CN107410825A - A kind of composite natral bacteriostatic agent and its preparation method and application - Google Patents
A kind of composite natral bacteriostatic agent and its preparation method and application Download PDFInfo
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- CN107410825A CN107410825A CN201710273606.6A CN201710273606A CN107410825A CN 107410825 A CN107410825 A CN 107410825A CN 201710273606 A CN201710273606 A CN 201710273606A CN 107410825 A CN107410825 A CN 107410825A
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- nisin
- chitosan oligosaccharide
- bacteriostatic agent
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- maillard reaction
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- 239000000022 bacteriostatic agent Substances 0.000 title claims abstract description 26
- 239000002131 composite material Substances 0.000 title claims abstract description 10
- 238000002360 preparation method Methods 0.000 title claims abstract description 8
- 239000004309 nisin Substances 0.000 claims abstract description 47
- 235000010297 nisin Nutrition 0.000 claims abstract description 38
- RQFQJYYMBWVMQG-IXDPLRRUSA-N chitotriose Chemical compound O[C@@H]1[C@@H](N)[C@H](O)O[C@H](CO)[C@H]1O[C@H]1[C@H](N)[C@@H](O)[C@H](O[C@H]2[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O2)N)[C@@H](CO)O1 RQFQJYYMBWVMQG-IXDPLRRUSA-N 0.000 claims abstract description 37
- NVNLLIYOARQCIX-MSHCCFNRSA-N Nisin Chemical compound N1C(=O)[C@@H](CC(C)C)NC(=O)C(=C)NC(=O)[C@@H]([C@H](C)CC)NC(=O)[C@@H](NC(=O)C(=C/C)/NC(=O)[C@H](N)[C@H](C)CC)CSC[C@@H]1C(=O)N[C@@H]1C(=O)N2CCC[C@@H]2C(=O)NCC(=O)N[C@@H](C(=O)N[C@H](CCCCN)C(=O)N[C@@H]2C(NCC(=O)N[C@H](C)C(=O)N[C@H](CC(C)C)C(=O)N[C@H](CCSC)C(=O)NCC(=O)N[C@H](CS[C@@H]2C)C(=O)N[C@H](CC(N)=O)C(=O)N[C@H](CCSC)C(=O)N[C@H](CCCCN)C(=O)N[C@@H]2C(N[C@H](C)C(=O)N[C@@H]3C(=O)N[C@@H](C(N[C@H](CC=4NC=NC=4)C(=O)N[C@H](CS[C@@H]3C)C(=O)N[C@H](CO)C(=O)N[C@H]([C@H](C)CC)C(=O)N[C@H](CC=3NC=NC=3)C(=O)N[C@H](C(C)C)C(=O)NC(=C)C(=O)N[C@H](CCCCN)C(O)=O)=O)CS[C@@H]2C)=O)=O)CS[C@@H]1C NVNLLIYOARQCIX-MSHCCFNRSA-N 0.000 claims abstract description 36
- 108010053775 Nisin Proteins 0.000 claims abstract description 36
- 238000006243 chemical reaction Methods 0.000 claims abstract description 13
- 235000013305 food Nutrition 0.000 claims abstract description 8
- 230000002421 anti-septic effect Effects 0.000 claims abstract description 5
- 239000003963 antioxidant agent Substances 0.000 claims abstract description 4
- 230000003078 antioxidant effect Effects 0.000 claims abstract description 4
- 239000003795 chemical substances by application Substances 0.000 claims abstract description 4
- 238000010438 heat treatment Methods 0.000 claims description 20
- YBJHBAHKTGYVGT-ZKWXMUAHSA-N (+)-Biotin Chemical compound N1C(=O)N[C@@H]2[C@H](CCCCC(=O)O)SC[C@@H]21 YBJHBAHKTGYVGT-ZKWXMUAHSA-N 0.000 claims description 14
- 238000000034 method Methods 0.000 claims description 13
- 239000011259 mixed solution Substances 0.000 claims description 13
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 12
- 239000012153 distilled water Substances 0.000 claims description 9
- 238000006116 polymerization reaction Methods 0.000 claims description 9
- 238000010792 warming Methods 0.000 claims description 9
- 229960002685 biotin Drugs 0.000 claims description 8
- 239000011616 biotin Substances 0.000 claims description 8
- 235000020958 biotin Nutrition 0.000 claims description 7
- 239000000843 powder Substances 0.000 claims description 6
- 238000004108 freeze drying Methods 0.000 claims description 5
- 238000004321 preservation Methods 0.000 claims description 5
- 241000251468 Actinopterygii Species 0.000 abstract description 11
- 241000588724 Escherichia coli Species 0.000 abstract description 5
- 230000015572 biosynthetic process Effects 0.000 abstract description 4
- 238000002474 experimental method Methods 0.000 abstract description 4
- 150000001875 compounds Chemical class 0.000 abstract description 3
- 239000000376 reactant Substances 0.000 abstract description 3
- 238000003786 synthesis reaction Methods 0.000 abstract 1
- 229920001661 Chitosan Polymers 0.000 description 18
- 239000000243 solution Substances 0.000 description 9
- 150000001408 amides Chemical class 0.000 description 5
