CN108094519B - Grape antagonistic yeast composite biological preservative and preparation method and application thereof - Google Patents

Grape antagonistic yeast composite biological preservative and preparation method and application thereof Download PDF

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CN108094519B
CN108094519B CN201810043121.2A CN201810043121A CN108094519B CN 108094519 B CN108094519 B CN 108094519B CN 201810043121 A CN201810043121 A CN 201810043121A CN 108094519 B CN108094519 B CN 108094519B
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grape
grapes
candida
antagonistic
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CN108094519A (en
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杨相政
贾连文
魏雯雯
王达
孙斐
郁网庆
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JINAN INSTITUTE OF FRUIT PRODUCTS CHINA COOP
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    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23BPRESERVING, e.g. BY CANNING, MEAT, FISH, EGGS, FRUIT, VEGETABLES, EDIBLE SEEDS; CHEMICAL RIPENING OF FRUIT OR VEGETABLES; THE PRESERVED, RIPENED, OR CANNED PRODUCTS
    • A23B7/00Preservation or chemical ripening of fruit or vegetables
    • A23B7/14Preserving or ripening with chemicals not covered by groups A23B7/08 or A23B7/10
    • A23B7/153Preserving or ripening with chemicals not covered by groups A23B7/08 or A23B7/10 in the form of liquids or solids
    • A23B7/154Organic compounds; Microorganisms; Enzymes
    • A23B7/155Microorganisms; Enzymes; Antibiotics
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23BPRESERVING, e.g. BY CANNING, MEAT, FISH, EGGS, FRUIT, VEGETABLES, EDIBLE SEEDS; CHEMICAL RIPENING OF FRUIT OR VEGETABLES; THE PRESERVED, RIPENED, OR CANNED PRODUCTS
    • A23B7/00Preservation or chemical ripening of fruit or vegetables
    • A23B7/14Preserving or ripening with chemicals not covered by groups A23B7/08 or A23B7/10
    • A23B7/144Preserving or ripening with chemicals not covered by groups A23B7/08 or A23B7/10 in the form of gases, e.g. fumigation; Compositions or apparatus therefor
    • A23B7/152Preserving or ripening with chemicals not covered by groups A23B7/08 or A23B7/10 in the form of gases, e.g. fumigation; Compositions or apparatus therefor in a controlled atmosphere comprising other gases in addition to CO2, N2, O2 or H2O ; Elimination of such other gases
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23BPRESERVING, e.g. BY CANNING, MEAT, FISH, EGGS, FRUIT, VEGETABLES, EDIBLE SEEDS; CHEMICAL RIPENING OF FRUIT OR VEGETABLES; THE PRESERVED, RIPENED, OR CANNED PRODUCTS
    • A23B7/00Preservation or chemical ripening of fruit or vegetables
    • A23B7/14Preserving or ripening with chemicals not covered by groups A23B7/08 or A23B7/10
    • A23B7/153Preserving or ripening with chemicals not covered by groups A23B7/08 or A23B7/10 in the form of liquids or solids
    • A23B7/154Organic compounds; Microorganisms; Enzymes

Abstract

The invention provides a grape antagonistic yeast composite biological preservative and a preparation method and application thereof. Every 100mL of grape antagonistic yeast composite biological fresh-keeping agent contains 1 × 108‑10Candida CFU (Candida oleophila), 1X108‑10CFU (Candida sake), 0.3g-0.8g of burdock fructo-oligosaccharide (BFO) and 0.8mL-1.0mL of hydrogen. The invention applies the two grape antagonistic yeasts to fruit preservation for the first time, and the invention is a novel grape antagonistic yeast composite biological preservative with obvious preservation effect. And the used raw materials are green components, so that the Chinese medicinal composition has no toxic or side effect on a human body and has no pollution to the environment. The compound preservative is simple to use and easy to popularize.

Description

Grape antagonistic yeast composite biological preservative and preparation method and application thereof
Technical Field
The invention belongs to the technical field of fruit preservation, and particularly relates to a grape antagonistic yeast composite biological preservative as well as a preparation method and application thereof.
