CN104351330A - Fresh-keeping agent for fresh edible mushrooms and preparation method thereof - Google Patents
Fresh-keeping agent for fresh edible mushrooms and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a fresh-keeping agent for fresh edible mushrooms, relates to a fresh-keeping agent for fresh edible mushrooms, prepared by compounding gloeostereum extract, auricularia auricula polysaccharide liquid and Nisin, and a preparation method thereof, belonging to the field of agricultural product processing, storage and fresh-keeping. The fresh-keeping agent comprises the following raw materials in percentage by mass of 6% of gloeostereum entity extract, 1.2% of auricularia auricula polysaccharide liquid, 1% of Nisin and 91.8% of distilled water by compounding. According to the invention, the fresh-keeping agent prepared by compounding several natural extracts has a good antibacterial effect, and inhibits escherichia coli and staphylococcus aureus and the like as the representatives of positive and negative spoilage microorganisms in food, so that the preservation period of edible mushrooms is prolonged, therefore, the fresh-keeping agent has a practical application value, and facilitates the improvement of the whole food preservation level.
Description
Technical field
The present invention relates to a kind of food preservative, be specifically related to fresh edible mushroom antistaling agent prescription of elm ear extract and Blackfungus polyhexose liquid and Nisin compound and preparation method thereof, belong to processing of farm products and keeping-freshness storage field.
Background technology
Edible mushroom, have high protein, low-fat feature, its aromatic flavour, mouthfeel is fabulous, the pure fresh perfume (or spice) of taste, nutritious, and having very high nutritive value and health-care effect, is the natural green food being conducive to human amino acid's nutrient balance.Edible mushroom also has very high medical value, and especially its active anticancer is higher.Carry out the extraction of cancer-resisting substance with its fructification, mycelium, zymotic fluid and make the cancer therapy drug of various formulation, there is very tempting prospect.Due to edible mushroom growth, gather restriction by the factors such as season, region, weather, be difficult to fresh-keeping and storing, and chemical industry antistaling agent causes food secondary pollution, food quality is declined, this is applied to the preservation of edible mushroom fresh goods with regard to impelling research and development from the antistaling agent of natural biological materials extraction.
Summary of the invention
Technical problem to be solved by this invention is: cannot the problem of long term storage for current edible mushroom fresh goods, provides fresh edible mushroom antistaling agent prescription of a kind of fungistatic effect better elm ear extract and Blackfungus polyhexose liquid and Nisin compound and preparation method thereof.
Technical solution of the present invention is: a kind of fresh antifaling agent of edible fungus, by mass percentage, elm ears or side handles of a utensil entity extraction liquid by 6%, the Blackfungus polyhexose extract of 1.2%, the Nisin of 1% and the distilled water of 91.8% are composited, described elm ears or side handles of a utensil entity extraction liquid is obtained by following steps: boil after soaking 24h under elm ears or side handles of a utensil entity is added 10 times of cold water normal temperature, continue 1.5h and add water to prevent dry paste, filter, filter residue adds 5 times of water boils, filter after continuing 0.5h, merged by twice water cooking liquid, being concentrated into water extract content after filtration under diminished pressure is 28%; Described black fungus Aqueous extracts is obtained by following steps: black fungus is soaked 3 h at temperature 70 DEG C, and pulverize, 100 mesh sieve are sieved, and add water mixing, ultrasonic disruption cell, and hot water extraction is centrifugal, gets supernatant, and it is 16% that vacuum rotary steam is concentrated into polyoses content.
Technique effect of the present invention is: the present invention takes several natural extract compound to make antistaling agent, there is good fungistatic effect, positives to food and negative spoilage microorganism representative Escherichia coli and staphylococcus aureus etc. suppress, extend fresh edible mushroom preservation term, there is actual application value, be conducive to the raising of whole food preservation level.
Concrete effect is as follows:
1, remarkable by elm ear aqueous extract, Blackfungus polyhexose liquid, Nisin compounded antifaling agent of edible fungus antibacterial effect, add the compositional liquor minimal inhibitory concentration of 0.2% natural antiseptic agent Nisin up to 15%, especially 95 IU/mL are brought up to the fungistatic effect of the gram-positive bacterias such as staphylococcus aureus, 67 more than IU/mL are also risen to the fungistatic effect of the Gram-negative bacterias such as Escherichia coli.Fully demonstrate bacteriostatic activity better, more shelf-stable.Bacteriostatic active ingredients in this compound preservative all has fabulous heat resistance, and still have good bacteriostatic activity after 121 DEG C of process 20min, relative bacteriostasis rate is up to more than 75%; 25 DEG C to 100 DEG C bacteriostatic activities are substantially constant, have good bacteriostatic activity from acidity to neutral compound preservative, illustrate that this compound preservative has good pH tolerance, can not tolerate the environment of highly basic, Mg in metal ion
2+to compound preservative bacteriostatic activity slightly booster action, Zn
2+, Fe
3+, Cu
2+ion suppresses the bacteriostatic activity of compound preservative.
