CN104031860B - A kind of spherical bacillus of high-yield thermostable zytase and application thereof - Google Patents
A kind of spherical bacillus of high-yield thermostable zytase and application thereof Download PDFInfo
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Abstract
A kind of spherical bacillus of high-yield thermostable zytase and application thereof, does is Classification And Nomenclature spherical lysine bacillus? (Lysinibacillus? sphaericus)? Xyn-1, being preserved in Chinese Typical Representative culture collection center C CTCC on May 5th, 2014, does is deposit number CCTCC? M? 2014183. Adopt Plackett-Burman design and response surface analysis method to determine the best fermentating enzyme-producing condition of bacterium shaking flask, can the fermentation level under this condition reach 5678.5? IU/mL. Utilize enzyme that this strain fermentation produces to be applied to the degraded of plant hemicellulose, can reach more than 60% through the xylobiose yield of hemicellulose enzymolysis.
Description
Technical field
The present invention relates to a kind of spherical bacillus and application thereof of high-yield thermostable zytase, belong to using microbeTechnical field.
Background technology
Zytase (XylanaseE.C3.2.1.8) is a kind of main xylan degrading enzyme, and it is by interior buttβ-Isosorbide-5-Nitrae wood sugar glycosidic bond in formula degradation of xylan, hydrolysate is mostly xylo-oligosaccharide, and with a small amount of wood sugar and ArabSugar. Zytase is with a wide range of applications, aspect paper industry: zytase can be used as bio-bleaching agent and substitutes traditionChemical bleaching, thereby reduce in a large number the consumption of chlorine, alleviate the pollution of papermaking to environment, zytase is wide as " slurrying enzyme "The general western developed country that is applied to, has brought good economic and social benefit; Zytase is widely used in food service industryThe preparation of the making food such as Flour product, fruit juice and wood oligose is produced; In fermentation industry, zytase is widely used in wheat etc.On the saccharification of fermentation raw material is processed; Feedstuff industry adds enzyme as a class feed, can effectively improve feed nutrition. ZytaseA staple product in becoming enzyme preparation production industry, market also presents the trend growing with each passing day for its demand.
In addition, can find by the integrated environment analysis to applying in the market zytase, zytase agentApplied environment is mostly in hot conditions, and for example paper-making process, needs high-temperature operation to remove delignification etc.; Food productionGenerally need the operations such as high-temperature sterilization. Research finds, the optimal reactive temperature of zytase is generally lower than 45 DEG C, therefore xylanEnzyme needs to lower the temperature in advance to improve the catalytic activity of enzyme before coming into operation in said process, these operations not only power consumption are also very bigGround hinders the application development of zytase. Therefore, obtain high yield heat resistant xylanase bacterial strain and devoted real attenuation lifeProduct has wide research and application prospect.
Summary of the invention
The object of this invention is to provide a kind of spherical bacillus of high-yield thermostable zytase and produce resistance in fermentationApplication in high-temperature xylanase, has very large promotional value.
The spherical bacillus of high-yield thermostable zytase, Classification And Nomenclature is spherical lysine bacillus Xyn-1(LysinibacillussphaericusXyn-1), in the center preservation of Chinese Typical Representative culture collection, deposit number is:CCTCCM2014183, preservation date is on May 5th, 2014, address: Wuhan, China Wuhan University.
The screening technique of bacterial strain of the present invention is as follows:
Screening and culturing based formulas: peptone 10g, yeast extract 3g, NaCl5g, xylan 4g, agar powder 15g, distilled water1000mL, pH8.0,121 DEG C of sterilizings, 20min.
Collection is positioned at some parts of Nanjing University of Technology's biological methane workshop agricultural straw compost diverse location aroundSample is inoculated in above-mentioned screening and culturing base, cultivates 24～48h, with 0.1% Congo red solution-treated 30min, then uses for 55 DEG C1mol/LNaCl solution decolouring, then chooses on culture medium the large and obvious bacterial strain of hydrolysis circle of transparent circle as primary dcreening operation bacterial strain;Subsequently, the enzymatic production ability of primary dcreening operation bacterial strain is verified, taking xylan as substrate, measured enzyme based on DNS method and live, choose productMeasure a highest strain strain X yn-1, its initial fermentation level is 3245.5IU/mL.
