CN110484462A - Protozoa category new species and its application - Google Patents
Protozoa category new species and its application Download PDFInfo
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- CN110484462A CN110484462A CN201910613434.1A CN201910613434A CN110484462A CN 110484462 A CN110484462 A CN 110484462A CN 201910613434 A CN201910613434 A CN 201910613434A CN 110484462 A CN110484462 A CN 110484462A
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B09—DISPOSAL OF SOLID WASTE; RECLAMATION OF CONTAMINATED SOIL
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- B09C1/00—Reclamation of contaminated soil
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- C12N9/14—Hydrolases (3)
- C12N9/24—Hydrolases (3) acting on glycosyl compounds (3.2)
- C12N9/2402—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1)
- C12N9/2405—Glucanases
- C12N9/2434—Glucanases acting on beta-1,4-glucosidic bonds
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- C12Y302/01—Glycosidases, i.e. enzymes hydrolysing O- and S-glycosyl compounds (3.2.1)
- C12Y302/01004—Cellulase (3.2.1.4), i.e. endo-1,4-beta-glucanase
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Abstract
The invention discloses protozoa category new species and its applications.The present invention provides Shinella glucanica J1A816, are CGMCC No.18003 in the deposit number of China Committee for Culture Collection of Microorganisms's common micro-organisms center.Bacterial strain J1A816 of the present invention has endoglucanase activity, can is one plant of functional protozoa category new species, will be with a wide range of applications in terms of sewage treatment, environmental improvement, soil improvement, bacterial manure preparation with degraded cellulose.
Description
Technical field
The present invention relates to microorganism fields, and in particular to protozoa category new species and its application.
Background technique
Protozoa category (Shinella) is under the jurisdiction of bacterium domain (Bacteria)-Proteobacteria (Proteobacteria)-
α-deformed rod Gammaproteobacteria (Alph aproteobacteria)-Rhizobiales order (Rhizobiales)-Rhizobiaceae
(Rhizobiaceae).1967, formation of the K.Crabtree and Elizabeth McCoy in research activated sludge flock
When, separation obtains one plant has the strains A TCC 19623 for forming flocculation ability of immigrants in sludge, special according to the phenotype of this plant of bacterium
Sign, preliminary designation are health and moving sticks bacteria (Zoogloea ramigera) (Crabtree, K.&McCoy, E. (1967)
.Zoogloea ramigera Itzigsohn,identification and description.Request for an
Opinion as to the status of the generic name Zoogloea.Int J Syst Bacteriol
17,1–10.).2006, Dong-Shan An etc. was from bacterial strain physiological and biochemical property and cytochemistry taxology angle to bacterial strain
Z.ramigera ATCC 19623 is analyzed, and is renamed as grain protozoa (Shinella granuli), and will
It establishes protozoa category (Shinella) (An, D.S., Im, W.T., Yang, H.C.&Lee, S.T. as type sepecies
(2006).Shinella granuli gen.nov.,sp.nov.,and proposal of the reclassification
of Zoogloea ramigera ATCC 19623as Shinella zoogloeoides sp.nov.Int J Syst
Evol Microbiol 56,443–448.).Currently, the category includes 8 effectively descriptions kind.
The source environment of protozoa category bacterial strain is extensive, such as destructor plant (Lee, M., Woo, S.G.&Ten, L.N.
