CN103815210B - The method that suppression Nidus collocaliae Nitrite or nitrite produce - Google Patents
The method that suppression Nidus collocaliae Nitrite or nitrite produce Download PDFInfo
- Publication number
- CN103815210B CN103815210B CN201210464527.0A CN201210464527A CN103815210B CN 103815210 B CN103815210 B CN 103815210B CN 201210464527 A CN201210464527 A CN 201210464527A CN 103815210 B CN103815210 B CN 103815210B
- Authority
- CN
- China
- Prior art keywords
- nidus collocaliae
- nitrite
- nitrate reductase
- reductase inhibitor
- nitrate
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
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
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
-
- 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
- A23L5/27—Removal of unwanted matter, e.g. deodorisation or detoxification by chemical treatment, by adsorption or by absorption
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Nutrition Science (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Mycology (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The present invention relates to a kind of method suppressing nitrite anions or nitrite to produce in containing nitrate anion or nitrate and the health product of nitrate reductase, food or medical material, suppress the activity of nitrate reductase in described health product, food or medical material including using health product, food or the acceptable nitrate reductase inhibitor of medical material.Particularly relate to a kind of method suppressing Nidus collocaliae Nitrite or nitrite to produce, including using the activity of nitrate reductase in Nidus collocaliae acceptable nitrate reductase inhibitor suppression Nidus collocaliae.Specifically, described method includes adding the acceptable nitrate reductase inhibitor of Nidus collocaliae of suppression effective dose to the Nidus collocaliae of fresh collection or adding the acceptable nitrate reductase inhibitor of Nidus collocaliae of suppression effective dose in Nidus collocaliae production process.Nitrate reductase inhibitor is selected from tungstates.The method of the present invention can effectively suppress the generation of Nidus collocaliae Nitrite or nitrite, reduces the edible risk of Nidus collocaliae.
Description
Technical field
This invention relates generally to a kind of use nitrate reductase inhibitor containing nitrate anion or nitrate with
And the health product of nitrate reductase, food or medical material suppress nitrite anions or the method for nitrite generation,
Particularly suppress the method that Nidus collocaliae Nitrite or nitrite produce, and nitrate reductase presses down
The purposes that preparation produces at suppression Nidus collocaliae Nitrite or nitrite.
Background technology
Nitrite belongs to high toxicity class chemicals, has many relevant research reports that its pathological symptom is described.
Wherein, baby is the most ripe due to digestive system and MHR system, is easier to suffer from nitrous
The methaemoglobinaemia that hydrochlorate causes, consequence the most then can be fatal.It addition, nitrite and nitric acid
Salt can produce compound nitrosamine with gastric acid effect.Nitrosamine is height carcinogen, has report to point out nitrosamine
Direct relation is all had with rectal cancer, gastric cancer and pancreatic cancer.Nitrite causes that the symptom of poisoning also includes chronic
Obstructive pulmonary disease, DNA mutation produce cerebroma with the child in anemia of pregnant woman.Therefore, health product, food and
Medical material nitrite must strictly control.
Nidus collocaliae is a kind of common health product, and it is to be belonged to (Collocalia) by Apodidae (Apodidae) esculent swift
The oozy saliva of multiple birds or the nest that builds up on overhanging cliff of saliva and its down condensation by mixing
Nest.It is Indonesia, Malaysia, Vietnam etc. main product.Record according to supplement to the Herbal: " Nidus collocaliae is sweet
Light flat, support greatly lung yin, preventing phlegm from forming and stopping coughing, mend and can be clear, for nursing one's health the panacea of asthenia, all diseases due to lung
Void, and lower passerby can not be eliminated, all can control it with this." traditional Chinese medical science thinks Nidus collocaliae: " nourishing YIN to relieve dryness, QI invigorating are mended
In, control deficient, expectoration is breathed heavily, spit blood, chronic dysentery, be suitable for having a delicate constitution, malnutrition, chronic dysentery chronic malaria,
Abundant expectoration cough, senile chronic bronchitis, bronchiectasis, emphysema, pulmonary tuberculosis, spit blood, spit blood and
Stomachache patients." modern medicine discovery, Nidus collocaliae can promote immunologic function, have delaying human body caducity, prolong life
The effect lengthened one's life.