The growth of physiological and biochemical property: strain X yn-1 is with OD600Absorbance represent, by bacterium logarithm period unitGrowth Dai Shiwei 3.0h is calculated in the growth of interior quantity. The molecular biology identification of bacterial strain shows, the 16SrRNA base of strain X yn-1Because sequence is KJ755851 at the number of registration of GenBank, with its phase of the most close kind LysinibacillussphaericusBeing 91% like property, is a new kind bacterium, the spherical lysine bacillus of called after Xyn-1 (LysinibacillusSphaericusXyn-1). This bacterial strain has been preserved in Chinese Typical Representative culture collection center, deposit number on May 5th, 2013Be respectively: CCTCCM2014183.
The spherical bacillus of described high-yield thermostable zytase produces the application of zytase in fermentation.
The application of the spherical bacillus of described high-yield thermostable zytase in degrading straw hemicellulose.
The concrete steps of application are as follows:
1) bacterial strain is carried out to inclined-plane cultivation, be placed at 37 DEG C, cultivate 48～72h, adopt beef-protein medium, toolBody formula: beef extract 3.0g, peptone 10.0g, NaCl5.0g, agar 20g, is dissolved in 1000mL distilled water pH7.4-7.6. In the LB culture medium of the bacterial strain streak inoculation sterilizing that above-mentioned inclined-plane is cultivated (peptone 10g, yeast extract 5g,NaCl10g, is dissolved in 1000mL distilled water pH7.0～7.2), under 37 DEG C of conditions, 200～250r/min cultivates 12h, utilizesCell counting count board is measured bacterial concentration, and is diluted to bacteria suspension concentration 5 × 106Individual/mL.
2) by step 1) the bacterium liquid that obtains is seeded in fermentation medium, and inoculum concentration is 4% (v/v), and culture medium consumption is25mL/250mL conical flask, 45 DEG C of cultivation temperature, shaking flask rotating speed 200r/min, fermentation time is 72h. Described fermentation mediumFor: xylan 10.7g, dusty yeast 5.6g, peptone 2g, NH4NO34g，(NH4)2SO46g，KH2PO40.9g，MgSO4·7H2O1g, NaCl5g, distilled water 1000mL.
3) fermentation prepare the enzymolysis of zytase for wheat stalk hemicellulose: by step 2) obtain zymotic fluid exist5500r/min, centrifugal 5min under 4 DEG C of conditions. Collect supernatant and obtain the fermentation fire resistant xylanase liquid that produces, and answeredFor the enzymolysis of Technique of Hemicellulose from Cornstalk. Enzymolysis process adds zytase with the ratio of 100～150IU/g hemicelluloseLiquid reacts 72h under 55 DEG C of conditions. After reaction finishes, filter and collect enzymolysis liquid, utilize HPLC method to measure respectively reduction in enzymolysis liquidSugar content, and calculate the enzymolysis efficiency of this process with this.
Beneficial effect
The invention provides a plant height and produce fire resistant xylanase strain X yn-1, its condition of culture is relatively gentle, screening bacteriumStrain is tested and appraised and belongs to spherical lysine bacillus, not yet carries out report before this for the zytase research of this bacterial strain,The present invention has further enriched the correlative study content of this kind fungus strain.
The fermentation level that the strain X yn-1 that screening obtains produces fire resistant xylanase reaches 5678.5IU/mL, relatively findsThe zytase production capacity of this bacterial strain is higher, and the fire resistant xylanase that fermentation obtains simultaneously has good heat endurance,Can be widely used in the industry such as papermaking, food.
Adopt statistical method to be optimized research to the production sweat of strain X yn-1, obtained best cultivationScheme, the zytase fermentation level of bacterial strain has had significant raising, is brought up to by the 3245.5IU/mL that initially ferments5678.5IU/mL。
Brief description of the drawings
The spherical lysine bacillus of Fig. 1 Xyn-1 flat-plate bacterial colony aspect graph.
Detailed description of the invention
Screening and the cultivation of embodiment 1 high-yield thermostable zytase bacterial strain
Enriched medium: NaNO30.5g、K2HPO4lg、MgSO4·7H2O0.5g、KCl0.5g、FeSO4·7H2O0.005g, distilled water 1000mL, pH nature, 121 DEG C of sterilizings, 20min.
Screening and culturing base: peptone 10g, yeast extract 3g, NaCl5g, xylan 4g, agar powder 15g,
Distilled water 1000mL, pH8.0,121 DEG C of sterilizings, 20min.
