CN103468610B - Fiber fiber microbe and active protein, and application thereof - Google Patents
Fiber fiber microbe and active protein, and application thereof Download PDFInfo
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Abstract
The invention discloses a fiber fiber microbe. The collection name of the microbe is 102807, the collection unit is CGMCC (China General Microbiological Culture Collection, the collection number is CGMCC NO:6794, and the collection date is November 9th, 2012. The invention also discloses application of the microbe in degrading aflatoxin M1 and a method for degrading the aflatoxin M1 by using the microbe. The method comprises the following steps: adding the microbe fermentation liquid into a solution containing aflatoxin M1, and standing at 15-35 DEG C for 24-72 hours. Besides, the invention also discloses an active protein for degrading aflatoxin M1, which is extracted from an extracellular solution of the microbe. The SDS-PAGE (sodium dodecyl sulfate polyacrylamide gel electrophoresis) test can determine that the molecular weight of the active protein is 150kDa or so.
Description
Technical field
The active substance that the present invention relates to microorganism and purposes and produced by microorganism, relates to the purposes of a kind of bacterial strain that fiber germ belongs to and this bacterial strain specifically, and the activated protein produced by this bacterial strain.
Background technology
Aflatoxins M1 (Aflatoxin M1, be called for short AFM1) is the extremely strong hepatotoxin of a kind of toxicity, can cause the liver injury of all animals, main manifestations is that liver lobule is downright bad, hydrocholecystis, under mucous membrane, hydrops under muscle layer and serous coat, causes because major Liver is hemorrhage and dead.AFM1 can also suppress the synthesis of phosphatide and cholesterol, affects the transport of lipid from liver, fat is deposited in liver, causes hepatomegaly.
AFM1 is the product formed through hydroxylation in vivo after animal takes in aflatoxin B1 (Aflatoxin B1, AFB1).An AFM1 part is discharged from urine and milk, a part is present in the edible portion of animal, as in breast, liver, eggs, kidney, blood and muscle, wherein common with breast, thus this also becomes the main source of AFM1 in human food prods or animal-feed, in addition due to the Main Foods that cow's milk and goods thereof are the mankind, particularly baby, so its hazardness is larger.Current prevention AFM1 pollution is a very difficult thing, and in order to the harm avoiding AFM1 to bring to the mankind as much as possible, scientific research personnel is being devoted to study the AFM1 toxin how removed in food or feed.
The conventional method removing aflatoxin has physical removal method and chemical removal method at present.Physical removal method, as extraction process, absorption method, heating method and radiation method, by being separated by aflatoxin, or making it to resolve into nontoxic or that toxicity is less molecule, eliminating the toxicity of aflatoxin.Chemical removal method mainly uses chemical reagent to remove aflatoxin, and using sodium sulfite solution or liming to carry out processing is two kinds of more common chemical detoxication methods.Although Physical and chemical method detoxification are very effective, inevitably bring the loss of food nutrition and higher cost, especially chemical residual problem causes the very big concern of people.Therefore, these methods are only applicable to the laboratory Study on degradation to agricultural-food usually at present.
The research of toxin minimizing technology is that the solution of food-safety problem is checked on by mainly relying on detection, eliminates the progress of harm to taking measures.But, a present situation that can not be ignored is, go abroad from domestic, no matter be that European Union is like this to the economic community that aflatoxin limitation regulation is strict, or the U.S., China, Australia, the large agricultural country that Brazil is such, when finding that in plant product, aflatoxin exceedes regulation limitation, its main processing mode transfers feed to, manufacture, extend the chain of an aflatoxin contamination, cause aflatoxin to the dual pollution of animals and humans, and due to animal derived product, special product as milk, obvious organoleptic feature (can not select moldy kernel as products such as peanuts) is not had after aflatoxin-contaminated, make the control of aflatoxin more complicated.Therefore, the degradation method of the Aflatoxins M1 existed in product is studied, find toxin of can degrading can not form to food the biological control measure polluted further again very meaningful.
Now studies have found that, many bacteriums and metabolic enzyme (as milk-acid bacteria, acetic bacteria, Bacillus subtilus) thereof all have certain Degradation to aflatoxin B1.Due to biological degradation have specificity strong, be not easy to produce the advantages such as by product, treatment condition gentleness, the nutritive ingredient do not destroyed in food, therefore by microorganism, aflatoxin is detoxified, it is made to be transformed into nontoxic or hypotoxic material, beyond doubt a kind of desirable mode.But the biological degradation had not yet to see for AFM1 is reported.
