CN109679868A - A kind of microorganism nitrosation inhibitor and preparation method thereof - Google Patents
A kind of microorganism nitrosation inhibitor and preparation method thereof Download PDFInfo
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- CN109679868A CN109679868A CN201811626173.9A CN201811626173A CN109679868A CN 109679868 A CN109679868 A CN 109679868A CN 201811626173 A CN201811626173 A CN 201811626173A CN 109679868 A CN109679868 A CN 109679868A
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- nitrosation
- microorganism
- inhibitor
- product
- nitrosamine
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- 230000002265 prevention Effects 0.000 description 1
- 230000017854 proteolysis Effects 0.000 description 1
- 239000012925 reference material Substances 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 235000021108 sauerkraut Nutrition 0.000 description 1
- 235000010344 sodium nitrate Nutrition 0.000 description 1
- 239000004317 sodium nitrate Substances 0.000 description 1
- 229940063673 spermidine Drugs 0.000 description 1
- 229940063675 spermine Drugs 0.000 description 1
- 235000013599 spices Nutrition 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 229960003732 tyramine Drugs 0.000 description 1
- 238000002604 ultrasonography Methods 0.000 description 1
- 210000004885 white matter Anatomy 0.000 description 1
- 235000020097 white wine Nutrition 0.000 description 1
- 235000013618 yogurt Nutrition 0.000 description 1
Classifications
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
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- 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
- A23L5/00—Preparation or treatment of foods or foodstuffs, in general; Food or foodstuffs obtained thereby; Materials therefor
- A23L5/20—Removal of unwanted matter, e.g. deodorisation or detoxification
- A23L5/28—Removal of unwanted matter, e.g. deodorisation or detoxification using microorganisms
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- 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
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
- A23V2400/165—Paracasei
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- 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
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
- A23V2400/179—Sakei
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Abstract
The invention belongs to food processing technology field, the lactic acid bacteria to solve the problems, such as living is not suitable for no zymotechnique product, provides a kind of microorganism nitrosation inhibitor and preparation method thereof.The microorganism nitrosation inhibitor is that staphylococcus xylosus, Lactobacillus saki and class lactobacillus plantarum composite fermentation culture are prepared.Solve the problems, such as that lactic acid bacteria living is not suitable for no zymotechnique product, the substance of N- nitrosamine formation is able to suppress by being directly made, avoiding lactic acid bacteria living influences product special flavour, and the microbial inhibitor not only inhibits the formation of N- nitrosamine, reduce nitrate residue, improve the security quality of product, the flavor of product is also improved while not influencing the qualities such as product color texture, application effect is good, it is equally applicable for the product such as burger of some no zymotechniques, sausage, in the products such as salted vegetables, improving to some easy product safety qualities for producing nitrosamine has far reaching significance.
Description
Technical field
The invention belongs to food processing technology fields, and in particular to a kind of microorganism nitrosation inhibitor and its preparation side
Method.
Background technique
N- nitrosamine is a kind of strong carcinogen, has carcinogenic, teratogenesis, mutagenicity, has greatly danger to human health
Evil, it mainly by nitrogen oxides and secondary amine reaction generate, for rich in protein, there are a certain amount of nitrite,
And holding putrefactive food easily to generate N- nitrosamine, and pH, temperature etc. is an important factor for influencing its generation.In butcher's meat system
In product processing, in order to play the role of color development, antibacterial, anti-oxidant, raising flavor, according to national standards, nitrite simultaneously
Maximum additive amount can reach 0.15 g/kg, and in processing, storage, the catabolite of protein can be N- nitrosamine in meat
Generation provide precursor, the influence of acid extraction is also greatly promoted the generation of N- nitrosamine in addition, so butcher's meat
The potential danger of N- nitrosamine formation is had in product.Therefore the content of N- nitrosamine in product how is reduced for food safety
Controlling party face has great importance.
Lactic acid bacteria is a kind of general name that can be acted on carbohydrate and generate the gram-positive bacteria of lactic acid, has enhancing
Immunity, prevention and treatment lactose intolerance, adjusting intestinal flora, inhibition pathogenic bacteria and other effects, are widely used in traditional fermented food
(such as Yoghourt, sauerkraut, ferment sausage), is improved the effect of product special flavour and nutrition.At present scholar using lactic acid bacteria act on come
Inhibit the formation spininess of N- nitrosamine in food for fermented food, be such as inoculated with viable bacteria in food production and ferment, to rise
To the inhibiting effect of degradation and amine substance formation to N- nitrosamine precursor such as nitrite, or directly inhibit N- nitrous
Amine is formed.But the product of no zymotechnique is less applicable in if in the processing such as bologna sausage.Early-stage study discovery, by what is pickled
Inoculating lactic acid bacterium leavening agent while auxiliary material is added in meat stuffing in de-airing mixer, first passes around the fermentation of 12 h after filling
Journey, according still further to bologna sausage processing technology be dried, boiling, baking, the techniques such as sootiness, test result shows that inoculating lactic acid bacterium is sent out
The formation that N- nitrosamine in bologna sausage can actually be reduced after ferment, can also reduce the residual quantity of nitrite, improve the safety of product
Property, but fermentation can extend the process-cycle of product, require process equipment and environmental sanitation higher, it is often more important that after fermented
Bologna sausage product has acescency, and consumer is not easily accepted by.
In view of the above problems, it is proposed that probing into its inhibition it is assumed that this special lactic acid bacteria strains can be cultivated in vitro
The mechanism that N- nitrosamine is formed extracts the substance for wherein effectively inhibiting N- nitrosamine, is then directly appended to similar bologna sausage, Baconic
In meat products Deng not fermenting step, the edible safety of Lai Tigao meat products, this will have vast potential for future development.
Summary of the invention
The present invention by using the advantage of lactobacillus-fermented technology in combination with causing to produce after lactobacillus-fermented in practical application
Product Production Time extends, unstable products, and big, the problems such as product is unstable is influenced on product special flavour, provides a kind of micro- life
Object nitrosation inhibitor and preparation method thereof.By way of In Vitro Fermentation culture, respectively in the degradation and suppression to nitrite
N- N-nitrosodimethylamine processed forms two aspects and carries out Stepwise Screening, and research PRO-MIX5 lactic acid bacteria fermenting agent inhibits N- nitrosamine
The mechanism of formation extracts the targeting substance wherein effectively to work, microorganism nitrosation inhibitor is made and is added into product, real
Can now inhibit the formation of N- nitrosamine, improve Product Safety, while not influencing the purpose of product sensory quality again, this for
Production safety, high-quality food have great importance.