Evaluate high and low Yu fine or not point of Nidus collocaliae traditionally and be six big indexs: the place of production, a head, integrated degree,
Cleanliness quality and mouthfeel.The place of production: with Vietnam can the Nidus collocaliae of peaceful product the most of fine quality, trade name is " pacifying
Swallow ".The Nidus collocaliae plucked on cavern and cliff is referred to as " hole swallow ".Pluck Yan Dong the most personal extremely hazardous,
The Nature is also robbed everything.For conservation of nature, swallow room arises at the historic moment.Nest in culture room in swallow room
On plank within doors and wall, swallow is attracted to live in concentrated communities with in kind or additive method, swallow room the Nidus collocaliae gathered
Cry " room swallow ".Commodity mostly are " room swallow " now, and it is because cleaning exquisiteness smooth, pure white, on commodity also referred to as
For " Bai Yan ".Cycle of separately gathering is longer, Nidus collocaliae band flavous " Huang Yan ", is India Ni Xi main product
Sub-and Malaysian." red swallow " is divided into " the red swallow in hole " and " the red swallow in room " two kinds.Red swallow may be because of the external world in hole
Environmental effect, outward appearance is oozed out red liquid exudation and moistening just as rock wall and is caused nest color from being pasted on the one of rock wall
While begin to change into blood red.The red swallow in room is that an Indonesia doctor going to moral to study abroad is in the nineties institute
Invention.Generational verctor is failed to understand, because of very much like with swallow hole environment, therefore may be with being affected by the external environment.Color
Pool is uniform compared with the red swallow in hole, and color also swallow redder than hole is deep.Owing to red swallow yield is few, commodity price is the most somewhat expensive.
Hole red swallow main product is in Thailand, and room red swallow then main product is in Indonesia.
20 August in year, Xi Jiangsheng industrial and commercial bureau announced in Nidus collocaliae containing substantial amounts of nitrite anions, average out to one by one
4,400ppm, Gao Gengda 11,000ppm are wherein the highest with red swallow content.National health department is stood subsequently
I.e. forbid that, from the Malaysia red swallow of import, other relevant Nidus collocaliae industries also receive the most serious strike.
These events attract domestic citizen's highest attention to Nidus collocaliae edible safety overseas.
It practice, nitrite is naturally occurring in surrounding, and can produce, greatly in animal and human endogenous
Vegetable also contains nitrite.It addition, sodium nitrite can be mainly used in salting down as food additive
Bacon based article etc..Mainly due to it, there is antibacterial action, particularly can prevent can be fatal meat poisoning shuttle
Shape bacillus cereus pollutes, but consumption may not exceed statutory standards.Food and Agricultural Organization of the United Nations/world health
The tissue combination food additive Committee of Experts (calling " Committee of Experts " in the following text) does not formulates the urgency of nitrite yet
Property reference dose.The every day of nitrite intake can be set to per kilogram of body weight 0 to 0.07 by the Committee of Experts
Milligram (calculates with nitrite ion).And state food code committee not yet with regard to nitrite as food
Pollutant formulate any food safety standard.According to state food code committee and the Laws of Hong Kong with regard to nitrous acid
Salt is used as the food additive of multiple cheese product and meat curing goods and formulates standard.According to GB
2760-2011 " food additive use standard " specifies, only sausage, bacon, western-style sausages, canned food
Can add nitrite etc. several based foods, other all foods the most do not allow to add.Wherein pickle cured meat product:
Nitrite standard max is 150ppm.And Hong Kong " preservative regulations in food " (132BD chapter)
Define the nitrite the highest allowance content of assigned foods.Hong Kong statutory standards: adding of non-heated process
Work meat product, nitrite standard max is 200ppm；The meat curing of heated process, nitrous acid
Salt standard max is 125ppm.But, " preservative regulations in food " the 3rd (10) article of regulation, these regulations
It is not suitable for the natural food containing any food additive.In a word, nitrite in cubilose content is in the world
Between the most not consistent reference and administrative standard.
For country in Southeast Asia, Nidus collocaliae has exported into one of national major income source.But, with at present
Nidus collocaliae treatment technology, Nidus collocaliae exported country is also merely able to the nitrite anions in Nidus collocaliae and nitrite to control
30ppm or following.China is the entrance state that Nidus collocaliae is maximum, and Hong Kong is also the Nidus collocaliae discrepancy that the whole world is maximum simultaneously
Mouth market, in order to ensure health and the reputation of country of its people, needs to improve Nidus collocaliae edible safety
Requirement, falls below detectable level the upper limit of Nidus collocaliae Nitrite and content of nitrite.This angle
On color, opposition and the problem of contradiction are that current Nidus collocaliae treatment technology institute is insurmountable.In recent years, some are relevant
Nidus collocaliae production method without nitrite anions and nitrite is mainly the most whole by repeating after constantly embathing Nidus collocaliae
Type.But, the nutritional labeling in Nidus collocaliae the percentage ratio repeating to embathe and run off during integer again and
The content that after process, nitrite anions and nitrite remain in Nidus collocaliae all can not get the support of data.
Therefore, need a kind of nitrite anions that can either efficiently control in Nidus collocaliae and nitrite in this area,
The method that can keep again cubilose nutritive composition.
Summary of the invention
In one embodiment, the present invention relates to a kind of containing nitrate anion or nitrate and nitrate also
The health product of protoenzyme, food or medical material suppress the method that nitrite anions or nitrite produce, including using
The acceptable nitrate reductase inhibitor of health product, food or medical material suppresses described health product, food or medicine
The activity of nitrate reductase in material.