Summary of the invention
An object of the present invention is to provide a kind of new bacterial strain, two of object is to provide the application of this bacterial strain a kind of in aflatoxin degradation M1, three of object is to provide a kind of method utilizing this strains for degrading Aflatoxins M1, and four of object is to provide the activated protein of a kind of energy aflatoxin degradation M1 utilizing this bacterial strain to produce.
The object of the invention is by following technical scheme realize:
New strains provided by the present invention is fiberoptic fiber germ, its preservation name is called 102807, be preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center (depositary institution CGMCC), deposit number is CGMCC NO:6794, preservation date on November 09th, 2012.Below described preservation name is called that the fiberoptic fiber germ of 102807 is referred to as 102807 bacterial strains.
102807 bacterial strains provided by the present invention, its bacterium colony and strain morphology are:
On nutrient agar, bacterium colony is light yellow, smooth surface, slightly projection, and translucent, the circular colonies of neat in edge, has gluey adhesion when provoking with inoculating needle.Observing after gramstaining, is the spherical thalline of Gram-positive, diameter 0.5 μm, single, paired or stack array.
Its biochemical reactions feature is as table 1:
Table 1
Based on the above results, fiberoptic fiber germ strain 102807 is accredited as fiberoptic fiber germ (Cellulosimicrobium cellulans), belong to actinomycetes door (Actinobacteria class), actinomycetales (Actinomycetales order), Micrococcineae (Micrococcineae suborder), fiber germ belongs to (Cellulosimicrobium sp.).
102807 bacterial strains of the present invention have AFM1 Degradation, and its fast growth, growing way are good, stationary phase is long, and also have Heat stability is good in degradation process, the feature of acid-fast alkali-proof, degradation rate level is high simultaneously.
The present invention completes the application of this bacterial strain in aflatoxin degradation M1 thus.
The present invention 102807 bacterial strain is used for the method for aflatoxin degradation M1, comprises the steps: the fermented liquid described preservation name being called the germ of 102807, joins in the material containing Aflatoxins M1, then places 24 ~ 72h under 15 ~ 35 DEG C of conditions.
The present invention 102807 bacterial strain is used for the method for aflatoxin degradation M1, simple to operate, the Aflatoxins M1 in food can not only be divided and take off, nor can cause further pollution, efficiently solve traditional detoxicating method Problems existing.
The activated protein of aflatoxin degradation M1 of the present invention, it obtains as follows: described activated protein obtains as follows: described preservation name is called the germ shake flask fermentation of 102807, then 0 ~ 6 DEG C, centrifugal under 5000 ~ 10000rpm condition, get centrifugal after supernatant liquor cross super filter tube concentrated after, carry out SDS-PAGE electrophoresis, then reclaim the albumen of molecular weight at 100 ~ 200kDa band place.
Described when carrying out shake flask fermentation fermention medium used be: often liter of described fermention medium is containing beef powder 3.0g, peptone 10.0g, glucose 1.0g, sodium-chlor 5.0g.
The condition of described shake flask fermentation is: under temperature 25 ~ 45 DEG C, pH value 5 ~ 9, rotating speed 110 ~ 140rpm condition, cultivate 24 ~ 72h.
The actual conditions of described high speed frozen centrifugation is: temperature 4 ~ 6 DEG C, rotating speed 5000rpm.
Described super filter tube specification is 10 ~ 100kDa.
After described SDS-PAGE electrophoresis, reclaim the albumen of molecular weight at 150kDa band place.
The activated protein prepared by 102807 bacterial strains provided by the invention, has the Degradation to AFM1 toxin, and detoxicating activity high, act on single-minded, other nutritive ingredient can not be destroyed, can be used for the removal of AFM1 in foodstuffs industry.
Accompanying drawing explanation
Fig. 1 is the bacterium colony picture cultivated in the present invention 102807 bacterial strain nutrient agar.
Fig. 2 is the bacterium colony picture after gramstaining cultivated in the present invention 102807 bacterial strain nutrient agar.
To be the present invention 102807 bacterial strain grow tree graph to the 16S rDNA sequential system of relevant kind to Fig. 3 composes.
Embodiment
Below in conjunction with drawings and the specific embodiments, the present invention is further illustrated.