The present invention is realized by following technical solution: a kind of microorganism nitrosation inhibitor, the microorganism nitrosation suppression
Preparation is 2 × 1011CFU staphylococcus xylosus, 2 × 1011CFU Lactobacillus saki and 2 × 1011The compound hair of CFU class lactobacillus plantarum
Ferment culture is prepared.
Specifically the preparation method comprises the following steps: using the MRS broth bouillon culture staphylococcus xylosus added with N- nitrosamine, pure mellow wine
To OD600=1.8-2.4, by 5000-7000g, 4 DEG C of 15 min of centrifugation obtain thallus, use for lactobacillus and class lactobacillus plantarum
The PBS of 25 mmol/L, pH=6 are rinsed thallus 1 ~ 2 time, then with the PBS with the dilution proportion of 1:3-10, addition final concentration of 1 ~ 5
After 30 DEG C of 2 ~ 5h of effect of lysozyme of mg/mL, 5 ~ 10 min of ultrasonic disruption, through 8000 ~ 12000 g, 4 DEG C of centrifugation 10-15
The precipitating that min is obtained is microorganism nitrosation inhibitor.
The staphylococcus xylosus, Lactobacillus saki, class lactobacillus plantarum mixed bacteria culture medium be added with 0.01 ~
The MRS broth bouillon of 0.5 μ g/mL concentration N- nitrosamine standard items.
Staphylococcus xylosus of the present invention, Lactobacillus saki, class lactobacillus plantarum mixed bacteria are PRO-MIX5
Commercial fermentation agent is manufactured by Italian Sa Ke company.
A kind of microorganism nitrosation inhibitor inhibits the application of N- nitrosamine, concrete application method in meat products
Are as follows: 0.02 ~ 1% microorganism nitrosation inhibitor of adding raw materials meat weight in meat products.
The invention has the following beneficial effects: the present invention is using the advantage of lactobacillus-fermented technology in combination with existing in practical application
The problem of, the lactic acid bacteria fermenting agent PRO- that N- nitrosamine is formed in bologna sausage can effectively be inhibited by screening to obtain using pre-stage test
MIX5(staphylococcus xylosus+ask wine lactobacillus+class lactobacillus plantarum), by way of In Vitro Fermentation culture, it is used respectively
Meat gruel culture medium and MRS culture medium culture are tested in simulated system, by different processing modes by nitrous acid in vitro
Salt degradation effect eliminates metabolite, endocellular enzyme, bacterium to the inhibiting effect Stepwise Screening directly formed to N- nitrosamine respectively
The effect of body, obtains the mechanism of action that lactic acid bacteria inhibits N- nitrosamine to be formed, and obtained one kind can be added directly into the system of product
In work, to inhibit the formation of N- nitrosamine, so that product is not required to fermentation and can obtain lactic acid bacteria bring beneficial effect, improve
The security quality of product, while the microorganism nitrosation inhibitor of product sensory quality is not influenced, solve zymotechnique to nothing
The adverse effect of fermented product such as requires high ask in the presence of destruction product original local flavor, extension process time, required production environment
Topic, also food-safe controlling party face has great importance.
The production of the microbial inhibitor solves the problems, such as that lactic acid bacteria living is not suitable for no zymotechnique product, passes through
The substance for being able to suppress the formation of N- nitrosamine is directly made, avoiding lactic acid bacteria living influences product special flavour, and micro- life
Object inhibitor not only inhibits the formation of N- nitrosamine, reduces nitrate residue, the security quality of product is improved, not
It influences to also improve the flavor of product while the qualities such as product color texture, application effect is good, is equally applicable for some nothings
The product of zymotechnique such as burger, sausage in salted vegetables product, have some easy product safety qualities raisings for producing nitrosamine
Far reaching significance.
Detailed description of the invention
Fig. 1 is the flow chart of screening production microorganism nitrosation inhibitor；
Fig. 2 is the effect of the FS and MFB of Different adding amount in different time to nitrite in simulated system, A, B, C figure difference
Indicate that addition FS and MFB 1.5mL, 3mL, 6mL, shoulder mark capitalization are indicated with the significant difference between processing group different time
(P<0.05)；
Fig. 3 is effect of the addition 20mLFS in different temperatures, different time to nitrite in simulated system, shoulder under different temperatures
Mark capitalization indicate between processing group different time significant difference (P<0.05)；
Fig. 4 is, to the operative condition of nitrite, different shoulder mark capitalizations indicate same anti-after PRO-MIX5 enzyme r e lease intracellular
Between seasonable between different disposal group significant difference (P<0.05), lowercase indicates between differential responses time same processing group
Significant difference (P<0.05)；
Fig. 5 is influence of the Different adding amount of microorganism nitrosation inhibitor to bologna sausage pH, and shoulder mark capitalization indicates different same
Between processing group significant difference (P<0.05)；
Fig. 6 is influence of the Different adding amount of microorganism nitrosation inhibitor to bologna sausage red scale value, and shoulder mark capitalization indicates not
With between same processing group significant difference (P<0.05)；
Fig. 7 is influence of the Different adding amount of microorganism nitrosation inhibitor to bologna sausage elasticity, and shoulder mark capitalization indicates different
With between processing group significant difference (P<0.05)；
The Different adding amount of Fig. 8 microorganism nitrosation inhibitor in bologna sausage to the influence of nitrite, shoulder mark capital letter matrix
Show it is different between processing group significant difference (P<0.05)；
Fig. 9 is influence of the Different adding amount of microorganism nitrosation inhibitor to sootiness Baconic pH, and shoulder mark capitalization indicates not
With between same processing group significant difference (P<0.05)；
Figure 10 is influence of the Different adding amount of microorganism nitrosation inhibitor to nitrite in sootiness Baconic, the capitalization of shoulder mark
Letter indicate it is different between processing group significant difference (P<0.05).