Background
The grapes are an ancient fruit variety, are native to Europe and Western Asia, and have been planted for over one thousand years in China. The grape is a fruit with delicious taste and high nutritive value, and the glucose content of the grape is up to 10 to 30 percent. The grape also contains various kinds of fruit acids which are helpful for digestion, and has the efficacy of strengthening the spleen and stomach. Therefore, the grapes are also ideal fruits for people with weak digestion ability. In addition, grapes are also rich in minerals and vitamins, such as calcium, potassium, phosphorus, iron, vitamin B1, vitamin B2, vitamin B6, vitamin C, vitamin P, and the like. Chinese medical science considers that the grapes are sweet in taste and neutral in nature, and have the effects of tonifying liver and kidney, replenishing qi and blood, stimulating appetite, promoting the production of body fluid and promoting urination. The grape is eaten frequently to relieve fatigue and alleviate symptoms such as neurasthenia. The raisin prepared from the grapes is a good tonic product for pregnant women, children and people with physical weakness and anemia.
The demand of the modern society for fresh grapes is increasing day by day, and after the fresh grapes are harvested, cells and tissues of the fresh grapes continue to breathe and transpire, so that the phenomena of shrinkage, weight loss, wilting, deterioration and the like are easily caused. Moreover, grape fruits have thin skins and high sugar acid content in the pulps, and are easy to break after picking to cause bacterial infection, so that the fruits are rotten. Therefore, the preservation of grapes is a very important problem in the production, storage and sale of grapes. How to control postharvest diseases and maintain fruit quality is a problem in the field of storage and preservation of grapes.
At present, the used fruit and vegetable fresh-keeping technology mainly comprises a low-temperature fresh-keeping method, a modified atmosphere fresh-keeping method, a low-pressure fresh-keeping method, a plastic film, a chemical fresh-keeping storage method and the like. The methods can keep the freshness of the fruits and vegetables to a certain extent, but have certain defects, such as large investment of low-temperature fresh-keeping and air-conditioning fresh-keeping methods and difficult popularization and application; the plastic film can inhibit the water loss of fruits and vegetables, but is easy to cause rottenness and peculiar smell; in the chemical fresh-keeping method (such as sulfite grape fresh-keeping agent), a large amount of organic bactericides and preservatives remain on the surfaces of fruits and vegetables and are toxic to human bodies. Therefore, the development of a grape fresh-keeping method which is environment-friendly, pollution-free, free of toxic and side effects on human bodies, simple in method, low in cost and easy to popularize and apply still remains a technical problem to be solved by technical personnel in the field.
Disclosure of Invention
Aiming at the defects of the prior art, the invention provides the grape antagonistic yeast composite biological preservative and the preparation method and the application thereof. The method is environment-friendly, pollution-free, free of toxic and side effects on human bodies, simple, low in cost and easy to popularize and apply.
In order to achieve the purpose, the invention adopts the technical scheme that: two screened grape antagonistic yeasts with the preservation effect are compounded with burdock fructo-oligosaccharide and hydrogen to form the composite preservative.
The grape antagonistic yeast composite biological preservative contains 1 multiplied by 10 per 100mL of grape antagonistic yeast composite biological preservative8-10CFU Candida Oligophila (Candida oleophila), 1 × 108-10CFU (Candida sake), 0.3g-0.8g of burdock fructo-oligosaccharide (BFO) and 0.8mL-1.0mL of hydrogen.
The preparation method of the grape antagonistic yeast composite biological preservative comprises the following steps: (1) fermenting and culturing Candida olivaceus (Candida oleophila) and Candida sake (Candida sake), and concentrating the fermentation liquid.
(2) Freeze drying burdock fructo-oligosaccharide and concentrated fermentation liquid together to make into powder or tablet.
(3) And preparing the hydrogen generation package.
And (2) concentrating the fermentation liquor obtained in the step (1) until the dry weight of the thallus is more than or equal to 200 g/L.
When in use, the hydrogen generating bag is dissolved in water to release hydrogen, and then the prepared powder or tablet is dissolved in the hydrogen generating bag (the final concentration of each component in the solution is as described above) and sprayed on grapes to be preserved.
The spraying amount is 8-10mL per kilogram of grapes and other fruits.
The invention also provides application of Candida utilis (Candida oleophila) and Candida sake (Candida sake) in preservation of grapes and other fruits.