Fermentation medium: peptone 10g, yeast extract 3g, NaCl5g, xylan 4g, distilled water 1000mL, pH nature, goes out121 DEG C of bacterium, 20min.
LB culture medium: peptone 10g, yeast extract 5g, NaCl10g, agar 20g, is dissolved in 1000mL distilled water,PH7.0～7.2,121 DEG C of sterilizings, 20min.
Collection is positioned at Nanjing University of Technology's biological methane workshop some duplicate samples of straw compost diverse location around5g, is placed in respectively 250mL conical flask by it, adds 50mL sterilized water, and 37 DEG C, 180r/min are cultivated 1h and prepared suspension, get 5mLSuspension joins in the triangular flask that fills 50mL enriched medium, after shaken cultivation 5～7d, gets nutrient solution and dilutes respectively 10-1～10-6, and respectively it is evenly applied on screening and culturing base, cultivate 24～48h for 55 DEG C, by 0.1% Congo red solution-treated30min, then decolour with 1mol/LNaCl solution. Observe the transparent circle situation of periphery of bacterial colonies, according to transparent circle and bacterium colony sizeSize can compare bacterial strain and produce the height of Xylanase activity, and carries out the primary dcreening operation of bacterial strain using this as index. Primary dcreening operation completesAfter, to the bacterium liquid dilution 10 of some primary dcreening operation bacterial strains-1～10-6, be inoculated into respectively 35 DEG C of cultivation 72h in fresh fermentation medium,Carrying out bacterial strain using zytase output in zymotic fluid as direct standard sieves again. So subculture is cultivated, superseded fermentation level a little less thanCulture, finally obtain a strain and keep higher xylanase ability and stable bacterial strain, utilize LB culture medium to enter itThe preservation of row bacterial strain purifying utilizes glycerine training method to carry out cold storage processing to purifying bacterial strain simultaneously.
The Biology identification of embodiment 2 bacteriums
1. the Morphological Identification of bacterial strain
Separation and purification inoculation, to solid LB culture medium, is cultivated to the flat board of 24h observation bacterial classification under 37 DEG C of conditionsColonial morphology. As Fig. 1, strain X yn-1, after the growth of solid LB culture medium, colony diameter 0.6cm, shape is regular projectionThing, can form endogenous spore, rounded, the opaque colony that is white in color on culture medium, smooth surface, colony edge rule.
2. the molecules of bacterial strain qualification
Screening bacterium is carried out to bacterial classification qualification, and bacterial classification is identified the bacterial 16 S rDNA sequence library based on ncbi database. ProfitExtract kit by bacterial genomes and extract the DNA of bacterial strain, and utilize bacterial 16 S rDNA universal primer to carry out pcr amplification, drawThing sequence is: 27F (5 '-AGAGTTTGATCMTGGCTCAG-3 '), 1492R (5 '-GGYTACCTTGTTACGACT-3 '). PCRReaction system is (25 μ L): template DNA 10ng, 10 × PCR buffer solution, 2.5 μ L, 25mmol/LMgCl21.5μL，dNTPMix2.5 μ L, the each 0.25 μ L of primer, 5U/ μ LrTaq polymerase 0.125 μ L. PCR response procedures is: 94 DEG C of denaturation 5min, withRear 35 circulations (72 DEG C are extended 2min for 94 DEG C of sex change 30s, 55 DEG C of annealing 1min), last 72 DEG C of reaction 10min. By after purifyingPcr amplified fragment censorship order-checking, and sequencing result is carried out to similitude comparison by the blast program of Genbank in NCBI.
Molecules qualification result: the 16SrDNA region sequence of bacterium Xyn-1 as shown in SEQIDNO:1, this sequenceBe committed to Genbank, number of registration is KJ755851, with its phase of the most close kind LysinibacillussphaericusBeing 91% like property, is a new kind bacterium, the spherical lysine bacillus of called after Xyn-1.
