Surfactant for cracking digestive organs of shrimps and shellfish

Abstract

The invention discloses and provides a surfactant for cracking digestive organs of shrimps and shellfish, which is long-chain fatty acid phosphonate: R-CH2-CO-P (= O) (OR ') 2, in which R is C28-C100 alkyl; r'is a soluble hydrophilic group. The surfactant can effectively improve the cracking efficiency of digestive organs of prawns and shellfishes, is used for fine processing of meat bodies of the prawns and shellfishes and recycling of the digestive organs, and has popularization and application values.

Description

Technical Field

[0001] The invention belongs to the field of biotechnology and relates to a surfactant for cracking shrimp and shellfish digestive organs, a cracking reagent containing the surfactant and application thereof. Background Art

[0002] In the field of aquaculture, shrimp and shellfish are both important products. However, the digestive organs of shrimp and shellfish, such as the intestines and stomach, face the problem of being difficult to completely lyse and remove during the processing and handling of shrimp / shellfish meat. Existing lysis methods mainly rely on high temperature, strong acid or enzymatic hydrolysis, but these methods often lead to degradation or loss of the fleshy body product. Therefore, the development of a new type of detergent that can efficiently lyse the digestive organs of shrimp while protecting the active ingredients is of great economic and technical significance.

[0003] On the other hand, tests have shown that the cleavage products of shrimp and shellfish digestive organs contain a large amount of peptides and nucleotides, which have certain nutritional and health value. If they can be recycled, they can also be used as nutritional additives and animal feed additives, realizing waste utilization and turning waste into treasure, but there is no relevant processing technology so far. Summary of the invention

[0004] In view of the above technical problems of shrimp and shellfish aquatic product processing in the prior art, we have developed a surfactant and a lysis reagent containing the surfactant, which can not only effectively degrade digestive organs such as the intestines and stomach of shrimp and shellfish, and remove them from shrimp and shellfish meat, but also can maintain the taste and nutritional value of the meat without damaging the quality of shrimp and shellfish meat; in addition, the lysis reagent has food safety. Specifically, the present invention includes the following technical solutions.

[0005] The first aspect of the present invention provides a surfactant for cracking shrimp and shellfish digestive organs, wherein the surfactant is a phosphate-based surfactant, which is a long-chain fatty acid phosphonate shown in Formula I:

[0006] R-CH2-CO-P(=O)(OR′)2I

[0007] Wherein, R is a long-chain alkyl group of C28-C100;

[0008] R' is a soluble hydrophilic group, which is related to the solubility and cleavage activity of the surfactant, and is selected from the following groups: carboxylic acid group (-COOH or its salt -COO-); sulfonic acid group (-SO3H or its salt -SO3-): alkyl sulfonic acid; alkylbenzene sulfonic acid; sulfuric acid group (-OSO3H or its salt -OSO3-): alkyl sulfate; amine group (-NH2, -NHR, -NR2): ester group (-COOR): polyvinyl alcohol (PEG) group: ethoxylate (-O-(CH2CH2O)nH); alcohol group (-OH): nitrile group (-C≡N): hydroxy acid residue: organic phosphate and phosphonate.

[0009] The above-mentioned alkyl group may be a straight-chain alkyl group or a branched-chain alkyl group, preferably a straight-chain alkyl group; the branch in the branched-chain alkyl group may be a 1-methylpentyl group (C6H13), that is, a 1-methylpentyl group (C6H13) is introduced into the long-chain main chain as a branch.

[0010] For example, the above-mentioned alkyl group is n-triacontane (C30H62), n-triacontane (C31H64), n-tetracontane (C40H82), n-pentacontane (C50H102), n-hexacontane (C60H126), etc.

[0011] The preparation method of the surfactant is very simple. For example, a long-chain fatty acid and a phosphonate compound are reacted under suitable conditions to prepare a phosphate-based surfactant. The optional reaction conditions are: a temperature of 80-120° C. and a reaction time of 2-4 hours.