On the one hand, the present invention relates to a kind of method suppressing Nidus collocaliae Nitrite or nitrite to produce,
Including using the activity of nitrate reductase in Nidus collocaliae acceptable nitrate reductase inhibitor suppression Nidus collocaliae.
Further, the method for the present invention includes adding suppression effective dose to the Nidus collocaliae of fresh collection
The acceptable nitrate reductase inhibitor of Nidus collocaliae or in Nidus collocaliae production process add suppression effective dose Nidus collocaliae
Acceptable nitrate reductase inhibitor.
Further, the method for the present invention farther includes to remove described nitrate reductase inhibitor.
Further, the suppression effective dose of nitrate reductase inhibitor can be about 100nM-1
μM/g Nidus collocaliae.
Further, described nitrate reductase inhibitor can be the tungsten that can produce free tungstate radicle
Hydrochlorate, such as sodium tungstate or potassium tungstate.
In another embodiment, the present invention relates to a kind of acceptable nitrate reductase inhibitor of Nidus collocaliae
In suppression Nidus collocaliae Nitrite or the aborning purposes of nitrite.
In another embodiment, the present invention relates to a kind of acceptable nitrate reductase inhibitor of Nidus collocaliae
Purposes in the compositions that preparation produces for suppressing Nidus collocaliae Nitrite or nitrite.
In an aspect, nitrate reductase inhibitor used in the present invention can be to produce free tungsten
The tungstates of acid group, such as sodium tungstate or potassium tungstate.
In another embodiment, the present invention relates to one through Nidus collocaliae acceptable nitrate reduction enzyme level
The Nidus collocaliae that agent processes, the content of its Nitrite and nitrite is suppressed.In an aspect, nitric acid
Salt reductase inhibitor is the tungstates that can produce free tungstate radicle, such as sodium tungstate or potassium tungstate.
In another embodiment, the present invention relates to one for suppressing Nidus collocaliae Nitrite or nitrous acid
The compositions that salt produces, it comprises the acceptable nitrate reductase inhibitor of Nidus collocaliae.In an aspect,
Nitrate reductase inhibitor is the tungstates that can produce free tungstate radicle, such as sodium tungstate or potassium tungstate.
In another embodiment, the present invention provides the Nidus collocaliae that the method according to the invention obtains.
Accompanying drawing explanation
Below with reference to accompanying drawing, the present invention is described in further detail.Those skilled in the art will manage
Solve, accompanying drawing and described in detail below be intended to exemplary only, should not be construed as in any way this
The restriction of bright scope.
Fig. 1 represents that the nitrate radical content gathering in Nidus collocaliae in commercially available Nidus collocaliae and the natural place of production (uses t inspection, *
Number expression has significance,statistical, and * * * represents the definitiveness of 99.99%).
Fig. 2 show in different Nidus collocaliae production environment (" Yan Dong " and " swallow room ") nitrate radical content and water sample away from
(using t inspection, * expression has significance,statistical to the relation of birds droppings distance, and * * * represents 99.99%
Definitiveness).
Fig. 3 is the electrophoresis pattern of the protein separated from the red swallow of the red swallow in hole and room respectively, wherein at 43kDa
Band represents the nitrate reductase of separation.
Fig. 4 is that in Fig. 3, protein at 43kDa is after Trypsin enzymolysis, during the flight of protein peptide extract
Between mass spectrum.
Fig. 5 shows that the nitrate anion in variety classes Nidus collocaliae or nitrate reduction and sodium tungstate are to nitrous acid
The inhibitory action that root or nitrite produce.
Detailed description of the invention
Unless otherwise defined herein, to have this area common for all technology used herein and scientific terminology
The implication that technical staff is generally understood.Although that similar or of equal value to approach described herein and material
A little methods and material can also be used in the practice of the present invention, it is preferable that method and material are the most in this article
It is been described by.Additionally, as used herein, numerical range includes the numeral defining this scope.The present invention's
Subhead is just to the purpose illustrated, and is not intended to limit the scope of the present invention.
As used herein, unless clearly indicated by the context, all percentage ratios are all calculated by weight.
The present invention is based on originating Nidus collocaliae Nitrite and nitrite generation and producing grinding of process
The following discovery studied carefully: naturally occur substantial amounts of nitrate anion or nitrate in various Nidus collocaliaes and there is nitrate
Reductase.During gathering at Nidus collocaliae, process or depositing, nitrate reductase is persistently by existence in Nidus collocaliae
Nitrate anion or nitrate reduction are nitrite anions or nitrite so that at Nidus collocaliae Nitrite or nitrous acid
The content of salt is constantly accumulated, thus can have a negative impact human body.
By the present invention in that with nitrate reduction enzyme specific inhibitor (such as tungstates), intercepted nitrous acid
The source that root and nitrite produce, thus eliminate the problem that the later stage repeats to be brought when embathing and (such as seek
The loss formed point), and reduce edible all kinds of Nidus collocaliaes (including but not limited to including Bai Yan, yellow swallow and red swallow)
Risk.Meanwhile, the present invention also without being modified natural Yan Dong or artificial swallow portion within doors, keeps natural
Yan Dong or artificial swallow natural ecological environment within doors, will create disturbances to reduce to minimum to esculent swift group.