The composite preservative is not only suitable for preserving grapes, but also suitable for preserving other fruits.
The grapes contain rich nutrient substances, the grape skin is thin and juicy, and the phenomena of rotting, dry branches, browning, threshing and the like are easy to occur in the storage process. The adoption of antagonistic microorganisms for controlling postharvest diseases of fruits and vegetables is a potential emerging technology. The saccharomycete has high antagonistic effect, produces no toxin and may be used together with chemical germicide to form the hot bacterial strain for biological disease preventing and controlling research. Different degrees of influence on the storage quality and antioxidant enzymes of grapes were investigated by treating grapes with pichia guilliermondii and hansenula polymorpha isolated and selected from strawberries. The candida utilis and candida sake are obtained by screening from the surface of the grape, and no relevant report exists when the candida utilis and the candida sake are applied to grape preservation.
Burdock fructo-oligosaccharide (BFO) is a water-soluble inulin-type oligosaccharide extracted from Burdock root, and is composed of furan-type fructose and furan-type glucose. At present, BFO activity researches find that the compound has wide physiological activity, can be used as an immunostimulant to improve the immunity of animals, improve the content of a plurality of immune factors and resist tumors; can also induce plants to generate systemic resistance, resist various diseases and increase the fruit yield. In the existing research, BFO is used for processing fruits at normal temperature, and the results show that BFO can activate the disease resistance of grapes to botrytis cinerea, apples to penicilliosis, bananas to anthracnose, kiwi fruits to botrytis cinerea, oranges to penicilliosis, strawberries and Chinese gooseberries to naturally occurring pathogens. The influence of BFO on the physiological activity of picked grapes is embodied in that the activities of CAT and SOD enzymes are improved, and the content of MDA is reduced. BFO is used as an effective inducer and can be used as a natural fruit and vegetable preservative to be applied to fruits.
It is also reported that the research group of life environment academy of sciences of university college of osaka university, japan found: the vegetables and fruits soaked in hydrogen-rich water have low temperature damage resistance. Fruits or vegetables which are easy to lose moisture and oxidize during cold storage of bananas, eggplants and the like are soaked in hydrogen-rich water for 10 minutes, naturally aired for 1 hour and then stored for 2 weeks in a cold storage mode. As a result, the deterioration of the pulp quality such as softening, discoloration and denting is reduced as compared with the pure water-soaked pulp. However, the same effect does not occur when the hydrogen-rich water is simply sprayed to the fruits or vegetables, so it is presumed that the natural airing process after the soaking has an effect.
The invention compounds the screened two grape antagonistic yeasts with the burdock fructo-oligosaccharide and the hydrogen-rich water for the first time to obtain the high-efficiency composite biological preservative. The grape antagonistic yeast composite biological preservative has the following beneficial effects:
(1) the invention relates to a novel grape antagonistic yeast composite biological preservative, which adopts green components as raw materials, has no toxic or side effect on human bodies, and has no pollution to the environment.
(2) The composite preservative has obvious effect on grape preservation. Can obviously maintain the quality of the grapes in the storage period and the shelf life and prolong the shelf life of the grapes. In addition, the grapes are inevitably damaged in the picking and transporting processes, and the composite preservative has a particularly remarkable effect on the preservation of the grapes with mechanical damage. The grape antagonistic yeast, BFO and the like can promote wound healing and can inhibit rapid deterioration induced by mechanical injury. Compared with other antistaling agents, the invention can better maintain the quality of the grapes.
(3) The compound preservative is simple to use and easy to popularize. The method is particularly suitable for the shipment of fruits and vegetables of farmhouses, and the fresh keeping of fruits and vegetables in logistics and stores or families.
Drawings
FIG. 1 is a graph showing the growth of 5 yeasts selected in experiment one.
FIG. 2 is a graph showing a comparison of the effect of 4 antagonistic yeasts on a stabbed grape stored for 10 days.
FIG. 3 is a graph showing a comparison of the effect of 4 antagonistic yeasts on whole grape bunch after storage for 10 days.
FIG. 4 is a graph showing a comparison of the effect of 4 antagonistic yeasts on the decay rate and the degranulation rate of grapes.