Specific embodiment
Embodiment 1: a kind of microorganism nitrosation inhibitor, the microorganism nitrosation inhibitor are 2 × 1011CFU xylose
Staphylococcus, 2 × 1011CFU Lactobacillus saki and 2 × 1011CFU class lactobacillus plantarum composite fermentation culture is prepared.Specifically
The preparation method comprises the following steps: being planted using MRS broth bouillon culture staphylococcus xylosus, Lactobacillus saki and class added with N- nitrosamine
Object lactobacillus is to OD600=1.8-2.4, and by 5000-7000g, 4 DEG C of 15 min of centrifugation obtain thallus, with 25 mmol/L, pH=
6 PBS is rinsed thallus 1 ~ 2 time, then adds the lysozyme of final concentration of 1 ~ 5 mg/mL with the PBS with the dilution proportion of 1:3-10
After 30 DEG C of 2 ~ 5h of effect, 5 ~ 10 min of ultrasonic disruption, through 8000 ~ 12000 g, 4 DEG C are centrifuged the precipitating that 10-15 min is obtained
As microorganism nitrosation inhibitor.
The staphylococcus xylosus, Lactobacillus saki, class lactobacillus plantarum mixed bacteria culture medium be added with 0.01 ~
The MRS broth bouillon of 0.5 μ g/mL concentration N- nitrosamine standard items.
Experimental material: fresh pork is purchased from the healthy and free from worry meat products Co., Ltd in Tianjin；Use lactic acid bacteria fermenting agent: PRO-
MIX5(staphylococcus xylosus+Lactobacillus saki+class lactobacillus plantarum), Italian Sa Ke company.
MRS broth bouillon, Beijing Suo Laibao Science and Technology Ltd；Methylene chloride (chromatographically pure), boric acid, potassium chloride, Asia
Sodium nitrate (NaNO2), Tianjin Fengchuan Chemical Reagent Science & Technology Co., Ltd..Dimethylamine hydrochloride (DMA HCl), Chinese medicines group
Learn reagent Co., Ltd.The mixed mark (N-Nitrosamines, NAS) of 9 kinds of N- nitrosamine, includes N- N-nitrosodimethylamine (N-
Nitrosodimethylamine, NDMA), N- diethylnitrosamine (N-nitrosodiethylamine, NDEA), N- methyl
Ethyl nitrosamine (N-nitrosomethylethylamine, NMEA), N- dibutyl nitrosamine (N-
Nitrosodibutylamide, NDBA), N- dipropyl nitrosamine (N-nitrosodipropylamine, NDPA), N- nitrous
Phenylpiperidines (N-nitrosopiperidine, NPIP), N- nitrosopyrolidine (N-nitrosopyrrolidine, NPYR), N-
Nitrosomorpholine (N-nitrosomorpholine, NMOR), N nitrosodiphenyl amine (N-nitrosodiphenylamine,
NDPheA), U.S. Supelco company.
Instrument and equipment: 7890A gas chromatograph, Agilent company of the U.S.；P/ACE MDQ capillary electrophoresis is equipped with purple
External detector, U.S. Beckman Kuerle company；PB-10 acidometer, German Sai Duolisi scientific instrument Co., Ltd；ZXSD-
B1090 constant incubator, the sincere experimental instruments and equipment limited of Shanghai intelligence；The universal high speed freezing centrifuge of Z323K, U.S.'s match is silent to fly
Generation that Science and Technology Ltd.；II D sonicator of SCIENTZ-, Xin Zhi bio tech ltd, Ningbo；HS07-314 is permanent
Warm water bath, Tianjin North China laboratory apparatus Co., Ltd；VXMNAL vortex oscillator, Ao Haosi company, the U.S..
The production of meat gruel culture medium: with lean pork: water=1:2 ratio homogenate, after 4 boiling water baths heat 10 mim, homogenate
In 121 DEG C of 20 min of high pressure sterilization, 4% glucose is added in gained liquid, obtains meat gruel culture medium, is used for desk study mistake
PRO-MIX5 lactic acid bacteria is cultivated in journey to use.
PRO-MIX5 lactic acid bacteria culture processing mode: extract thallus: 5000 g, 4 DEG C of 15 min of centrifugation discard supernatant
Obtain thallus；Rinse thallus: with the phosphate buffer (Phosphate buffer solution, PBS) of 25 pH=6 mM
5000 g, 4 DEG C of 15 min of centrifugation are rinsed thallus 2 times；Broken thallus: the lysozyme of final concentration of 1 mg/mL of addition is at 30 DEG C
2 h are acted on, then with ultrasonic cell disruption instrument with the power of 200 W, 2 s of ultrasound stop 2 s totally 5 min.Extract bacterial chip:
10000 g, 4 DEG C of 10 min of centrifugation.
NDMA simulated system preparation: with reference to " Inhibition of N-nitrosation of such as Kuniyuki
Secondary amines in vitro by tea extracts and catechins " method, and be modified slightly, make
NaNO2 and DMA HCl final concentration is respectively 20 mmol/L and 10 mmol/L in reaction system, and the above reagent is with concentration
After the system is reacted 1 h in 80 DEG C of water-baths, 6 mol/L are added in 25 mmoL/L, the phosphate buffered saline of pH=6
NaOH adjusts pH to 13 to terminate nitrosation reaction, is placed in ice bath after cooling down rapidly and 5 mL methylene chloride are added, fully shake 1
Min is centrifuged 5 min, the methylene chloride of suction foot after mixing well methylene chloride and sample under the conditions of 4 DEG C, 10000 g
The limpid part of layer is crossed after 0.45 μm of filter membrane in the bottle for being transferred to gas chromatograph to upper machine.
Index determining method: the measurement of nitrite: referring to " HPCE method detect in fruits and vegetables and meat products simultaneously nitrate with
Content of nitrite " in method measure nitrate residue.
The measurement of N- nitrosamine: referring to GB 5009.26-2016 " measurement of N- nitrosamine compound in food " to sample
N- nitrosamine in product extracts, extracting and purifying, crosses film after concentration.GC conditions used in this test are as follows: sample volume: 1
μL；Injector temperature is 250 DEG C；Column temperature rise gradient is 50 DEG C of 4 min of holding, and 10 DEG C/min speed rises to 180 DEG C and keeps 2
Min, 20 DEG C/min rise to 220 DEG C, keep 10 min, rear operation is with 235 DEG C of 2 min of holding；Nitrogen phosphorous detector temperature: 330 DEG C；
6 mL/min of 2 mL/min of hydrogen flow rate, air velocity 60 mL/min, carrier gas N2 flow velocity.The sample gas phase after film will be crossed
Chromatographic condition measures 9 kinds of N- nitrosamine, while analyze quantitative.