2, treated edible fungus fresh-keeping effect is all improved, and wherein elm ear, black fungus, Nisin compound preservative effect are better than positive control chemical preservative, safer.The blank group shelf-life is the shortest, 15 days time, substantially lose commodity value, softening at the end generation water soaking of edible mushroom, and produces with tart flavour.The chemically treated edible fungus fresh-keeping phase reaches 18 days, mainly because benzoic acid can suppress or the physiological activity of killing microorganisms, development of microorganisms is slowed down or stops.1., the enzyme system of interference microorganism the antiseptic principle of chemical preservative roughly has following 3 kinds:, destroys that it is metabolic normally, the activity of inhibitory enzyme.2., make the albumen of microorganism query solid and sex change, disturb its existence and breeding; 3., change cytoplasm permeability of the membrane, make the enzyme in its body and metabolite effusion cause its inactivation.When the edible mushroom 21 days of elm ear compound preservative process, still commodity value is had, because the polysaccharide material in elm ear all has inhibition in various degree to Gram-negative bacteria and gram-positive bacteria, especially to proteus, Escherichia coli, salmonella, staphylococcus aureus, hay bacillus etc., extended the edible mushroom shelf-life.
3, edible mushroom fresh goods antistaling agent Be very effective of the present invention, bacteriostatic activity are good, shelf-stable.Each component, all from wholefood, all to human body and environmentally friendly, without complicated chemical composition, also can not cause chemical residual, can not bring safety problem, fill up the market vacancy.
4, use after each component mixing of the present invention, the function of each component obtains complementation and coordination, edible mushroom fresh goods preservation term can being made to significantly improve, avoiding the shortcoming that these materials effect when using separately is bad, unstable.
5, in antifaling agent of edible fungus, active component concentration is too low can not play its fresh-keeping effect, excessive concentration then can have a negative impact to edible fungus fresh-keeping and also waste resource, increase cost, preferably, antifaling agent of edible fungus of the present invention in use, add water according to 6% elm ears or side handles of a utensil entity extraction liquid, 1.2% Blackfungus polyhexose extract, 1%Nisin formula join 25mL compound preservative, namely add 0.25gNisin, 1.5g elm ears or side handles of a utensil entity extraction liquid, 0.3g Auricularia polysaccharide extract mixed dissolution in 25mL distilled water is compound preservative, 4 DEG C of preservations are for subsequent use.
6, in order to improve edible mushroom preservation effect, antistaling agent application is preferably low-temperature atomizing and is evenly sprayed on edible mushroom surface.
Accompanying drawing explanation
Fig. 1 is that variable concentrations elm ear Aqueous extracts of the present invention and anticorrisive agent compositional liquor are to Escherichia coli Growth amount influence curve figure;
Fig. 2 be variable concentrations elm ear Aqueous extracts of the present invention and anticorrisive agent compositional liquor antibacterial after viable count curve map;
Fig. 3 is Nisin canonical plotting of the present invention;
Fig. 4 is that elm ear Aqueous extracts of the present invention is to the inhibitory potency figure of two kinds of bacterium;
Fig. 5 is the inhibitory potency figure of auricularia auriculajudae Aqueous extracts of the present invention to two kinds of bacterium;
Fig. 6 is the inhibitory potency figure of Nisin of the present invention to two kinds of bacterium;
Fig. 7 be under treatment of different temperature of the present invention compound preservative to the relative bacteriostasis rate of indicator bacteria;
Fig. 8 is that the lower compound preservative of the different pH process of the present invention is to the relative bacteriostasis rate of indicator bacteria;
Fig. 9 is compound natural preservative of the present invention fungistatic effect figure to staphylococcus aureus under different metal ionization;
Figure 10 be compound natural preservative of the present invention under different metal ionization to colibacillary fungistatic effect figure;
Figure 11 is the weight-loss ratio change during mushroom preservation process of the present invention;
Figure 12 is the impact of Preservation Treatment of the present invention on mushroom cell membrane integrity;
Figure 13 is the impact of Preservation Treatment of the present invention on mushroom soluble solid content;
Figure 14 is the impact of Preservation Treatment of the present invention on mushroom brown stain degree.
Detailed description of the invention
A kind of fresh edible mushroom antistaling agent prescription, by mass percentage, add water according to 6% elm ears or side handles of a utensil entity extraction liquid, 1.2% Blackfungus polyhexose extract, 1%Nisin formula join 25mL compound preservative, namely add 0.25gNisin, 1.5g elm ears or side handles of a utensil entity extraction liquid, 0.3g Auricularia polysaccharide extract mixed dissolution in 25mL distilled water is compound preservative, 4 DEG C of preservations are for subsequent use.Atomizing type is adopted to spray fresh edible mushroom.
Edible mushroom fresh goods antistaling agent of the present invention is for the preservation of edible mushroom fresh goods as antistaling agent.
The preparation method of edible mushroom fresh goods antistaling agent of the present invention, comprises the steps:
Project process route:
1, the preparation of antifaling agent of edible fungus:
The preparation of 1.1 elm ear Aqueous extracts: get elm ears or side handles of a utensil entity clear water and wash away floating ash, to put in thermostatic drying chamber 60 DEG C to dry to constant weight, take the fructification of 200g drying, place 24h under adding 10 times of cold water normal temperature to be placed on electromagnetic oven and to boil, timing is started during to seethe with excitement, boil 1.5h, the later stage needs a small amount of constantly adding water repeatedly, and agitation as appropriate is to prevent dry paste.Use double gauze filters, and gained filter residue adds 5 times of poach to boiling, and after continuing 0.5h, double gauze filters.To merge with second time water cooking liquid first time, simmer down to 200mL after filtration under diminished pressure, is elm ear Aqueous extracts, and measuring water extract content is 28%.
