CN103436576B - Staphylococcus xylosus A2 can transform the novelty teabag that metmyoglobin generates nitrosomyoglobin - Google Patents
Staphylococcus xylosus A2 can transform the novelty teabag that metmyoglobin generates nitrosomyoglobin Download PDFInfo
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Abstract
Staphylococcus xylosus A2 can transform the novelty teabag that metmyoglobin generates nitrosomyoglobin, belongs to Food science/technical field of meat science, relates to the purposes of staphylococcus xylosus A2.The microbe fermentation method nitrite substituted in meat product is a brand-new research field, the invention provides a strain and can transform the staphylococcus xylosus A2 that metmyoglobin generates nitrosomyoglobin, it can transform metmyoglobin and generate nitrosomyoglobin in MSA substratum, i.e. butcher's meat haematochrome, thus provide a possible solution route for fermentation method substitutes Nitrite in Meat Products.Staphylococcus xylosus A2 of the present invention possesses conversion and generates nitrosomyoglobin, the i.e. ability of butcher's meat haematochrome.Thus, utilize staphylococcus xylosus A2 to ferment, the colour generation effect of Nitrite in Meat Products can be substituted, reduce its usage quantity, reduce the carinogenicity that product is possible.
Description
Technical field
The invention belongs to Food science/technical field of meat science, relate to the purposes of staphylococcus xylosus A2, particularly relate to staphylococcus xylosus A2 and can transform the novelty teabag that metmyoglobin generates nitrosomyoglobin.
Background technology
Color is one of major criterion weighing meat and meat products quality.The color of meat depends primarily on the wherein content of myohaemoglobin and the state residing for it.Ground state myohaemoglobin (Myoglobin, Mb) (is Fe in porphyrin ring
2+) not in conjunction with any subunit, and meeting and water molecules.In the presence of oxygen, myohaemoglobin can be combined with the oxygen molecule of 1 molecule, forms oxymyoglobin (Oxymyoglobin, MbO
2), thus become shiny red.Now, iron is still with ferrous ion (Fe
2+) exist.But oxygen and other oxygenants, can by two.Valency Fe forms is ferric iron, finally forms metmyoglobin (Metmyoglobin, Met-Mb), for brown.Ground state myohaemoglobin, oxymyoglobin and metmyoglobin generally exist in meat simultaneously.In the meat of long-time placement, metmyoglobin content is higher, is rendered as brown color.
Meat is pickled middle interpolation nitrite and can be contributed to it and form stable redness pool.Why nitrite can play colour generation effect in butcher's meat, is because they can form nitrous acid in acid condition.Nitrous acid is unstable, when reducing substances exists, disproportionation reaction occurs, transforms and generate nitrogen protoxide (NO).Nitrogen protoxide and myohaemoglobin reaction, finally generate nitrosomyoglobin (Nitrosylmyoglobin, NO-Mb).
Sodium Nitrite is the key ingredient that current meat is pickled, its effect in meat product mainly contains: suppress spoilage microorganisms growth, delay food spoilage, anti-oxidant, produce the typical flavor of butcher's meat, and with pigment composition in meat---myohaemoglobin effect, produce nitrosomyoglobin, form the red pool of stable butcher's meat.Wherein, the colour generation effect of nitrite is the major reason that it uses in meat product.But due to himself toxicity and potential carinogenicity, its application is restricted.Therefore, people carry out the use of the nitrite in alternative meat product in the urgent need to a kind of new method.
Summary of the invention
The microbe fermentation method nitrite substituted in meat product is a brand-new research field, the object of this invention is to provide a strain and can transform the staphylococcus xylosus A2 that metmyoglobin generates nitrosomyoglobin, it can transform metmyoglobin and generate nitrosomyoglobin in MSA substratum, i.e. butcher's meat haematochrome, thus provide a possible solution route for fermentation method substitutes Nitrite in Meat Products.
The present invention is by a large amount of microbe to screen, and Late Cambrian one staphylococcus xylosus A2, it has and transforms the ability that metmyoglobin generates nitrosomyoglobin, possesses the potentiality of alternative meat nitrite colour generation effect.