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical group CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 5
- 235000013372 meat Nutrition 0.000 description 5
- 230000003115 biocidal effect Effects 0.000 description 4
- 239000013078 crystal Substances 0.000 description 4
- 230000001408 fungistatic effect Effects 0.000 description 4
- 238000002329 infrared spectrum Methods 0.000 description 4
- 239000000203 mixture Substances 0.000 description 4
- 230000000844 anti-bacterial effect Effects 0.000 description 3
- 238000005452 bending Methods 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 239000000178 monomer Substances 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 108010005094 Advanced Glycation End Products Proteins 0.000 description 2
- 241000194035 Lactococcus lactis Species 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 235000014897 Streptococcus lactis Nutrition 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 230000000845 anti-microbial effect Effects 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 239000001257 hydrogen Substances 0.000 description 2
- 229910052739 hydrogen Inorganic materials 0.000 description 2
- 235000014655 lactic acid Nutrition 0.000 description 2
- 239000004310 lactic acid Substances 0.000 description 2
- 235000013622 meat product Nutrition 0.000 description 2
- 239000002504 physiological saline solution Substances 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- 238000001228 spectrum Methods 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 1
- 108010062877 Bacteriocins Proteins 0.000 description 1
- 108090000317 Chymotrypsin Proteins 0.000 description 1
- 241001478240 Coccus Species 0.000 description 1
- 229920000742 Cotton Polymers 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 241000191967 Staphylococcus aureus Species 0.000 description 1
- 238000002441 X-ray diffraction Methods 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- -1 amine group oligosaccharides Chemical class 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 229960002376 chymotrypsin Drugs 0.000 description 1
- 239000013066 combination product Substances 0.000 description 1
- 229940127555 combination product Drugs 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 238000005238 degreasing Methods 0.000 description 1
- 230000001934 delay Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 238000004821 distillation Methods 0.000 description 1
- 229940088598 enzyme Drugs 0.000 description 1
- 235000013373 food additive Nutrition 0.000 description 1
- 239000002778 food additive Substances 0.000 description 1
- 235000019249 food preservative Nutrition 0.000 description 1
- 239000005452 food preservative Substances 0.000 description 1
- 125000000524 functional group Chemical group 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 125000000896 monocarboxylic acid group Chemical group 0.000 description 1
- 229920001542 oligosaccharide Polymers 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 101150101567 pat-2 gene Proteins 0.000 description 1
- 239000008363 phosphate buffer Substances 0.000 description 1
- 150000003141 primary amines Chemical class 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 238000005057 refrigeration Methods 0.000 description 1
- 238000007634 remodeling Methods 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- YNJBWRMUSHSURL-UHFFFAOYSA-N trichloroacetic acid Chemical compound OC(=O)C(Cl)(Cl)Cl YNJBWRMUSHSURL-UHFFFAOYSA-N 0.000 description 1
- 238000004879 turbidimetry Methods 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L3/00—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs
- A23L3/34—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals
- A23L3/3454—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals in the form of liquids or solids
- A23L3/3463—Organic compounds; Microorganisms; Enzymes
- A23L3/3526—Organic compounds containing nitrogen
-
- 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
- A23B4/00—General methods for preserving meat, sausages, fish or fish products
- A23B4/14—Preserving with chemicals not covered by groups A23B4/02 or A23B4/12
- A23B4/18—Preserving with chemicals not covered by groups A23B4/02 or A23B4/12 in the form of liquids or solids