The preparation of 1.2 black fungus Aqueous extracts: black fungus 200g drying (70 DEG C, 3 h) → pulverize → 100 mesh sieve to sieve → add water 200mL mixing → ultrasonic disruption cell → hot water extraction → centrifugal (3 000 r/min, 15 min) → get supernatant → vacuum rotary steam simmer down to 200mL → mensuration polyoses content of powder are 16%.
1.3 Nisin solution: Nisin is mixed with solution for standby with sterile distilled water.
1.4 add water according to 6% elm ears or side handles of a utensil entity extraction liquid, 1.2% Blackfungus polyhexose extract, 1%Nisin formula join 25mL compound preservative, namely add 0.25gNisin, 1.5g elm ears or side handles of a utensil entity extraction liquid, 0.3g Auricularia polysaccharide extract mixed dissolution in 25mL distilled water is compound preservative, 4 DEG C of preservations are for subsequent use.
2, edible mushroom preservation
2.1 edible mushroom collections to choose the same day full, any surface finish is disease-free, in the same size, color evenly, the edible mushroom that has no mechanical damage is stand-by.
2.2 take edible mushroom by every part of 150g, are placed in open tray, adopt low-temperature atomizing to spray antifaling agent of edible fungus, as edible mushroom parachute-opening, first that cap is downward, spray lamella side, then above the sprinkling cap that turns over.In 3 ~ 5 DEG C of precooling 4h, after packing with the polyethylene preservative film membranes of thick 13 μm, 41 group, boxes, are placed in the polyethylene plastic bag of thick 25 μm, put into antistaling box, store under the environment of 3 ~ 5 DEG C.Carry out under the condition of all operations below 4 DEG C.
Example 1:
1, the fungistatic effect of elm ear Aqueous extracts and compound preservative is to when bacteriostasis mode
The bacteriostatic activity of 1.1 elm ear Aqueous extracts and compositional liquor
1.1.1 the bacteriostatic activity of elm ear Aqueous extracts
The test data of elm ear Aqueous extracts is obtained as table 1 through repeating test repeatedly.
Table-1 elm ears or side handles of a utensil entity Aqueous extracts tiring to different bacterium
| Bacterial classification | Antibacterial circle diameter mean value (mm) | Elm ear Aqueous extracts tires (IU/mL) |
| Escherichia coli | 9.1 | 23.65172 |
| Salmonella | 10.1 | 29.34781 |
| Staphylococcus aureus | 19.3 | 88.20657 |
| Bacillus subtilis | 15.7 | 62.89098 |
| Bacillus cereus | 6.6 | 0.79571 |
Wider by the antimicrobial spectrum of table-1 known elm ears or side handles of a utensil entity Aqueous extracts, to the inhibitory potency of staphylococcus aureus up to 88.20657 IU/mL, tiring also up to 62.89098 IU/mL to bacillus subtilis, but about 20 IU/mL are only had to the fungistatic effect of Escherichia coli and salmonella.In the bacterial strain that test adopts, staphylococcus aureus and bacillus subtilis are typical gram-positive bacteria, Escherichia coli and salmonella are typical Gram-negative bacteria, so can find out that the action effect of elm ear Aqueous extracts to gram-positive bacteria is better than the action effect to Gram-negative bacteria.This result can to provide as the application and development field of biological bacteriostatic agent at elm ears or side handles of a utensil entity and test support accordingly.
1.1.2 the bacteriostatic activity of compositional liquor
The test data of anticorrisive agent compositional liquor is obtained as table-2 through repeating test repeatedly.
Table-2 anticorrisive agent compositional liquors tiring to different bacterium
| Bacterial classification | Antibacterial circle diameter mean value (mm) | Tire (IU/mL) of elm ear compositional liquor |
| Escherichia coli | 16.1 | 67.32121 |
| Salmonella | 16.7 | 71.75143 |
| Staphylococcus aureus | 20.5 | 95.80125 |
| Bacillus subtilis | 19.1 | 86.30792 |
| Bacillus cereus | 15.2 | 62.25808 |
As can be seen from table-2, very strong at the compositional liquor fungistatic effect adding seldom amount natural antiseptic agent, obvious bacteriostasis can be shown when low concentration.Especially to the fungistatic effect of the gram-positive bacterias such as staphylococcus aureus up to 95 IU/mL, be promoted to 67 more than IU/mL to the fungistatic effect of the Gram-negative bacterias such as Escherichia coli, be not that the bacillus cereus fungistatic effect very with fungistatic effect rises to 62.25808 IU/mL by elm ear Aqueous extracts itself simultaneously yet.The bacteriostatic activity fully demonstrating the elm ears or side handles of a utensil entity extraction liquid composite with natural antiseptic agent is better, more shelf-stable.
The MIC of 1.2 elm ear Aqueous extracts and compositional liquor
1.2.1 the MIC of elm ears or side handles of a utensil entity Aqueous extracts
MIC result through test determination elm ears or side handles of a utensil entity Aqueous extracts is as shown in table 3.
The MIC of table-3 elm ears or side handles of a utensil entity Aqueous extracts
-expression has no colony growth; + indicate a small amount of colony growth; ++ for bacterium colony is more but be less than plate area 1/3; +++ bacterium colony reach plate area 1/2 and more than.
When being 20% by test data known elm ears or side handles of a utensil entity Aqueous extracts concentration, Escherichia coli, staphylococcus aureus, bacillus subtilis and bacillus cereus bacterium colony have a small amount of growth, the not long bacterium of salmonella.When Aqueous extracts concentration all bacterium 25% time originally have no growth, so show that elm ears or side handles of a utensil entity Aqueous extracts has good bacteriostatic activity, and showing that minimum inhibitory concentration is 20%, is best to the bacteriostatic activity of various bacterium.
