CN109868232A - A kind of teas bacillus and its application - Google Patents
A kind of teas bacillus and its application Download PDFInfo
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- CN109868232A CN109868232A CN201811086378.2A CN201811086378A CN109868232A CN 109868232 A CN109868232 A CN 109868232A CN 201811086378 A CN201811086378 A CN 201811086378A CN 109868232 A CN109868232 A CN 109868232A
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- 239000008103 glucose Substances 0.000 description 1
- 229940116332 glucose oxidase Drugs 0.000 description 1
- 235000019420 glucose oxidase Nutrition 0.000 description 1
- 229930182470 glycoside Natural products 0.000 description 1
- 210000005255 gram-positive cell Anatomy 0.000 description 1
- 150000002431 hydrogen Chemical class 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 239000003262 industrial enzyme Substances 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 238000005304 joining Methods 0.000 description 1
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- BJHIKXHVCXFQLS-FUTKDDECSA-N keto-L-fructose Chemical compound OC[C@H](O)[C@H](O)[C@@H](O)C(=O)CO BJHIKXHVCXFQLS-FUTKDDECSA-N 0.000 description 1
- 239000010985 leather Substances 0.000 description 1
- TWRXJAOTZQYOKJ-UHFFFAOYSA-L magnesium chloride Substances [Mg+2].[Cl-].[Cl-] TWRXJAOTZQYOKJ-UHFFFAOYSA-L 0.000 description 1
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- 229960002160 maltose Drugs 0.000 description 1
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- 235000013372 meat Nutrition 0.000 description 1
- 238000002844 melting Methods 0.000 description 1
- 230000008018 melting Effects 0.000 description 1
- 235000013336 milk Nutrition 0.000 description 1
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- 229910052757 nitrogen Inorganic materials 0.000 description 1
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- 230000020477 pH reduction Effects 0.000 description 1
- 238000004806 packaging method and process Methods 0.000 description 1
- 238000012856 packing Methods 0.000 description 1
- 239000001814 pectin Substances 0.000 description 1
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- 230000035699 permeability Effects 0.000 description 1
- 238000013081 phylogenetic analysis Methods 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 229940041153 polymyxins Drugs 0.000 description 1
- 239000000256 polyoxyethylene sorbitan monolaurate Substances 0.000 description 1
- 235000010486 polyoxyethylene sorbitan monolaurate Nutrition 0.000 description 1
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 description 1
- 229920000053 polysorbate 80 Polymers 0.000 description 1
- 230000008092 positive effect Effects 0.000 description 1
- 229910052700 potassium Inorganic materials 0.000 description 1
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- MUPFEKGTMRGPLJ-ZQSKZDJDSA-N raffinose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO[C@@H]2[C@@H]([C@@H](O)[C@@H](O)[C@@H](CO)O2)O)O1 MUPFEKGTMRGPLJ-ZQSKZDJDSA-N 0.000 description 1
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- 229910000030 sodium bicarbonate Inorganic materials 0.000 description 1
- 235000017557 sodium bicarbonate Nutrition 0.000 description 1
- 229910000029 sodium carbonate Inorganic materials 0.000 description 1
- SUKJFIGYRHOWBL-UHFFFAOYSA-N sodium hypochlorite Chemical compound [Na+].Cl[O-] SUKJFIGYRHOWBL-UHFFFAOYSA-N 0.000 description 1
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- UQZIYBXSHAGNOE-XNSRJBNMSA-N stachyose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO[C@@H]2[C@@H]([C@@H](O)[C@@H](O)[C@@H](CO[C@@H]3[C@@H]([C@@H](O)[C@@H](O)[C@@H](CO)O3)O)O2)O)O1 UQZIYBXSHAGNOE-XNSRJBNMSA-N 0.000 description 1
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Landscapes
- Enzymes And Modification Thereof (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The present invention relates to a kind of teas bacillus and its applications.The bacterium be teas bacillus (Paenibacillus theae) YN15, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3 China Committee for Culture Collection of Microorganisms's common micro-organisms center is preserved on March 1st, 2017, deposit number is CGMCC No.13439.The discovery of the novel species and using enrich we using microbial resources.Series bacillus novel bacterial provided by the present invention has alpha-galactosidase, beta galactosidase and tryptic activity, the angle excited from food safety and enzyme activity in situ, microorganism resource is provided for the processing of Pu'er tea and other industrial applications of protease, with good application prospect.