- A23B4/20—Organic compounds; Microorganisms; Enzymes
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L3/00—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs
- A23L3/34—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals
- A23L3/3454—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals in the form of liquids or solids
- A23L3/3463—Organic compounds; Microorganisms; Enzymes
- A23L3/3544—Organic compounds containing hetero rings
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L3/00—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs
- A23L3/34—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals
- A23L3/3454—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals in the form of liquids or solids
- A23L3/3463—Organic compounds; Microorganisms; Enzymes
- A23L3/3562—Sugars; Derivatives thereof
-
- 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
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- Polymers & Plastics (AREA)
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- Chemical Kinetics & Catalysis (AREA)
- Food Science & Technology (AREA)
- Microbiology (AREA)
- Health & Medical Sciences (AREA)
- Nutrition Science (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Food Preservation Except Freezing, Refrigeration, And Drying (AREA)
Abstract
The invention discloses a kind of compound Maillard reaction natural bacteriostatic agent and its preparation method and application.Composite natral bacteriostatic agent provided by the invention, it is the derivative that chitosan oligosaccharide nisin is prepared by Maillard reaction.Above-mentioned bacteriostatic agent provided by the invention can be used as antiseptic, antistaling agent and antioxidant in food service industry.The present invention has synthesized NEW TYPE OF COMPOSITE natural bacteriostatic agent using chitosan oligosaccharide with the used Maillard reaction of nisin.By bacteriostasis rate check experiment, the synthesis of derivatives moderately reacted improves more than 10 times for the bacteriostasis of Escherichia coli.Proved in the fresh-keeping experiment of plum fish, the reactant moderately synthesized can effectively extend the freshness date of plum fish, can effectively delay TVB N, TBA growth.
Description
Technical field
The present invention relates to a kind of composite natral bacteriostatic agent, is that chitosan oligosaccharide-nisin is prepared by Maillard reaction
Derivative, belong to biological technical field.
Background technology
Nisin（Nisin）Also referred to as nisin, it is to be closed by some streptococcus lactis in metabolic process
Into the bacteriocin with stronger antibacterial action with secretion, can be formed by the chymotrypsin degradation in human body in enteron aisle
Harmless amino acid, it is a kind of universally acknowledged safe natural biological food preservative and antiseptic.But due to lactic acid chain
Coccus element is applied to that dissolubility during food is poor, susceptible to hydrolysis and food composition（Such as protein, lipid）The deficiencies of reacting,
Cause the reduction of Nisin antibacterial activities, shorten its antibacterial time.
Chitosan oligosaccharide (Chitooligosaccharide, COS) is chitosan main chain after the degraded fracture of physics, chemistry or enzyme
The obtained degree of polymerization is 2-20 low molecule amount basic amine group oligosaccharides, has good water solubility, function scope wider, biological
The advantages that activity is high, in food service industry as antiseptic, antistaling agent and antioxidant by extensive concern.Chitosan has certain
Antibiotic property, but narrower antimicrobial spectrum and poor dissolubility limit its application in food.Chitosan oligosaccharide dissolubility is compared with shell
Glycan is stronger, but biocidal property is weaker.Nisin is good natural bacteriostatic agent, but its dissolubility is poor.
The content of the invention
The technical problems to be solved by the invention are：A kind of composite natral bacteriostatic agent is provided, is by chitosan and lactic acid
The combination of streptostacin, improves the dissolubility and biocidal property of combination product, and then improves its application in food.