1.2.2 the MIC of compositional liquor
Through test determination compositional liquor MIC result as table-4 shown in.
The MIC of table-4 compositional liquors
-expression has no colony growth; + indicate a small amount of colony growth; ++ for bacterium colony is more but be less than plate area 1/3; +++ bacterium colony reach plate area 1/2 and more than.
Observe known compositional liquor concentration by table-4 and have a small amount of growth at 15% pair of Escherichia coli, salmonella and bacillus cereus bacterium colony, the not long bacterium of staphylococcus aureus, bacillus subtilis, but when Aqueous extracts concentration all bacterium colonies 20% time have no growth, so show that compositional liquor has good bacteriostatic activity, and the minimum inhibitory concentration of compositional liquor of obtaining a result is 15%.
The mode of action that 1.3 compound natural preservatives are antibacterial
1.3.1 elm ear Aqueous extracts and compositional liquor are on the impact of indicator bacteria growth curve
Under cultivating at 37 DEG C, to be first elm ear Aqueous extracts and E. coli broth timing sampling in 48h of 1g/mL containing 10%, 20% concentration, again by containing 10%, 15% compositional liquor respectively with E. coli broth in 48h timing sampling, survey absorbance draw growth curve see Fig. 1.
As shown in Figure 1: Initial stage of culture 0 ~ 18h, under the effect of compositional liquor, the bacterium number of indicator bacteria declines fast, the absorbance of compositional liquor is all below 0.1, and compositional liquor concentration is higher, the amplitude that bacterium number declines is larger, describes compositional liquor and has very strong bacteriostasis.Late stage of culture 18 ~ 48h, the sample OD value containing compositional liquor maintains less than 0.1, can find out that the compositional liquor bacteriostasis of adding natural antiseptic agent is very stable, be conducive to the preservation of sample.
Under cultivating at 37 DEG C, to be first elm ear Aqueous extracts and E. coli broth timing sampling in 48h of 1g/mL containing 10%, 20% concentration, again by containing 10%, 15% compositional liquor respectively with E. coli broth in 48h timing sampling, utilize dull and stereotyped writing-method measure viable count see Fig. 2.
As shown in Figure 2, under the effect of the compositional liquor of 10% and 15%, in 6h, viable count is by 10
6cfu/mL drops to 10
1cfu/mL, and along with the prolongation of time, colibacillary viable count is also in obvious decline, the culture medium containing the compositional liquor of 15% after 12h does not almost have the growth of bacterium, and this illustrates that compositional liquor has stronger bactericidal action.
1.3.2 the cultivating again of elm ear Aqueous extracts and compositional liquor mode of action agar block
By step measurements viable count, result uses the elm ear Aqueous extracts fluid nutrient medium containing 20% to cultivate, and before cultivating, dull and stereotyped bacteria colony count is 317cfu/mL, gets sterile letheen block cultivation 24h rear plate counting method and increases to 65cfu/mL.Illustrate after certain hour is cultivated, the agar block of not long bacterium still has viable bacteria remain, this sub-fraction bacterium should be suppressed, so the mode of action of elm ear extract to bacterium is inhibitory action.
According to the research to elm ear Aqueous extracts, further by step measurements compositional liquor viable count, result uses the compositional liquor fluid nutrient medium containing 15% to cultivate, and before cultivating, dull and stereotyped bacteria colony count is 74cfu/mL, and getting sterile letheen block cultivation 24h rear plate counting method does not almost have.Illustrate that the agar block of not long bacterium does not almost have viable bacteria to remain, so elm ear compound natural preservative is strictly antibacterial to the mode of action of bacterium after certain hour is cultivated.
The development of example 2 compound natural preservative
The measurement result of 2.1 Nisin inhibitory potencies
Cylinder-plate method is adopted to test Nisin, with slide measure Measurement accuracy antibacterial circle diameter, the value recorded is substituted into formula Y=a × lg(X)+b, linear between the logarithm that standard is tired and antibacterial circle diameter, regression equation is that y=6.3289x+4.0322, Y represent the logarithm value of tiring, X represents antibacterial circle diameter (not containing aperture), coefficient correlation 0.9922, meets further test requirements document, as shown in Figure 3.
2.2 variable concentrations elm ear Aqueous extracts are to the inhibition of different strains
Elm ear Aqueous extracts all has inhibitory action to Escherichia coli, staphylococcus aureus.When increasing progressively between the concentration of elm ear Aqueous extracts is from 7%, 14%, 28%, bacteriostasis strengthens gradually.When the concentration of elm ear Aqueous extracts reaches 7%, it has been greater than the Nisin effect of 2% to Escherichia coli bacteriostasis, and inhibitory potency is 15IU/mL.When concentration is 14%, present stronger bacteriostasis, respectively 35IU/mL is reached to the inhibitory potency of Escherichia coli, staphylococcus aureus, 20IU/mL.And it is also less than the Nisin effect of 2% to staphylococcus aureus bacteriostasis when the concentration of elm ear Aqueous extracts reaches 28%, but present stronger bacteriostasis to Escherichia coli, inhibitory potency reaches 60IU/mL.Show that elm ear Aqueous extracts is better than staphylococcus aureus colibacillary fungistatic effect.