Description
Technical field
The present invention relates to a kind of teas bacillus (Paenibacillus) novel bacterial and its cultural method and application.
Background technique
Using morphology as basis of classification, series bacillus is included into bacillus by early stage research.1993, Ash et al.
It is stood to share 231 kinds so far after bacillus genus, sees http://www.bacterio.net/
paenibacillus.html.Type sepecies be mostly viscous class gemma bars (PaenibacilluspolymyxaATCC 842T).Class bud
The separation source of spore Pseudomonas is very extensive, is isolated from the environment such as rhizosphere soil, air, water body and food.1996, by
Heyndrickx etc. corrects the correlation properties of class gemma Pseudomonas.The growth of aerobic or amphimicrobian, produces gemma, and cell is in bar
Shape, gram-positive cell wall, moves by periflagellum, medium temperature, and main fatty acid is anteiso- formula saturated fatty acid C15:0.Class
Gemma Pseudomonas G+C content range is 39 ~ 59 mol%.
The concept of polyphase sort was initially proposed by Colwell in 1970, was referred to using a variety of different information of microorganism,
Including phenotype, genotype and phylogenetic information integrates the process of microorganisms classification and phyletic evolution.
Microorganism in bacillus genus can produce various bioactivators, be with a wide range of applications.Such as
Type sepecies Paenibacillus polymyxa can produce polymyxins, can destroy the permeability of gram negative bacterial cell plasma membrane, cause
Intracellular organic matter leakage has huge market and application prospect to play bactericidal effect.It is isolated from the class gemma bar of Pu'er tea
Bacterium provides microorganism resource for the processing of Pu'er tea and other industrial applications of protease, with good application prospect.
Summary of the invention
One of the objects of the present invention is to provide a kind of teas bacillus.The strain has trypsase and beta galactose
Glycosides enzymatic activity provides strain money in industrial applications such as industry, food, medicine and agriculturals for beta galactosidase and trypsase
Source.
The second object of the present invention is to provide the preparation method of the strain.
The third object of the present invention is to provide the application of the strain.
In order to achieve the above object, the present invention adopts the following technical scheme:
A kind of teas bacillus, it is characterised in that the deposit number of the strain is CGMCC No.13439.
The 16S rDNA of above-mentioned teas bacillus is base sequence shown in SEQ ID NO.1.
A method of preparing above-mentioned teas bacillus, it is characterised in that the specific steps of this method are as follows: by class bud
Spore bacillus is inoculated in culture medium, and aerobic culture is carried out under conditions of 25-45 DEG C, pH 6.7-8.2.
Above-mentioned cultivation temperature is 37 DEG C, and pH is 6.8 ~ 7.3.
There are also the NaCl that mass percent is no more than 8 % in above-mentioned culture medium.
A kind of bacterial strain of above-mentioned teas bacillus answering in liquid fermentation production galactosidase and trypsase
With.
On the basis of common knowledge of the art, above-mentioned each optimum condition, can any combination to get each preferable reality of the present invention
Example.The reagents and materials used in the present invention are commercially available.
The positive effect of the present invention is that: the present invention provides a novel species for teas bacillus, the bacterial strains
Classification position bePaenibacillusSp., it according to the naming method of the international bacterial systematics committee, is intended to order this kind
It is entitledPaenibacillustheaesp.nov..The discovery of the novel species and using enrich we using microbial resources,
Certain contribution is preferably made that using series bacillus later to us.Series bacillus YN15 of the invention can extract
Beta galactosidase and trypsase are widely used in industries such as food, medicine and chemical industry.