Main agents in embodiment and test kit: high performance liquid chromatography (HPLC) (VICAM company of U.S. Aflatoxins M1 immune affinity column (IAC-AFLM1)); Tonka bean camphor (Shenyang City's reagent three factory); Aflatoxins M1 standard substance (Sigma company).
Key instrument equipment in embodiment: biochemical cultivation case (Guangdong Medical Apparatus and Instruments Factory); Isothermal vibration incubator (the bright damp physics and chemistry apparatus company limited of sky tin); High performance liquid chromatograph (Waters, US).The starting material of other undeclared manufacturers and reagent are common market marketing channel and buy.
Universal method in embodiment:
A, degradation experiment method:
The aseptic triangular flask of long-pending 150mL of trying to please loads 50mL pure milk (monarch Le Bao), adds Aflatoxins M1 (AFM1) the standard substance 0.125mL that concentration is 200ppb, to final concentration 0.5ppb.The bacterium liquid that 1mL ferments is added, 37 DEG C, 110rpm shaking culture 72h in every bottle.Each bacterial strain do two parallel, simultaneously using one bottle of milk sample not adding AFM1 and bacterium liquid as blank, one bottle with the addition of AFM1 but the milk sample not adding bacterium liquid as negative control.
B, AFM1 measuring method:
The sample 20.00g(that taking needs to detect AFM1 content is accurate to 0.01g) be placed in 50mL centrifuge tube, 10000rpm, 4 DEG C of centrifugal 10min; Supernatant liquor glass fiber filter paper after centrifugal is filtered in funnel, filtrate crosses AFM1 immune affinity column (crossing column flow rate 2 ~ 3mL/min), then dry up with after the ultrapure washing post of 10mL, then 1mL elution is used, then elutriant ultrapure water is settled to 2mL, vortex vibration 1min, then crosses 0.45 μm of filter membrane, upper machine testing AFM1 content.(typical curve establishes 4 points: 0.5,1.0,5.0,10.0.Unit: ng/mL);
The elutriant more than used is the methyl alcohol/acetonitrile mixture of volume ratio 4:5.
The calculating of c, degradation rate:
The mensuration of d, growth curve:
By inoculation in nutrient broth liquid nutrient medium, at 37 DEG C, shaking culture under 110rpm condition, every 4h samples coated plate, carries out enumeration, METHOD FOR CONTINUOUS DETERMINATION 48h.
The screening of embodiment 1 Aflatoxins M1 degradation bacteria strains:
Parting material: the fecal sample of milk cow in grain sample mouldy in the soil sample in the environment such as farmland, surrounding area, Shijiazhuang, pond, grain depot, dairy cow farm.
Substratum:
Primary dcreening operation substratum (g/L): KH
2pO
40.25g, NH
4nO
31.0g, MgSO
47H
2o0.25g, FeSO
40.001g, agar 17.0g, coumarin 1 .0g, pH7.0, tonka bean camphor needs filtration sterilization, other 121 DEG C of steam sterilizing 15min.
Seed culture medium (g/L): extractum carnis 3g, peptone 5g, NaCl 10g, 121 DEG C of steam sterilizing 15min.
Multiple sieve and fermention medium (nutrient broth) (g/L): beef powder 3.0g, peptone 10.0g, glucose 1.0g, sodium-chlor 5.0g, pH7.5 ± 0.1,121 DEG C of steam sterilizing 15min.
Strain store medium (nutrient agar medium) (g/L): beef extract 3.0g, peptone 10.0g, sodium-chlor 5.0g, agar 15.0g, pH7.5 ± 0.1,121 DEG C of steam sterilizing 15min.
Concrete screening method:
Strains separation and screening route: from parting material, collected specimens → degraded tonka bean camphor bacterial strain is separated → bacterial strain screening (taking tonka bean camphor as sole carbon source cultured continuously three generations) → carry out multiple sieve → object bacterial strain with AFM1.
(1) primary dcreening operation of degraded AFM1 bacterial strain:
The sample gathered in parting material is loaded disposable sterilized sampler bag process:
The sample 1g getting collection, in sterilizing 18mm × 180mm test tube, adds sterilized water 10ml, and with tampon sealing, vibration 24h, Aspirate supernatant 1ml, successively by 10
-1, 10
-2, 10
-3, 10
-4, 10
-5dilution, then in each test tube, tonka bean camphor solution is added, (tonka bean camphor and the independent wiring solution-forming filtration sterilization of sterilized water, tonka bean camphor is water insoluble, make it be uniformly distributed in water so will first heat, be finally added in test tube), make its ultimate density reach 0.1%, mix, make sample liquid.Get 0.2mL sample liquid respectively, coat the flat board of primary dcreening operation substratum, can ensure that tonka bean camphor becomes sole carbon source and the energy like this, cultivate 7d for 37 DEG C, observations, to the picking list bacterium colony of growth phenomenon be had, number respectively and nutrient agar medium of transferring (i.e. strain store medium) inclined-plane after line separation and purification.