The optimization of embodiment 3 fermentation culture conditions
Adopt single-factor analysis therapy to be optimized strain X yn-1 condition of culture, determine best condition of culture: culture mediumConsumption 25mL/250mL conical flask, 45 DEG C of cultivation temperature, shaking flask rotating speed 200r/min, bacteria suspension concentration is 5 × 106Individual/mL, everyBottle inoculum concentration is 1.0mL, initial pH7.0, and fermentation time is 72 hours. The optimization of culture medium composition:
Determine relevant 9 factors of culture medium of producing enzyme to Xyn-1 by single-factor analysis therapy. Adopt Plackett-BurmanThe enzyme material impact factor is produced in design screening, and the design of selecting test number (TN) N=12 gathers the lactose in culture medium, maltose, wood9 factors such as sugar, dusty yeast, peptone, sodium chloride, magnesium sulfate, ammonium sulfate and potassium dihydrogen phosphate (%) are investigated, and the results are shown inTable 1～2, on the order of producing enzyme impact are: xylan > dusty yeast > potassium dihydrogen phosphate > peptone.
Table 1 strain X yn-1 cultivates factor Plackett-Burman design experiment
| Experimental group | A | B | (C) | D | E | F | G | H | I | J | (K) | Enzyme (IU/mL) alive |
| 1 | 1 | -1 | 1 | 1 | -1 | 1 | -1 | -1 | -1 | 1 | 1 | 3279.24 |
| 2 | -1 | 1 | 1 | 1 | -1 | 1 | 1 | -1 | 1 | -1 | -1 | 3391.19 |
| 3 | -1 | 1 | 1 | -1 | 1 | -1 | -1 | -1 | 1 | 1 | 1 | 3267.78 |
| 4 | -1 | -1 | -1 | 1 | 1 | 1 | -1 | 1 | 1 | -1 | 1 | 3011.58 |
| 5 | -1 | 1 | -1 | -1 | -1 | 1 | 1 | 1 | -1 | 1 | 1 | 3311.02 |
| 6 | 1 | -1 | -1 | -1 | 1 | 1 | 1 | -1 | 1 | 1 | -1 | 3138.08 |
| 7 | 1 | 1 | -1 | 1 | -1 | -1 | -1 | 1 | 1 | 1 | -1 | 3395.20 |
| 8 | 1 | -1 | 1 | -1 | -1 | -1 | 1 | 1 | 1 | -1 | 1 | 3084.53 |
| 9 | -1 | -1 | 1 | 1 | 1 | -1 | 1 | 1 | -1 | 1 | -1 | 3154.11 |
| 10 | -1 | -1 | -1 | -1 | -1 | -1 | -1 | -1 | -1 | -1 | -1 | 3178.16 |
| 11 | 1 | 1 | -1 | 1 | 1 | -1 | 1 | -1 | -1 | -1 | 1 | 3239.72 |
| 12 | 1 | 1 | 1 | -1 | 1 | 1 | -1 | 1 | -1 | -1 | -1 | 3161.27 |
To three key factors, i.e. xylan, dusty yeast, potassium dihydrogen phosphate, has carried out the center based on Box-BehnkenModular design, has designed totally 17 groups of experimental points of 3 levels of 3 factors, completes response surface analysis optimization, as shown in table 2.
The experiment of table 2 strain X yn-1 fermentation condition Box-Behnken modular design
In analytic process, set up Quadratic response surface regression model, to analyze optimal response factor level, obtained matchingQuadratic regression equation is as follows:
Secondary respective face regression model is significant (R2=0.9884), models fitting degree is fine, illustrates that this model returnsReturn significantly, so the theoretical prediction that this model can be optimized for strain X yn-1 enzyme ferment condition. By data analysis, mouldType proposes the Optimal compositions of fermentation medium condition of this process: xylan 10.7g, dusty yeast 5.6g, peptone 2g, NH4NO34g，(NH4)2SO46g，KH2PO40.9g，MgSO4·7H2O1g, NaCl5g, distilled water 1000mL, the enzymatic production of model prediction is the highestLevel reaches 5653.11IU/mL, compared to the 3245.5IU/mL of initial fermentation level, utilizes this optimization method can significantly improve bacteriumThe enzymatic production ability of strain Xyn-1.
Fire resistant xylanase is produced in embodiment 4 fermentations
Slant medium: adopt beef-protein medium, specifically formula: beef extract 3.0g, peptone 10.0g,NaCl5.0g, agar 20g, is dissolved in 1000mL distilled water pH7.4-7.6.
LB culture medium: peptone 10g, yeast extract 5g, NaCl10g, agar 20g, is dissolved in 1000mL distilled water,PH7.0～7.2,121 DEG C of sterilizings, 20min.