Biological deposits material proves:It is micro- to be deposited in China on March 1st, 2017 by teas Bacillus YN15 of the invention
Biological inoculum preservation administration committee common micro-organisms center (CGMCC), preservation address: BeiChen West Road, Chaoyang District, BeiJing City 1
Institute 3, Institute of Microorganism, Academia Sinica, postcode: 100101.The deposit number of the bacterial strain are as follows: CGMCC No. 13439.
The classification naming of the bacterial strain is class BacillusPaenibacillustheaeYN15。
Detailed description of the invention
Fig. 1 shows the 16S rRNA phylogenetic evolution tree of series bacillus novel species bacterial strain YN15 of the present invention.
Specific embodiment
The present invention is further illustrated below by the mode of embodiment, but does not therefore limit the present invention to the reality
It applies among a range.In the following examples, the experimental methods for specific conditions are not specified, according to conventional methods and conditions, or according to quotient
The selection of product specification.Heretofore described room temperature refers to the temperature for the operation room tested, generally 25 DEG C.
Embodiment 1, new strains YN15 of the present invention are isolated and purified
Pu'er cooked tea leachate is taken, by its dilution spread in TSA solid medium, 37 DEG C are cultivated 2-3 days, picking single colonie,
Then scribing line purifying obtains.
It is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center (CGMCC), preservation is compiled
Number be CGMCC No.13439.
The appearance features of embodiment 2, new strains YN15 of the present invention
1. colony characteristics
The single colonie for taking bacterial strain YN15 is transferred on TSA solid medium (agar), cultivated in 37 DEG C of constant incubators for 24 hours,
36h and 48h, the features such as observing the size, color, edge, protrusion, smoothness, viscosity, transparency of its bacterium colony respectively.As a result it shows
Show, neat in edge, microprotrusion is smooth, opaque, milky bacterium colony, diameter about 1mm.
2. cell morphology characteristic
Bacterial strain YN15 is Gram-positive bacillus, and top round blunt, atrichia, production gemma, cell size is about 1.0 μ m, 3.1 μ
m；The life of cell thallus list or fasciation.
The growth characteristics of embodiment 3, new strains YN15 of the present invention
Picking cultivates the fresh cultured object of 24 h on TAS solid medium (agar), is inoculated into TSA fluid nutrient medium, and 37 DEG C
20 ~ 24 h of shaking table culture, as seed.
The composition (1000ml) of liquid TSA culture medium: tryptone 15 g, soy peptone 5g, yeast extract 2.5g,
Glucose 5g, starch 4g, distilled water are settled to 1000ml.
1. growth temperature:
In the aseptic inoculation TSA fluid nutrient medium for transferring fresh by 1% (v/v) inoculum concentration by the YN15 seed cultivated, mix.
It is respectively placed in 20 DEG C, 25 DEG C, 30 DEG C, 37 DEG C, 40 DEG C, 45 DEG C, 50 DEG C of water-bath culture, measures its growing state.Obtain bacterial strain
YN15 growth temperature range is 25-45 DEG C, 37 DEG C of optimum temperature.
2. growing NaCl tolerance
The YN15 seed cultivated by 2% (v/v) inoculum concentration switching NaCl concentration is respectively 0.0%, 1.0%, 2.0%, 3.0%,
4.0%, 5.0%, 6.0% and 8.0% TSA culture medium, 37 DEG C of cultures, keeps a record to growth conditions.Bacterial strain YN15 as the result is shown
NaCl tolerance is 0-5%, optimum concentration 0%.