(2011).Shinella daejeonensis sp.nov.,a nitrate-reducing bacterium isolated
from sludge of a leachate treatment plant.Int J Syst Evol Microbiol 61,2123–
2128.), soil (Matsui, T., Shinzato, N., Tamaki, H., Muramatsu, M.&Hanada, S. (2009)
.Shinella yambaruensis sp.nov.,a 3-methyl-sulfolane-assimilating bacterium
Isolated from soil.Int J Syst Evol Microbiol 59,536-539.), the particle of water filtering element
(An,D.S.,Im,W.T.,Yang,H.C.&Lee,S.T.(2006).Shinella granuli gen.nov.,sp.nov.,
and proposal of the reclassification of Zoogloea ramigera ATCC 19623as
Shinella zoogloeoides sp.nov.Int J Syst Evol Microbiol56,443-448.), house refuse
(Vaz-Moreira,I.,Faria,C.,Lopes,A.R.,Svensson,L.A.,Moore,E.R.B.,Nunes,O.C.&
Manaia,C.M.(2010).Shinella fusca sp.nov.,isolated from domestic waste
Compost.Int J Syst Evol Microbiol 60,144-148.), plant dross (Lin, D.X., Wang, E.T.,
Tang,H.,Han,T.X.,He,Y.R.,Guan,S.H.&Chen,W.X.(2008).Shinella kummerowiae
sp.nov.,a symbiotic bacterium isolated from root nodules of the herbal legume
Kummerowia stipulacea.Int J Syst Evol Microbiol 58,1409-1413.), hydrocarbon pollution
Sandy beach (SubhashY.andLeeS.S.(2016).Shinellacurvatasp.nov.,isolated from
hydrocarbon-contaminated desert sands.Int J Syst Evol Microbiol 66,3929–3934)
Equal environment.In general, protozoa majority is isolated from organic matter environment abundant, therefore, to organic matter
Degradation capability and its application be also concerned.Such as: to the degradation of 4- amino phenyl sulfonyl hydrochlorate (Biala, S., Chadha, P.,
Saini,H.S.(2014).Biodegradation of 4-aminobenzenesulfonate by indigenous
isolate Shinella yambaruensis SA1 and its validation by genotoxic analysis.B
Iotechnol.Bioprocess.Eng.19,1034-1041.), to the degradation of chlorobenzene phthalein (Liang, B., Wang, G., Zhao,
Y.,Chen,K.,Li,S.,&Jiang,J.(2011).Facilitation of bacterial adaptation to
chlorothalonil-contaminated sites by horizontal transfer of the
chlorothalonil hydrolytic dehalogenase gene.Appl.Environ.Microbiol.77,4268–
4272.), degradation (Sepehr S., Shahnavaz B., Asoodeh the A.&Karrabi M (2019) of Pyrogentisinic Acid
.Biodegradation of phenol by cold-tolerant bacteria isolated from alpine
soils of Binaloud Mountains in Iran.J Environ Sci Health ATox Hazard Subst
Environ Eng54,367-379.), to promise nicotine (Qiu J., Li N., Lu Z., Yang Y., Ma Y., Niu L., He
J.&Liu W(2016).Conversion of nornicotine to 6-hydroxy-nornicotine and 6-
hydroxy-myosmine by Shinella sp.strain HZN7.Appl Microbiol Biotechnol 100,
Degradation 10019-10029.) and degradation (Bai, Y., Sun, Q., Zhao, C., Wen, D.&Tang, the X. to pyridine
(2009).Aerobic degradation of pyridine by a new bacterial strain,Shinella
Zoogloeoides BC026.J.Ind.Microbiol.Biotechnol.36,1391-1400.), and not yet find about Shen
The report of Salmonella degraded cellulose.
Cellulose is the most abundant one kind reproducibility carbohydrate sources in nature, and structure is sufficiently complex, micro-
Biology plays key effect in its biomass transformation process.The endoglucanase that microorganism generates is cellulose degradation and turns
The crucial enzyme system of change.Endoglucanase producing strains energy directly degradation modification cellulose, therefore, using CMC-Na conduct
Substrate can quickly screen bacterial strain (Zhou X, Chen H, the Li Z.CMCase activity of tool endoglucanase activity
assay as a method for cellulase adsorption analysis.Enzyme Microbiol Technol,
2004,35(5):455-459.)。
Summary of the invention
The object of the present invention is to provide a protozoa category new species and its applications.
In a first aspect, claimed protozoa category new species.
Present invention protozoa category new species claimed are specially Shinella glucanica J1A816.Its
It is CGMCC No.18003 in the deposit number of China Committee for Culture Collection of Microorganisms's common micro-organisms center.
Second aspect, a kind of claimed microbial inoculum.
The active constituent of present invention microbial inoculum claimed is the Shinella glucanica J1A816.