By the inoculation of separation and purification in primary dcreening operation substratum, be placed in 37 DEG C of incubators and cultivate one week, observe the growing state of bacterial strain.For avoiding the carbon source interference of cell in former environment, picking has the bacterial strain of growth phenomenon, and in kind cultivate for continuous three times, finally preserving those can take tonka bean camphor as the bacterial strain that the primary dcreening operation substratum of sole carbon source grows.Finally, through primary dcreening operation, obtain 37 strain bacterial classifications altogether.
(2) the multiple sieve of degraded AFM1 bacterial strain:
AFM1 Degrading experiment: the 37 strain bacterial classifications obtained in primary dcreening operation are inoculated into fermention medium respectively, 37 DEG C of shake flask fermentations 24 hours.Respectively get fermented liquid 1mL to join 50mL respectively and be added with in the milk sample of Aflatoxins M1 (0.5ppb), be placed on 37 DEG C, in 110rpm shaking culture case after shaking culture 72h, measure AFM1 content in milk, calculate degradation rate.Milk sample same for the 50ml not adding fermented liquid and AFM1 is done blank simultaneously.
The result of above AFM1 Degrading experiment is as table 2, more than 50%, three strains are had to AFM1 degradation rate in 37 strain bacterial classifications, be respectively 102807 bacterial strains, 102812 bacterial strains, 102815 bacterial strains, wherein the degradation rate of 102807 bacterial strains to AFM1 is the highest, being that 69.2%, 102815 bacterial strains take second place, is 62.4%, 102812 bacterial strains are minimum, are 54.7%.
Table 2 is degraded the multiple sieve result of AFM1 bacterial strain
The qualification of embodiment 2 102807 bacterial strain
Bacterial classification solid medium (nutrient agar medium) (g/L): beef extract 3.0g, peptone 10.0g, sodium-chlor 5.0g, agar 15.0g, pH7.5 ± 0.1,121 DEG C of steam sterilizing 15min.
Major equipment: biochemical cultivation case (Guangdong Medical Apparatus and Instruments Factory), Automated microbiology analyzer (French Mei Liai biotech firm), high performance liquid chromatograph (Waters, US), fluorescence are just putting microscope (Japanese OLYMPUS company), isothermal vibration incubator (the bright damp physics and chemistry apparatus company limited of sky tin).
Main agents: biochemical identification Reagent Tube (Beijing overpass Technology Co., Ltd.), API biochemical identification carton (French Mei Liai biotech firm), gram staining liquid (French Mei Liai biotech firm), VITEK biochemical identification carton (French Mei Liai biotech firm).
(1) strain morphology is observed and gramstaining qualification:
Bacterium colony and strain morphology are observed: on nutrient agar plate, observe colonial morphology, color and luster after 37 DEG C of cultivation 24h, examine under a microscope cell shape after gramstaining.
(2) Physiology and biochemistry detects:
Picking cultivates the bacterial strain of 24h on nutrient agar plate, mixes in the brine tube of aseptic 0.45%, makes the bacteria suspension of 0.5 Maxwell unit, inoculation biochemical reaction assessor, by observations after the explanation cultivation corresponding time of each biochemical tube.Simultaneous vaccination API identification card and VITEK fully-automatic analyzer loading comparison biochemical results.
(3) molecular biology identification:
16S rDNA gene sequencing: check order after 16S rDNA gene order pcr amplification, sequencing result uses Blast comparison at NCBI, the recognised standard sequence data with bacterial strain 16S rDNA homology is obtained from GenBank database, use MEG4.1 computed in software sequence similarity after alignment, and do Phylogenetic Analysis.This step test is undertaken by Chinese industrial Microbiological Culture Collection administrative center and is submitted to report the test.
The result of above three identification experiments is as follows:
As shown in Figure 1,102807 bacterial strains bacterium colony on nutrient agar is light yellow, smooth surface, slightly projection, and translucent, the circular colonies of neat in edge, has gluey adhesion when provoking with inoculating needle.Observing after gramstaining, as shown in Figure 2, is the spherical thalline of Gram-positive, diameter 0.5 μm, single, paired or stack array.