3. growing pH range
Prepare the sodium carbonate liquor of 0.1M, the sodium bicarbonate solution of 0.1M, 1/15M disodium phosphate soln, 1/15M biphosphate
Potassium solution, the disodium phosphate soln of 0.2M, the citric acid solution of 0.1M as mother liquor, with mother liquor pH value be respectively 3.0,
4.0,5.0,6.0,7.0,8.0,9.0, the YN15 seed cultivated is accessed by 1% inoculum concentration, 37 DEG C of cultures, to growth conditions
It keeps a record.The pH range for obtaining bacterial strain YN15 growth is 6.7 ~ 8.2, and optimal pH is 6.8 ~ 7.3.
The physio-biochemical characteristics of embodiment 4, new strains YN15 of the present invention
Utilize API ZYM and API20E identification systems (production firm: bioM é rieux) and the physiological and biochemical test method of routine
(east show pearl, Cai Miaoying etc., 2001) carries out physiological and biochemical property identification to bacterial strain YN15 and the relevant bacterial strain that belongs to.
Wherein API ZYM is the micro method system an of sxemiquantitative, is aimed at designed by research enzyme activity.The technology pair
Various samples (tissue, cell, biological fluid, washing water, soil, oil, etc.) be all suitable for.It system and can rapidly study 19
The activity of kind enzyme.It is few with sample amount.Experimental procedure is as follows:
1) preparation of samples: minimum volume dilute sample: can with 2 ml sterile distilled waters or it is other such as common physiological saline not
Need buffer.A bacteria suspension is prepared, turbidity is between McFarland No 5 and No 6.From inclined-plane or centrifugation meat soup
The pure culture of culture can be used to prepare bacteria suspension.
2) preparation of test bar: prepare a culture plate and lid.Remember sample number in the side of disk.A plastics can be used
Wash bottle, packing about 5ml distilled water is in being able to maintain certain humidity in culture plate, when in order to cultivate.It is taken out from the packaging of sealing
API ZYM test bar, sets in culture plate.
3) inoculation of test bar: being inoculated with suction pipe, and 2 drops sample (65 μ l) are accessed in each cup of test bar.
4) test bar culture: after inoculation, plastic closure is put on pallet, 37 DEG C are cultivated 4 hours.All determination condition (when
Between, temperature, culture medium, suspension concentration) to be consistent.The test bar of inoculation is kept in dark place.
5) the result observation of test bar: after culture, 1 drop ZYM A reagent and 1 is added to drip ZYM B reagent.Add lustre to after five minutes, such as
Fruit be it is positive, it is 10 seconds lower that test bar is placed in intense light source (1000 watts of light bulbs), light bulb is placed on cup upper 4 seconds.This is to disappear
Except the yellow of strong orchid extra in cup.Negative reaction becomes colorless after exposure.Test bar is set again after daylight lower a few minutes, so that it may
Generate comparison result.
6) record reaction: shown in reaction result following table
| Alkaline phosphatase | + | Guang ammonia calculates arylaminopeptidase | – | Beta galactosidase | + |
| Esterase (C4) | + | Trypsase | + | Beta-glucuronidase enzyme | – |
| Lipoid esterase (C8) | w | Chymotrypsin | + | Alpha-Glucosidase | + |
| Lipoidase (C14) | – | Acid phosphatase | + | β-glucosyl enzym | – |
| Leucine arylaminopeptidase | + | Naphthols-AS-BI- phosphohydrolase | + | N- acetyl-glucosaminidase | – |
| Valine arylaminopeptidase | – | Alpha-galactosidase | + | Alpha-Mannosidase | – |
| β-fucosidase | + |
The major physiological biochemical character of bacterial strain YN15: facultative aerobic growth.NaCl tolerance is 0%-5%, is grown at 0% best.