Therefore, one of the object of the invention is to provide a kind of suppression Nidus collocaliae Nitrite or nitrite to produce
Method, described method includes suppressing the activity of nitrate reductase in Nidus collocaliae.The method of the present invention can include
The acceptable nitrate reductase inhibitor of Nidus collocaliae of suppression effective dose is added or swallow to the Nidus collocaliae of fresh collection
Nest production process adds the acceptable nitrate reductase inhibitor of Nidus collocaliae of suppression effective dose.
As used herein, it is Asia that term " nitrate reductase " refers to nitrate or reducing nitrate radical
Nitrate or the enzyme of nitrite anions.In one embodiment, nitrate reductase can be from taking root slowly
The pericentral siphon nitrate reductase of tumor bacterium.In the main the place of production such as Indonesia and Malaysia of Nidus collocaliae,
Generally there are root nodule bacteria in swallow room or swallow hole environment, it can secrete nitrate reductase so that gathers or processes
Nidus collocaliae in there is nitrate reductase.
As used herein, including can be effective for term " the acceptable nitrate reductase inhibitor of Nidus collocaliae "
Suppress the activity of nitrate reductase and significant adverse will not be brought to the quality of Nidus collocaliae and/or edible safety
The inhibitor of impact.In one aspect, the acceptable nitrate reductase inhibitor of Nidus collocaliae can include having
Imitate the activity of suppression nitrate reductase and remove without further or facilitate after a procedure and easily removing
Inhibitor.
Those skilled in the art also will it is well known that, any activity that can suppress nitrate reductase, and
Human body will not be brought relatively nitrite anions or the bigger adverse effect of nitrite and will not be to the nutrition of Nidus collocaliae
Value etc. brings the inhibitor of significant impact can be applied to the present invention, is used for suppressing Nidus collocaliae Nitrite
Or the generation of nitrite.Such as, suppression nitrate reductase inhibitor can be selected from tungstates.At one
In embodiment, the nitrate reductase inhibitor of the present invention can be specificity nitrate reductase inhibitor,
The activity of its exclusive suppression nitrate reductase.
Preferably, nitrate reductase inhibitor is the tungstates that can produce free tungstate radicle.Further,
Tungstates can be selected from sodium tungstate or potassium tungstate.
In one embodiment, tungstates can be sodium tungstate, and it is a kind of nitre used in plant experimental
Hydrochlorate reductase inhibitor, suppresses it to live by producing chemical reaction with the molybdenum ion in nitrate reductase
Property.Sodium tungstate colorless and odorless, does not also confirm harmful.On the contrary, sodium tungstate can be by strengthening pancreas merit
Can come anti-diabetic.Furthermore, sodium tungstate has certain water solublity, therefore, if it is desired to, added
Trace sodium tungstate can embathe technology by tradition Nidus collocaliae and conveniently remove.Relative to nitrite anions and nitrous
The high toxicity of hydrochlorate, sodium tungstate inhibits the generation of Nidus collocaliae Nitrite and nitrite, significantly lowers
The risk of edible Nidus collocaliae.
Without being bound by theory, tungstate radicle in sodium tungstate by with the molybdenum ion phase interaction in nitrate reductase
With thus inhibit the activity of nitrate reductase, and then inhibit the generation of nitrite anions or nitrite.
According to the teachings of the present invention, skilled person can easily appreciate that, human body will not be produced not by other
Acceptable adverse effect, the tungstates that can produce free tungstate radicle are equally applicable to the present invention, such as tungsten
Acid potassium.
Further, the nitric acid of other non-tungstates being suitable for it is easily envisaged that according to those skilled in the art
Salt reductase inhibitor, for suppression Nidus collocaliae Nitrite and the generation of nitrite, as long as it can
Suppression nitrate reductase activity and be that Nidus collocaliae is acceptable.
As used herein, term " suppression effective dose " refers to nitrate reduction in any effective suppression Nidus collocaliae
The amount of enzyme.Those skilled in the art will it is well known that, the suppression effective dose of nitrate reductase inhibitor is can
Determine according to the content of nitrate reductase in Nidus collocaliae, and SC service ceiling is not to human body generation injury to be
Preferably.Those skilled in the art can determine nitrate reductase inhibitor according to the types and sources etc. of Nidus collocaliae
Effective dose.
Preferably, the suppression effective dose usage amount of nitrate reductase inhibitor of the present invention can be about 100nM
-1 μM/g Nidus collocaliae, e.g., from about 200-800nM/g Nidus collocaliae, all such as from about 400-600nM/g Nidus collocaliaes, about 450
-550nM/g Nidus collocaliae, about 500nM/g Nidus collocaliae etc..