FIG. 5 shows the comparison of the fresh-keeping effect of the compound fresh-keeping agent, burdock fructo-oligosaccharide and hydrogen-rich water on grapes in experiment II.
FIG. 6 shows the effect of the three-compound antistaling agent experiment on the content of ascorbic acid.
FIG. 7 shows the effect of the compound preservative of the three experiments on the activity of CAT enzyme.
FIG. 8 shows the effect of the compound antistaling agent of experiment three on SOD enzyme activity.
Detailed Description
The technical solution and the technical effects thereof are further illustrated by the following specific examples, experiments and drawings, and the following description is only for explaining the present invention but not limiting the present invention in any way, and any changes or substitutions based on the present invention shall fall within the protection scope of the present invention. The methods of the present invention are conventional in the art unless otherwise specified.
Experiment I, screening and identification of grape antagonistic yeast and research on antibacterial effect
1.1 antagonistic Yeast isolation
The inventor collects Crimson seedless grapes (Crimson grapes) from vineyards of south Villa of Ricey, Qingdao and used for separating and screening antagonistic yeasts. Cutting grape epidermis into 1cm × 1cm pieces, adding into 90ml triangular flask of PDB liquid culture medium, dripping several drops of 0.1% streptomycin sulfate, shake culturing in constant temperature shaking table (25 deg.C, 180rpm) for 30 min, and performing gradient dilution (10)-5,10-6,10-7) Spreading 100 μ L of the extract on PDA plates, culturing at 25 deg.C for 2-3d, primarily selecting colony with similar morphology, combining and counting, and purifying and storing after microscopic examination to confirm as yeast.
Selecting single colonies with different culture characteristics, combining and counting similar colonies by an observation method, primarily screening 5 yeast strains which are relatively dominant in the fruit epidermal micro-ecological environment, measuring the light absorption value of the yeast to determine the growth curve (figure 1) of the yeast at 25 ℃ in 5 of GY1, GY7, GY15, GY18 and GY19, wherein the fastest growth speed is GY15 and GY7, the next GY19 and the slower growth speed is GY1 and GY18 according to the time of entering a stabilization period. Because GY18 has slow growth speed and has peculiar smell after fermentation, four yeasts with the numbers of GY1, GY7, GY15 and GY19 are selected to carry out antagonistic experimental study on the preservation of grapes.
1.2 antagonistic yeast identification of grape
In the China general microbiological culture Collection center, the strain identification is carried out on GY1, GY7, GY15 and GY19, the strain identification results are shown in Table 1, four kinds of yeast are registered in CGMCC, but the antagonistic fresh-keeping effect in grapes is the first research.
TABLE 1 Yeast identification Classification
Figure GDA0002980416610000051
1.3 antagonistic yeast bacteria inhibition effect research of grape
(1) Antagonistic yeast for fresh-keeping effect of stabbing grapes
After the grapes of Kerissen are picked and are pre-cooled in a cold storage for 48 hours, the equator position of the fruits is stabbed by aseptic toothpicks, 1 wound is formed on each fruit, and the depth of the wound is 3 mm. Spray-treating grape with yeast suspension in physiological saline, all yeast concentrations used in the test were diluted to 1 × 108one/mL, control group was normal saline. After the spray treatment, the grapes were placed in a plastic basket packaging bag with a humidity of 85%, and the grapes were observed for deterioration at room temperature (20 ℃).
As shown in FIG. 2, after the treated grapes were stored at room temperature for 10 days, a large amount of tissue fluid exuded from the wound of the control grapes, the surface of the entire basket of grapes was wet and sticky, and the fruits began to rot. The wounds of the grapes treated by GY1 and GY7 exude more interstitial fluid, and the grapes are generally seen to have fruit cracking and rot more seriously than CK. The grape wound treated by GY15 and GY19 and the periphery thereof are dry, have healing-like characteristics, and have clean whole grape surface, no wet and sticky tissue fluid and no rotten phenomenon. Therefore, the GY15 and GY19 yeasts have obvious fresh-keeping effect on grapes due to mechanical injury, and the main mechanisms of the yeasts can consume nutrient components at wounds, promote wound healing and inhibit rapid deterioration induced by mechanical injury.