The pH range of growth is 6.7 ~ 8.2, and optimal pH is 6.8 ~ 7.3.Catalase is negative；Oxidizing ferment characteristic measurement is feminine gender；Starch water
Solution is positive, and hydrolyzes Tween 60 and aesculin；Tween 20, Tween 80, casein, sodium cellulosate, xylan are not hydrolyzed
Equal substances；It is detected using BiologGenlll microwell plate, YN15 can utilize dextrin, D-Maltose, D- trehalose, D- fiber two
Sugar, gentiobiose, sucrose, D- turanose, stachyose, gossypose, α-D- lactose, melibiose, α-D- glucose, D-MANNOSE, D-
Fructose, L- fructose, D-Fructose -6- phosphoric acid, pectin, D- galacturonic acid, D- glucuronic acid etc. are used as carbon source.
Embodiment 5, new strains YN15(CGMCC No.13439 of the present invention) 16S rRNA gene PCR amplification and sequence
Measurement and 16S rRNA phylogenetic character.
1. extracting genomic DNA
By series bacillusPaenibacillusSp. YN15(CGMCC No.13439) it is inoculated in TSA fluid nutrient medium, it will give birth to
The long fermentation liquid to late log phase, 12000 revs/min are centrifuged 1 minute, remove supernatant；With TES (50 mM Tris, 50 mM
EDTA-Na2, 50mMNaCl, pH 8.0-8.2) and solution washes 3 times；Thallus is mixed with 0.4 mL TES solution, is added appropriate molten
Bacterium enzyme, 37 DEG C keep the temperature 1 hour；0.04 mL20%SDS is added, 60 DEG C keep the temperature 30 minutes；0.18 mL 5M NaClO is added4, mix
It is even；Isometric chloroform-isoamyl alcohol (24:1) is added, gently shakes up 1 minute or so, is centrifuged (12000 revs/min, 10 minutes),
Aspirate supernatant；Supernatant is added 37 DEG C of 20 μ l, 0.2 % RNA enzyme and keeps the temperature 30 minutes, chloroform-isoamyl alcohol (24:1, v/v)
Processing one time；20 μ l Proteinase Ks (50-70 μ g/mL) is added in supernatant, and 37 DEG C keep the temperature 1 hour, chloroform-isoamyl alcohol (24:
1, v/v) it handles one time；2 times of volume ice ethanol precipitations of supernatant, 70% ice alcohol solution dipping 5 minutes, 12000/ minute from
The heart 5 minutes.It is dissolved in after drying in sterile water as template DNA.
The PCR amplification and sequencing of 2.16S rRNA gene
Forward primer for PCR amplification is 5 '-AGAGTTTGATCCTGGCTCAG-3 ' (nt 8-27), reverse primer 5 '-
AAGGAGGTGATCCAGCC-3 ' (nt 1541-1557), correspond respectively to the 16S rRNA gene of Escherichia coli 8-27 and
1541-1557 base.PCR reaction system (20 μ l) are as follows: 10 × buffer, 2 μ l, 25 mmol/L MgCl22μL、10 mmol/
1.5 μ l of L dNTPs, each 1 μ l of 30 pmol/L primers, ddH213.4 μ l of O, 1 μ l of Taq DNA enzymatic, 1 μ l of template.PCR reaction
Condition are as follows: 95 DEG C of 10 min, 95 DEG C of 1 min, 55 DEG C of 1 min, 72 DEG C of 1 min30s, 30 circulations；72 DEG C of 10 min, 4
DEG C save.
The sequencing of PCR product using ABI BigDye3.1 sequencing kit (Applied Biosystems) and
Automatic dna sequencer (model ABI3730; Applied Biosystems).
Sequencing result shows bacterial strain YN15(CGMCC No.13439) 16S rRNA gene order length be 1574bp.
16S rRNA sequence as described above using software MEGA version6.0.software package draw into
Change relational tree.It is calculated using neighbor-joining, and with maximum-likelihood and maximum-parsimony
Verifying calculating is carried out, bootstrap is set as 1000 circulations.As a result as shown in the figure.
By the 16S rRNA gene order and phylogenetic analysis of bacterial strain YN15, primarily determine that YN15 is class gemma bar
A member of Pseudomonas, and withPaenibacilluscontaminansCKOBP-6T16S rRNA gene similarity degree highest, together
Source property is up to 93%, the boundary value 97% far smaller than planted.