Fermentation medium: xylan 10.7g, dusty yeast 5.6g, peptone 2g, NH4NO34g，(NH4)2SO46g，KH2PO40.9g，MgSO4·7H2O1g, NaCl5g, distilled water 1000mL, pH nature;
First strain X yn-1 is seeded to slant medium, is placed at 37 DEG C, cultivate 48～72h. Choose single bacterium from inclined-planeDrop down onto in 250mL conical flask, LB culture medium is housed in conical flask, liquid amount is 25mL/250mL conical flask, under 37 DEG C of conditionsBe placed in the shaking table of 200～250r/min, cultivate 12h, obtain first order seed. By the counting to first order seed, controlled fermentationInoculum concentration be that (bacteria suspension concentration is 5 × 10 to 1mL6Individual/mL).
Producing enzyme fermentation condition of culture is: fermentation medium consumption 25mL/250mL conical flask, and 45 DEG C of cultivation temperature, shaking table turnsSpeed 200r/min, bacteria suspension concentration is 5 × 106Individual/mL, every bottle of inoculum concentration is 1mL, initial pH7.0, fermentation time is 72h, sends outFerment finishes, and measures the enzyme of fire resistant xylanase in zymotic fluid and lives. Zytase (Xylanase) assay method reference literature(BaileyMJ,PeterB,andKaisaP.InterlaboratorytestingofmethodsforassayOfxylanaseactivity[J] .JournalofBiotechnology, 1992,23 (3): 257-270.), xylanEnzyme activity unit (IU) is defined as 1min hydrolyzed xylan substrate and generates the required enzyme amount of 1 μ mol wood sugar and be defined as an enzyme activityUnit. Found that fermentation level is up to 5678.5IU/mL, by zymotic fluid centrifugal treating 5min under the rotating speed of 5500r/min,Collect supernatant and be fermenting enzyme liquid.
The application that embodiment 5 strain X yn-1 produce zytase degrading maize straws hemicellulose
1. the extraction of Technique of Hemicellulose from Cornstalk
The maize straw reclaiming carries out pulverization process, sieves and obtains the stalk that particle mean size is 2mm. By after treatmentStraw sample washing and drying. Take 10g and dry straw sample, add wherein 1% NaOH solution 100mL, and at 65 DEG C of barsUnder part, stir 60min. After reaction finishes, add ethanol/water (50/50 volume ratio) solvent, and by filtered and recycled filtrate. RegulateFiltrate pH to 6.0, and be placed in 4 DEG C of standing 30min, by solution centrifugal 5min under 5000r/min condition, remove supernatant subsequentlyLiquid, reclaims precipitation washing and drying and obtains experiment Technique of Hemicellulose from Cornstalk sample.
2. strain X yn-1 produces zytase degrading maize straws hemicellulose
Based on the research of the above-mentioned sweat for strain X yn-1, determine that this bacterial strain produces the best cultivation of zytaseCondition, and the application of producing enzyme with this. According to the process described in embodiment 4, strain X yn-1 is produced to enzyme and cultivate and obtainCorresponding fermenting enzyme liquid, the enzyme work of measuring fire resistant xylanase in zymotic fluid reaches 5678.5IU/mL. Take jade prepared by 0.5gRice stalk hemicellulose sample, and add zytase liquid with the enzymolysis ratio of 100～150IU/g hemicellulose, at 55 DEG C of barsUnder part, carry out enzymolysis 72h. After reaction finishes, filter and collect enzymolysis liquid, utilize HPLC method to measure each content of reducing sugar in enzymolysis liquid,And calculate the enzymolysis efficiency of this process with this. Experimental result shows, utilizes the enzymolysis liquid of strain X yn-1 fermentation preparation to carry out cornThe degraded of stalk hemicellulose, its xylobiose yield nearly 60%.
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Claims (8)
1. a plant height produces the spherical bacillus of fire resistant xylanase, and its Classification And Nomenclature is spherical lysine bacillus(Lysinibacillussphaericus) Xyn-1, is preserved in Chinese Typical Representative culture collection on May 5th, 2014Heart CCTCC, deposit number is CCTCCNO:M2014183.
2. the spherical bacillus of high-yield thermostable zytase claimed in claim 1 produces the application of zytase in fermentation.
3. the spherical bacillus of high-yield thermostable zytase claimed in claim 1 answering in degrading straw hemicelluloseWith.