[0012] The preparation method of the above surfactant is: reacting a long-chain fatty acid with a phosphonate compound under suitable conditions to obtain a phosphate-based surfactant. The reaction conditions are: temperature of 80-120° C. and reaction time of 2-4 hours.

[0013] The digestive organs to which the above-mentioned surfactant is applicable particularly refer to the intestines, stomach, liver, pancreas and other internal organs of shrimps and shellfishes.

[0014] A second aspect of the present invention provides a lysis reagent for lysis of shrimp and shellfish digestive organs, comprising the phosphate-based surfactant described above.

[0015] Preferably, the above-mentioned lysis reagent comprises the following components, preferably consists of the following components:

[0016] Tris-Cl buffer: provides a stable pH environment;

[0017] Triazole: With its good stabilizer ability, it helps stabilize nucleic acids and proteins;

[0018] The phosphate-based surfactant according to any one of claims 1 to 3: replacing the conventional surfactant in the prior art, and being able to further improve the cleavage efficiency;

[0019] Polyethylene glycol such as PEG 4000: used to stabilize and protect proteins, so that the shrimp and shellfish flesh bodies maintain nutrition and taste during the lysis process;

[0020] NaCl: Used to regulate osmotic pressure and helps prevent protein aggregation.

[0021] In one embodiment, the amounts of the components in the above lysis reagent are respectively:

[0022] The concentration of Tris buffer is 10-200 mM, pH 7.0-9.5, preferably about 50 mM, about pH 8.0;

[0023] The concentration of triazole is 1-8% (v / v), preferably about 5%;

[0024] The concentration of the phosphate-based surfactant as described above is 0.5-2% (v / v), for example, the phosphate-based surfactant in which R is triacontyl and R' is carboxyl -COOH is preferably about 1%;

[0025] The concentration of polyethylene glycol, such as PEG 4000, is 5-15% (w / v), and the concentration of PEG 4000 is preferably about 10%;

[0026] The NaCl concentration is 10 to 1000 mM, preferably about 150 mM.

[0027] It should be understood that when expressing numerical characteristics in this article, the terms "approximately", "about" or "around" mean that the number represented may have an error range or floating range of ±10%, ±8%, ±6%, ±4% or ±2%.

[0028] The third aspect of the present invention provides the use of the above-mentioned lysis reagent in clearing shrimp and shellfish digestive organs, or in preparing shrimp and shellfish digestive organ lysis liquid.

[0029] In one application embodiment, the digestive organs of shrimp and shellfish are cleaned by the following method:

[0030] soaking the shelled / peeled fleshy bodies in an aqueous solution of a lysis reagent, wherein the concentration of the surfactant as described above is 0.5%-2% (w / v);

[0031] Keep warm at 40-70°C, preferably 60°C for 2-4 hours, shaking gently if necessary;

[0032] The fleshy body is separated from the liquid to obtain shrimp meat / shellfish meat with the internal organs such as the shrimp and shellfish intestines, stomach, liver, and pancreas removed.

[0033] In another application embodiment, shrimp and shellfish digestive organ lysate is prepared by the following method:

[0034] The digestive organs of shrimp and shellfish, including the intestines and stomach, are immersed in an aqueous solution of a lysis reagent, wherein the concentration of the surfactant described above is 0.5% to 2% (w / v);

[0035] Incubate at 40-70°C, preferably 60°C for 0.5-1.0 hour, shaking or stirring if necessary;

[0036] Centrifuge and collect the supernatant to obtain a lysate. If necessary, further freeze-dry the lysate to obtain a lyophilized powder.

[0037] The lysate obtained by the above method or the lyophilized powder of the lysate mainly contains polypeptides and is rich in nucleotides, and can be used as a nutritional supplement and feed additive.