The measurement of pH value: it is measured referring to the method for GB 5009.237-2016 " measurement of food pH value ".
Data analysis: calculating average value and standard deviation with Microsoft Excel 2010, with Statistix 8.1 into
Row significance analysis, SigmaPlot 10.0 map.
Example 1: PRO-MIX5 is cultivated in meat gruel culture and makes microbial inhibitor
Since PRO-MIX5 is the advantage milk fermentation agent for obtaining being able to suppress the formation of N- nitrosamine using screening in bologna sausage, and
Ferment 12 h when effect it is best, for keep making in vitro as far as possible when products therefrom it is consistent, this test connects lactic acid bacteria fermenting agent
Kind 12 h(35 DEG C of liquid meat gruel culture medium fermentation obtained into preceding method).
Degradation of the fermentating metabolism product to nitrite: often adding nitrite in meat products processing, and nitrous
Hydrochlorate is the precursor to form carcinogenic substance N- nitrosamine.It is contemplated that whether sub- to reduce N- by reducing nitrate residue
The formation of nitramine.Lactic acid bacteria metabolite is normally present in fermented supernatant fluid, wherein containing a large amount of lactic acid, lactein, peroxide
Change the substances such as hydrogen, in conjunction with trial test as a result, guess may is that the fermentating metabolism product of lactic acid bacteria reduces content of nitrite.In
It is to test using the preferred PRO-MIX5 commercial fermentation agent of pre-stage test as research object, be seeded in liquid meat gruel culture medium and train
Obtained mixed fermentation liquid (Mixed fermentation broth, MFB) is supported, through 5000 g, 4 DEG C of 15 min of centrifugation are sent out
Ferment supernatant (Fermentation supernatant, FS), the as metabolite of the commercial fermentation agent.It is this to understand
Whether activity, operative temperature and the action time of MFB and FS can remove the nitrite in simulated system, devise 2
A test, the operative temperature of testing program one are set as 4 DEG C (marinated temperature of simulation meat), add respectively in 50 mL centrifuge tubes
Enter each 1.5 mL of MFB and FS, 3 mL, 6 mL, NaNO is added with final concentration of 0.15 mg/mL2Standard solution adds distilled water
To 30 mL, the action time of MFB and FS are respectively 0 h, 8 h, 16 h, the nitrous in stipulated time immediately measurement system
It is as shown in Figure 2 to obtain result for phosphate content.Fig. 2 test result shows the extension with action time and adds the increasing of concentration
Add, FS and MFB cannot content of nitrite significantly in degradation system.
In testing program two, for the effect of further verifying metabolite (FS), the addition concentration of FS is improved, takes 20
NaNO is added in 50 mL centrifuge tubes, with final concentration of 0.15 mg/mL in mL FS2Standard solution, addition distilled water to 30 mL,
Be respectively placed in 4 DEG C (simulate marinated temperature) and 35 DEG C of (simulation fermentation temperature) environment and act on, effect 0,8,21,32h stands
Nitrite i.e. in sampling and measuring system, as a result as shown in figure 3, from figure 3, it can be seen that being added under the conditions of 4 DEG C and 35 DEG C
Content of nitrite does not significantly reduce with the extension of reaction time in the simulated system of FS, it is seen that with the increasing of FS additive amount
Add, the raising of reaction temperature and the metabolite for extending thallus fermentation in reaction time are still without significant degrading nitrite
Effect, to sum up result strongly suggests that the substance that degrading nitrite can be played in fermentation is not present in the hair of lactic acid bacteria
In ferment metabolite.
To the degradation of nitrite after lactic acid bacteria enzyme r e lease intracellular: the report such as Zhang Xinyue, nitrous of degrading in LCR6013
Hydrochlorate is mainly the effect of nitrite reductase, and the enzyme belongs to endocellular enzyme, cell homogeneity gap is primarily present in, to nitrous
Hydrochlorate degradation is significant, is secondly cytoplasm enzyme and cell fragment, in fermentating metabolism product to the degradation of nitrite
Experiment results proved lactic acid bacteria metabolite it is not significant to nitrite degradation, this promotes it is contemplated that may is that endocellular enzyme
In action.So by meat gruel culture medium culture 12 h of the PRO-MIX5 lactic acid bacteria added with nitrite (10 μ g/mL)
Streptococcus acidi lactici fermented solution is obtained after (35 DEG C), the purpose for adding nitrite here is to promote degrading nitrite as inducer
Substance generates, and its additive amount is larger to degradation effect correlation.4 are obtained according to the pretreatment mode (1) (2) (3) (4) of table 1
Group mixed liquor carries out bacteriolyze enzyme effect to thallus and clasmatosis is handled, and gained liquid is respectively after so that it is released endocellular enzyme
MFB(1)；MPW(2) (mixture of precipitation and water)；MPP(3) (mixture of
Precipitation and PBS)；LA(4) (lactic acid) takes 20 mL in 50 mL centrifuge tubes, respectively with final concentration of
NaNO2 standard solution is added in 0.1 mg/mL, and addition distilled water to 30 mL is placed at 30 DEG C and reacts, and measures the 0th h, 1 respectively
H, in 2 h reaction systems nitrite residual quantity, as a result as shown in Figure 4.Test result shows as made from PRO-MIX5
MFB(1), MPW(2), MPP(3), LA(4) with the extension of reaction time, degradation (P < can be played to nitrite
0.05), and Different treatments to the descending sequence of nitrite degradation rate successively are as follows: (2) > (1) > (4) > (3), wherein
(1) and (2) processing mode is specifically as shown in table 2 to nitrite degradation rate during insulation reaction, illustrates clasmatosis
After releasing endocellular enzyme, nitrite degradation is acted on significantly, but processing mode (2) degradation effect is due to intracellular better than (1)
It is unstable after enzyme r e lease, thereby increases and it is possible to its activity of function influence by such as hydrogen peroxide of metabolite in supernatant, and use water
Broken thallus after fermented supernatant fluid is replaced then so that endocellular enzyme and bacterial chip is played a role, when keeping the temperature 2 h to Asia
The degradation rate of nitrate can not preferably maintain nitrite up to 53.49%, Xiang Fanyong, 0.01 mol/L, the PBS of pH=7.4
The stability of reductase, is greatly reduced enzymatic activity instead, this result and consulted reference materials result are inconsistent, it may be possible to due to strain
Difference, the tolerance of pH and concentration to PBS is also different.