Described staphylococcus xylosus A2 (Staphylococcus xylosus A2) extracts and the bacterial classification of isolation identification from fermenting air-dry sausage, be deposited in China Committee for Culture Collection of Microorganisms's common micro-organisms center (CGMCC), address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City institute of microbiology of the Chinese Academy of Sciences, its culture presevation is numbered: CGMCC No.5049, preservation date is on July 8th, 2011, and the Classification And Nomenclature of bacterial classification is wood sugar glucose coccus (Staphylococcus xylosus).
Staphylococcus xylosus A2 in the present invention has following biological characteristics:
Morphological specificity: staphylococcus xylosus A2 gramstaining result is positive, arranges in irregular botryoidalis, without brood cell, without pod membrane, atrichia.
Colony characteristics: this bacterium nutritional requirement is not high, well-grown on General nutrition agar plate.On MSA agar plate, through 32 DEG C, 48h cultivation, form diameter about 1 ~ 2mm, circular, protruding, white colony.Evenly muddy growth in liquid medium within.
Staphylococcus xylosus A2 of the present invention possesses conversion and generates nitrosomyoglobin, the i.e. ability of butcher's meat haematochrome.Thus, utilize staphylococcus xylosus A2 to ferment, the colour generation effect of Nitrite in Meat Products can be substituted, reduce its usage quantity, reduce the carinogenicity that product is possible.
Accompanying drawing explanation
Fig. 1 is that inoculation staphylococcus xylosus A2 is on the impact of metmyoglobin-MSA system colour-change;
Fig. 2 is that inoculation staphylococcus xylosus A2 is on the impact of metmyoglobin-MSA system 350 ~ 450nm place spectrogram;
Fig. 3 is that inoculation staphylococcus xylosus A2 is on the impact of metmyoglobin-MSA system 450 ~ 700nm place spectrogram;
Fig. 4 is the ESR spectrum figure of two groups of metmyoglobin-MSA systems.
Embodiment
Below in conjunction with accompanying drawing, the present invention is further described.
One, test design
Myohaemoglobin is dissolved in the 50mM sodium phosphate buffer of pH6.5, is configured to the solution of 20mg/mL.Heating 30min is to remove the red enzyme of residual high ferro flesh of may degrading, and under 4 DEG C of conditions, the centrifugal 1min of 10,000g, removes the protein of sex change.Metmyoglobin solution is obtained after 4 DEG C of placement 7d.After metmyoglobin solution being used the sterilizing of 0.45tm membrane filtration, join in MSA (Man Rogosa and Sharp) liquid nutrient medium, make final metmyoglobin concentration be 2mg/mL.Staphylococcus xylosus A2 is inoculated in this substratum, makes its final concentration be 10
6cFU/mL.After 32 DEG C of Anaerobic culturel 18h, by centrifugal removing mycetocyte.By colour-difference meter, its colour-change is measured, and measure substratum system ultraviolet/visible (UV-Vis) absorption spectrum and the free resonance spectrum of electronics (ESR), identify wherein myohaemoglobin existence.
Two, MSA (Manntilo Salt Agar) culture medium prescription
Extractum carnis 1.0g, shows peptone 10.0g, PEARLITOL 25C 10.0g, sodium-chlor 75.0g, distilled water 1000mL, pH7.2 ~ 7.6.
Three, color difference measurement
Colour-difference meter transmission mode (transmittance mode) is selected in the color measurenent of Fluid simulation system, instrument is through self-inspection and zero point, standard (L*=95.26, a*=-0.89, b*=1.18), after correcting, sample is filled 2mm cuvette and is placed on liquid load sample platform and measures.
Four, ultraviolet-visible (UV-Vis) spectral scan
The UV-Vis absorption spectrum of different myohaemoglobin derivative is different.Can be identified the conversion situation of myohaemoglobin by spectral scan and conversion process tracking.Utilize ultraviolet-visible spectrophotometer to carry out spectral scan to obtained sample, with substratum adjustment System baseline, sweep limit is 350 ~ 700nm, and scanning interval is 1nm.