Nitrate reductase inhibitor can with any applicable form such as solid, liquid known in the art,
Powder, suspension etc. the most such as apply, soak or spraying etc. is added to Nidus collocaliae.Nitrate
Reductase inhibitor may be added to that the Nidus collocaliae of fresh collection, or can each stage in the Nidus collocaliae course of processing
Add to Nidus collocaliae, to suppress the activity of nitrate reductase, thus nitrite anions or nitrous are decreased or even eliminated
The generation of hydrochlorate.
The present invention also provides for a kind of compositions for suppressing Nidus collocaliae Nitrite or nitrite to produce, its
Comprise the nitrate reductase inhibitor of the present invention.
The present invention further provides the Nidus collocaliae that a kind of nitrate reductase inhibitor through the present invention processes, through this
The Nidus collocaliae processed is compared with the Nidus collocaliae processed without this, and the content of its nitrite anions or nitrite is greatly reduced
Even do not contain nitrite anions or nitrite.
The present invention also provide for the nitrate reductase inhibitor of a kind of present invention at suppression Nidus collocaliae Nitrite or
The aborning purposes of nitrite.
The present invention also provides for the nitrate reductase inhibitor of a kind of present invention in preparation for suppressing the Nidus collocaliae Central Asia
Purposes in the compositions that nitrate anion or nitrite produce.
Embodiment
The present invention is further illustrated below by embodiment.But, it is real that the scope of the present invention is not limited to these
Execute example.Unless stated otherwise, all reagent and equipment are the most commercially available.
Containing substantial amounts of nitrate anion or nitrate in the natural surroundings in embodiment 1. Nidus collocaliae and the place of production thereof
According to China " mensuration of national food safety standard-nitrite in food " GB5009.33-2010
Described in method, the commercially available and natural all kinds of Nidus collocaliaes gathered are detected.Inventor have detected commercially available
The content of nitrate anion in the Nidus collocaliae gathered with the natural place of production, the most commercially available hole swallow, the red swallow in room and room Huang swallow are divided
Not from ground (being shown in Table 1) such as Indonesia, Vietnam, Malaysia and Thailand, and what gathered in the natural place of production
Hole swallow, the red swallow in room and room Huang swallow gather respectively from Malaysia Ge Mandong hole and Indonesia Java Island on swallow
Room.It was found that the most commercially available or natural Nidus collocaliae gathered all contains a large amount of nitrate anion or nitrate (is shown in Table
1 and Fig. 1).
NO3-N and NO2-N content in the different Nidus collocaliae kind of table 1
It follows that according to China " mensuration of national food safety standard-nitrite in food " GB5009.33
-2010 methods, have detected nitrate anion in the birds droppings sample collected by different Nidus collocaliae production environments and water sample
Or the content of nitrate.Result is shown in table 2-3, shows at the birds droppings sample collected by different Nidus collocaliae production environments
With water sample all contains nitrate.Further, the nitrate anion in water sample or nitrate content and collection place
Distance dependent (see Fig. 2) away from birds droppings.Owing to birds droppings containing a large amount of nitrate anions, without being bound by theory, send out
A person of good sense speculates, nitrate anion enters the production environment of Nidus collocaliae by water.
NO3-N and NO2-N content in the birds droppings sample that the different Nidus collocaliae production environment of table 2 is collected
NO3-N and NO2-N content in the water sample that table 3 Nidus collocaliae production environment is collected
-ND represents and is not detected by.
Containing nitrate reductase in embodiment 2. Nidus collocaliae
Inventor passes through SDS-PAGE and Coomassie blue stain technology from the Malaysia red swallow in hole, Sabah and India
The specific protein band respectively being isolated by a treaty 43kDa in the red swallow in room, Nicaea Java Island (sees figure
3).Carry out above-mentioned specific proteins band decolouring, Trypsin enzymolysis and desalination process, afterwards for protein peptide
Extract utilizes liquid phase time of-flight mass spectrometer to be analyzed.Described egg is extrapolated according to protein peptide mass discrepancy
The aminoacid sequence of leucorrhea, this aminoacid sequence comprises fragment APLLGCGCGNKGCMDNYLSGR
(seeing Fig. 4).Warp and the protein sequence comparison in Protein Data Bank SWISSPROT, confirm that this tool is special
The protein band of the opposite sex is the pericentral siphon nitrate reductase coming from slow raw root nodule bacteria.Concrete grammar and step under
Literary composition describes in detail.
SDS-PAGE analyzes
For the edible Nidus collocaliae (EBN) of every kind (the Malaysia red swallow in hole, Sabah and Indonesia Java Island
The red swallow in room), weigh about 40mg, and by containing 0.5%Triton X100 and 10 μ g/mL leupeptins
High salt lysis buffer (1M NaCl, 10mM HEPES, 1mM EDTA, 1mM EGTA, pH 7.5)
Extract protein.At 4 DEG C of vortex 30min and in frozen water after ultrasonic 30, by protein extract lyophilizing
And be resuspended in 200 μ l MilliQ water.After quantitatively, 12 μ g of every kind of EBN sample are extracted albumen and bears
It is loaded in 8% polyacrylamide gel and carries out electrophoresis at 60V.After Separation of Proteins, pass through coomassie
Blue reagent dyeing gel.After gel decolours, gel is placed in viewbox acquisition photograph.