4. the spherical bacillus of high-yield thermostable zytase according to claim 2 produces answering of zytase in fermentationWith, it is characterized in that comprising the steps:
1) inclined-plane is cultivated: spherical lysine bacillus Xyn-1 is seeded on slant medium, at 37 DEG C, cultivates 48 ~ 72h；
2) seed culture: the single bacterium colony on picking step 1) inclined-plane, in LB culture medium, is placed in 200 ~ 250 under 37 DEG C of conditionsIn the shaking table of r/min, cultivate 12h, obtain seed liquor;
3) enzymatic production: by step 2) in seed liquor be seeded in fermentation medium, inoculum concentration is for 1 ~ 10% connecing by volumeKind, 45 DEG C of bottom fermentation 72h preparations contain the zymotic fluid of fire resistant xylanase.
5. the spherical bacillus of high-yield thermostable zytase according to claim 4 produces answering of zytase in fermentationWith, it is characterized in that: described in step 1), slant medium is: beef extract 3.0g, peptone 10.0g, NaCl5.0g, fine jadeFat 20g, is dissolved in 1000mL distilled water pH7.4-7.6.
6. the spherical bacillus of high-yield thermostable zytase according to claim 4 produces answering of zytase in fermentationWith, it is characterized in that: step 2) described LB culture medium is: peptone 10g, yeast extract 5g, NaCl10g, agar 20G, is dissolved in 1000mL distilled water pH7.0 ~ 7.2.
7. the spherical bacillus of high-yield thermostable zytase according to claim 4 produces answering of zytase in fermentationWith, it is characterized in that: described in step 3), fermentation medium is: xylan 10.7g, dusty yeast 5.6g, peptone 2g, NH4NO34g，(NH4)2SO46g，KH2PO40.9g，MgSO4·7H2O1g, NaCl5g, distilled water 1000mL, pH nature.
8. the spherical bacillus of high-yield thermostable zytase according to claim 3 is in degrading straw hemicelluloseApplication, it is characterized in that: spherical lysine bacillus Xyn-1 fermentation is obtained to the zymotic fluid containing fire resistant xylanase,Enzymolysis ratio with 100 ~ 150IU/g hemicellulose adds the zymotic fluid containing fire resistant xylanase, enzymolysis under 55 DEG C of conditions72h。
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| CN105567605A (en) * | 2016-02-19 | 2016-05-11 | 黑龙江省科学院大庆分院 | Lysinibacillus sp. and application thereof, degumming auxiliary containing lysinibacillus sp. and preparation method of degumming auxiliary |
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| CN101402963A (en) * | 2008-11-14 | 2009-04-08 | 南开大学 | Fire resistant xylanase XynA1, gene for encoding the enzyme and uses thereof |
| CN102321558A (en) * | 2011-09-22 | 2012-01-18 | 天津科技大学 | High-yield strain of high temperature resistant 1,4-beta-D-xylanase, method for producing high temperature resistant 1,4-beta-D-xylanase through fermentation of high-yield strain, and high temperature resistant 1,4-beta-D-xylanase |
| CN103667151A (en) * | 2013-12-13 | 2014-03-26 | 湖南鸿鹰生物科技有限公司 | Bacillus thermophilus capable of producing high-temperature and alkali resistant xylanase and application of bacillus thermophilus |
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2014
- 2014-05-27 CN CN201410228680.2A patent/CN104031860B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101402963A (en) * | 2008-11-14 | 2009-04-08 | 南开大学 | Fire resistant xylanase XynA1, gene for encoding the enzyme and uses thereof |
| CN102321558A (en) * | 2011-09-22 | 2012-01-18 | 天津科技大学 | High-yield strain of high temperature resistant 1,4-beta-D-xylanase, method for producing high temperature resistant 1,4-beta-D-xylanase through fermentation of high-yield strain, and high temperature resistant 1,4-beta-D-xylanase |
| CN103667151A (en) * | 2013-12-13 | 2014-03-26 | 湖南鸿鹰生物科技有限公司 | Bacillus thermophilus capable of producing high-temperature and alkali resistant xylanase and application of bacillus thermophilus |
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| Title |
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| Lysinibacillus xylanilyticus sp. nov., a xylandegrading bacterium isolated from forest humus;Chang Soo Lee等;《International Journal of Systematic and Evolutionary Microbiology》;20090803;第60卷;281-286页 * |
| 中国新分离碱性耐热芽孢杆菌的木聚糖酶产酶特征;易霞 等;《生物技术》;20071031;第17卷(第5期);31-35页 * |
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