In order to solve the above technical problems, the technical solution adopted by the present invention is：
Composite natral bacteriostatic agent provided by the invention, it is the derivative that chitosan oligosaccharide-nisin is prepared by Maillard reaction
Thing.
Above-mentioned bacteriostatic agent provided by the invention can be used as antiseptic, antistaling agent and antioxidant in food service industry.
The present invention also provides a kind of method for preparing described bacteriostatic agent, comprises the following steps：
（1）Chitosan oligosaccharide is with nisin with 1-10:1 mass ratio is dissolved in distilled water（Total mass concentration control is 2%）,
And adjust pH to 2-3 with food-grade HCL.
（2）Mixed solution is stirred at room temperature 2 hours to ensure fully dissolving, then by mixed solution at 40-70 DEG C
Heat 40-50h.
（3）Sample after the completion of heating cools down rapidly, and then freeze-drying is into powder, the preservation at -18 DEG C.
Preferably, in order to reach optimal combination ratio, step（1）Chitosan oligosaccharide is with nisin with 5:1 quality
Than being dissolved in distilled water.
Applicant has found that the combination of the degree of polymerization of chitosan oligosaccharide on chitosan oligosaccharide and nisin influences very by studying
Greatly, the degree of polymerization of one group of preferable chitosan oligosaccharide provided by the invention is 9-16.
In the combination of the Maillard reaction of chitosan oligosaccharide and nisin, the influence of temperature is most important, and temperature can not
It is too low can not be too high, it is too low if both reaction it is not thorough, if too high, the fungistatic effects of both combination products by
Suppress.Choosing, step of the present invention（2）In, mixed solution heats 48h at 60 DEG C.
Further, in order to reach optimal combination effect, step（2）In, heating passes through gradient-heated method, specific stream
Cheng Shi:40 DEG C of heating 8-10h, 50 DEG C of heating 12-16h are then warming up to, continue to be warming up to 60 DEG C of heating 16-20h, be warming up to 70
DEG C heating 4h.
Applicants have found that biotin is added in mixed solution can improve the knot of chitosan oligosaccharide and nisin
Conjunction rate.Preferably, it is 1-3 to add the weight of biotin and the ratio of nisin:1, promote shell few adding biotin
When sugar is combined with nisin, the degree of polymerization of optimal chitosan oligosaccharide is 6-11.
Beneficial effects of the present invention：
The present invention has synthesized NEW TYPE OF COMPOSITE natural bacteriostatic agent using chitosan oligosaccharide with the used Maillard reaction of nisin.Pass through
Bacteriostasis rate experiment understands that the synthetic moderately reacted improves 10 times for the bacteriostasis of Escherichia coli than nisin
More than.Proved in the experiment of plum fish, the reactant moderately synthesized can effectively extend the freshness date of plum fish by 4 days, Ke Yiyou
Effect delays TVB-N, TBA growth.
The present invention is successfully prepared chitosan oligosaccharide-breast using nisin and chitosan oligosaccharide as raw material by Maillard reaction
Acid streptococci element derivative, and its anti-microbial property is have studied, provided fundamental basis for the exploitation of novel and multifunctional food additives.
Brief description of the drawings
The embodiment of the present invention is described in further detail below in conjunction with the accompanying drawings.
The infrared spectrogram of Fig. 1 nisins (a), chitosan oligosaccharide (b) and its Mei Lade derivatives (c).
The X-ray diffractogram of Fig. 2 nisins (a), chitosan oligosaccharide (b) and its Mei Lade derivatives (c).
Fig. 3 differences bacteriostatic agent compares for the fungistatic effect of Escherichia coli.
Total plate count of Fig. 4 plum fishes in cold storage procedure.
The output of Fig. 5 plum fishes VBN in cold storage procedure.
TBA value of Fig. 6 plum fishes in cold storage procedure.
Embodiment
Embodiment 1
The preparation method for chitosan oligosaccharide-nisin that the present embodiment provides, comprises the following steps：
（1）Chitosan oligosaccharide is with nisin with 5:1 mass ratio is dissolved in distilled water, and total mass concentration control is 2%, is used in combination
Food-grade HCL adjusts pH to 2.5.