Physiology and biochemistry detected result is as shown in table 3:
Table 3 bacterial strain 102807 biochemical reaction feature
Molecular biology identification (16S rDNA gene sequencing) result is as shown in Fig. 3 and table 4:
The 16S rDNA sequential analysis table of table 4102807 bacterial strain
| Sequence alignment: 102807 |
| 100.00％Cellulosimicrobium funkei ATCC BAA-886 T(AY501364) |
| 99.70％Cellulosimicrobium cellulans DSM43879 T(X83809) |
| 97.43％Luteimicrobium subarcticum R19-04 T(AB489904) |
| 97.43％Cellulosimicrobium terreum DS-61 T(EF076760) |
| 96.60％Promicromonospora flava CC0387 T(AM992980) |
| 96.52％Isoptericola variabilis MX5 T(AJ298873) |
| 96.45％Isoptericola nanjingensis H17 T(HQ222356) |
| 96.45％Sanguibacter antarcticus KOPRI21702 T(EF211071) |
| 95.99％Isoptericola dokdonensis DS-3 T(DQ387860) |
| 95.99％Isoptericola jiangsuensis CLG T(EU852101) |
Comprehensive above test-results, bacterial strain 102807 is accredited as fiberoptic fiber germ (Cellulosimicrobiumcellulans), belong to actinomycetes door (Actinobacteria class), actinomycetales (Actinomycetalesorder), Micrococcineae (Micrococcineae suborder), fiber germ belongs to (Cellulosimicrobium sp.).
Embodiment 3 102807 bacterial strain is at different temperatures to AFM1 degradation effect:
102807 inoculation embodiment 1 selected are to fermention medium, 37 DEG C of fermentation 24h, be prepared into fermented liquid, then being inoculated in by fermented liquid with the addition of in the milk of AFM1, adjustment pH is 7, rotating speed 110rpm, and temperature is set to from 25 DEG C to 45 DEG C, every 5 DEG C is five hydraulic tests of a gradient, measures and calculate AFM1 degradation rate after shaking culture 72h.
Replace 102807 bacterial strains to be contrast experiment with 102812 bacterial strains and 102815 bacterial strains simultaneously.
Result shows, 102807 bacterial strains degradation rate 30 DEG C time is the highest, reaches 59.7%, still keeps the comparatively high degradation rate of 56.9% when 35 DEG C; 102812 bacterial strains and the degradation rate of 120815 bacterial strains to AFM1 are more than 50%.
Embodiment 4 102807 bacterial strain is at various ph values to AFM1 degradation effect:
102807 inoculation embodiment 1 selected are to fermention medium, 37 DEG C of fermentation 24h, be prepared into fermented liquid, then being inoculated in by fermented liquid with the addition of in the milk of AFM1, milk pH value is adjusted to 5 ~ 9, and every 0.5 is nine hydraulic tests of a gradient, and then adjusting temperature is 37 DEG C, rotating speed 110rpm, measures after shaking culture 72h and calculates AFM1 degradation rate.
Replace 102807 bacterial strains to be contrast experiment with 102812 bacterial strains and 102815 bacterial strains simultaneously.
Result shows, 102807 bacterial strains are all more stable to the degradation rate of AFM1 in the scope of pH5 ~ 9, and reach maximum 78.6% when pH8.5; 102812 bacterial strains and 120815 bacterial strains poor and low to AFM1 degradation rate at different pH stability inferior.
The result of comprehensive above embodiment 3 and embodiment 4 is analyzed, and 102807 strain growth speed are fast, growing way good, stationary phase is long, and Heat stability is good in degradation process, acid-fast alkali-proof is all better, and AFM1 degradation rate level is high.
The preparation of the activated protein of the degraded AFM1 that embodiment 5 102807 bacterial strain produces
A) by 102807 inoculation to fermention medium, in 30 DEG C, pH8.5,110rpm condition bottom fermentation 24h is prepared into fermented liquid:
B) by fermented liquid at 4 DEG C, under 5000r/min condition after frozen centrifugation 20min, get supernatant liquor for subsequent use as the outer crude extract of born of the same parents;
C) preliminary purification of activated protein:
By outer for the born of the same parents of b) step crude extract respectively with the molecular weight 10kDa that dams, the super filter tube of 30kDa, 50kDa and 100kDa carries out ultrafiltration concentration, and the filter membrane then concentrated solution being crossed 0.22 μm is degerming, obtains the protein solution of 4 kinds of molecular weight;
D) protein SDS-PAGE electrophoresis:
The protein solution of 4 middle-molecular-weihydroxyethyl c) step obtained carries out SDS-PAGE electrophoresis respectively, finds and reclaims the albumen of molecular weight at about 150kDa band place.