Five, spectrum (ESR) measures
Electronic self-rotary resonant technology (ESR) in order to lone-pair electron analysis, and containing lone-pair electron in NO molecule, therefore, can utilize ESR technology, can distinguish oxymyoglobin (MbO
2) and the red myohaemoglobin derivative of nitrosomyoglobin (NO-Mb) two kinds.ESR measuring method is as follows: get about 0.3mL sample, is directly drawn in ESR kapillary, immerses in liquid nitrogen and cools rapidly.Use the Bruker ECS106 electron spin resonanceapparatus being equipped with cryogenic sample pond to measure, location parameter is as follows: modulating frequency 100kHz; Modulation amplitude 1.0mT; Microwave power 2.0mW; Microwave frequency 9.44GHz; Temperature 150K; Time of response 30s.When it should be noted that carrying out ESR measures, in MSA substratum, paramagnetic substance manganous sulfate can not be added.
Six, results and analysis
Be inoculated in MSA liquid nutrient medium by staphylococcus xylosus A2, do blank simultaneously, lower 32 DEG C of anaerobic condition cultivates 18h, measures its value of chromatism and spectrum change situation.
By vision, can observe very clearly, cultivate through 18h, the metmyoglobin-MSA system of inoculation staphylococcus xylosus A2 presents shiny red, and control group then presents brown pool.Transmission aberration is adopted to carry out quantitative analysis to simulated system color, result as shown in Figure 1, the a*-value (red scale value) of staphylococcus xylosus A2 group is significantly higher than control group (P < 0.05), this show this group sample will " redder " some; And more another two groups lower of its L*-value (brightness value), but difference is not significantly (P > 0.05).Also significantly do not distinguish between the b*-value (yellow value degree) of each group.
Meanwhile, carry out UV-Vis spectral scan to each treatment group, result as Figure 2-3.As seen from the figure, blank group has charateristic avsorption band in 410,505 and 635nm place, and this is the feature of metmyoglobin; And the absorption spectrum curve inoculating staphylococcus xylosus A2 group changes, there is charateristic avsorption band in 420,540 and 580nm place, the typical collection of illustrative plates of this red myohaemoglobin derivative just.Red myohaemoglobin derivative is divided into oxymyoglobin (MbO
2) and nitrosomyoglobin (NO-Mb) two kinds.Due to the two absorption curve and characteristic peaks very close, so be difficult to distinguish it only by this kind of index.
As shown in Figure 4, there is the signal of paramagnetic substance in inoculation staphylococcus xylosus A2 system, and control group system ESR Signal aspects is not for exist paramagnetic substance, and this shows to there is nitric oxide molecule in this system.Research finds that this system g value is 1.9870, shows further, and detection obtains nitrosomyoglobin (NO-Mb), and the main component of its butcher's meat pigment exactly.This shows that metmyoglobin can be converted into nitrosomyoglobin by this staphylococcus xylosus, and this provides a possible approaches for microbe fermentation method substitutes nitrite.
Claims (3)
1. staphylococcus xylosus A2 can transform the purposes that metmyoglobin generates nitrosomyoglobin, and described staphylococcus xylosus A2 deposit number is: CGMCC No.5049.
2. staphylococcus xylosus A2 according to claim 1 can transform the purposes that metmyoglobin generates nitrosomyoglobin, it is characterized in that described staphylococcus xylosus A2 transforms metmyoglobin and generates nitrosomyoglobin in MSA substratum.
3. staphylococcus xylosus A2 according to claim 2 can transform the purposes that metmyoglobin generates nitrosomyoglobin, it is characterized in that described MSA culture medium prescription is: extractum carnis 1.0g, show peptone 10.0g, PEARLITOL 25C 10.0g, sodium-chlor 75.0g, distilled water 1000mL, pH 7.2 ~ 7.6.
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| CN113234612B (en) * | 2021-02-05 | 2023-07-25 | 善恩康生物科技（苏州）有限公司 | Lactobacillus fermentum ZS 40A/A preparation for preventing colitis |
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| CN101717742B (en) * | 2009-12-25 | 2012-07-11 | 扬州大学 | Staphylococcus xylosus and application thereof in producing fermented segmental pork |
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