In above-mentioned microbial inoculum, the microbial inoculum can also include carrier.The carrier can be solid carrier or liquid-carrier.It is described
Solid carrier can be mineral material, vegetable material or high-molecular compound；The mineral material can for clay, talcum, kaolin,
At least one of montmorillonite, white carbon, zeolite, silica and diatomite；The vegetable material can be in corn flour, bean powder and starch
At least one；The high-molecular compound can be polyvinyl alcohol and/or polyglycols.The liquid-carrier can for organic solvent,
Vegetable oil, mineral oil or water；The organic solvent can be decane and/or dodecane.In the microbial inoculum, the active constituent can be with
It is deposited in the form of the fermentation liquid of the living cells, living cells that are cultured, the filtrate of cell culture or cell and the mixture of filtrate
In.The microbial inoculum dosage form can be a variety of dosage forms, such as liquor, emulsion, suspending agent, pulvis, granule, wettable powder or water dispersion
Granula.
As needed, surfactant (such as polysorbas20, Tween 80), adhesive, stabilization can be also added in the microbial inoculum
Agent (such as antioxidant), pH adjusting agent.
The third aspect, the claimed Shinella glucanica J1A816 or the microbial inoculum are as follows
Application in any:
(A1) endoglucanase is prepared；
(A2) preparation has the product of endoglucanase activity.
Fourth aspect, the claimed Shinella glucanica J1A816 or the microbial inoculum are as follows
Application in any:
(B1) degraded cellulose；
(B2) preparation is used for the product of degraded cellulose.
5th aspect, the claimed Shinella glucanica J1A816 or the microbial inoculum are as follows
Application in any:
(C1) sewage treatment；
(C2) environmental improvement；
(C3) soil improvement；
(C4) bacterial manure preparation produces.
6th aspect, claimed following any substance:
(D1) endoglucanase or tool prepared using the Shinella glucanica J1A816 or the microbial inoculum
There is the product of endoglucanase activity；
(D2) utilize prepared by the Shinella glucanica J1A816 or the microbial inoculum for degraded cellulose
Product.
Further, in (D1), the active constituent of the product can be the Shinella glucanica
J1A816, or can be the endoglucanase prepared using the Shinella glucanica J1A816 or the microbial inoculum.
Further, in (D2), the active constituent of the product can be the Shinella glucanica
J1A816, or can be the endoglucanase prepared using the Shinella glucanica J1A816 or the microbial inoculum.
7th aspect, the claimed Shinella glucanica J1A816 is in preparing the microbial inoculum
Application.
Eighth aspect, claimed following bacterial strain or its application:
The bacterial strain is bacterial strain in Shinella glucanica strain；Bacterium in the Shinella glucanica strain
The 16S rRNA gene order of strain at least has 98.7% or more similitude compared with SEQ ID No.1；The Shinella
Bacterial strain is Gram-negative aerobic bacteria in glucanica strain；Bacterial strain is 28 in the Shinella glucanica strain
At DEG C, after cultivating 72 hours on R2A culture medium, wet, smooth and cream-colored colonies can be formed；The Shinella
The growth tolerance range of bacterial strain is 4-40 DEG C, 0-3%NaCl and pH 6.0-8.0, optimum growing condition in glucanica strain
It is 28 DEG C, 0-1%NaCl, pH 7；The oxidizing ferment test positive of bacterial strain, nitric acid in the Shinella glucanica strain
Salt reduction is negative；It can be sole carbon source using maltose, L- rhamnose, sucrose and cellobiose；The Shinella
The main fatty acid of bacterial strain is C in glucanica strain16:0、C18:1ω7c 11-methyl、C19:0Cyclo ω 8c and Summed
features 8(C181ω 7c and/or C18:1ω6c)；The polar lipid ingredient of bacterial strain in the Shinella glucanica strain
Including diphosphatidylglycerol (DPG), phosphatidyl glycerol (PG), phosphatidyl-ethanolamine (PE), hydroxyl phosphatidyl-ethanolamine (OH-
PE), phosphatidyl choline (PC).
The application be the bacterial strain it is following it is any in application:
(A1) endoglucanase is prepared；
(A2) preparation has the product of endoglucanase activity；
(B1) degraded cellulose；
(B2) preparation is used for the product of degraded cellulose；
(C1) sewage treatment；
(C2) environmental improvement；
(C3) soil improvement；
(C4) bacterial manure preparation produces.