（2）Mixed solution is stirred at room temperature 2 hours to ensure fully dissolving, then heats mixed solution at 60 DEG C
48h。
（3）Sample after the completion of heating cools down rapidly, and then freeze-drying is into powder, the preservation at -18 DEG C.
The degree of polymerization of chitosan oligosaccharide is 13.
Embodiment 2
The preparation method for chitosan oligosaccharide-nisin that the present embodiment provides, comprises the following steps：
（1）Chitosan oligosaccharide is with nisin with 5:1 mass ratio is dissolved in distilled water, and total mass concentration control is 2%, is used in combination
Food-grade HCL adjusts pH to 2.7.
（2）Mixed solution is stirred at room temperature 2 hours to ensure fully dissolving, then heats mixed solution, heating is logical
Gradient-heated method is crossed, idiographic flow is:40 DEG C of heating 8h, 50 DEG C of heating 12h are then warming up to, continue to be warming up to 60 DEG C of heating
16h, it is warming up to 70 DEG C of heating 4h.
（3）Sample after the completion of heating cools down rapidly, and then freeze-drying is into powder, the preservation at -18 DEG C.
In this implementation, the degree of polymerization of the chitosan oligosaccharide of selection is 11.
Embodiment 3
The preparation method for chitosan oligosaccharide-nisin that the present embodiment provides, comprises the following steps：
（1）Chitosan oligosaccharide is with nisin with 5:1 mass ratio is dissolved in distilled water, and total mass concentration control is 2%, is used in combination
Food-grade HCL adjusts pH to 2.3.
（2）Mixed solution is stirred at room temperature 2 hours to ensure fully dissolving, then adds biotin, adds biotin
Weight and the ratio of nisin be 2:1, mixed solution is then heated into 48h at 60 DEG C.
（3）Sample after the completion of heating cools down rapidly, and then freeze-drying is into powder, the preservation at -18 DEG C.
In this implementation, the degree of polymerization of the chitosan oligosaccharide of selection is 8.
The structural characterization of embodiment 4
Utilize infrared spectrum and X-ray diffraction measure monomer lactic acid streptostacin（Nisin）, chitosan oligosaccharide（COS）And nisin
The structure change of rhzomorph-chitosan oligosaccharide maillard reaction product (MRPs).Infra-red sepectrometry uses KBr tablettings, with Tensor27 types
Infrared spectrometer records infrared spectrum, scanning wavelength scope 4000-400cm-1.XRD spectrum is recorded with D8 types x-ray diffractometer,
Cu K alpha rays source, the kV of tube voltage 40, the mA of tube current 40,2 °/min of sweep speed, scope is strafed as 10-80 °.
Infrared spectrum analysis is to study the indispensable instrument of microstructure of high-molecular compound, each on collection of illustrative plates
Absworption peak corresponds to atom or the situation of functional group's vibration in molecule.
Fig. 1 gives the infrared light of chitosan oligosaccharide-nisin prepared by nisin, chitosan oligosaccharide, embodiment 1
Spectrogram.In the infrared spectrum of nisin（A in Fig. 1）, 3301. 9 cm-1The broad peak at place belongs to streptococcus lactis
The COOH of element O-H stretching vibration peaks, 2963.2 cm-1The broad peak at place belongs to its COOH C-H stretching vibration peaks；1653.9
cm-1Belong to the absworption peak of acid amides band, 1535.5 cm-1Belong to the bending peak of primary amine；1449.6 cm-1Belong to COO-'s
Symmetrical stretching vibration peak.In the infrared spectrogram (b in Fig. 1) of chitosan oligosaccharide, 3444.7cm-1Having for place belongs to compared with strong absworption peak
- the NH in chitosan oligosaccharide2With-OH stretching vibration peak；2874.1cm-1The strong absworption peak at place belongs to-CH on chitosan oligosaccharide residue3
With the stretching vibration peak of-CH keys；In 1631.7cm-1、1526.3cm-1And 1394cm-1Place's absworption peak is attributed to C=O and stretched respectively
Contracting vibration (acid amides I), N-H in-plane bending vibration (acid amides II), and N-H out-of-plane bending vibration (acid amides III),
1147.58 cm-1For the asymmetric stretching vibration peak of C-O-C oxygen bridges.In the infrared light of nisin-chitosan oligosaccharide derivative
In spectrogram (c in Fig. 1), 3385.2 cm-1The strong absworption peak at place corresponds to Nisin N-H and the O-H vibration peaks of chitosan oligosaccharide,
The peak, there occurs blue shift, shows the formation of hydrogen bond between COS and Nisin compared with nisin.And it is located at 2925.5 cm-1Place
Nisin C-H stretching vibration peaks and positioned at 1632.7 cm-1The acid amides band peak at place is then there occurs red shift, this turn
Become the formation for then having further demonstrated that hydrogen bond between nisin and COS.
