CN115553334A - Application of L-lysine or L-threonine as food low-temperature preservative - Google Patents
Application of L-lysine or L-threonine as food low-temperature preservative Download PDFInfo
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- CN115553334A CN115553334A CN202211297851.8A CN202211297851A CN115553334A CN 115553334 A CN115553334 A CN 115553334A CN 202211297851 A CN202211297851 A CN 202211297851A CN 115553334 A CN115553334 A CN 115553334A
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- KDXKERNSBIXSRK-YFKPBYRVSA-N L-lysine Chemical compound NCCCC[C@H](N)C(O)=O KDXKERNSBIXSRK-YFKPBYRVSA-N 0.000 title claims abstract description 90
- AYFVYJQAPQTCCC-GBXIJSLDSA-N L-threonine Chemical compound C[C@@H](O)[C@H](N)C(O)=O AYFVYJQAPQTCCC-GBXIJSLDSA-N 0.000 title claims abstract description 69
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Images
Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23B—PRESERVING, e.g. BY CANNING, MEAT, FISH, EGGS, FRUIT, VEGETABLES, EDIBLE SEEDS; CHEMICAL RIPENING OF FRUIT OR VEGETABLES; THE PRESERVED, RIPENED, OR CANNED PRODUCTS
- A23B7/00—Preservation or chemical ripening of fruit or vegetables
- A23B7/14—Preserving or ripening with chemicals not covered by groups A23B7/08 or A23B7/10
- A23B7/153—Preserving or ripening with chemicals not covered by groups A23B7/08 or A23B7/10 in the form of liquids or solids
- A23B7/154—Organic compounds; Microorganisms; Enzymes
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23B—PRESERVING, e.g. BY CANNING, MEAT, FISH, EGGS, FRUIT, VEGETABLES, EDIBLE SEEDS; CHEMICAL RIPENING OF FRUIT OR VEGETABLES; THE PRESERVED, RIPENED, OR CANNED PRODUCTS
- A23B7/00—Preservation or chemical ripening of fruit or vegetables
- A23B7/04—Freezing; Subsequent thawing; Cooling
-
- 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
Abstract
The invention provides an application of L-lysine (L-lysine) or L-threonine (L-threonine) as a low-temperature food preservative. The invention also provides a low-temperature preservation method of straw mushrooms, which comprises the steps of soaking the straw mushrooms in the solution of L-lysine or L-threonine, draining water and refrigerating. According to the invention, L-lysine or L-threonine is proportioned into a solution, and a straw mushroom fruiting body soaking experiment is carried out, so that the L-lysine or L-threonine can obviously delay the cold injury of straw mushrooms at low temperature, can be used as a low-temperature preservative for food such as straw mushrooms and the like, and can be used for developing functional nutritional type straw mushroom food rich in L-lysine or L-threonine.
Description
Technical Field
The invention belongs to the field of food, and relates to a low-temperature food preservation technology, in particular to application of L-lysine or L-threonine as a low-temperature food preservative.
Background
Straw mushroom (Volvariella volvacea) is called "Chinese mushroom" and is an important edible fungus. The straw mushroom not only has delicious taste, but also has important medical care effects, such as oxidation resistance, immunity regulation, tumor resistance and the like. The content of the essential amino acid of the straw mushroom fruiting body accounts for 40.47-44.47% of the total amount of the amino acid, and is obviously higher than the recommended value of the essential amino acid of FAO/WHO. The growth cycle of the straw mushroom is short (about 10 days), and the straw mushroom becomes edible mushroom with high economic benefit. However, the growth and fruiting of the hypha of the straw mushroom require relatively high temperature (28-35 ℃), especially the hypha and the fruiting body are not resistant to low temperature (0-10 ℃), even under the conventional low temperature (4 ℃), the hypha and the fruiting body can be softened, liquefied and even rotten in a short time, namely the hypha and the fruiting body are generally called as low-temperature autolysis. The uniqueness of the straw mushroom 'low-temperature autolysis' provides a preservation problem. Therefore, the shelf life of the straw mushrooms is obviously shorter than that of other mushrooms all the time, so that the cultivation and the popularization of the straw mushrooms are greatly limited, and the economic benefit of degrading the straws is indirectly influenced. Although there are some solutions for cryopreservation of volvariella volvacea, these solutions often present high cost and food safety issues.