It is demonstrated experimentally that bacterial strain J1A816 of the present invention represents a new species of protozoa category, it is named as Shinella
glucanica.Bacterial strain of the present invention is demonstrated simultaneously with endoglucanase activity, can be one plant of function with degraded cellulose
Property protozoa category new species, will have in terms of sewage treatment, environmental improvement, soil improvement, bacterial manure preparation wide
General application prospect.
Preservation explanation
It is recommended that classification naming: Shinella glucanica
Join the biomaterial (strain) of Ju: J1A816
Preservation mechanism: China Committee for Culture Collection of Microorganisms's common micro-organisms center
Preservation mechanism abbreviation: CGMCC
Address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3
Preservation date: on June 19th, 2019
Collection is registered on the books number: CGMCC No.18003
Detailed description of the invention
Fig. 1 is the polar lipid composition detection result of bacterial strain J1A816.
The endoglucanase activity that Fig. 2 is bacterial strain J1A816 screens picture.
Fig. 3 is the 16S rRNA gene order phylogenetic tree construction based on related to Rhizobiaceae kind of bacterial strain J1A816.
Phylogenetic tree is with Bradyrhizobium japonicum USDA 6T(GenBank accession no.AB231927) is
Outer group.
Specific embodiment
Experimental method used in following embodiments is conventional method unless otherwise specified.
The materials, reagents and the like used in the following examples is commercially available unless otherwise specified.
The separation and identification of embodiment 1, bacterial strain J1A816
One, the separation of bacterial strain J1A816
Bacterial strain J1A816 of the present invention is isolated from the pedotheque of Inner Mongol Badanjilin Desert, and specific lock out operation is as follows:
Use R2A culture medium as isolation medium, specific ingredient are as follows: glucose 0.5g, soluble starch 0.5g,Peptone
0.5g, yeast extract 0.5g, sour hydrolyzed casein 0.5g, Sodium Pyruvate 0.3g, dipotassium hydrogen phosphate 0.3g, bitter salt
0.05g, agar 15g plus ddH2O is settled to 1L, pH value 7.2.
Separating screening method: taking 2g soil sample in 18 physiological saline, after oscillation 30min or so, mixed liquor is taken to be diluted to
10-4Afterwards, a small amount of bacterium (0.2-0.3mL) suspension is taken to be coated on R2A culture medium flat plate, 30 DEG C are cultivated 2-3 weeks or so.The micro- life of picking
The single colonie that wherein lawn is complete and morphological feature is different is inoculated in the flat lining out of R2A and trained by the preferable plate of object diversity
It supports, pure culture is obtained, so as to follow-up study.Pure bacterial strain obtained is carried out using 20% (v/v) glycerol as protective agent simultaneously
Liquid nitrogen preservation and -80 DEG C of freezings.Obtain one plant and frozen glue bacterium sample Shen Salmonella Shinella zoogloeoides DSM
287TThe novel species of nearest edge (16S rRNA gene similitude is 98.2%), strain number J1A816.
Two, the identification of bacterial strain J1A816
Bacterial strain J1A816 is grown on R2A culture medium (U.S.'s BD culture medium) under the conditions of 28 DEG C, carries out shape to this plant of bacterium
The research of state, Physiology and biochemistry, cytochemistry and gene level, other special circumstances will be described.
1, the cellular morphology observation and physiological and biochemical property detection of bacterial strain J1A816
The growth temperature detection range of bacterial strain J1A816 is 4,15,28,30,37,40 and 45 DEG C；It grows salinity (NaCl)
Detection range is several (0,1,2,3,4,5,6,7,8,9,10,15) concentration gradients of 0-15% (0-15g/100ml)；Grow pH
Several (4,5,6,7,8,9,10,11) gradients of the detection range between pH4-11.The physiological and biochemical property of bacterial strain uses API
The detection of the detection kits such as 50CH, API ZYM and BiOLOG GEN III carbon source.Other bacterial strain physiological characteristics, including leather are blue
Albert'stain Albert attribute, oxygen demand, contact enzymatic activity, oxidase active, Gelatinolytic activity, starch hydrolysing activities and cellulose hydrolysis
Activity, carried out referring especially to " actinomyces system identification handbook " (XuLH.Actinomycetesystematics:
principles,methodsand practices[M].Beijing:Science Press,2007,93-108.)。
Qualification result shows that bacterial strain J1A816 is gram-negative bacterium, aerobic.Bacterial strain is at 28 DEG C, R2A culture
After cultivating 72 hours on base, wet, smooth and cream-colored colonies can be formed.The growth tolerance range of bacterial strain is 4-40 DEG C, 0-3%
NaCl and pH 6.0-8.0, optimum growing condition are 28 DEG C, 0-1%NaCl, pH 7.