The testing program of the production two kinds of leavening degrading nitrite liquid of PRO-MIX5 and LC of table 1
2 MFB(1 of table) and MPW(2) during insulation reaction to the degradation rate of nitrite
MFB(1 the influence that) and MPW(2) NDMA is formed: the degradation of nitrite is made in conjunction with after lactic acid bacteria enzyme r e lease intracellular
With obtaining the MFB(1 that can be effectively reduced content of nitrite) and MPW(2), but the final purpose of this test is to can reduce
The formation of N- nitrosamine, and due to only providing NDMA residual quantity no more than 3 μ g/kg in meat products in current national standard, with
The simulated system for forming NDMA further screens the production of microbial inhibitor.It is added in the simulated system for generating NDMA
MFB(1) and MPW(2) each 5 mL is in the NDMA simulated system that total volume is 20 mL, 80 DEG C of heating after 30 DEG C of 2 h of insulation reaction
1 h, blank control group (control check, CK group) then directly heat reaction, measure the production quantity of NDMA.Obtain result such as
Shown in table 3, MFB(1) and MPW(2) it is greatly promoted the formation of NDMA, this may be since this step is using meat gruel
Culture medium, and after inoculum fermented, protein degradation makes to mention in system containing second level amine substance for the formation of N- nitrosamine
A large amount of precursor is supplied.Unlike but, MFB(1) MPW(2 is significantly higher than to the NDMA promotion ability formed) (P<0.05),
And its promotion ability is almost MPW(2) twice, MPW(2) and difference (see Table 2) MFB(1) be that MPW(2) be centrifuged
Supernatant is removed, thus illustrates that there may be the substances that NDMA can be promoted to be formed in the metabolite of the leavening, into one
There may be the substances for being able to suppress NDMA formation in the prepared fermentation liquid precipitate of step guess, and the substance is likely to be present in bacterium
In body.
3 MFB(1 of table) and the MPW(2) influence to NDMA formation
Note: different shoulder mark capitalizations indicate the significant difference (P < 0.05) of NDMA production quantity between different disposal group.
Example 2. makes microbial inhibitor with MRS culture medium culture PRO-MIX5
The precursors such as amine substance are provided to exclude meat gruel culture medium as the formation of N- nitrosamine, and further energy in verifying 2.1.3
Inhibit the active principle of NDMA to be present in the guess in thallus, PRO-MIX5 is seeded to MRS broth bouillon in this step
In cultivated.
The influence that the thallus of Different treatments forms NDMA: this step will be in the MRS for being added with 10 μ g/mLNaNO2
With viable bacteria, (thallus that centrifugation MRS culture medium obtains rinses thallus with PBS to the PRO-MIX5 thallus of culture medium amplification cultivation respectively
It is added twice) with the state of dead bacterium (by the thallus after flushing in 100 DEG C of 15 min of heating water bath) with the additive amount of 0.1 g of weight in wet base
20 mL are formed in the simulated system of NDMA, and have screened the different modes of action: direct 80 DEG C of heating are anti-after simulated system is added
Answer 1 h；First 1 h is reacted in 80 DEG C of heating again after 2 h of insulation effect at 30 DEG C, while doing blank control.Obtain result such as 4 institute of table
Show: available 4 kinds of application modes to some extent promote NDMA formed, but facilitation effect far below 2.1.3's as a result, its
Middle viable bacteria keeps the temperature 2 h NDMA forming amounts in simulated system and is apparently higher than other three groups, and adds the direct nitrosation reaction of viable bacteria
NDMA forming amount illustrates to react during viable bacteria keeps the temperature 2 h and in 80 DEG C, viable bacteria may also above dead bacterium group in group
Produce some substances that NDMA can be promoted to be formed.And dead bacterium and dead bacterium keep the temperature 2 h in simulated system and promote to imitate to NDMA
Fruit is almost consistent, and promotion rate is lower, compares viable bacteria result, thus it is speculated that may contain the object for inhibiting N- nitrosamine to be formed in thallus
Matter, and the substance is not a kind of active protease, has certain heat resistance.
The influence that the different thallus states of table 4 and different application mode form NDMA
Note: in table different shoulder mark capitalizations indicate NDMA production quantity between different disposal groups significant difference (P<0.05).
The targeting substance that can effectively inhibit NDMA to be formed in positioning thallus: further to verify guess, centrifugation amplification training
MRS fluid nutrient medium after supporting PRO-MIX5 obtains thallus, after rinsing thallus twice with PBS, with thallus: the ratio of PBS=1:4
Dilution, after bacterial cell disruption, the supernatant being centrifuged is enzyme extract intracellular, is precipitated as bacterial chip.Respectively by supernatant,
Precipitating, mixture (supernatant+precipitating), which are added in the NDMA simulated system of total volume 20mL, to react, while blank control is arranged
Group, measurement NDMA final forming amount.Obtain that the results are shown in Table 5: precipitating group, i.e. bacterial chip (the packet of addition PRO-MIX5
Containing substances such as cell wall fragments, cell membrane fragments, organelle, inclusion bodys) there is significant inhibitory effect to the formation of NDMA, it can
Can there is certain suction-operated to N- nitrosamine due to bacterial chip, but its specific mechanism still needs to further further investigate；
And supernatant and mixture then promote NDMA and are formed, wherein mixture group facilitation effect is noticeably greater than supernatant, illustrates to precipitate
The presence of object bacterial chip may will increase the facilitation that supernatant forms NDMA, and the mechanism specifically to interact is urgently
Research.
The influence that the product of 5 PRO-MIX5 thallus of table different location after crushing forms NDMA
Note: in table different shoulder mark capitalizations indicate NDMA production quantity between different disposal groups significant difference (P<0.05).