The shortage of protein resources has become a worldwide problem. How to develop new high-quality protein products to meet the increasing nutritional needs of human beings becomes the subject of urgent research. The straw mushroom 'low-temperature autolysis' is an abnormal metabolic activity. Recently, we have interfered the ubiquitin-conjugated E2 enzyme (UBEV 2) mediated autolytic ubiquitination pathway to develop the low-temperature preservation technology of the straw mushroom, and the antifreeze phenotype of the straw mushroom stored for a long time (at least 48 h) at low temperature is obtained by using the inhibitor compound L345-0044 of the UBEV2 to spray in the growth period of the button of the straw mushroom (patent number: 110202124029.0). The total amount of soluble sugar and free amino acid of the straw mushroom sprayed by L345-0044 is remarkably up-regulated (DOI: 10.13346/j. Mycosystema.220069), and the interference of abnormal metabolism of the straw mushroom is suggested to change the quality of the straw mushroom. The antifreezing phenotype of the straw mushroom is obtained by regulating and controlling the process by utilizing the abnormal metabolism capability of the straw mushroom under the cold action, and meanwhile, the novel functional nutritional food of the straw mushroom can be possibly developed.
Disclosure of Invention
The invention aims to provide application of L-lysine or L-threonine as a low-temperature food preservative, and aims to solve the technical problems of poor low-temperature preservation pertinence, unobvious effect, high cost and food safety of straw mushrooms in the prior art.
The invention provides an application of L-lysine or L-threonine as a low-temperature food preservative.
Furthermore, the L-lysine or L-threonine is an essential amino acid for human body.
Further, the food is straw mushroom.
The invention also provides a low-temperature preservation method of straw mushrooms, which comprises the steps of soaking the straw mushrooms in the solution of L-lysine or L-threonine, draining water and refrigerating.
Preferably, the refrigeration temperature is 0 to 8 ℃.
Preferably, the soaking time is 2 to 4 minutes.
Furthermore, the L-lysine or L-threonine is an essential amino acid for human body.
Furthermore, the concentration of the human body essential amino acid L-lysine or L-threonine solution is 200-300 MuM.
Preferably, the concentration of the L-lysine solution is 200 μ M; the concentration of the L-threonine solution was 300. Mu.M.
On the basis of analyzing the molecular mechanism of the anti-freezing phenotype of the long-time low-temperature fresh-keeping of the straw mushrooms under the spraying effect of L345-0044 (shown in figure 1), the invention utilizes the principle that enhanced lysine metabolism provides acetyl coenzyme A (acetyl-CoA) to promote citric acid circulation, and further develops a functional nutritional type low-temperature-resistant fresh-keeping technology of the straw mushrooms.
According to the invention, the straw mushroom fruiting body soaking experiment is carried out by using the solution prepared from the L-lysine or the L-threonine, and the amino acid can obviously delay the frostbite of the straw mushroom at low temperature and has the potential of becoming a low-temperature straw mushroom preservative.
Compared with the prior art, the invention has the advantages of positive and obvious technical effect. The invention relates to a method for low-temperature-resistant preservation of straw mushrooms by adopting L-lysine or L-threonine, which has the following advantages:
(1) The straw mushroom treated by the method can effectively delay the rotting, running and sinking of the mushroom body under a low-temperature anti-freezing experiment, the fruiting body is hard and solid, and the good commodity appearance is maintained;
(2) The method for soaking by adopting L-lysine or L-threonine has simple operation and low treatment cost, and can prolong the low-temperature fresh-keeping of the straw mushroom to at least 2 days;
(3) The method lays a foundation for further developing a low-temperature-resistant preservation technology of functional nutritional straw mushrooms rich in human essential amino acid L-lysine or L-threonine.