Bacterial strain J1A816 oxidizing ferment test positive, nitrate reduction negative；Maltose, L- rhamnose, sucrose can be utilized
It is sole carbon source with cellobiose.
Bacterial strain J1A816 and nearly edge strain representative strain S.zoogloeoides DSM 287T、S.yambaruensis
DSM 18801T、S.daejeonensis JCM 16236T、S.kummerowiae JCM 14778T、S.curvata JCM
31239T、S.granuli JCM 13254TThe physiological and biochemical property difference of representative strain is shown in Table 1.
1 bacterial strain J1A816 of table and protozoa belong to nearly edge control strain physiological and biochemical property difference table
Note: in table ,+it is expressed as the positive ,-it is expressed as feminine gender.
As shown in Table 1 result as it can be seen that bacterial strain J1A816 of the present invention with the nearly edge bacterial strain of protozoa delivered in part
There are notable differences on physiological and biochemical property.
2, the cytochemistry feature detection of bacterial strain J1A816
Fatty acid, the quinones of bacterial strain J1A816 are detected by GC gas-chromatography, HPLC liquid chromatogram and TLC thin-layer chromatography
The cytochemistries such as type, polar lipid component (Sasser M.Identification of bacteria by gas
ghromatography of cellular fatty acids,MIDI Technical Note 101.Newark,DE:MIDI
inc；1990.Minnikin DE,O'Donnell AG,Goodfellow M,Alderson G,Athalye M et al.An
integrated procedure for the extraction of bacterial isoprenoid quinones and
polar lipids.J Microbiol Methods 1984；2:233–241.).The fatty acid composition of bacterial strain J1A816 is shown in Table
2, the main fatty acid of bacterial strain J1A816 is C as the result is shown16:0、C18:1ω7c 11-methyl、C19:0Cyclo ω 8c and
Summed features 8(C181ω 7c and/or C18:1ω 6c) (table 2).In bacterial strain J1A816, polar lipid ingredient includes two
Phosphatidyl glycerol (DPG), phosphatidyl glycerol (PG), phosphatidyl-ethanolamine (PE), hydroxyl phosphatidyl-ethanolamine (OH-PE), phosphatide
Phatidylcholine (PC), as shown in Figure 1.
The fatty acid composition of 2 bacterial strain J1A816 of table
3, the endoglucanase activity detection of bacterial strain
The endoglucanase activity of bacterial strain J1A816, using sodium cellulose glycolate CMC-Na as unique in plate
Carbon source progress plate screening (Reinhold-Hurek B, Hurek T, Claeyssens M, et al.Cloning,
expression in Escherichia coli,and characterization of cellulolytic enzymes
of Azoarcus sp.,a root-invading diazotroph.J Bacteriol,1993,175(21):7056-
7065.).Specific step is as follows: preparing CMC-Na screening and culturing medium (formula: (NH4)2SO44g, NaCl 0.1g, MgSO4·
7H20 0.1g, calcium chloride 0.1g, yeast extract 0.5g, Fe (III) EDTA0.033g, CMC-Na 2g, agar 15g use ddH2O
Complement to 1L, pH 7.0), it will be in the bacterial strain J1A816 of logarithmic growth phase, picking single colonie is inoculated in CMC-Na screening and culturing
On base, after culture 3 days, detected using endoglucanase activity of the Congo red dye liquor to bacterial strain J1A816, as shown in Figure 2
Apparent transparent circle is produced around bacterial strain J1A816, illustrates that bacterial strain J1A816 has preferable endoglucanase activity.And
Secondary screening verifying is carried out using identical experimental method and finds experimental result (Teather R M, Wood P identical with primary dcreening operation
J.Use of Congo red-polysaccharide interactions in enumeration and
characterization of cellulolytic bacteria from the bovine rumen.Appl Environ
Microbiol,1982,43(4):777-80.)。
4, the determination of bacterial strain phyletic evolution status
The genomic DNA for extracting bacterial strain J1A816 is sequenced, and by 16S rRNA gene order (SEQ ID therein
No.1 it) is compared online in internal authority systematic bacteriology analytical database (http://www.ezbiocloud.net/)
(Kim OS,Cho YJ,Lee K,et al.2012,Introducing EzTaxon-e:a prokaryotic 16S rRNA
gene sequence database with phylotypes that represent uncultured species.Int
J Syst Evol Microbiol,62:716-721.).The strain that bacterial strain J1A816 of the present invention and protozoa belong to as the result is shown
Similitude highest.The 16S rRNA gene order and known strain (S.zoogloeoides DSM of bacterial strain J1A816 of the present invention
287T) highest similitude be 98.2%, the value (threshold values for dividing bacterial species) lower than 98.7%.According to from reference text
Stackebrandt E, Ebers J.2006, offer " Taxonomic parameters revisited:tarnished gold
standards.Microbiol Today,33:152-155.".Illustrate that bacterial strain J1A816 is 1 novel species that protozoa belongs to.
