that: in the S2, the sage extract is added into the distilled water.
8. The preparation method of the flavored squid nutritional biological product according to claim 7, wherein the preparation method comprises the following steps: the weight concentration of the sage extract is 0.2-2%.
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