Zhou Jianshu etc. report that elm ears or side handles of a utensil entity decocting liquid and elm fungus fermentation liquor cause germ to have good inhibitory action to shigella dysenteriae, Pseudomonas aeruginosa, Escherichia coli, staphylococcus aureus and salmonella enteritidis, be a kind of rare collection eat and medicine for the Precious Edible Fungi kind of one.Deep discussion has been carried out to the active ingredient of elm ear bacteriostasis, elm ears or side handles of a utensil entity decocting liquid has good inhibitory action to pathogenic bacteria such as shigella dysenteriae, Pseudomonas aeruginosa, Escherichia coli, staphylococcus aureus and salmonella enteritidis, and the active ingredient of its bacteriostasis is a kind of glucosides class material.It is a kind of good natural antiseptic agent compared with wide spectrum.Substantially identical with this experiment conclusion, as shown in Figure 4.
2.3 variable concentrations black fungus Aqueous extracts are to the inhibition of different strains
Variable concentrations gradient auricularia auriculajudae Aqueous extracts has bacteriostasis in various degree to these two kinds of bacterium.When the concentration of Aqueous extracts progressively increases, bacteriostasis obviously strengthens.When the concentration of auricularia auriculajudae Aqueous extracts is 8%, have partial inhibition to Escherichia coli, be greater than the Nisin effect of 2%, inhibitory potency is 15IU/mL.When the concentration of auricularia auriculajudae Aqueous extracts is 15%, to the inhibitory action of staphylococcus aureus, be still less than the Nisin effect of 2%, have stronger inhibitory action to Escherichia coli, be greater than the Nisin effect of 2%, inhibitory potency is 29IU/mL.Result shows that auricularia auriculajudae Aqueous extracts is better than staphylococcus aureus to colibacillary fungistatic effect simultaneously.
Diao little Qin etc. think that the Blackfungus polyhexose of variable concentrations has inhibitory action in various degree to Escherichia coli, staphylococcus aureus.Along with the increase of Blackfungus polyhexose concentration, its bacteriostatic activity strengthens, and starts to show bacteriostasis, and be better than staphylococcus aureus to colibacillary inhibition under same concentrations when concentration reaches 10 mg/mL, substantially identical with this conclusion.The people such as Zhang Tangwei think that Nostoc commune polysaccharide all has certain inhibitory action to bacillus subtilis, Escherichia coli, and minimum-bacteriostat mass concentration MIC is between 125 ~ 250 μ g/mL.And it is not obvious to the inhibitory action of staphylococcus aureus.Substantially identical with this experiment conclusion, as shown in Figure 5.
2.4 variable concentrations Nisin are to the inhibition of different strains
Nisin has stronger bacteriostasis to golden staphylococci, and when the concentration of Nisin is 1%, inhibitory potency reaches 20IU/mL.When the concentration of Nisin increases, staphylococcic fungistatic effect is significantly increased.When the concentration of Nisin is 2%, inhibitory potency reaches 53IU/mL.And to Escherichia coli without obvious suppression, and increase not obvious with the increase fungistatic effect of concentration.Then illustrate that Nisin is better than Escherichia coli to golden staphylococci fungistatic effect, has stronger bacteriostasis to golden staphylococci, to Escherichia coli without obvious inhibitory action.
At present, more to the antifungal mechanism research of Nisin, the bacteriostasis of Nisin based on it to the strong suction-operated of cell surface and then cause cytoplasmic release to realize.Nisin is positively charged LALLFWL, and thus it can act on anion component electronegative on gram-positive bacteria cell wall, as LTA, teichuronic acid, acidic polysaccharose and phosphatide.Interactional result forms tubular structure with cell membrane, and the cell constituent of small-molecular-weight is leaked from duct.Thus Nisin has stronger inhibitory action to large-scale gram-positive bacterium, to Gram-negative bacteria, yeast and mould all without effect.Substantially identical with this experiment conclusion, as shown in Figure 6.
2.5 each natural antiseptic agent orthogonal building optimum results
Shitosan, Nisin, elm ear and auricularia auriculajudae Aqueous extracts have different inhibitory action to Escherichia coli, staphylococcus aureus, and the variable concentrations of each anticorrisive agent and the difference accordingly between fungistatic effect are also found.Nisin only has stronger inhibitory action to staphylococcus aureus, and when concentration is 2%, inhibitory potency reaches 53IU/mL, to Escherichia coli without obvious inhibitory action.Shitosan and elm ear and auricularia auriculajudae Aqueous extracts antimicrobial spectrum wider, fungistatic effect is stronger.When chitosan concentration is 2%, Escherichia coli fungistatic effect is greater than to the Nisin of 2%, inhibitory potency is 21IU/mL.When the concentration of auricularia auriculajudae Aqueous extracts is greater than 10%, there is stronger inhibitory action to Escherichia coli, be greater than the Nisin effect of 2%.When the concentration of elm ear Aqueous extracts is greater than 10%, it has been greater than the Nisin effect of 2% to Escherichia coli bacteriostasis.Therefore by they compounds by a certain percentage, respective bacteriostasis is played.