LC-MS/MS Q-TOF analyzes
Prepared by sample
The excision polyacrylamide gel in the corresponding position in Coomassie blue stain region is analyzed.Clean
This gel slice, decolouring and passing through 25ng/ μ l trypsin carries out digested in-gel 30min on ice.So
Pass through the peptide of each gel slice of 50%ACN/0.1% formic acid extraction afterwards and be submitted to Q-TOF-
By ZipTip desalination before LC/MS system.
LC-MS/MS Q-TOF system and data analysis
Use the Nanoflow LC being coupled to Nanospray-hybrid Q-TOF Premier tandem mass spectrometer
(nano-Acquity) (Waters Q-TOF Premier configures Waters NanoAcquity UPLC) is carried out
LC-MS/MS.Use program MassLynx (4.1 version) carries out data acquisition and instrument controlling.Use 180
μm X 20mm Symmetry C18 trapping column and 75 μm X 250mm BEH130C18 analytical columns.Stream
Dynamic is 0.1% formic acid/water (A) and 0.1% formic acid/acetonitrile (B) mutually.LC condition of gradient elution is as follows: initial
1-60%B (120min), 99%B (120-150min), then initial concentration 1%B (150-180min),
Flow velocity is 200nL/min.Following basic parameter is used to run this mass spectrograph with positive ion mode: ion source temperature
Degree is 80 DEG C, and capillary voltage is 2.4KV, and sample introduction cone voltage is 35V, and collision cell airflow rate is
0.50mL/min.Impact energy can change according to z and m/z during MS/MS scans, and its exact value
From shop instruction.Data dependency analysis is provided that 1-s MS m/z 250-2000 and the highest 3 seconds
MS/MS m/z 50-2000 (continuous mode), the 30-second dynamically gets rid of.Three are selected in each MS/MS scans
+ 2 ,+3 or+4 charged ions that individual abundance is the highest, its intensity is increased to 40 countings/more than s.Use
ProteinLynx 2.2.5 (smooth 3/2Savitzky Golay and center 4 passage/80% barycenter collection (centroid))
Process initial data, from European Bioinformatics institute (European Bioinformatics Institute
(EBI) the MS/MS data set obtained by SwissProt and Gallus gallus protein database search).
For generally containing nitrate reductase in Nidus collocaliae, without being bound by theory, inventor speculates, swallow
The ground such as the main place of production Indonesia of nest and Malaysia, nitrogen source relative deficiency, in the production environment of Nidus collocaliae
In generally there are slow raw root nodule bacteria to promote fixed nitrogen, while slowly raw root nodule bacteria also secrete nitrate reductase, and
Enter in Nidus collocaliae product.
Nitrate anion in embodiment 3. variety classes Nidus collocaliae or nitrate reduction and sodium tungstate are to nitrite anions
Or the inhibitory action that nitrite produces
Weigh 10 milligrams of sodium tungstate (CAS#:10213-10-2；SIGMA-ALDRICH) it is placed in 100mL
In volumetric flask, add 80 milliliters of pure water, the most ultrasonic 5 minutes, afterwards, add water to scale, i.e.
Obtain the sodium tungstate aqueous solution as inhibitor.
Three kinds of Nidus collocaliaes (the i.e. white swallow in Indonesia room, Indonesia of above-mentioned sodium tungstate inhibitor will be scribbled
The red swallow of room Huang swallow and Indonesia room) and be immersed in containing 1 without the comparison Nidus collocaliae of sodium tungstate inhibitor is whole
In the test tube of M potassium nitrate solution or pure water, and it is placed in 37 DEG C of water-baths 15 days.After 15 days,
According in China " mensuration of national food safety standard-nitrite in food " GB5009.33-2010
The method described, measures the nitrite anions in all test tubes or content of nitrite.Discovery scribbles sodium tungstate and presses down
Nitrite anions in the Nidus collocaliae group of preparation or content of nitrite are without significant change, and are not coated with sodium tungstate suppression
The Nidus collocaliae matched group Nitrite of agent or nitrite then dramatically increase (seeing Fig. 5 B and 5C).With 10
When the purification nitrate reductase of mU/mL replaces Nidus collocaliae to add in different test tubes, all experimental results with
It is consistent (seeing Fig. 5 A) during use Nidus collocaliae.
As it will be apparent to those skilled in the art that, many amendments of the present invention can be made and become
Change without departing from its spirit and scope.Specific embodiments described herein is given the most by way of example,
And the gamut of the equivalent being only awarded together with these claim with claims limits this
Bright.
Claims (10)
1. suppress the method that Nidus collocaliae Nitrite or nitrite produce, can accept including using Nidus collocaliae
Nitrate reductase inhibitor suppression Nidus collocaliae in the activity of nitrate reductase, wherein said nitrate reduction
Enzyme inhibitor is the tungstates that can produce free tungstate radicle.