The content of fatty acid feature of the new microbe of the present invention of embodiment 6
The measurement of the total fatty acid content of bacterial strain YN15.
Configure following solution: I, 45g sodium hydroxide are dissolved in 150ml methanol and 150ml distilled water；The dense salt of II, 190ml
Acid, 275ml methanol are dissolved in 135ml distilled water；III, 200ml n-hexane are uniformly mixed with 200ml ether；IV, 10.8 grams
Sodium hydroxide is dissolved in 900ml distilled water；V, saturated sodium chloride solution.
1) appropriate bacterial cultures is taken, is placed in 8ml screw socket glass tube, 1ml solution is added, tighten blind nut, boiling water bath
5min takes out oscillation 5-10 seconds, tightens blind nut once again, continues boiling water bath 25min；
2) after sample cell is cooling, 2ml solution is added, oscillation is covered tightly, 80 ± 1 DEG C of water-bath 10min, ice bath are then accurately controlled
It is cooling；This step needs strict temperature control and time, in case carboxylic acid and ring type fatty acid are destroyed；
3) 1.25ml solution is added in cooling sample cell, quick oscillation 10min or so discards lower layer's water phase；
4) 3ml solution is added in remaining organic phaseAnd a few drop solution, quick oscillation 5min or so takes on 2/3rds
Layer organic phase is set spare in gas-chromatography sample bottle.
6890 gas chromatograph of HP is equipped with shunting/Splitless injecting samples mouth, flame ionization ditector (FID) and HP gas
Phase chromatography chem workstation (HP CHEMSTATION ver A 5.01)；Chromatographic column is Ultra-2 column, long 25m, internal diameter
0.2mm, 0.33 μm of thickness of liquid film；Furnace temperature is second order temperature programming: 170 DEG C of initial temperature, every min5 DEG C rises to 260 DEG C, then
310 DEG C are risen to 40 DEG C/min, maintains 1.5min；250 DEG C of injector temperature, carrier gas is hydrogen, and flow velocity 0.5ml/min is shunted
Sample introduction mode, split ratio 100:1,2 μ l of sample volume；300 DEG C of detector temperature, hydrogen flow rate 30ml/min, air velocity
216ml/min supplements gas (nitrogen) flow velocity 30ml/min.
The results show that the main cell fatty acid of bacterial strain YN15 is anteiso- formula saturated fatty acid anteiso-C15:0, percentage
Content is 43%.Meet the main fatty acid type of bacillus genus.And with equal on similar strain fatty acid species and content
It is variant, judge that similar strain belongs to variety classes with this.
G+C mol% the content characteristics of the new microbe of the present invention of embodiment 7
The G+C mol% assay of bacterial strain YN15 genomic DNA.
Using melting temperature (Tm) method, with escherichia coli (E.coliK12, AS1.365) it is control comparisons, instrument used
Device is Agilent Technologies company Cary Series UV-Vis Spectrophotometer, with PTP-1 number
Temperature controller temperature control.Steps are as follows:
1) DNA sample to be measured is diluted to OD with 0.1 × SSC260Nm value is between 0.3 ~ 0.4；
2) the OD value for recording 25 °C first in wavelength 260nm, then sets temperature program, to 95 °C since 30 DEG C, therebetween often
Minute increases 1 DEG C；
3) OD value, which rises, indicates that denaturation starts, and records colorimetric utensil temperature and OD value, until the denaturation of OD value invariant representation finishes；
4) it according to thermal denaturation curve, obtains molten chain temperature (Tm), calculates G+C mol% content.
The calculation formula in 0.1 × SSC solution are as follows:
+ 2.08 (Tm of G+C mol%=G+Cmol% (AS1.365)It is unknown- Tm AS1.365)
Test measurementE.coli K12 AS1.365Tm be 79.07 °C and 51.2 mol%, the Tm value and G of strain to be tested YN15
+ Cmol% is respectively 82.02 °C and 57.7 mol%.