(2) Antagonistic yeast fresh-keeping effect on whole bunch of grapes
After the grapes of Kerisen are picked and precooled in a refrigeration house for 48 hours, the whole bunch of grapes with similar weight is used for experiments. Spray-treating grape with yeast liquid suspended in physiological saline, and collecting yeast concentration for all testsDilution of 1X108one/mL, control group was normal saline. After the spray treatment, the grapes were placed in a plastic basket packaging bag with a humidity of 85%, and the grapes were observed for deterioration at room temperature (20 ℃).
As shown in fig. 3, when the treated grapes are stored at room temperature for 10 days, the preservation effect of the yeasts on the grapes is very remarkable. The fruit stalks of the control group have serious browning, the browning is already spread to the joints of the grapes, the grapes have serious browning and shrinking phenomena, and the threshing is more obvious. The main grape stalks processed by GY15 and GY19 are partially browned, the parts of the main grape stalks keep good emerald green color, the freshness of the grape stalks is kept well, the grape fruits are less browned, full without shrinkage, less threshed and remarkable in preservation effect. This may be associated with antagonism of yeast on nutrient consumption of fruit stalks, stalks and threshing wounds, promotion of wound healing, inhibition of rapid deterioration induced by mechanical damage, thereby showing a preservation effect on whole grapes.
(3) Antagonistic influence of yeast on rot rate and threshing rate of grapes
As shown in figure 4, after the treated grapes are stored for 10 days at room temperature, the fruit removing rate of the whole bunch of grapes can be reduced remarkably by GY15, GY19, GY1 and GY7, the rotting rate of the punctured grapes is reduced remarkably by GY15 and GY19, the reduction range is 50%, and GY1 and GY7 cannot reduce the rotting rate of the punctured grapes but increase the rotting rate of the punctured grapes. Therefore, the GY15 and GY19 yeasts have remarkable effects on inhibiting the rot and degranulation of grapes.
Through the experiments, the grape fresh-keeping effect of the two strains GY15(Candida oleophila) and GY19(Candida lake) is obvious, and the grape rot and degranulation can be effectively inhibited. Thus, the present invention ultimately identifies two antagonistic yeasts: candida olivaceus (Candida oleophila) and Candida sake (Candida sake) are used for preparing the grape antagonistic yeast composite biological preservative.
Experiment II, research on synergistic effect of the antagonistic saccharomycete composite preservative on grape preservation
The existing research shows that the burdock fructo-oligosaccharide and hydrogen-rich water also have a certain fresh-keeping effect, but the effect is still to be improved. Due to the fact thatThe invention further researches and prepares a composite preservative by compounding the two screened antagonistic yeasts with the burdock fructo-oligosaccharide and the hydrogen-rich water for grape preservation, and the preservation effect can be expected to be greatly improved. In the experiment, the grape antagonistic yeast composite biological preservative (composite preservative for short) is used, and every 100mL of the composite biological preservative contains 1 × 108CFU Candida Oligophila (Candida oleophila), 1 × 108CFU sake Candida (Candida sake), 0.3g Burdock fructo-oligosaccharide (BFO) and 0.8mL hydrogen.
Selecting Kerison grapes which are free from diseases, insect pests, mildew and rot, mechanical damage and have full fruit grains and neat and compact spike shapes, pre-cooling the picked Kerison grapes in a cold storage for 48 hours, and respectively using a compound preservative (TR1) and 1x108CFU/mL Candida Olivarum (Candida oleophila) and 1X108CFU/mL Candida sake (TR2), 50% saturated hydrogen-rich water (TR3), 0.5% (0.5g/100mL) Burdock fructo-oligosaccharide BFO (TR4) spray-treated grapes, and the control group was physiological saline (CK).
After the spray treatment, the grapes were placed in a plastic basket packaging bag with a humidity of 85%, and the grapes were observed for deterioration at room temperature (25 ℃).
2.1 research on synergistic effect of the composite antistaling agent on grape
As a result: as shown in FIG. 5, after 10 days of storage at room temperature, the grapes in the control group (CK) begin to mildew, soften and rot, and the grapes lose water and rot seriously, while the grapes treated by the compound preservative (TR1) have cleaner surfaces, no wet and sticky tissue fluid and almost no rot. Grapes processed by TR2, TR3 and TR4 by 0.5% of burdock fructo-oligosaccharide BFO processed by two yeasts can inhibit grape rot to different degrees, but the preservation effect is far less obvious than that of TR1 processed by the compound preservative.