Fig. 2 give Mei Lade derivatives (c) and the nisin (a) of the preparation of embodiment 1, chitosan oligosaccharide (b) and its
X-ray diffractogram, it is shown that the crystal property of differential responses thing and product.Nisin monomer gives typically
The crystal formation of NaCl crystal.Chitosan oligosaccharide shows stronger diffraction maximum 2 θ=12.3 ° and 19.9 °, shows two in raw material chitosan oligosaccharide
Individual diffraction maximum is crystal I, II characteristic peak respectively.From nisin and the maillard reaction product of chitosan oligosaccharide（MRPs）'s
Diffraction pattern can be seen that the diffraction maximum of the sample, and there occurs a fixed width；Compared with chitooligose monomer, the characteristic peak of product is to low
Angle is offset, and shows that Maillard reaction can reduce the crystallinity of reactant.
The biocidal property of embodiment 3
Standard strain（Escherichia coli and staphylococcus aureus）Adjusted through overactivation and with turbidimetry to fixed initial concentration
Afterwards, ten times of gradient dilutions are to suitable concn.Trained after 4.5ml bacterium solutions are mixed with 0.5ml bacteriostatic agents or physiological saline at 37 DEG C
Support one hour.Then extract 1ml mixed liquors and carry out flat board culture.Flat board cultivates 48 ± h at 37 DEG C.Bacteriostatic agent be by by it
It is preceding it is lyophilized after powder dissolve in 20Mm phosphate buffers（PH 6.0）In, concentration is (50 ug/ml).
Fig. 3 is fungistatic effect of the different bacteriostatic agents for Escherichia coli.It is nisin to scheme A bacteriostatic agents, and figure B is
Chitosan oligosaccharide, figure C are chitosan oligosaccharide-nisin mixture, and figure D is that chitosan oligosaccharide-nisin that embodiment 1 makes is multiple
Compound, figure E are chitosan oligosaccharide-nisin compound that embodiment 2 makes, and figure F is chitosan oligosaccharide-breast that embodiment 3 makes
Acid streptococci element-biotin composite.
Can intuitively it find out very much from Fig. 3, the fungistatic effect difference of different bacteriostatic agents is clearly.
The concrete application example of embodiment 4-plum fish is fresh-keeping
1. sample treatment
By fresh plum fish with being divided into 4 groups, it is respectively put into different disposal group solution and soaks 30 minutes（Solution concentration is 1.5%, PH
It is worth for 6.0 ± 0.01）, taking-up dries, weighs and load valve bag, is then placed into refrigerating in 4 ± 1 DEG C of refrigerators.Sample every
It takes out detection.
The condition of 4 kinds of different disposal groups is：It is prepared by distilled water, chitosan oligosaccharide-nisin mixture, embodiment 2
Bacteriostatic agent prepared by bacteriostatic agent and embodiment 3.
Wherein, in accompanying drawing 4-5 legend, what Open circle represented is chitosan oligosaccharide-nisin mixture；Evil mind is justified
What shape represented is distilled water；What blank square represented is bacteriostatic agent prepared by embodiment 2；What black squares represented is to implement
Bacteriostatic agent prepared by example 3.