Screening to lactic acid bacteria training method, bacterial chip additive amount: obtaining bacterial chip according to result above is PRO-
The active principle that MIX5 inhibits NDMA to be formed, in conjunction with inducer in culture medium in 1 addition to the importance of degradation effect, respectively
(inducer is not added using four kinds of MRS fluid nutrient mediums；Add 10 μ g/mL NaNO2；Add 0.04 μ g/mL NAS；Addition
10 μg/mL NaNO2+ 0.04 μ g/mL NAS) amplification cultivation PRO-MIX5, it handles to obtain thallus through upper step same procedure broken
Piece is added in the simulated system for be formed NDMA respectively with 0.2%, 0.5%, 1%, 1.5%, 2% ratio according to weight in wet base and is reacted, simultaneously
Do blank control, the influence that final different training methods and additive amount form NDMA, the results are shown in Table 6, wherein
Using the MRS culture medium amplification cultivation PRO-MIX5 of 0.04 μ g/mL NAS of addition, obtained bacterial chip forms NDMA
Inhibitory effect is best, relatively stable to the inhibitory effect of NDMA with the variation of additive amount, and in several additive amount models of screening
Enclose that interior additive amount is lower, the forming amount of NDMA is lower, can be used additive amount range wider, in this, as what is selected for this testing sieve
The substance that inhibitory effect preferably inhibits NDMA to be formed, further applies in the processing of actual product.
The influence that bacterial chip and Different adding amount obtained by the different training methods of table 6 form NDMA
It is final the preparation method is as follows:
The bacterial chip of PRO-MIX5 has the effect of inhibiting the formation of N- nitrosamine, and uses 0.04 μ g/mL concentration NAS of addition
MRS broth bouillon culture PRO-MIX5, by 5000 g, 4 DEG C of 15 min of centrifugation obtain thallus, with 25 mmol/L, pH=6
PBS rinse thallus 2 times, then with the PBS with the dilution proportion of 1:4, after the bacteriolyze enzyme effect for adding final concentration of 1 mg/mL,
5 min of ultrasonic disruption, through 10000 g, the precipitating (bacterial chip) that 4 DEG C of 10 min of centrifugation are obtained is that can effectively inhibit N-
The substance that nitrosamine is formed, as microorganism nitrosation inhibitor.
Embodiment 2: application of the prepared microorganism nitrosation inhibitor in bologna sausage:
The production of bologna sausage: marinated: by taking 400 g pork hind leg muscles as an example, to add 10 g of cooking wine, 14 g of salt, 2 g of glucose, white sugar 2
G, 2 g of composite phosphate, 0.275 g of sodium ascorbate, 0.075 g(0.15 g/kg additive amount of sodium nitrite), 20 mL of water is in 4
20 h are pickled in DEG C refrigerator；
It mixes filling, bowel lavage: show condition 100g, 40 g of starch, 10 g of mashed garlic, 1.2 g of white pepper powder, egg 40 being added into pickled meat
G, 80 g of ice water mixes well rear bowel lavage with de-airing mixer；
Shortening: by the intestines filled after warm water rinse, exhaust the shortening in Fumigator, shortening parameter are as follows: dry 70 DEG C 60
75 DEG C of 60 min, 85 DEG C of 60 min of boiling, 85 DEG C of 60 min of sootiness, baking min.
By microorganism nitrosation inhibitor respectively in the 0.05%(1 group of meat weight), 0.25%(2 group), 0.5%(3 group) ratio adds
It is added in the meat stuffing after pickling, is mixed filling, bowel lavage, shortening and obtain bologna sausage product, while 0% microorganism nitrosation suppression will be added
Preparation is blank control group (Control Check, CK group), measures sense organ, pH, red scale value, elasticity, the nitrite, 8 of product
The indexs of correlation such as kind biogenic amine, 9 kinds of N- nitrosamine, to analyze the microorganism nitrosation inhibitor of addition different proportion to bologna sausage sense
The influence of official and security quality.
Index determining method: subjective appreciation method: selection 10 subjective appreciation personnel to bologna sausage finished product according to 1 standard of table into
Row evaluation:
Subjective appreciation standard of the table 7 to bologna sausage finished product
The measurement of pH value: it is measured referring to the method for GB 5009.237-2016 " measurement of food pH value ".
The measurement of color difference: the white show condition in bologna sausage is rejected, and remainder is minced to be measured in color difference meter.
Elasticity measurement: bologna sausage is cut into 1 cm3The small square of volume, it is elastic using the P-35 probe measurement of Texture instrument,
Specific location parameter are as follows: speed before surveying: 1 mm/s；Speed in survey: 1 mm/s；Speed behind side: 1 mm/s；Shift length: 5 mm；
Displacement time: 5 s；Trigger force: 5 g.
The measurement of nitrite: it is measured referring to GB 5009.33-2016 " measurement of nitrite in food and nitrate "
Nitrate residue.
The measurement of biogenic amine: it is measured referring to Du Zhihui " method of the different fermentations agent to the research of ferment sausage qualitative effects "
8 kinds of biogenic amines in sample: tryptamines, phenyl ethylamine, putrescine, cadaverine, histamine, tyrasamine, spermine and spermidine.
The measurement of N- nitrosamine: consistent with N- nitrosamine measuring method in index determining method described in embodiment 1.
Data analysis: consistent with data analysing method described in embodiment 1.
As a result with analysis: influence of the Different adding amount of microorganism nitrosation inhibitor to bologna sausage organoleptic quality: such as 8 institute of table
Show, 1 group of bologna sausage flavor and flavour are best, and whole acceptance level is higher, and total score is even higher than CK group, illustrate that appropriate thallus is added
After fragment, mouthfeel not only will not influence, can be improved the flavor of product instead, increase acceptable degree；And 2 groups and 3 groups of sense organs are commented
Score value is below CK group, shows that smell and flavour are significant lower, this is because added microorganism nitrosation inhibitor is in
Slightly sour state (pH 6), additive amount height can make product mouthfeel meta-acid, therefore reduce the person of judging to the acceptance of product.
Influence of the Different adding amount of 8 microorganism nitrosation inhibitor of table to bologna sausage sense organ
Note: shoulder mark capitalization indicate it is different between processing group significant difference (P<0.05).
Influence of the Different adding amount of microorganism nitrosation inhibitor to bologna sausage pH: as shown in figure 5, with microorganism nitrous
Change inhibitor adding proportion increase, pH be in slow decreasing trend, wherein CK group with 1 group difference not significantly (P > 0.05).But
2 groups and 3 groups of pH be substantially less than CK group (P< 0.05), cause product mouthfeel meta-acid, this result and Analyses Methods for Sensory Evaluation Results one
It causes, but entirety pH is 5.9 or more.Therefore microorganism nitrosation inhibitor is in products application it should be noted that additive amount, prevents from adding
Add excess and influences the organoleptic quality of product.From the point of view of pH result of variations, influence phase of 0.05% additive amount (1 group) to product pH
To smaller.