Protozoa is chosen to belong in all effective kind of bacterial strains and Rhizobiaceae with protozoa category adjacent to category part kind bacterium
The 16S rRNA gene order of strain constructs systematic evolution tree (Fig. 3).
In conclusion bacterial strain J1A816 of the present invention and existing Shen Bordetella strain have the apparent feature difference in part, including
Phenotype, Physiology and biochemistry and cytochemistry component etc..The phylogenetic analysis based on gene level further illustrates simultaneously
Bacterial strain J1A816 can be different from each effective kind of existing Shen Bordetella, sufficiently demonstrate bacterial strain J1A816 of the present invention and represent Shen Shi
A new species for Pseudomonas, are named as Shinella glucanica.Simultaneously by producing the detection of endoglucanase ability, also demonstrate,prove
Bacterial strain of the present invention, which is illustrated, has cellulose degradation ability, is with a wide range of applications in terms of cellulose degradation.It can be used for dirt
Water process, environmental improvement, soil improvement, the production of bacterial manure preparation etc..
Bacterial strain Shinella glucanica J1A816 is preserved in China General Microbiological on June 19th, 2019
Culture presevation administrative center, deposit number are CGMCC 18003.
<110>Inst. of Medicinal Biological Technology, Chinese Academy of Medical Sciences
<120>protozoa category new species and its application
<130> GNCLN191597
<160> 1
<170> PatentIn version 3.5
<210> 1
<211> 1315
<212> DNA
<213> Shinella glucanica
<400> 1
gcgtgggaat ctacccatct ctacggaata actcagggaa acttgtgcta ataccgtata 60
cgcccttcgg gggaaagatt tatcggagat ggatgagccc gcgttggatt agctagttgg 120
tggggtaaag gcctaccaag gcgacgatcc atagctggtc tgagaggatg atcagccaca 180
ttgggactga gacacggccc aaactcctac gggaggcagc agtggggaat attggacaat 240
gggcgcaagc ctgatccagc catgccgcgt gagtgatgaa ggccctaggg ttgtaaagct 300
ctttcaccgg tgaagataat gacggtaacc ggagaagaag ccccggctaa cttcgtgcca 360
gcagccgcgg taatacgaag ggggctagcg ttgttcggaa ttactgggcg taaagcgcac 420
gtaggcgggt atttaagtca ggggtgaaat cccagagctc aactctggaa ctgcctttga 480
tactgggtac ctagagtatg gaagaggtga gtggaattcc gagtgtagag gtgaaattcg 540
tagatattcg gaggaacacc agtggcgaag gcggctcact ggtccattac tgacgctgag 600
gtgcgaaagc gtggggagca aacaggatta gataccctgg tagtccacgc cgtaaacgat 660
gaatgttagc cgtcggcatg catgcatgtc ggtggcgcag ctaacgcatt aaacattccg 720
cctggggagt acggtcgcaa gattaaaact caaaggaatt gacgggggcc cgcacaagcg 780
gtggagcatg tggtttaatt cgaagcaacg cgcagaacct taccagccct tgacatcccg 840
atcgcggaca gtggagacat tgtccttcag ttaggctgga tcggagacag gtgctgcatg 900
gctgtcgtca gctcgtgtcg tgagatgttt gggttaagtc ccgcaacgag cgcaaccctc 960
gcccttagtt gccagcattc agttgggcac tcttaagggg actgcccggt gataagccca 1020
aaaggaaagg tggggatgac gtcaagtcct catggccctt acgggctggg ctacacacgt 1080
gctacaatgg tggtgacagt gggcagcgag acagcgatgt cgagctaatc tccaaaagcc 1140
atctcagttc ggattgcact ctgcaactcg agtgcatgaa gttggaatcg ctagtaatcg 1200
cggatcagca tgccgcggtg aatacgttcc cgggccttgt acacaccgcc cgtcacacca 1260
tgggagttgg ttttacccga aggcgatgcg ctaaccgcaa ggaggcagtc gacca 1315
Claims (10)
1.Shinella glucanica J1A816, in China Committee for Culture Collection of Microorganisms's common micro-organisms
The deposit number of the heart is CGMCC No.18003.