As can be seen from Table 2, after the grapes are stored for 10 days at room temperature, the rotting rate of the grapes treated by the compound preservative is only 1.7%, while the rotting rates of TR2, TR3 and TR4 are 5.2%, 6.3% and 7.5% respectively, while the rotting rate of the grapes treated by the compound preservative is high and reaches 47.2% compared with that of the grapes treated by the control treatment.
TABLE 2 Effect of antagonistic yeast composite antistaling agent on grape rotting rate
Figure GDA0002980416610000071
And (4) conclusion: the compound preservative has obvious effect on preserving grapes at room temperature in shelf life. Compared with other components which are used independently, the composite microbial agent has lower rotting rate and better preservation, and shows that the composite microbial agent has a synergistic effect after two kinds of saccharomycetes, hydrogen-rich burdock fructooligosaccharide and the like are compounded.
2.2 Effect of the composite antistaling agent of the invention on pathogenic microorganisms of grapes
According to the national food safety standard GB 29921-2013 pathogenic bacteria limit in food, the postharvest pathogenic bacteria of the grapes are detected. The results are shown in Table 3: for stabbed grapes, the escherichia coli exceeded national safety standards on day 4 without treatment with a preservative, while staphylococcus aureus began to exceed national standards on day 7. The Escherichia coli on the fourth day and the Staphylococcus aureus on the 7 th day of the group treated by the preservative are controlled within the national standard. Therefore, the protection effect of the compound preservative on the grapes with the stabbing wound is supposed, and the inhibition effect of the rot is probably related to the control of the proliferation of the bacterial microorganisms.
TABLE 3 Effect of the composite antistaling agent of the invention on grape pathogenic bacteria
Figure GDA0002980416610000072
Figure GDA0002980416610000081
And judging the bacteriostatic effect by referring to national food safety standard 'pathogenic bacteria limit in food' GB 29921-2013.
Experiment III, the research of the mechanism of the composite antistaling agent for grape fresh keeping
3.1 Effect of the composite antistaling agent of the invention on the content of malondialdehyde in grape
In order to further research the preservation mechanism of the compound preservative, the inventor researches the relationship between the content of the grape malondialdehyde directly related to the wound degree and the use of the compound preservative.
As shown in Table 4, the results show that the compound preservative (same as the formula in the second experiment) has a very significant inhibiting effect on the increase of the MDA content of the stabbed grapes. The expression is as follows: on the fourth day after treatment, the MDA content of the compound preservative treatment group is 13 times that of the control group, and on the third day is 10 times. Therefore, the target of the composite preservative on the grape preservation effect is supposed to be possible to control the wound.
TABLE 4 Effect of the composite antistaling agent of the invention on grape MDA
Figure GDA0002980416610000082
3.2 Effect of the composite antistaling agent of the invention on the antioxidant system of the grape
The 3.1 research shows that the inhibition effect of the compound preservative on MDA of grapes is probably related to the integrity of grape cell protection. And an antioxidant system consisting of ascorbic acid, CAT, SOD and the like is beneficial to removing free radicals and protecting the integrity of cell membranes, so that the MDA content is reduced, and the local necrosis caused by wounds is inhibited.
Therefore, the invention further researches the relationship between the compound preservative and the content of ascorbic acid, CAT and SOD. The test is divided into two groups, one group is stabbed grapes, the control group is treated by normal saline to be CK1, and the test group is treated by the compound preservative disclosed by the invention to be TR 1. One group is grapes without mechanical damage, a control group is CK2 treated by normal saline, and a test group is TR2 treated by the compound preservative of the invention.
After treatment, the grapes are placed in a plastic basket packaging bag with the humidity of 85 percent and are placed at room temperature (20 ℃) for detecting the content of ascorbic acid, CAT and SOD in 0 day, 1 day, 4 days, 7 days and 10 days respectively. The results are shown in FIGS. 6 to 8.