[0038] The present invention develops a surfactant specially used for cracking the digestive organs of shrimp and shellfish and a cracking liquid composition containing the surfactant. The cracking reagent has food safety, can effectively degrade the digestive organs in shrimp meat and shellfish meat, and does not damage the quality of the shrimp meat and shellfish meat, thereby maintaining the taste and nutritional value of the meat body; the product of cracking the digestive organs of shrimp and shellfish using the cracking reagent contains a large amount of polypeptides and nucleotides, can be used as a nutritional supplement and feed additive, turning waste into treasure, and has a prospect of promotion and application in the field of aquatic product processing. BRIEF DESCRIPTION OF THE DRAWINGS

[0039] Figure 1 A photograph of DNA agarose gel electrophoresis in the Examples is shown. DETAILED DESCRIPTION

[0040] Shrimp and shellfish are important shelled aquatic products. Shrimp includes freshwater shrimp and marine shrimp. Common shellfish include clams, oysters, scallops, mussels: mussels, clams, abalone, cockles, etc. In the food industry, the processing of these shelled aquatic products is to separate the meaty body, namely shrimp meat and shellfish meat, from the shell.

[0041] Since the digestive organs of shrimp and shellfish, including the intestines, stomach, liver, pancreas and other internal organs, have a fishy smell and affect the appearance quality of shrimp and shellfish meat, it is generally necessary to remove the digestive organs from the flesh. Most of the digestive organs can be peeled off by mechanical / physical methods, but some of them still remain in the flesh and are difficult to separate by mechanical / physical methods. At present, the flesh is generally treated by lysis methods such as high temperature, strong acid or enzymatic hydrolysis to remove the remaining digestive organs, but these methods will also cause degradation and loss of meat, and will destroy the nutritional value and / or taste of the meat.

[0042] In addition, digestive organs including the intestines, stomach, liver, pancreas and other internal organs are generally thrown away as waste, however, digestive organs are not without use value. Biological tests have found that the lysate of digestive organs including the intestines, stomach, liver, pancreas and other internal organs is rich in peptides and nucleotides, which can be used as nutritional supplements, feed additives, etc. For example, the role of nucleotides in genetic nutrition and activating human cell functions has attracted more and more attention from nutritionists. On the one hand, nucleotides can stimulate the activity of macrophages, and on the other hand, they can locate and find and cut viral gene chains in time, quickly engulf viral cells, prevent the transcription and translation of viral genes, and protect normal cells from damage. Nucleotide supplement foods are available on the market in many countries such as the United States, Canada, European countries and Japan.

[0043] The phosphate-based surfactants and lysis reagents we developed are food-safe and can maintain the nutritional value and taste of the flesh when lysis of digestive organs, thus improving the quality of shrimp and shellfish processed products.

[0044] The phosphate-based surfactant is a long-chain fatty acid phosphonate, and has the structural formula I:

[0045] R-CH2-CO-P(=O)(OR′)2I

[0046] Wherein, R is a C28-C100 alkyl group, which can be selected from the following group:

[0047] C30 to C50 alkyl groups, for example, triacontan (C30H62), trihexane (C31H64), etc., can be further extended to C40 (such as tetracontan);

[0048] C50 to C100 ultra-long chain alkyl, for example, n-pentacontane (C50H102), n-hexacontane (C60H126), etc.

[0049] The structure of the candidate R chain: The long chain can contain straight chain and branched chain structures. It is possible to consider introducing a branch chain, such as 1-methylpentyl (C6H13) as a branch chain, combined with the long chain main chain.

[0050] R' is a soluble group, which is related to the solubility and cleavage activity of the surfactant and can be selected from the following groups:

[0051] Carboxylic acid groups (-COOH and its salts -COO-), for example, acetate, lactate, etc.;

[0052] Sulfonic acid groups (-SO3H and its salts -SO3-), for example, alkylsulfonic acids, alkylbenzenesulfonic acids;

[0053] Sulfate groups (-OSO3H and salts thereof, such as -OSO3-), for example, alkyl sulfates;

[0054] Amine groups (-NH2, -NHR, -NR2), e.g., alkylamines, quaternary ammonium salts;

[0055] Ester groups (-COOR), e.g., fatty acid esters;

[0056] Polyvinyl alcohol (PEG) groups, e.g., ethoxylate (-O-(CH2CH2O)nH);

[0057] alcohol group (-OH), e.g., long-chain alcohols;

[0058] a nitrile group (-C≡N), for example, an alkylnitrile;

[0059] Hydroxy acid residues, e.g., citric acid, malic acid;

[0060] Organic phosphates and phosphonates, for example, groups derived from phosphoric acid and phosphonic acid.