Influence of the Different adding amount of microorganism nitrosation inhibitor to bologna sausage redness and elasticity: as shown in Figure 6, Figure 7, micro-
The red scale value and elasticity of product obtained by three test groups and CK group of biological nitrosation inhibitor Different adding amount are without significant
Difference (P>0.05).Illustrate three kinds of additive amounts of this test institute microbe nitrosation inhibitor to nitrous acid in bologna sausage product
The elasticity of salt color development and product does not make a significant impact.
Influence of the Different adding amount of microorganism nitrosation inhibitor to nitrite in bologna sausage: as shown in figure 8, with micro-
The increase of biological nitrosation inhibitor additive amount, nitrate residue significantly reduce, between each group difference up to the level of signifiance (P< 0.05), three kinds of additive amounts are respectively 18.92%, 37.83%, 51.19% to nitrite degradation rate, illustrate that microorganism nitrosation presses down
Preparation has the effect of reducing nitrate residue, and degradation effect and additive amount are proportional.It, may in conjunction with pH situation of change
It is since acid effect promotes the degradation of nitrite.But scholar reports that, as pH < 4.5, the degradation of nitrite is mainly
Acid effect；As pH > 4.5, predominantly enzyme effect, and this test each group pH illustrates that microorganism nitrosation inhibits 5.9 or more
The substance with degrading nitrite effect other than deacidification may be contained in agent, mainly include that cell wall is broken in bacterial chip
The substances such as piece, cell membrane fragments, organelle, inclusion body still have its mechanism for specifically playing degradation to nitrite
Wait further investigate.
Influence of the Different adding amount of microorganism nitrosation inhibitor to biogenic amine in bologna sausage: biogenic amine is a kind of fat
Race, aromatic series or heterocyclic low molecule nitrogenous compound, it is mainly since microbial action protein makes it be degraded to amino
Acid, then reacted and generated by amino acid decarboxylase.After adding microorganism nitrosation inhibitor it can be seen from table 9, in bologna sausage product
Tryptamines and histamine are produced, this may be that the subacidity of added microorganism nitrosation inhibitor promotes amino acid decarboxylases
Activity also may be the entrained biogenic amine of microorganism nitrosation inhibitor itself.Since biogenic amine is adjusting DNA, RNA and egg
There is important role in terms of white matter synthesis and biological membrane stability.And microorganism nitrosation inhibitor added by this research
In containing the substances such as cell membrane, nucleic acid, thus infer from external source carry and make a possibility that Content of Biogenic Amines increases in bologna sausage compared with
Greatly.With histamine toxicity maximum, followed by tyrasamine in common biogenic amine.Three kinds of microorganism nitrosation inhibitor additive amounts can be shown
Write the formation for reducing tyrasamine, and between three groups difference it is not significant (P>0.05).In general, additive amount is lower 1 group (0.05%)
Relatively preferably.Limit standard is had no for the Content of Biogenic Amines in bologna sausage at present, but according to current some correlation standards, such as
Histamine and tyramine content must not exceed 100 mg/kg and 100~800 mg/kg respectively in European Union's regulation food, and the country is only upper
Histamine must not exceed 100 mg/kg in prescribed ferment meat products in extra large provincial standard.This test is added microorganism nitrosation and is inhibited
Histamine contained in each group bologna sausage and tyrasamine are far below these relevant criterions after agent, will not do harm to huamn body.
The influence that the Different adding amount of 9 microorganism nitrosation inhibitor of table forms biogenic amine in bologna sausage
Note: four groups of putrescine are not detected, and "/" expression is not detected in figure, and the above Content of Biogenic Amines unit is (mg/kg), shoulder mark
Capitalization indicate between processing group different time significant difference (P<0.05)
The influence that the Different adding amount of microorganism nitrosation inhibitor forms 9 kinds of N- nitrosamine in bologna sausage: since NDMA exists
It in more food, and is more toxic, only regulation NDMA content must not exceed 3 μ g/kg in national standard, and as shown in table 10,0.05% is micro-
Biological nitrosation inhibitor (1 group) and 0.5% microorganism nitrosation inhibitor (3 groups) can significantly inhibit the shape of NDMA in bologna sausage
At inhibiting rate can reach 52.87% ~ 63.60%, and inhibitory effect is obvious；And 2 groups of (0.25% microorganism nitrosation inhibitor) NDMA
Forming amount and CK group difference not significantly (P>0.05).But three kinds of microorganism nitrosation inhibitor additive amounts can significantly reduce bologna sausage
The total amount that 9 kinds of N- nitrosamine are formed in product, wherein best with the inhibitory effect of 0.05% additive amount (1 group), to NDMA, NMEA,
NPIP, NMOR, NDPheA inhibitory effect are very significant, and the inhibiting rate formed to 9 kinds of N- nitrosamine can reach 41.04%,
0.25% microorganism nitrosation inhibitor (2 groups) and the 0.5% corresponding inhibiting rate of microorganism nitrosation inhibitor (3 groups) additive amount
Respectively 16.13% and 13.48%, the two inhibitory effect difference it is not significant (P>0.05).Illustrate to add PRO-MIX5 lactic acid bacteria bacterium
Body fragment (microorganism nitrosation inhibitor) can significantly inhibit the formation of N- nitrosamine, and can play compared under few additive
Preferable effect, improves the safety of bologna sausage product, and application effect is good.This may be to have due to bacterial chip to N- nitrosamine
There is certain suction-operated, also may be to contain certain substances that N- nitrosamine can be blocked to be formed in bacterial chip mixture,
Its specific mechanism still needs further to be furtherd investigate.
The influence that 10 Different adding amount microorganism nitrosation inhibitor of table forms 9 kinds of nitrosamine in bologna sausage
Note: the above content of nitrosamines unit is (μ g/kg), and shoulder mark capitalization is indicated between processing group different time
Significant difference (P<0.05)
According to the analysis of the above sense organ and security quality indices, when microorganism nitrosation inhibitor additive amount is larger,
Bologna sausage can generate slightly sour mouthfeel, but the addition of on the whole microorganism nitrosation inhibitor is to the pH, red scale value and bullet of bologna sausage
Property influence and little, or even the flavor of product can be improved after appropriate addition；Can more importantly play degrading nitrite with
And the effect for inhibiting N- nitrosamine to be formed, the security quality of bologna sausage is improved, although will increase certain biogenic amines (tryptamines and histamine)
Content, but content is relatively low, can't do harm to huamn body, and biogenic amine is the essential substance of human body, such as color
Amine is adjustable blood pressure, histamine can participate in local immunity reaction, adjust intestinal physiology function, adjust white blood corpuscle and some protein
The physiological functions such as quantity.All in all, addition microorganism nitrosation inhibitor can improve bologna sausage while not influencing mouthfeel
Product Safety, and apparent compared with that can serve under few additive.