The purposes of the new microbe of the present invention of embodiment 8
It is measured through API ZYM identification systems (production firm: bioM é rieux), bacterial strain YN15 has beta galactosidase and pancreas egg
White enzyme.
Beta galactosidase is commonly called as lactase, can be catalyzed beta galactose glycosidic bond and hydrolyze.Currently, the application of lactase
Field is very wide.In dairy industry, lactase can be used for producing Low lactose milk product and galactooligosaccharide.In Dairy Processing, adopt
It, can be to avoid the problems such as crystallization of lactose, whey are precipitated in freeze concentration dairy products with lactose enzyme hydrolysis lactose.In the life of Yoghourt
In production, acidification reaction can be accelerated and improve fermentation efficiency, make Yoghourt that there is distinctive olibanum flavor.It, can be in cheesemaking
Accelerate the maturation of cheese.In terms of analysis, glucose oxidase and lactase are used in combination and prepare biosensor, directly surveyed
Determine the content of lactose in the dairy products such as cow's milk, the method is simple and efficient, low-cost.In terms of environmental protection, lactase can divide
The lactose in whey is solved, higher BOD pollution caused by water after wheys draining is slowed down.As people are to lactase
Further research, lactase will be used wider and wider, and microbe-derived lactase is paid close attention to by people.Separate sources
Lactose enzyme viability it is different, there is also difference, the microbial resources for obtaining more galactopoiesis carbohydrases can mention application range
The adaptability of high enzyme.
Protease is the class of enzymes that protein peptide bond is catalytically decomposed, closely bound up with the life of the mankind.In all industry
It is proteolytic enzyme in enzyme preparation 75%, and protease is that the maximum enzyme of ratio is occupied in industrial enzymes, accounts about the whole world
60% or so of annual total sales volume.Since microbial protease is ectoenzyme and microbial cells are easy to cultivate, with
Animal and plant protease is compared, and has downstream processing technology relatively easy, the advantage with industrial mass production.Protease
Have in multiple industries such as food, brewing, medicine, weaving, leather, detergents and cosmetic detergent, feed and aquatic products processings and answers extensively
With playing an important role to the development of national economy.
It is measured through API ZYM identification systems (production firm: bioM é rieux), bacterial strain YN15 has the β-half of greater activity
Lactoside enzyme and trypsase (specific qualification process is shown in embodiment 4).As microbe-derived beta galactosidase and pancreas egg
White enzyme has in industries such as industry, food, medicine and agriculturals to be widely applied.
Sequence table
<110>Shanghai University
<120>a kind of teas bacillus and its application
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1599
<212> DNA
<213> Paenibacillus sp.