[0061] In one example, R in the molecular structure of the phosphate-based surfactant is a triacontyl group, and R' is a carboxyl group -COOH.

[0062] It is easy for a person skilled in the art to understand that the dosage / concentration of each component of Tris-Cl, Triazole, polyethylene glycol such as PEG 4000, phosphate-based surfactant and NaCl in the lysis reagent provided by the present invention can be determined through simple experiments according to the species of shrimps and shellfish to be treated.

[0063] It should be understood that the specific reaction conditions when using lysis reagents to treat shrimp meat, shellfish meat, and digestive organs can also be determined through simple experiments based on the species of shrimp and shellfish to be treated.

[0064] For example, the steps applied to shrimp digestive organ lysis include:

[0065] Sample preparation: Take fresh shrimp samples, remove the shells and keep the digestive organs;

[0066] Lysis treatment: placing the shrimp digestive organs in a lysis solution containing the phosphate-based surfactant at a concentration of 0.5%-2% (w / v), and incubating at 60° C. for 30 minutes;

[0067] Post-treatment: After the lysis is completed, the insoluble residue is removed by centrifugation or filtration, and the supernatant is collected to obtain the lysate.

[0068] To make the present invention more clearly understandable, preferred embodiments are described in detail below with reference to the accompanying drawings. Those skilled in the art should understand that the following embodiments are only used to illustrate the present invention and are not intended to limit the present invention.

[0069] Example

[0070] This article involves the addition amount, content and concentration of various substances, and the percentages mentioned here refer to the mass percentages unless otherwise specified.

[0071] In the examples herein, the temperature when the shrimp digestive organs are lysed using a lysing agent is about 55-60° C. If no specific description is given for a particular operating temperature, the temperature generally refers to room temperature (15-35° C.).

[0072] DL2000 marker used in the examples was purchased from Thermo Fisher Scientific.

[0073] Two animal tissue DNA extraction kits: two domestic reagent companies.

[0074] Other reagents were purchased from Aladdin or MacLean.

[0075] Example 1: Shrimp digestive organ lysis and nucleotide analysis of lysate

[0076] Preparation of lysis reagent

[0077] The formula of the lysis reagent is: Tris-Cl (pH 8.0) 50 mM, Triazole 5%, phosphate-based surfactant (R is triacontyl, R' is carboxyl -COOH) 1%, PEG 4000 10%, and NaCl 150 mM.

[0078] Shrimp digestive organs lysis

[0079] The shrimp digestive organs (liver, pancreas and stomach) were placed in a lysis reagent and incubated at 60°C for 30 minutes. After the lysis reaction was completed, the insoluble residue was removed by centrifugation or filtration, and the supernatant was collected to obtain the shrimp digestive organ lysate for analysis.

[0080] For comparison, two animal tissue DNA extraction kits from two other domestic reagent companies were used to extract nucleic acid from shrimp digestive organs.

[0081] After the cleavage reaction and kit extraction, agarose gel electrophoresis was performed according to the conventional method. The results are shown in Figure 1 The analytes corresponding to the lines from left to right are shown below.

[0082] Line1 DL2000marker Line2 Shrimp digestive organ lysate sample 1 Line3 Shrimp digestive organ lysate sample 2 Line4 Nucleic acid extraction samples of animal tissue DNA extraction kit from reagent company No. 1 Line5 Nucleic acid extraction samples of animal tissue DNA extraction kit from reagent company No. 2

[0083] from Figure 1 It can be seen from the electrophoresis image that the brightness of lanes 2 and 3 (shrimp digestive organ lysate samples) is high, indicating that the nucleotide DNA concentration is high; the brightness of lanes 4 and 5 (shrimp digestive organ DNA extraction kit extract) is low, indicating that the DNA concentration is lower than the nucleotide concentration in the lysate sample of the present invention. This result shows that the lysing reagent containing a specific phosphate-based surfactant of the present invention can efficiently lyse shrimp digestive organs, turn waste into treasure, prepare biological products with health value, and improve the economic benefits of shrimp and shellfish aquatic product processing enterprises.