Drawings
FIG. 1 Multi-component integrated analysis of L345-0044 spray-applied Volvariella volvacea fruiting bodies under cold pressing treatment. (A) The L345-0044 spray-applied straw mushroom fruiting body is preserved at 4 deg.C for 0h (P0 h), 24h (P24 h), 48h (P48 h). The inhibitor compound L345-0044 of UBEV2 is utilized to carry out spraying effect in the growth and development period of the button of the straw mushroom, and the specific content is shown in the patent number: 202110124029.0 ". (B) a flow chart of multi-group chemical analysis of straw mushroom fruiting bodies. (C) GST-pulldown results of GST-SSB2 (VVO _ 07431) and cold-pressed 4h of the total protein of Volvariella volvacea were analyzed by SDS-PAGE. (D) STRING interaction network analysis of NIP 1. (E) analysis of the heat map of protein translation. (F) KEGG enrichment analysis of differentially expressed genes in P0_ vs _ P24. (G) KEGG enrichment analysis of differentially expressed genes in P0_ vs _ P48.
FIG. 2 non-targeted metabolome analysis of L345-0044 spray-acted Volvariella volvacea fruiting bodies under cold pressure treatment. (A) VENN analysis of significantly different metabolites. (B) Heat map analysis of significantly different metabolites overlapping in P0_ vs _ P24 and P0_ vs _ P48. (C) Pathway enrichment analysis of overlapping significantly different metabolites in P0_ vs _ P24 and P0_ vs _ P48. (D) Correlation thermogram analysis of overlapping significantly different metabolites in P0_ vs _ P24 and P0_ vs _ P48.
FIG. 3 Combined metabolome and transcriptome analysis of L345-0044 spray-acted Volvariella volvacea under cold pressing treatment. (A) Thermographic analysis of lysine biosynthesis (KEGG module: M00030) and lysine degradation (KEGG module: M00032) under cold pressure treatment. (B) Thermographic analysis of the first carbon oxidation (KEGG module: M00010) and the second carbon oxidation (KEGG module: M00011) in citric acid cycles under cold pressing treatment. (C) Lysine mediated acetyl-coa synthesis cycles including M00010, M00030 and M00032. (D) Verification of the antifreeze phenotype of the transgenic yeast (pYES 2-NTB-VVO _ 02032). pYES2-NTB transformed yeast was used as a control group. For details of the method, see "patent No.: 202211030798.5 ".
FIG. 4 shows the antifreeze test of straw mushroom. (A) Straw mushrooms soaked in L-lysine 200. Mu.M or L-threonine 300. Mu.M solutions were stored at 4 ℃ for 24 hours and 48 hours. (B) the water-soaked Volvariella volvacea is stored at 4 ℃ for 24 hours and 48 hours. (C) The water, L-lysine and L-threonine soaked volvariella volvacea was stored at 4 ℃ for 24 hours. Lys represents lysine. Tyr represents L-threonine.
Detailed Description
Example 1
A method for low-temperature-resistant preservation of straw mushrooms based on L-lysine comprises the following steps:
(1) L345-0044 spray-acted straw mushroom fruiting bodies treated by cold pressing are collected (figure 1A), and absolute quantitative transcriptomics and non-target metabolome combined analysis is carried out to excavate key target pathways and metabolites (figure 1B). CO-IP experiments confirmed that SSB2 (VVV07431) is an interacting protein of UBEV2 (DOI: 10.1016/j. Jprot.2020.103668). SSB2 is a ribosome-associated chaperone involved in folding newly synthesized polypeptide chains (DOI: 10.1016/j. Cell.2017.06.038). GST-pulldown experiment found eukaryotic translation initiation factor 3 subunit C (NIP 1) to be an interacting protein of SSB2 (FIG. 1C). The STRING enrichment assay further confirmed that the NIP1 interacting protein was mainly a protein translation initiation factor gene (fig. 1D). Overexpression of genes involved in translation initiation indicated that protein translation was enhanced in the L345-0044 spray-applied Volvariella volvacea fruiting body during cold stress (FIG. 1E). The differential expression genes obtained by transcriptomics are adopted to carry out KEGG enrichment analysis, and the result shows that the activity of a protein processing in the endoplasmic reticulum pathway of P0_ vs _ P24 and P0_ vs _ P48 is enhanced (figures 1F and 1G), so that the enhancement of protein processing activities such as protein synthesis, protein degradation, protein operation, amino acid metabolism and the like is helpful for the anti-freezing effect of the straw mushroom.