2. the method for claim 1, it includes adding suppression effective dose to the Nidus collocaliae of fresh collection
The acceptable nitrate reductase inhibitor of Nidus collocaliae or in Nidus collocaliae production process add suppression effective dose Nidus collocaliae
Acceptable nitrate reductase inhibitor.
3. method as claimed in claim 2, it farther includes to remove described nitrate reductase inhibitor.
4. method as claimed in claim 2, wherein said suppression effective dose is 100nM-1 μM/g Nidus collocaliae.
5. the method as described in any one of claim 1-4, wherein said tungstates is sodium tungstate or potassium tungstate.
6. the acceptable nitrate reductase inhibitor of Nidus collocaliae is at suppression Nidus collocaliae Nitrite or nitrite
Aborning purposes, wherein said nitrate reductase inhibitor is the tungstates that can produce free tungstate radicle.
7. the acceptable nitrate reductase inhibitor of Nidus collocaliae preparation be used for suppressing Nidus collocaliae Nitrite or
Purposes in the compositions that nitrite produces, wherein said nitrate reductase inhibitor is for can produce trip
Tungstates from tungstate radicle.
Purposes the most as claimed in claims 6 or 7, wherein said tungstates is sodium tungstate or potassium tungstate.
9., for the compositions suppressing Nidus collocaliae Nitrite or nitrite to produce, it comprises Nidus collocaliae can
The nitrate reductase inhibitor accepted, wherein said nitrate reductase inhibitor is for can produce free tungsten
The tungstates of acid group.
10. compositions as claimed in claim 9, wherein said tungstates is sodium tungstate or potassium tungstate.
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210464527.0A CN103815210B (en) | 2012-11-16 | 2012-11-16 | The method that suppression Nidus collocaliae Nitrite or nitrite produce |
| HK14110855A HK1197162A1 (en) | 2012-11-16 | 2014-10-29 | Method for inhibiting production of nitrite in edible birds nest |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210464527.0A CN103815210B (en) | 2012-11-16 | 2012-11-16 | The method that suppression Nidus collocaliae Nitrite or nitrite produce |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN103815210A CN103815210A (en) | 2014-05-28 |
| CN103815210B true CN103815210B (en) | 2016-12-21 |
Family
ID=50750739
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201210464527.0A Active CN103815210B (en) | 2012-11-16 | 2012-11-16 | The method that suppression Nidus collocaliae Nitrite or nitrite produce |
Country Status (2)
| Country | Link |
|---|---|
| CN (1) | CN103815210B (en) |
| HK (1) | HK1197162A1 (en) |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| SG10201601905RA (en) * | 2016-03-11 | 2017-10-30 | Kah Meng Lim | Extract And Method of Extraction |
| CN107853691A (en) * | 2017-11-01 | 2018-03-30 | 金腰燕（广州）食品有限公司 | The method for removing nitrite in cubilose |
| CN113262303B (en) * | 2020-06-10 | 2022-02-01 | 南方医科大学珠江医院 | Use of nitrate reductase inhibitors in the manufacture of a medicament |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1895088A (en) * | 2006-04-12 | 2007-01-17 | 王锡洪 | Curing-food additive and use thereof |
| CN101595952A (en) * | 2009-06-19 | 2009-12-09 | 四川省食品发酵工业研究设计院 | The preparation method of pickle natural compound nitrite inhibitor |
| CN101692880A (en) * | 2009-10-09 | 2010-04-14 | 江苏大学 | Method for removing nitrites in cubilose |
| CN101971990A (en) * | 2010-10-21 | 2011-02-16 | 北京同仁堂健康药业（福州）有限公司 | Method for cleaning and processing bird's nest |
| CN102362637A (en) * | 2011-09-30 | 2012-02-29 | 邓毛程 | Method for removing nitrite in edible bird's nest by using microbial viable bacteria suspension and/or crude enzyme liquid and application |
| CN102406099A (en) * | 2011-11-08 | 2012-04-11 | 北京同仁堂健康药业（福州）有限公司 | Method for removing nitrite from bird's nest |
-
2012
- 2012-11-16 CN CN201210464527.0A patent/CN103815210B/en active Active
-
2014
- 2014-10-29 HK HK14110855A patent/HK1197162A1/en unknown
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1895088A (en) * | 2006-04-12 | 2007-01-17 | 王锡洪 | Curing-food additive and use thereof |
| CN101595952A (en) * | 2009-06-19 | 2009-12-09 | 四川省食品发酵工业研究设计院 | The preparation method of pickle natural compound nitrite inhibitor |
| CN101692880A (en) * | 2009-10-09 | 2010-04-14 | 江苏大学 | Method for removing nitrites in cubilose |
| CN101971990A (en) * | 2010-10-21 | 2011-02-16 | 北京同仁堂健康药业（福州）有限公司 | Method for cleaning and processing bird's nest |
| CN102362637A (en) * | 2011-09-30 | 2012-02-29 | 邓毛程 | Method for removing nitrite in edible bird's nest by using microbial viable bacteria suspension and/or crude enzyme liquid and application |