By four kinds of natural antiseptic agent shitosans, Nisin, elm ear Aqueous extracts and auricularia auriculajudae Aqueous extracts, show employing cylinder-plate method by table 2-3, carry out 4 factor 3 horizontal quadrature tests, calculate orthogonal building sample and tire and carry out variance analysis.By the primary and secondary order of extreme difference size decisive factor, average size determines that level is good and bad.Staphylococcus aureus and colibacillary antibacterial interpretation of result are obtained: concerning two kinds of bacterium, optimization formula all: with elm ear Aqueous extracts for principal element, and being optimum with level 3, be secondly Nisin, is optimum with level 2.Be black fungus Aqueous extracts again, being optimum with level 3, is finally shitosan, is optimum with level 1.Show that anticorrosion composite preferred plan is: elm ear Aqueous extracts 20%, black fungus Aqueous extracts 20%, Nisin 1%, shitosan 0% thus.And be greater than F critical value by F ratio and obtain: there is remarkable impact with or without the fungistatic effect of elm ear Aqueous extracts on compositional liquor.As table-5, table-6, table-7, shown in table-8.
Table-5 orthogonal building liquid are to the antibacterial results and analysis of staphylococcus aureus
| Factor | Elm ear | Black fungus | nisin | Shitosan | Tire (IU/mL) |
| Experiment 1 | 1 | 1 | 1 | 1 | 0 |
| Experiment 2 | 1 | 2 | 2 | 2 | 7.58 |
| Experiment 3 | 1 | 3 | 3 | 3 | 8.78 |
| Experiment 4 | 2 | 1 | 2 | 3 | 14.07 |
| Experiment 5 | 2 | 2 | 3 | 1 | 22.58 |
| Experiment 6 | 2 | 3 | 1 | 2 | 29.1 |
| Experiment 7 | 3 | 1 | 3 | 2 | 317.79 |
| Experiment 8 | 3 | 2 | 1 | 3 | 179.6 |
| Experiment 9 | 3 | 3 | 2 | 1 | 535 |
| Average 1 | 5.453 | 110.62 | 69.567 | 185.86 | |
| Average 2 | 21.917 | 69.92 | 185.55 | 118.157 | |
| Average 3 | 344.13 | 190.96 | 116.383 | 67.483 | |
| Extreme difference | 338.677 | 121.04 | 115.983 | 118.377 |
Table-6 orthogonal building liquid are to the antibacterial result analysis of variance table of staphylococcus aureus
| Factor | Deviation square | The free degree | F ratio | F critical value | Conspicuousness |
| Elm ear | 218794.358 | 2 | 10.2 | 5.14 | * |
| Black fungus | 22761.687 | 2 | 1.061 | 5.14 | |
| nisin | 20427.962 | 2 | 0.952 | 5.14 | |
| Shitosan | 21164.563 | 2 | 0.987 | 5.14 | |
| Error | 64354.21 | 6 |
Table-7 orthogonal building liquid are to the antibacterial results and analysis of Escherichia coli
| Factor | Elm ear | Black fungus | nisin | Shitosan | Tire (IU/mL) |
| Experiment 1 | 1 | 1 | 1 | 1 | 0 |
| Experiment 2 | 1 | 2 | 2 | 2 | 7 |
| Experiment 3 | 1 | 3 | 3 | 3 | 9.78 |
| Experiment 4 | 2 | 1 | 2 | 3 | 18.15 |
| Experiment 5 | 2 | 2 | 3 | 1 | 22.58 |
| Experiment 6 | 2 | 3 | 1 | 2 | 7.58 |
| Experiment 7 | 3 | 1 | 3 | 2 | 46.75 |
| Experiment 8 | 3 | 2 | 1 | 3 | 37.57 |
| Experiment 9 | 3 | 3 | 2 | 1 | 54.07 |
| Average 1 | 5.593 | 21.633 | 15.05 | 25.55 | |
| Average 2 | 16.103 | 22.383 | 26.407 | 20.443 | |
| Average 3 | 46.13 | 23.81 | 26.37 | 21.833 | |
| Extreme difference | 40.537 | 2.177 | 11.357 | 5.107 |
Table-8 orthogonal building liquid are to the antibacterial result analysis of variance table of Escherichia coli
| Factor | Sum of square of deviations | The free degree | F ratio | F critical value | Conspicuousness |
| Elm ear | 2655.282 | 2 | 20.081 | 6.94 | * |
| Black fungus | 7.336 | 2 | 0.055 | 6.94 | |
| nisin | 257.118 | 2 | 1.945 | 6.94 | |
| Shitosan | 41.824 | 2 | 0.316 | 6.94 | |
| Error | 264.45 | 4 |
The physical and chemical stability research of example 3 elm ear compound natural preservative
The measurement result of 3.1 heat endurances and analysis
Through the heat endurance of measuring compound preservative, experimental result is as Fig. 7,2%Nisin can be conversed by regression equation of tiring and 77.80IU/mL and 690.25IU/mL is respectively to staphylococcus aureus and colibacillary inhibitory potency, obvious compound preservative processes compound preservative bacteriostatic activities without significant change through-20 DEG C,-20 DEG C to 4 DEG C bacteriostasis rates are the highest, after 4 DEG C, bacteriostatic activity starts to reduce, and reach 100 DEG C of activity and tend to be steady, 121 DEG C of relative bacteriostasis rates are still more than 75%; It can thus be appreciated that this compound preservative has good heat endurance and is applicable to low-temperature preservation, and this conclusion has very large meaning in the Application and Development of Combined-preservative.