Embodiment 3: application of the prepared microorganism nitrosation inhibitor in sootiness Baconic
The production of sootiness Baconic: marinated agent prescription: 10 jin of pig brisket, salt 90g, 15 g of composite phosphate, sodium nitrite
0.6g, sodium ascorbate 2.5 g, niacinamide 1g, egg liquid 20g, 50 g of white sugar, glucose 20 g, carragheen 5g, greatly
Beans separated protein powder 8g, white wine 10mL, 7.0 g of monosodium glutamate, 2 jin of the spice water (water cooked with following condiment: capsicum 2
G, 2 g of Chinese prickly ash, 1 g of pepper, ginger 6 is g).
Process flow: pig brisket → finishing → preparation pickling liquid → salt water injection → vacuum tumbling 4h → sootiness (55 DEG C,
Sootiness 8h).
Microorganism nitrosation inhibitor is added into curing agent in the 0% of meat weight, 0.05%, 0.25%, 0.5% ratio respectively
In, sootiness Baconic is made through step in 1, measures the indexs of correlation such as the pH, nitrite, 9 kinds of N- nitrosamine of product, is added with analysis
The influence for adding the microorganism nitrosation inhibitor of different proportion to form Baconic's quality and N- nitrosamine.
Index determining method pH, nitrite, nitrosamine measuring method are identical in embodiment 1.
As a result with analysis: as shown in figure 9, addition 3 kinds of different proportions microorganism nitrosation inhibitor to sootiness Baconic's
Effect of Acidity On Absorption is little, will not make its mouthfeel meta-acid, influence flavor.
As shown in Figure 10, the microorganism nitrosation inhibitor of Different adding amount can significantly reduce nitrous acid in sootiness Baconic
Salt residual quantity, degradation effect are descending successively are as follows: and 0.5% > 0.05% > 0.2%, application effect is good.
The inhibition situation formed to 9 kinds of N- nitrosamine in sootiness Baconic is as shown in table 11, compares CK group, adds in Baconic
0.05% effect with microorganism nitrosation inhibitor inhibition N- nitrosamine is relatively preferable, is to the NDMA inhibiting rate formed
27.58%, the inhibiting rate for forming total amounts to 9 kinds of N- nitrosamine is 13.83%, and 0.25% and 0.5% additive amount is without significantly inhibiting effect
Fruit.Illustrate that obtained bacterial chip can not only inhibit the formation of N- nitrosamine in product, and compared with energy under few additive
Inhibiting effect is played, application effect is good.
The influence that 11 Different adding amount microorganism nitrosation inhibitor of table forms 9 kinds of nitrosamine in Baconic
According to embodiment 1 and embodiment 2, illustrate that the microorganism nitrosation inhibitor is added into the production of no fermented product really
It can be improved product safety quality, can inhibit the formation of N- nitrosamine to some extent, and do not influence the flavor of product and add
Work technique, while also there are some additional effects, the residual quantity of nitrite is such as reduced, the flavor etc. of product is improved.This is micro-
Biological nitrosation inhibitor be expected to apply some no zymotechniques but be easy to produce N- nitrosamine other products in, such as meat
The products such as ball, sausage, salted vegetables.
Claims (5)
1. a kind of microorganism nitrosation inhibitor, it is characterised in that: the microorganism nitrosation inhibitor is 2 × 1011CFU xylose
Staphylococcus, 2 × 1011CFU Lactobacillus saki and 2 × 1011CFU class lactobacillus plantarum composite fermentation culture is prepared.
2. a kind of microorganism nitrosation inhibitor according to claim 1, it is characterised in that: the microorganism nitrosation suppression
Preparation specific is the preparation method comprises the following steps: use the MRS broth bouillon culture staphylococcus xylosus added with N- nitrosamine, pure mellow wine cream
Bacillus and class lactobacillus plantarum are to OD600=1.8-2.4, and by 5000-7000g, 4 DEG C of 15 min of centrifugation obtain thallus, with 25
The PBS of mmol/L, pH=6 are rinsed thallus 1 ~ 2 time, then add final concentration of 1 ~ 5 mg/ with the PBS with the dilution proportion of 1:3-10
After 30 DEG C of 2 ~ 5h of effect of lysozyme of mL, 5 ~ 10 min of ultrasonic disruption, through 8000 ~ 12000 g, 4 DEG C of centrifugation 10-15 min
Obtained precipitating is microorganism nitrosation inhibitor.
3. a kind of microorganism nitrosation inhibitor according to claim 2, it is characterised in that: the staphylococcus xylosus,
Lactobacillus saki, class lactobacillus plantarum mixed bacteria culture medium be added with 0.01 ~ 0.5 μ g/mL concentration N- nitrosamine mark
The MRS broth bouillon of quasi- product.
4. a kind of microorganism nitrosation inhibitor according to claim 1 or 2, it is characterised in that: the xylose grape ball
Bacterium, Lactobacillus saki and class lactobacillus plantarum are PRO-MIX5 leavening.
5. a kind of microorganism nitrosation inhibitor of any of claims 1 or 2 inhibits the application of N- nitrosamine in meat products,
It is characterized in that: concrete application method are as follows: 0.02 ~ 1% microorganism nitrosation inhibitor of adding raw materials meat weight in meat products.
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| CN111398265A (en) * | 2020-04-15 | 2020-07-10 | 天津农学院 | Establishment method of in-vitro nitrosation simulation system for blocking NPYR generation by CSEO proportion in salted meat product |
| CN112617156A (en) * | 2020-12-30 | 2021-04-09 | 天津百世耕食品有限公司 | Preparation process of pickled salted vegetable capable of inhibiting nitrosation reaction |
| CN113974052A (en) * | 2021-11-01 | 2022-01-28 | 宜宾郭满堂生态食品有限公司 | Color fixative and application thereof to pressed salted duck |
| CN114134085A (en) * | 2021-12-17 | 2022-03-04 | 海信（山东）冰箱有限公司 | Paenibacillus provirus capable of inhibiting production of nitrosamine and application thereof |
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