(2) The VENN analysis using the significantly different metabolites between P0_24 and P0_48 yielded 30 overlapping significantly different metabolites (P value. Ltoreq.0.05, VIP. Ltoreq.1) (FIG. 2A). Most of the 30 significantly different metabolites were up-regulated in expression, including L-glutamine, L-lysine, D-glucose-1, 5-lactone (D-Glucono-1, 5-lactone) (FIG. 2B). Pathway enrichment analysis of 30 significantly different metabolites yielded enriched pathways (P < 0.05), including lysine biosynthesis (lysine biosynthesis), glycine, serine, threonine metabolism (Glycine, serine and threonine metabolism) and Aminoacyl-tRNA biosynthesis (Aminoacyl-tRNA biosynthesis) (fig. 2C). Our recent studies demonstrated that the additive D-glucose-1, 5-lactone can improve the cryopreservation quality of volvariella volvacea (DOI: 10.1016/j. Postharvbio.2021.111784). The overlapping significantly different metabolite correlation heatmap results indicate that both L-lysine and D-glucose-1, 5-lactone have strong correlations (fig. 2D), suggesting that lysine has a positive role in cold stress tolerance of volvariella volvacea.
(3) Heatmap analysis showed that lysine biosynthesis (KEGG module: M00030) and lysine degradation (KEGG module: M00032) were upregulated during cold pressing treatment in L345-0044 spray samples (fig. 3A). The expression profile shows a reduced activity of the first carbon oxidation in the citric acid cycle (KEGG module: M00010) (FIG. 3B). M00030 (lysine biosynthesis) was linked to M00032 (lysine degradation) providing that acetyl coa participates in M00010 (first carbon oxidation) (fig. 3C). Yeast amino acid dehydrogenase is responsible for the biosynthesis and catabolism of lysine (DOI: 10.3389/fpls.2020.00587 and DOI: 10.1002/jobm.201900189). Yeast antifreeze phenotype experiments demonstrated that Saccharomyces cerevisiae amino acid dehydrogenase (VVO _ 02032) is a cold tolerance gene (FIG. 3D; patent No.: 202211030798.5), suggesting that enhanced lysine metabolism is critical to cold stress resistance. Enhanced lysine biosynthesis and catabolism linked to M00010, forming a lysine-mediated acetyl-coa synthesis cycle to maintain normal functioning of the citrate cycle (fig. 3C).
(4) The above results indicate that the acquisition of cold resistance by volvariella volvacea can benefit from the accumulation of lysine and its lysine-mediated enhancement of the acetyl-coa synthesis cycle. Therefore, the research on whether the cold resistance of the straw mushroom can be effectively improved by adding exogenous lysine is carried out. Selecting straw mushrooms V23 harvested in the same batch, which are similar in size, free of parachute opening and obvious in damage;
(5) 300ml of L-lysine solution with the concentration of 200 mu M is prepared;
(6) Soaking the straw mushrooms in the L-lysine solution for 2 minutes, and then draining;
(7) Directly putting the treated straw mushrooms at 4 ℃ for a cold storage test for 0,24,48 hours;
(8) The shape, color and hardness of the L-lysine solution infiltrated straw mushroom fruiting body under cold treatment are evaluated.
As can be seen from FIG. 4A, the straw mushroom fruiting body soaked by the L-lysine solution is full in shape, bright in color and hard and solid in hardness under cold treatment (2-48 hours), and is significantly superior to the control treatment state in all aspects. Has better sensory evaluation effect, and the frost resistance of the straw mushroom is obviously improved. The processed straw mushroom can be preserved for at least 2 days.