| CN102406099A (en) * | 2011-11-08 | 2012-04-11 | 北京同仁堂健康药业（福州）有限公司 | Method for removing nitrite from bird's nest |
Also Published As
| Publication number | Publication date |
|---|---|
| HK1197162A1 (en) | 2015-01-09 |
| CN103815210A (en) | 2014-05-28 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| JP6548520B2 (en) | Insecticide having various effects and production method thereof | |
| CN106397629B (en) | Method, the chondroitin sulfate by this method extraction and the application of chondroitin sulfate are extracted from sturgeon fish-bone | |
| Taufek et al. | Performance of mycelial biomass and exopolysaccharide from Malaysian Ganoderma lucidum for the fungivore red hybrid Tilapia (Oreochromis sp.) in Zebrafish embryo | |
| CN103815210B (en) | The method that suppression Nidus collocaliae Nitrite or nitrite produce | |
| CN104351752A (en) | Production method of proteoglycan protein compound functional capsule product from edible fungi/alga | |
| Fan et al. | Recent advances in edible bird's nests and edible bird's nest hydrolysates | |
| CN107125498A (en) | A kind of feed for improving Tilapia mossambica immunity and preparation method thereof | |
| TW201221133A (en) | A medical compositions for inhibition of influenza Hemagglutinin and process for the preparation thereof | |
| TW201408772A (en) | Method of producing Cordyceps sinensis using Zophobas morio as host | |
| Sleman et al. | Evaluation of blood, biochemical and biological effects of microalgae Chlorella and germinated barley powder as a source of prebiotic on common carp Cyprinus carpio L. | |
| CN106381274B (en) | A kind of Staphylococcus pasteuri and its fermentation marinated pig trotters and preparation method of preparation | |
| CN104757537A (en) | Moringa oleifera fine tablet preparation method | |
| CN103961398B (en) | Four gentleman are dispersed in preparation and improve the application in the medicine of rabbit immune performance | |
| RU2170040C1 (en) | Canned meat | |
| CN106084007A (en) | The isolation identification of a kind of tenebrio molitor natural anti-oxidation octapeptide and functional analysis | |
| CN105076234A (en) | Preparation process of botanical insecticide capable of preventing and treating maize weevil | |
| Jin et al. | The nutritional roles of zinc for immune system and COVID-19 patients | |
| Percário et al. | Antioxidant activity of edible blushing wood mushroom, Agaricus sylvaticus Schaeff.(Agaricomycetideae) in vitro | |
| Adinath et al. | Chromium induced alterations in different individual microalga and their consortia | |
| CN107484829A (en) | A kind of DHA algal oil goat milk piece and preparation method thereof | |
| Wasito | Effects of Red Onion Extract (Allium ascalonicum L.) in Pulmonary Histopathological Lesions of Layer Chickens: A Possible Herbal Medicine Solution to Overcome Avian Influenza and Newcastle Disease Viruses | |
| KR101911398B1 (en) | Mass isolation method of hemolymph from cricket | |
| Singh et al. | Biochar led induced resistance to control Fusarium oxysporum f. sp. lycopersici | |
| Ramli et al. | Beyond tradition: a novel approach for edible bird nest cleaning and its processing | |
| CN105859859B (en) | A kind of feed addictive for disease preventing and treating |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| REG | Reference to a national code |
Ref country code: HK Ref legal event code: DE Ref document number: 1197162 Country of ref document: HK |
|
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| REG | Reference to a national code |
Ref country code: HK Ref legal event code: GR Ref document number: 1197162 Country of ref document: HK |
|
| TR01 | Transfer of patent right |
Effective date of registration: 20180913 Address after: Hongkong, Tongluowan, China Patentee after: Yin Fei Ni Ti SGC (Hongkong) Co.,Ltd. Address before: Stanley, Hongkong, China Co-patentee before: Dong Tingxia Patentee before: Zhan Huaqiang Co-patentee before: Chen Jialun Effective date of registration: 20180913 Address after: Stanley, Hongkong, China Co-patentee after: Dong Tingxia Patentee after: Zhan Huaqiang Co-patentee after: Chen Jialun Address before: Hongkong, Kowloon, Clear Water Bay, China Patentee before: THE HONG KONG University OF SCIENCE AND TECHNOLOGY |
|
| TR01 | Transfer of patent right | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20191107 Address after: Guangdong city of Shenzhen province Qianhai Shenzhen Hong Kong cooperation zone before Bay Road No. 1 building 201 room A (located in Shenzhen Qianhai business secretary Co. Ltd.) Patentee after: Jingqi Biotechnology (Shenzhen) Co.,Ltd. Address before: Hongkong, Tongluowan, China Patentee before: Yin Fei Ni Ti SGC (Hongkong) Co.,Ltd. |
|
| TR01 | Transfer of patent right |