2. total plate count
With reference to national standard GB 47892-2010, the refrigeration plum fish of tail and physiological saline is taken to be put into homogenizing bag and pat 2 minutes（Pay attention to
Record every group of fish gross weight）, leachate is diluted to 3 suitable concentrations, respectively takes 1ml to be separately added into culture dish, is mixed,
（30±1）Cultivated 72 hours at DEG C.As a result such as Fig. 4.
3. VBN
According to GB/T 5009.44-2003《Meat and the analysis method of meat products sanitary standard》] in Micro-kjoldahl method survey
It is fixed, determine 3 times, average.As a result such as Fig. 5.
4.PH values
According to GB/T 9695.5-2008《Meat determines with meat products pH》, take the g of meat sample 10 to shred every time, add 90 mL distillations
Water, 30 min of vibration are mixed, pH value determination, are repeated 3 times, results averaged.
5.TBA
10 g meat samples are taken to add 20mL20% solution of trichloroacetic acid.3 min are beaten in beater.5 minutes are stood, double-deck degreasing
Cotton is filtered once, is pipetted 5 mL supernatants with pipette, is added in 20 mL test tubes, add 5 mL TBA solution, be stoppered bottle
Mouthful.Test tube cools down 30min after the min. of water-bath 40 takes out at 100 DEG C in room temperature water.Take supernatant colorimetric at 532 nm.
Record A values and calculate TBA values with below equation.Amount (TBA) with the material of TBA reactions is with the mg numbers of the MDA in every kg meat
To represent.As a result such as Fig. 6.
TBA values (mg/kg)=A532 x 41.5
In formula：A532 is absorbance of the solution at 532 nm；41.5 be per conversion coefficients of the mL containing content of MDA.
The above described is only a preferred embodiment of the present invention, being not the limitation for making other forms to the present invention, appoint
What those skilled in the art changed or be modified as possibly also with the technology contents of the disclosure above equivalent variations etc.
Imitate embodiment.But it is every without departing from technical solution of the present invention content, the technical spirit according to the present invention is to above example institute
Any simple modification, equivalent variations and the remodeling made, still fall within the protection domain of technical solution of the present invention.
Claims (8)
1. a kind of composite natral bacteriostatic agent, it is the derivative that chitosan oligosaccharide and nisin are prepared by Maillard reaction.
2. application of the bacteriostatic agent described in claim 1 in food service industry as antiseptic, antistaling agent and antioxidant.
3. the bacteriostatic agent described in claim 1, it is characterised in that preparation method comprises the following steps：
（1）Chitosan oligosaccharide is with nisin with 1-10:1 mass ratio is dissolved in distilled water（Total mass concentration control is 2%）,
And adjust pH to 2-3 with food-grade HCL；
（2）Mixed solution is stirred at room temperature 2 hours to ensure fully dissolving, then heats mixed solution at 40-70 DEG C
40-50h；
（3）Sample after the completion of heating cools down rapidly, and then freeze-drying is into powder, the preservation at -18 DEG C.
4. prepare the method described in claim 3, it is characterised in that step（1）Chitosan oligosaccharide is with nisin with 5:1 matter
Amount ratio is dissolved in distilled water.
5. according to the method for claim 3, it is characterised in that the degree of polymerization of chitosan oligosaccharide is 9-16.
6. according to the method for claim 3, it is characterised in that step（2）In, heating passes through gradient-heated method, specific stream
Cheng Shi:40 DEG C of heating 8-10h, 50 DEG C of heating 12-16h are then warming up to, continue to be warming up to 60 DEG C of heating 16-20h, be warming up to
70 DEG C of heating 4h.
7. according to the method for claim 3, it is characterised in that step（2）In, after mixed solution fully dissolves, add in addition
Enter biotin, it is 1-3 to add the weight of biotin and the ratio of nisin:1.
8. according to the method for claim 7, it is characterised in that the degree of polymerization of chitosan oligosaccharide is 6-11.
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| CN111543577A (en) * | 2020-04-07 | 2020-08-18 | 舟山昌国食品有限公司 | Fishy smell removing method for small cuttlefish product |
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