Comparative example 1
(1) Selecting straw mushrooms which are similar in size, have no parachute opening and have no obvious damage from the straw mushrooms harvested in the same batch as that in the example 1;
(2) 300ml of a control aqueous solution was used;
(3) Soaking the straw mushrooms in a control aqueous solution for 2 minutes, and then draining;
(4) Directly putting the treated straw mushrooms at 4 ℃ for a cold storage test for 0,24,48 hours;
(5) The shape, color and hardness of the straw mushroom fruiting body under cold treatment were evaluated.
Comparative example 2
(1) L-threonine is metabolized to produce acetyl CoA (DOI: 10.1042/bj1560449 and DOI: 10.1126/science.1173288), which in turn is involved in the citric acid cycle. KEGG enrichment analysis showed up-regulation of Glycine, serine and threonine metabolism (Glycine and threonine metabolism) pathways in P0_ vs _ P24 (fig. 1F), indicating enhanced threonine metabolic activity under cold press treatment. Therefore, we studied whether exogenous addition of threonine can also maintain the activity of first carbon oxidation in the citric acid cycle by producing acetyl CoA, provide energy supply, and effectively improve the cold tolerance of volvariella volvacea;
(2) Selecting straw mushrooms V23 harvested in the same batch, which are similar in size, free of umbrella opening and obvious in damage;
(3) 300ml of L-threonine solution with the concentration of 300 MuM is prepared;
(4) Soaking the straw mushrooms in a control L-threonine solution for 2 minutes, and then draining;
(5) Directly putting the harvested straw mushrooms at 4 ℃ for a refrigeration test for 0,24,48 hours;
(6) The shape, color and hardness of the straw mushroom fruiting body under cold treatment were evaluated.
The results show that the straw mushroom fruiting body soaked by the L-lysine solution treated by the method of example 1 is plump in shape, bright in color and hard in hardness under cold treatment (figure 4A), and is remarkably superior to the state treated by the comparative example 1 in all aspects (figure 4B). The result of fig. 4 shows that the color of the cross-sectional view of the L-threonine solution fruiting body treated in the comparative example 2 still keeps certain brightness under cold treatment (2-48 hours), and the brightness degree is equivalent to the state of the L-lysine soaked straw mushroom fruiting body treated by the method in the example 1, and is obviously better than the state of the straw mushroom fruiting body treated in the comparative example 1. The results in FIG. 4 further confirm that the antifreeze property of the straw mushroom can be remarkably improved by adding L-lysine or L-threonine to stimulate the synthesis of acetyl CoA and further participate in the citric acid cycle.
Various other modifications and changes may be made by those skilled in the art based on the above-described technical solutions and concepts, and all such modifications and changes should fall within the scope of the claims of the present invention.
Claims (7)
- The use of L-lysine or L-threonine as a low-temperature food preservative.
- 2. Use according to claim 1, characterized in that: the L-lysine or L-threonine is an essential amino acid for human body.
- 3. The method for preserving volvariella volvacea at a low temperature according to claim 2, wherein the concentration of the solution of the essential amino acid L-lysine or L-threonine in the human body is 200 to 300 μ M respectively.
- 4. Use according to claim 1, characterized in that: the food is straw mushroom.
- 5. A low-temperature preservation method of straw mushrooms is characterized by comprising the following steps: soaking the straw mushroom in a solution of L-lysine or L-threonine for 2 to 4 minutes, draining water, and refrigerating at the refrigerating temperature of 0 to 8 ℃.
- 6. The method for keeping volvariella volvacea fresh at a low temperature as claimed in claim 4, wherein said L-lysine or L-threonine is an essential amino acid for human body.
- 7. The low-temperature refreshing method for straw mushrooms according to claim 4, characterized in that the concentration of the L-lysine solution is 200 to 300 μ M; the concentration of the L-threonine solution is 200 to 300 mu M.
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