2. a kind of microbial inoculum, active constituent is Shinella glucanica J1A816 described in claim 1.
3. Shinella glucanica J1A816 described in claim 1 or microbial inoculum as claimed in claim 2 are following any
In application:
(A1) endoglucanase is prepared；
(A2) preparation has the product of endoglucanase activity.
4. Shinella glucanica J1A816 described in claim 1 or microbial inoculum as claimed in claim 2 are following any
In application:
(B1) degraded cellulose；
(B2) preparation is used for the product of degraded cellulose.
5. Shinella glucanica J1A816 described in claim 1 or microbial inoculum as claimed in claim 2 are following any
In application:
(C1) sewage treatment；
(C2) environmental improvement；
(C3) soil improvement；
(C4) bacterial manure preparation produces.
6. following any substance:
(D1) it is prepared using Shinella glucanica J1A816 described in claim 1 or microbial inoculum as claimed in claim 2
Endoglucanase or product with endoglucanase activity；
(D2) it is prepared using Shinella glucanica J1A816 described in claim 1 or microbial inoculum as claimed in claim 2
The product for degraded cellulose.
7. substance according to claim 6, it is characterised in that: in (D1), the active constituent of the product is described
Shinella glucanica J1A816 is to be prepared using the Shinella glucanica J1A816 or the microbial inoculum
Endoglucanase；Or
In (D2), the active constituent of the product is the Shinella glucanica J1A816 or is using described
Shinella glucanica J1A816 or the endoglucanase of microbial inoculum preparation.
8. Shinella glucanica J1A816 described in claim 1 answering in the microbial inoculum described in preparation claim 2
With.
9. bacterial strain, it is characterised in that: the bacterial strain is bacterial strain in Shinella glucanica strain；The Shinella
The 16S rRNA gene order of bacterial strain at least has 98.7% or more phase compared with SEQ ID No.1 in glucanica strain
Like property；Bacterial strain is Gram-negative aerobic bacteria in the Shinella glucanica strain.