[0084] The above description of the embodiments is to facilitate the understanding and use of the invention by those skilled in the art. It is obvious that those skilled in the art can easily make various modifications to these embodiments and apply the general principles described herein to other embodiments without creative work. Therefore, the present invention is not limited to the above embodiments, and improvements and modifications made by those skilled in the art based on the tips of the present invention without departing from the scope of the present invention should be deemed to be still within the protection scope of the present invention.

Claims

1. A surfactant for cracking the digestive organs of shrimp and shellfish, characterized in that: The surfactant is a long-chain fatty acid phosphonate shown in Formula I: R-CH2-CO-P(=O)(OR′)2I Wherein, R is a C28-C100 alkyl group; R' is a soluble hydrophilic group selected from the following groups: carboxylic acid group (-COOH or its salt -COO-); sulfonic acid group (-SO3H or its salt -SO3-): alkyl sulfonic acid; alkylbenzene sulfonic acid; sulfuric acid group (-OSO3H or its salt -OSO3-): alkyl sulfate; amine group (-NH2, -NHR, -NR2): ester group (-COOR): polyvinyl alcohol (PEG) group: ethoxylate (-O-(CH2CH2O)nH); alcohol group (-OH): nitrile group (-C≡N): hydroxy acid residue: organic phosphate and phosphonate.

2. The surfactant according to claim 1, characterized in that The alkyl group is a linear alkyl group or a branched alkyl group, preferably a linear alkyl group.

3. The surfactant according to claim 1, characterized in that The alkyl groups are triacontandecane (C30H62), hexahedrondecane (C31H64), tetracontandecane (C40H82), pentacontandecane (C50H102), and hexacontandecane (C60H126).

4. The surfactant according to claim 1, characterized in that The long-chain fatty acid and the phosphonate compound are reacted under appropriate conditions to obtain a phosphate-based surfactant.

5. A lysis reagent for lysis of shrimp and shellfish digestive organs, characterized in that: Contains the surfactant according to any one of claims 1 to 3.

6. The lysis reagent according to claim 1, characterized in that The method comprises the following components, and preferably consists of the following components: Tris-Cl buffer, Triazole, the surfactant according to any one of claims 1 to 3, polyethylene glycol PEG4000, and NaCl.

7. The lysis reagent according to claim 1, characterized in that The dosage of the components is respectively: Tris buffer concentration is 10-200 mM, pH 7.0-9.5; Triazole concentration is 1-8% (v / v); The surfactant concentration of any one of claims 1 to 3 is 0.5-2% (v / v); Polyethylene glycol, such as PEG 4000, at a concentration of 5-15% (w / v); The NaCl concentration is 10~1000mM.

8. Use of the lysis reagent according to any one of claims 6 to 7 in removing shrimp and shellfish digestive organs, or in preparing shrimp and shellfish digestive organ lysis liquid.

9. The use according to claim 8, characterized in that Clean the digestive organs of shrimp and shellfish by the following methods: soaking the shelled / peeled fleshy bodies in an aqueous solution of a lysis reagent, wherein the concentration of the surfactant as claimed in claim 1 is 0.5%-2% (w / v); Keep warm at 40-70°C, preferably 60°C for 2-4 hours; Separating the flesh from the liquid to obtain shrimp / shellfish meat from which the intestines, stomach and viscera of the shrimp / shellfish have been removed; or Shrimp and shellfish digestive organ lysate was prepared by the following method: Soaking the digestive organs of shrimp and shellfish including the intestines, stomach and viscera in an aqueous solution of a lysis reagent, wherein the concentration of the surfactant as claimed in claim 1 is 0.5%-2% (w / v); Incubate at 40-70°C, preferably 60°C for 0.5-1.0 hour; Centrifuge and collect the supernatant to obtain the lysate.

10. The use according to claim 9, characterized in that The obtained lysate or the lyophilized powder of the lysate is used as a nutritional supplement or a feed additive.

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