The measurement result of 3.2 pH stability and analysis
Through measuring compound preservative pH stability result as shown in Figure 8, 2%Nisin can be conversed by regression equation of tiring and 77.80IU/mL and 690.25IU/mL is respectively to staphylococcus aureus and colibacillary inhibitory potency, through measuring, the pH value of compound preservative is about 6.0, after regulating the pH value of compound preservative, measure bacteriostatic activity, the relative bacteriostasis rate of each sample is as shown in figure-8, as pH from 5 to 8 along with the rising of pH, the bacteriostatic activity of compound preservative is reduction trend, when pH is greater than 7, the reduction of bacteriostasis rate becomes obvious, when pH equals 8 and 9, bacteriostatic activity is almost lost, under this pH visible, compound preservative loses rejection ability to for examination bacterium, after compound preservative sample pH value is now transferred to 6.0 again, bacteriostatic activity still can not recover, can infer that in compound preservative, some bacteriostatic active ingredients there occurs irreversible inactivation in the basic conditions, the distilled water of control experiment is when pH is less than 4, and bacteriostatic activity is remarkable, and under this pH, the bacteriostatic activity of compound preservative is compound preservative active component and the coefficient result of solution acidity, when pH is between 5-7, along with the increase of acidity, the bacteriostatic activity of compound preservative strengthens gradually, and when pH equals 5, compound preservative bacteriostatic activity is the highest, it can thus be appreciated that the antibacterial pH of the best of this compound preservative is 5.
The measurement result of 3.3 metal ion effect and analysis
Different metal ion through measuring compound preservative (being labeled as compositional liquor in figure) affects experimental result as shown in Figure 9, Figure 10.
2%Nisin can be conversed by regression equation of tiring and 77.80IU/mL and 690.25IU/mL is respectively to staphylococcus aureus and colibacillary inhibitory potency.From figure-9 and figure-10, the antibacterial substance in compound preservative is to there being Mg
2+, K
+, Ca
2+, Mn
2+have good bacteriostasis property when plasma exists, because metal ion experimental temperature is 25 DEG C, compare discovery in conjunction with normal temperature compound preservative in temperature experiment (25 DEG C) inhibition zone size and only have Mg
2+compound preservative is tired and is greater than normal temperature compound preservative and tires, and wherein Mg is described
2+there is the effect strengthening bacteriostatic activity, K
+, Ca
2+, Mn
2+on the almost no impact of compound preservative activity.Add Zn
2+, Fe
3+, Cu
2+the compound preservative sample of ion is substantially the same with the inhibition zone of the solion sample of contrast, visible Zn
2+, Fe
3+, Cu
2+ion suppresses the activity of certain antipathogenic composition in compound preservative, wherein Fe
3+inhibition is slightly weak, and this feature will contribute to this compound preservative and obtain more dynamical utilization in antibacterial field.
Example 4 mushroom preservation is tested
The subjective appreciation of 4.1 edible mushrooms
As can be seen from following table, different disposal is on the fresh-keeping impact of edible mushroom, and from then on table can be found out, treated edible fungus fresh-keeping effect is all improved, wherein elm ear, black fungus, Nisin compound preservative effect are better than positive control chemical preservative, safer.The blank group shelf-life is the shortest, 15 days time, substantially lose commodity value, softening at the end generation water soaking of edible mushroom, and produces with tart flavour.The chemically treated edible fungus fresh-keeping phase reaches 18 days, mainly because benzoic acid can suppress or the physiological activity of killing microorganisms, development of microorganisms is slowed down or stops.The antiseptic principle of chemical preservative roughly has following 3 kinds: the enzyme system 1. disturbing microorganism, destroys that it is metabolic normally, the activity of inhibitory enzyme.2. make the albumen of microorganism query solid and sex change, disturb its existence and breeding; 3. change cytoplasm permeability of the membrane, make the enzyme in its body and metabolite effusion cause its inactivation.When the edible mushroom 21 days of elm ear compound preservative process, still commodity value is had, because the polysaccharide material in elm ear all has inhibition in various degree to Gram-negative bacteria and gram-positive bacteria, especially to proteus, Escherichia coli, salmonella, staphylococcus aureus, hay bacillus etc., extended the edible mushroom shelf-life.
Table-9 different disposal are to the results of sensory evaluation of mushroom different storage fresh-keeping effect
| Period of storage | Index | Contrast | Chemical preservative benzoic acid | Elm ear compound preservative |
| Color and luster | 4 | 4 | 4 | |
| 3d | Hardness | 4 | 4 | 4 |
| Local flavor | 4 | 4 | 4 | |
| Color and luster | 4 | 4 | 4 | |
| 6d | Hardness | 4 | 4 | 4 |
| Local flavor | 3 | 4 | 4 | |
| Color and luster | 3 | 4 | 4 | |
| 9d | Hardness | 4 | 3 | 4 |
| Local flavor | 3 | 3 | 3 | |
| Color and luster | 3 | 3 | 3 | |
| 12d | Hardness | 3 | 3 | 4 |
| Local flavor | 2 | 3 | 3 | |
| Color and luster | 2 | 3 | 3 | |
| 15d | Hardness | 2 | 3 | 3 |
| Local flavor | 2 | 2 | 3 | |
| Color and luster | 1 | 2 | 2 | |
| 18d | Hardness | 2 | 2 | 3 |
| Local flavor | 1 | 2 | 2 | |
| Color and luster | 1 | 1 | 2 | |
| 21d | Hardness | 1 | 2 | 3 |
| Local flavor | 1 | 1 | 2 |
4.2 compound preservative Preservation Treatment are on the impact of mushroom weight-loss ratio
The main source of edible mushroom mass loss is exactly moisture, the content of moisture and the form of existence affect microbial growth breeding, thus affect the shelf stability of food and disease resistance, also the quality of food quality is therefore related to, there is a variety of change in the existence form of duration of storage moisture, as moisture absorption and desorb, moisture evaporate, water transport and redistributing.