Embodiment 6 102807 bacterial strain fermentation liquor is for the method for AFM1 in milk of degrading
Experimental group: by 102807 inoculation to fermention medium 30 DEG C, pH8.5,110rpm speed conditions bottom fermentation 28h makes fermented liquid, then fermented liquid is joined in the milk containing AFM1 (0.5ppb), after placing 24h under 25 DEG C of conditions, detect the content of AFM1 in milk, and calculate degradation rate.
Negative control group: that does not add fermented liquid contains AFM1 milk (0.5ppb) group.
In the present embodiment milk, AFM1 content carries out according to the general AFM1 measuring method provided above.
Embodiment 7 102807 bacterial strain fermentation liquor is for the method for AFM1 in lean pork of degrading
Experimental group: by 102807 inoculation to fermention medium 37 DEG C, pH8,140rpm speed conditions bottom fermentation 36h makes fermented liquid, then fermented liquid is joined (0.5ppb) in the lean pork (lean pork is broken into minced meat shape) containing AFM1 to mix, after placing 36h under 15 DEG C of conditions, detect the content of the AFM1 in lean pork, and calculate degradation rate.
Negative control group: that does not add fermented liquid contains AFM1 lean pork (0.5ppb) group.
In the present embodiment lean pork, the content of AFM1 is undertaken by the national examination criteria mensuration of Aflatoxins M1 and B1 (in the GB5009.24-2010 food).
Result shows: contain the milk of AFM1 in embodiment 6 after 102807 bacterial strain fermentation liquor process, its AFM1 content is reduced to 0.157ppb by original 0.5ppb, and degradation rate is 70.6%; Contain the lean pork of AFM1 in embodiment 7 after 102807 bacterial strain fermentation liquor process, its AFM1 content is reduced to 0.173ppb by original 0.5ppb, and degradation rate is 65.4%.
Claims (3)
1. a fiberoptic fiber germ (
cellulosimicrobium cellulans), it is characterized in that the preservation name of described germ is called 102807, depositary institution is CGMCC, and deposit number is CGMCC NO:6794, preservation date on November 09th, 2012.
2. the application of fiberoptic fiber germ according to claim 1 in aflatoxin degradation M1.
3. the method for an aflatoxin degradation M1, it is characterized in that it comprises the steps: the fermented liquid of germ preservation name being called 102807, join in the material containing Aflatoxins M1, then under 15 ~ 35 DEG C of conditions, 24 ~ 72h is placed, described preservation name is called that the deposit number of the germ of 102807 is CGMCC NO:6794, preservation date on November 09th, 2012.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101381708A (en) * | 2007-09-04 | 2009-03-11 | 中国科学院大连化学物理研究所 | Fibrized cellulomonas cartae, hydrolase and application thereof in taxane conversion aspect |
| CN101691554A (en) * | 2009-06-30 | 2010-04-07 | 广西科学院 | Cellulosimicrobium cellulans capable of producing acrylic acid |
| CN102634470A (en) * | 2012-04-10 | 2012-08-15 | 宋建民 | Cellulosimicrobium cellulans and method for producing trehalose through penetration fermentation of cellulosimicrobium cellulans |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101381708A (en) * | 2007-09-04 | 2009-03-11 | 中国科学院大连化学物理研究所 | Fibrized cellulomonas cartae, hydrolase and application thereof in taxane conversion aspect |
| CN101691554A (en) * | 2009-06-30 | 2010-04-07 | 广西科学院 | Cellulosimicrobium cellulans capable of producing acrylic acid |
| CN102634470A (en) * | 2012-04-10 | 2012-08-15 | 宋建民 | Cellulosimicrobium cellulans and method for producing trehalose through penetration fermentation of cellulosimicrobium cellulans |
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| 陈燕红.一株纤维化纤维微细菌的生物学特性及其对几种苯环类化合物的利用研究.《微生物学通报》.2008,第35卷(第7期),1021-1027. * |
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