10. bacterial strain described in claim 9 it is following it is any in application:
(A1) endoglucanase is prepared；
(A2) preparation has the product of endoglucanase activity；
(B1) degraded cellulose；
(B2) preparation is used for the product of degraded cellulose；
(C1) sewage treatment；
(C2) environmental improvement；
(C3) soil improvement；
(C4) bacterial manure preparation produces.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114085790A (en) * | 2021-11-11 | 2022-02-25 | 怀化学院 | Bacillus scherzekii JC11, method for degrading phenol-containing waste liquid and application |
| CN114774330A (en) * | 2022-05-27 | 2022-07-22 | 中国医学科学院医药生物技术研究所 | Novel species of cellulolytic female phytophthora and application thereof |
| KR20230030488A (en) * | 2021-08-25 | 2023-03-06 | 주식회사 씨디아이 | Novel Shinella granuli CK-4 strain with high capability of 1,4-dioxane decomposition and method for treating 1,4-dioxane-containing wastewater using the same |
Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1436243A (en) * | 2000-05-22 | 2003-08-13 | 明治制果株式会社 | Endoglucanase NCE5 and cellulase preparations contg. same |
| CN101225377A (en) * | 2008-01-23 | 2008-07-23 | 广西大学 | Endoglucanase as well as encoding gene and use thereof |
| CN102272297A (en) * | 2008-12-30 | 2011-12-07 | 生化酶股份有限公司 | Fungal endoglucanases, their production and use |
| CN103451170A (en) * | 2007-03-12 | 2013-12-18 | 明治制果药业株式会社 | Endoglucanase PPCE and cellulase preparation containing the same |
| RU2013112839A (en) * | 2013-03-15 | 2014-10-27 | Ирина Анатольевна Афти | BIOLOGICAL PRODUCT FOR CLEANING ENVIRONMENTAL OBJECTS FROM HYDROCARBON POLLUTION, METHOD FOR ITS PRODUCTION AND APPLICATION |
| CN105483101A (en) * | 2016-01-14 | 2016-04-13 | 云南师范大学 | Low-temperature salt-tolerant product-inhibition-resistant beta-N-acetyl glucosamine enzyme JB10NagA |
| CN106350469A (en) * | 2016-11-09 | 2017-01-25 | 上海交通大学 | Bacillus with high temperature resistance and cellulose degradation capacity and application thereof |
| CN107974441A (en) * | 2017-12-18 | 2018-05-01 | 浙江大学 | Endoglucanase, its encoding gene cel5A-h37 and its application |
-
2019
- 2019-07-09 CN CN201910613434.1A patent/CN110484462B/en active Active
Patent Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1436243A (en) * | 2000-05-22 | 2003-08-13 | 明治制果株式会社 | Endoglucanase NCE5 and cellulase preparations contg. same |
| CN103451170A (en) * | 2007-03-12 | 2013-12-18 | 明治制果药业株式会社 | Endoglucanase PPCE and cellulase preparation containing the same |
| CN101225377A (en) * | 2008-01-23 | 2008-07-23 | 广西大学 | Endoglucanase as well as encoding gene and use thereof |
| CN102272297A (en) * | 2008-12-30 | 2011-12-07 | 生化酶股份有限公司 | Fungal endoglucanases, their production and use |
| RU2013112839A (en) * | 2013-03-15 | 2014-10-27 | Ирина Анатольевна Афти | BIOLOGICAL PRODUCT FOR CLEANING ENVIRONMENTAL OBJECTS FROM HYDROCARBON POLLUTION, METHOD FOR ITS PRODUCTION AND APPLICATION |
| CN105483101A (en) * | 2016-01-14 | 2016-04-13 | 云南师范大学 | Low-temperature salt-tolerant product-inhibition-resistant beta-N-acetyl glucosamine enzyme JB10NagA |
| CN106350469A (en) * | 2016-11-09 | 2017-01-25 | 上海交通大学 | Bacillus with high temperature resistance and cellulose degradation capacity and application thereof |
| CN107974441A (en) * | 2017-12-18 | 2018-05-01 | 浙江大学 | Endoglucanase, its encoding gene cel5A-h37 and its application |
Non-Patent Citations (2)
| Title |
|---|
| 朱年青等: "里氏木霉纤维素酶的分离纯化及酶学性质", 《生物加工过程》 * |
| 高培基等: "绿色木霉的内切葡聚糖酶在纤维素水解中作用机制的研究", 《自然科学进展》 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20230030488A (en) * | 2021-08-25 | 2023-03-06 | 주식회사 씨디아이 | Novel Shinella granuli CK-4 strain with high capability of 1,4-dioxane decomposition and method for treating 1,4-dioxane-containing wastewater using the same |
| CN114085790A (en) * | 2021-11-11 | 2022-02-25 | 怀化学院 | Bacillus scherzekii JC11, method for degrading phenol-containing waste liquid and application |
| CN114085790B (en) * | 2021-11-11 | 2023-07-18 | 怀化学院 | Shen's bacillus JC11, method for degrading phenol-containing waste liquid and application thereof |
| CN114774330A (en) * | 2022-05-27 | 2022-07-22 | 中国医学科学院医药生物技术研究所 | Novel species of cellulolytic female phytophthora and application thereof |
| CN114774330B (en) * | 2022-05-27 | 2023-07-21 | 中国医学科学院医药生物技术研究所 | Novel species of cellulolytic female phytes and application thereof |
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