<400> 1
agagtttgat catggctcag gacgaacgct ggcggcgtgc ctaatacatg caagtcgagc 60
gggcttgttc cttcggggac aagctagcgg cggacgggtg agtaacacgt aggcaacctg 120
cccatgggac tgggataacg tcgggaaacc gatgctaata ccggatacgt gattccttcg 180
catgagggaa tctggaagga cggcgcaagc tgtcaccgat ggatgggcct gcggcgcatt 240
agctagttgg tggggtaacg gctcaccaag gcgacgatgc gtagccgacc tgagagggtg 300
atcggccaca ctgggactga gacacggccc agactcctac gggaggcagc agtagggaat 360
cttcggcaat gggcgaaagc ctgaccgagc aacgccgcgt gagtgatgaa ggctttcggg 420
tcgtaaaact ctgttgccag agaagaacgg taggtagagt aactgctatc tatgtgacgg 480
tatctgagaa gaaagccccg gctaactacg tgccagcagc cgcggtaata cgtagggggc 540
aagcgttgtc cggaattatt gggcgtaaag cgcgcgcagg cggttgttta agtctggggt 600
ttaagttcgg ggctcaaccc cgtatcgccc tggaaactgg ggaactggag tgtaggagag 660
gaaagtggaa ttccacgtgt agcggtgaaa tgcgtagaga tgtggaggaa caccagtggc 720
gaaggcgact ttctggccta taactgacgc tgaggcgcga aagcgtgggg agcaaacagg 780
attagatacc ctggtagtcc acgccgtaaa cgatgtacgc taggtgttgg gggtttcgat 840
accctcggtg ccgaagttaa cacattaagc gtaccgcctg gggagtacgc tcgcaagagt 900
gaaactcaaa ggaattgacg gggacccgca caagcagtgg agtatgtggt ttaattcgaa 960
gcaacgcgaa gaaccttacc aggtcttgac atcgggatga ccgcttcaga gatggagctt 1020
tccttcggga catcccagac aggtggtgca tggttgtcgt cagctcgtgt cgtgagatgt 1080
tgggttaagt cccgcaacga gcgcaaccct tgatcttagt tgccagcgcg taaaggtggg 1140
ccctctaaga tgactgccgg tgacaaaccg gaggaaggtg gggatgacgt caaatcatca 1200
tgccccttat gacctgggct acacacgtac tacaatggcc ggtacaacgg gaagcgaagg 1260
agcgatccgg agcgaatcct aagaagccgg tctcagttcg gattgcaggc tgcaacccgc 1320
ctgcatgaag tcggaattgc tagtaatcgc ggatcagcat gccgcggtga atacgttccc 1380
gggtcttgta cacaccgccc gtcacaccac gagagtttac aacacccgaa gtcggtgagg 1440
taacctgaga gtttgcggag caaacttgct tcgtaagcgt tgaccggggg ccagccgccg 1500
aaggtggggt agatgattgg ggtgaagtcg taacaaggta gccgtatcgg aaggtgcggc 1560
tggatcacct cctt 1599
Claims (6)
1. a kind of teas bacillus, it is characterised in that the deposit number of the strain is CGMCC No.13439.
2. teas bacillus according to claim 1, it is characterised in that the 16S rDNA of the teas bacillus is SEQ
Base sequence shown in ID NO.1.
3. a kind of method for preparing teas bacillus according to claim 1, it is characterised in that the specific step of this method
Suddenly are as follows: series bacillus is inoculated in culture medium, aerobic culture is carried out under conditions of 25-45 DEG C, pH 6.7-8.2.
4. according to the method described in claim 3, pH is 6.8 ~ 7.3 it is characterized in that the cultivation temperature is 37 DEG C.
5. the method according to claim 3 or 4, it is characterised in that there are also mass percents in the culture medium not
More than the NaCl of 8 %.
6. a kind of bacterial strain of teas bacillus described in claim 1 produces galactosidase and trypsase in liquid fermentation
In application.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103740618A (en) * | 2013-12-31 | 2014-04-23 | 光明乳业股份有限公司 | Novel bacillus-like strain as well as culture method and application thereof |
| CN104877942A (en) * | 2015-06-11 | 2015-09-02 | 上海大学 | New strain of Paenibacillus sp.YN2 as well as culture method and application of new strain |
| CN105802879A (en) * | 2016-03-25 | 2016-07-27 | 浙江工业大学 | Paenibacillus elgii D7 and application thereof to cellulose degradation |
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2018
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| CN103740618A (en) * | 2013-12-31 | 2014-04-23 | 光明乳业股份有限公司 | Novel bacillus-like strain as well as culture method and application thereof |
| CN104877942A (en) * | 2015-06-11 | 2015-09-02 | 上海大学 | New strain of Paenibacillus sp.YN2 as well as culture method and application of new strain |
| CN105802879A (en) * | 2016-03-25 | 2016-07-27 | 浙江工业大学 | Paenibacillus elgii D7 and application thereof to cellulose degradation |
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