Fig. 6 shows that the composite antistaling agent can improve the ascorbic acid content of grapes in stabbing groups and non-stabbing groups, and the improvement effect in the stabbing groups is obviously better than that in the non-stabbing groups. It is speculated that the increase in ascorbic acid is related to the protection of ascorbic acid by the reduction of hydrogen molecules on the one hand, and on the other hand, BFO and yeast can both act as inducers to increase ascorbic acid synthesis. The ascorbic acid content of the stabbed fruits is higher than that of the non-stabbed fruits, probably because the yeast of the stabbed fruits obtains more nutrition, and the induction effect of the yeast growth on the stabbed fruits is more obviously related.
As shown in 7 and 8, the compound preservative has obvious inhibiting effect on the reduction of enzyme activity of CAT and SOD stabbing fruits. This may cause the compound preservative to inhibit the MDA content of the stabbed grapes and protect the whole fruits. The composite preservative has little influence on the enzyme activities of CAT and SOD of non-stabbed fruits, mainly because the enzyme activities of CAT and SOD of a non-stabbed fruit control group are not related to the obvious change of the enzyme activities in the early stage after grape harvest. According to the results, the protection effect of the compound preservative on the stabbed grapes is presumed to have a significant relation with the reduction of the inhibition of the enzyme activity of CAT and SOD.
Example 1 grape antagonistic yeast composite biological preservative, every 100mL grape antagonistic yeast composite biological preservative contains 1 × 108CFU Candida Oligophila (Candida oleophila), 1 × 108CFU sake Candida (Candida sake), 0.3g Burdock fructo-oligosaccharide (BFO) and 0.8mL hydrogen.
The preparation method comprises the following steps: (1) fermenting and culturing Candida olivaceus (Candida oleophila) and Candida sake (Candida sake), wherein the fermentation method is conventional in the field. Concentrating the fermentation liquor until the dry weight of the thallus is more than or equal to 200 g/L.
(2) Freeze drying burdock fructo-oligosaccharide and concentrated fermentation liquor, and tabletting.
(3) And preparing the hydrogen generation package.
Example 2 grape antagonistic yeast composite biological preservative, every 100mL grape antagonistic yeast composite biological preservative contains 1 × 1010CFU Candida Oligophila (Candida oleophila), 1 × 1010CFU sake Candida (Candida sake), 0.8g Burdock fructo-oligosaccharide (BFO) and 1.0mL hydrogen.
The preparation method is the same as example 1.
Example 3A grape antagonistic yeast composite biological antistaling agent, every 100mL grape antagonistic yeast composite biological antistaling agent contains 1 × 109CFU Candida Oligophila (Candida oleophila), 1 × 109CFU sake Candida (Candida sake), 0.5g Burdock fructo-oligosaccharide (BFO) and 0.9mL hydrogen.
The preparation method is the same as example 1.
Example 4A grape antagonistic yeast composite biological antistaling agent, every 100mL grape antagonistic yeast composite biological antistaling agent contains 1 × 108CFU Candida Oligophila (Candida oleophila), 1 × 109CFU sake Candida (Candida sake), 0.4g Burdock fructo-oligosaccharide (BFO) and 0.8mL hydrogen.
The preparation method is the same as example 1.

Claims (2)

1. The application of the grape antagonistic yeast composite biological preservative in fruit preservation is characterized in that the fruits are grapes;
every 100mL of grape antagonistic yeast composite biological fresh-keeping agent contains 1 × 108-10CFU Candida Oligocarpa, 1 × 108 -10CFU sake candida, 0.3g to 0.8g of burdock fructo-oligosaccharide and 0.8mL to 1.0mL of hydrogen; the CGMCC number of the candida olivaceus is CGMCC 2.1734; the number of the Candida sake is CGMCC 2.2695;
the preparation method of the grape antagonistic yeast composite biological preservative comprises the following steps: (1) fermenting and culturing candida olivaceus and candida sake, concentrating fermentation liquor, and concentrating the fermentation liquor until the dry weight of the bacteria is more than or equal to 200 g/L; (2) freeze drying burdock fructo-oligosaccharide and concentrated fermentation liquor together to obtain powder or tablet; (3) and preparing the hydrogen generation package.
2. The use of claim 1, wherein 8-10mL of grape antagonistic yeast composite biological preservative is sprayed on each kilogram of fruits.
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