As seen from the figure, between preservation term, the moisture of mushroom is all on a declining curve, the speed that the moisture of blank group declines is obviously faster than other groups, 92.75% is reached at the weight-loss ratio of the 21st day mushroom, chemical preservative can alleviate weightlessness, but does not have the Be very effective of elm ear anticorrisive agent, and when 21 days, the weight-loss ratio of mushroom compound preservative process is respectively 85%.Figure 11 is shown in weight-loss ratio change during mushroom preservation process.
4.3 compound preservative Preservation Treatment are on the impact of mushroom cell membrane integrity
Electrical conductivity is the inverse of resistivity, represent the ability that material transmits electric current, the integrality of plant and fruits and vegetables cell membrane system normally leakagely carries out assessing by what detect cell membrane ion, and the electrical conductivity that the leak magnitude of cell membrane is water extract by detecting tested plant tissue realizes, relative conductivity is low, illustrate that cell membrane system integrality is better, otherwise, then illustrate that cell membrane system is seriously impaired.
As seen from the figure, along with fresh keeping time extends, mushroom relative conductivity is in rising trend, and use more blank group of the edible mushroom rate of climb of elm ear compound preservative and chemical preservative process slow, the rate of climb of elm ear compound preservative process is the slowest.Show that elm ear compound preservative is best to mushroom cell membrane integrity protectiveness.Figure 12 is shown in the impact of Preservation Treatment on mushroom cell membrane integrity.
4.4 compound preservative Preservation Treatment are on the impact of edible fungi soluble solid content
The height of mushroom soluble solid content, can to represent in tissue sugar and other solid content number, also relevant with histiocytic retention ability, content and the Premeabilisation of cells of soluble solid are pressed into direct ratio, can prevent moisture penetration from arriving beyond cell membrane, thus water holding capacity is strong, and this is significant to preventing their nutriment loss.
As seen from the figure, soluble solid is all on a declining curve, illustrate that the retentiveness of cell is in decline, glucides etc. are all in reduction, its empty group soluble solid content declines the most obvious, and the edible fungi soluble solid content of chemical preservative process declines slightly slow, and the edible mushroom of elm ear compound preservative process declines the slowest, as can be seen here, elm ear compound preservative is conducive to keeping soluble solid content.Figure 13 is shown in the impact of Preservation Treatment on mushroom soluble solid content.
4.5 compound preservative Preservation Treatment are on the impact of edible mushroom brown stain degree
After edible mushroom collection, cell still carries out the metabolic activity enlivened, under normal circumstances, phenol wine elder brother oxidation-reduction system for respiratory carrier in complete tissue remains dynamic equilibrium, when histocyte is impaired, oxygen invades in a large number, and awake class is oxidative polymerization into melanin more further, and now brown stain occurs.
As seen from the figure, from overall browning degree, along with fresh keeping time extends, brown stain degree heighten degree, blank process brown stain degree is the most serious, compared with blank, positive chemical preservative process alleviates mushroom brown stain degree, but compared with the process of positive group chemical preservative, elm ear compound preservative prevents mushroom brown stain better effects if, whole freshness date, the mushroom brown stain degree of compound preservative process is all lower than chemical preservative processed group, show that elm ear compound preservative is for control brown stain more remarkable effect, and elm ear composite biological preservative is more safer than chemical preservative, therefore have more develop and spread to be worth.Figure 14 is shown in the impact of Preservation Treatment on mushroom brown stain degree.
Claims (1)
1. a fresh antifaling agent of edible fungus, it is characterized in that, by mass percentage, elm ears or side handles of a utensil entity extraction liquid by 6%, the Blackfungus polyhexose extract of 1.2%, the Nisin of 1% and the distilled water of 91.8% are composited, described elm ears or side handles of a utensil entity extraction liquid is obtained by following steps: boil after soaking 24h under elm ears or side handles of a utensil entity is added 10 times of cold water normal temperature, continue 1.5h and add water to prevent dry paste, filter, filter residue adds 5 times of water boils, filter after continuing 0.5h, merged by twice water cooking liquid, being concentrated into water extract content after filtration under diminished pressure is 28%; Described black fungus Aqueous extracts is obtained by following steps: black fungus is soaked 3 h at temperature 70 DEG C, and pulverize, 100 mesh sieve are sieved, and add water mixing, ultrasonic disruption cell, and hot water extraction is centrifugal, gets supernatant, and it is 16% that vacuum rotary steam is concentrated into polyoses content.
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| CN106900840A (en) * | 2017-03-06 | 2017-06-30 | 蒙阴万华食品有限公司 | A kind of edible antistaling agent and its application |
| CN110419565A (en) * | 2019-08-05 | 2019-11-08 | 厦门绿帝共享实业有限公司 | A kind of fresh-keeping fermentation liquid of novel meat deodorization and preparation method thereof |
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| CN105995958A (en) * | 2016-05-19 | 2016-10-12 | 丽江和融生物种植有限公司 | Toadstool processing method |
| CN106900840A (en) * | 2017-03-06 | 2017-06-30 | 蒙阴万华食品有限公司 | A kind of edible antistaling agent and its application |
| CN110419565A (en) * | 2019-08-05 | 2019-11-08 | 厦门绿帝共享实业有限公司 | A kind of fresh-keeping fermentation liquid of novel meat deodorization and preparation method thereof |
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