CN114774330A

CN114774330A - Novel species of cellulolytic female phytophthora and application thereof

Info

Publication number: CN114774330A
Application number: CN202210585141.9A
Authority: CN
Inventors: 张玉琴; 韩雪飞; 邓阳; 姜竹鸣; 苏静; 余利岩; 赵莉莉
Original assignee: Institute of Medicinal Biotechnology of CAMS
Current assignee: Institute of Medicinal Biotechnology of CAMS
Priority date: 2022-05-27
Filing date: 2022-05-27
Publication date: 2022-07-22
Anticipated expiration: 2042-05-27
Also published as: CN114774330B

Abstract

The invention discloses a new species of cellulose female phytophthora and application thereof. The invention provides an antibacterial female cellulose bacterium (Herbiconiux cellulolyticus) IMB220419, which is registered with the accession number of CGMCC No.24729 in the China general microbiological culture Collection center. The cellulolytic female bacteria (Herbiconiux cellulolyticus) IMB220419 provided by the invention is a new species of the female bacteria, has cellulose degradation capability and has wide application prospect in the aspect of cellulose degradation. Can be used for sewage treatment, environmental management, soil improvement, bacterial manure preparation production and the like.

Description

Novel species of cellulolytic female phytophthora and application thereof

Technical Field

The invention relates to the field of microorganisms, in particular to a novel species of cellulose-decomposing female phytophthora and application thereof.

Background

The genus hermiconix (herbiconicux) belongs to the family Microbacteriaceae (Microbacteriaceae) (Park, y.h., Suzuki, k.i., Yim, d.g., Lee, k.c., Kim, e.g., Yoon, j.s., Kim, s.j., Kho, y.h., Goodfellow, M).&(1993) Supergenetic classification of peptidoglycan group B microorganisms by nucleic acid sequencing of 5S ribosomal RNA Antonie Van Leeuwenhok 64, 307. sup. 313.) the model species of the genus Herbicinux ginseng is reclassified by Leifsonia ginseng (Behredt, U.S.Schumann, P.G., Hamada, M., Suzuki, K.,

C.&ulrich, A. (2011). Recoverification of Leifsonia ginsengen.nov., comb.nov.and description of Herbiconi.sp.nov., an actinobacterium associated with the phenyl siloxane of Solanum tuberose L.int J.Syst Evol.Microbiol 61, 1039-1047.). To date, the female species has received a total of 4 valid descriptive species: herbiconicixginsengi, Herbiconiciux solani (Behrent, U.S., Schumann, P., Hamada, M., Suzuki, K.,

C.&Ulrich,A.(2011).Reclassification of Leifsonia ginsengi(Qiu etal.2007)as Herbiconiux ginsengi gen.nov.,comb.nov.and description of Herbiconiux solani sp.nov.,an actinobacterium associated with the phyllosphere of Solanum tuberosum L.Int J Syst EvolMicrobiol 61,1039-1047.)，Herbiconiux flava(Hamada,M.,Komukai,C.,Tamura,T.,Evtushenko,L.I.,Vinokurova,N.G.&Suzuki,K.(2012).Description of Herbiconiux flava sp.nov.and emended description of the genus Herbiconix. int J Syst Evol Microbiol 62,795-799.) and Herbiconiux molar picolia (Kim, B.C., Park, D.S., Kim, H.Oh, H.W., Lee, K.H., Shin, K.S.&Bae, K.S (2012). Herbicoux moechopic sp. nov., a xylolytic bacterial isolated from the gut of fire-bound needles, Moechopa diphysis (Pascoe.) Int Syst ethanol Microbiol 62,90-95.), isolated from plants and animals, respectively.

The female Phyllomycete strain does not produce hypha, the cell is in a short rod shape, the cell wall contains L-/D-Dab, Gly, Ala and threo-3-hydroxyglutaminic acid, and belongs to B2 gamma type cell wall, the main polar lipid components comprise Diphosphatidylglycerol (DPG), Phosphatidylglycerol (PG) and Glycolipid (GL), dominant respiratory quinone is MK-11, and a small amount of MK-10 is also contained; the dominant fatty acid contains ai-C_15:0。

Cellulose is the most abundant renewable carbohydrate resource in nature, the structure of the cellulose is very complex, and microorganisms play a key role in the biomass conversion process. Microbially produced cellulases are a key enzyme system for cellulose degradation and conversion. The cellulase-producing strain can directly degrade modified cellulose CMC-Na, so that a strain with cellulose degradation capability can be rapidly screened by using CMC-Na as a substrate (Zhou, X., Chen, H. & Li, Z. (2004). CMCase activity assay as a method for cellulose adsorption and analysis. enzyme Microbiol Technol,35, 455-) 459.).

Disclosure of Invention

The invention aims to provide a novel species of cellulose-degrading female phytophthora and application thereof.

In a first aspect, the present invention claims a new species of the genus estetrum.

The new species of the female phytophthora claimed in the present invention is a cellulolytic female phytophthora (Herbiconiux cellulolyticus) IMB220419 with accession number of CGMCC No.24729 in the common microbiology center of the chinese committee for culture collection of microorganisms.

The cellulolytic female bacteria (Herbiconiux cellulolyticus) IMB220419 is gram-positive bacteria and aerobic. The strain can form wet, smooth and milky colony after being cultured on R2A culture medium at 28 ℃ for 72 hours. The strain has growth tolerance range of 4-37 deg.C, 0-5% NaCl and pH of 6.0-11.0, and optimal growth conditions of 28 deg.C, 0-1% NaCl and pH of 8.0-9.0. The strain has positive gelatin liquefaction reaction, positive cellulose hydrolysis and negative nitrate reduction reaction. The 16S rRNA sequence of the strain is shown in SEQ ID No. 1.

In a second aspect, the invention claims a culture.

The culture claimed in the present invention is a culture of the female cellulolytic bacterium (herbiconicux cellulolyticus) IMB220419 described in the first aspect above, which is a substance obtained by culturing the said female cellulolytic bacterium (herbiconicux cellulolyticus) IMB220419 in a bacterial culture medium (all substances in the culture vessel).

In the above culture, the substance includes metabolites of female cellulolytic bacteria (Herbiconicux cellulolyticus) IMB220419 (cell itself) and female cellulolytic bacteria (Herbiconicius cellulolyticus) IMB 220419.

In the above culture, the bacterial culture medium may be a solid culture medium or a liquid culture medium.

The term "culture" refers to a general term for liquid or solid media on which a microorganism population grows after being artificially inoculated and cultured. I.e. a product obtained by growing and/or amplifying a microorganism, which may be a biologically pure culture of the microorganism, or which may contain a certain amount of a culture medium, metabolite or other component produced during the cultivation. The term "culture" also includes a subculture obtained by passaging a microorganism, which may be a generation of the culture or a mixture of several generations.

In a specific embodiment of the present invention, the bacterial culture medium is specifically R2A culture medium.

In a third aspect, the invention claims a metabolite.

The metabolites claimed by the present invention are the metabolites of the aforementioned cellulolytic female bacteria (Herbiconicux cellulolyticus) IMB 220419.

The term "metabolite" refers to a primary metabolite and/or a secondary metabolite produced during metabolism of a microorganism. Primary metabolism refers to the process in which microorganisms absorb various nutrients from the outside and produce substances and energy for maintaining life activities through catabolism and anabolism. The primary metabolic product is primary metabolic product, such as monosaccharide or monosaccharide derivative, nucleotide, vitamins, amino acids, fatty acid, etc., and various macromolecular polymers composed of them, such as protein, nucleic acid, polysaccharide, lipid, etc. The secondary metabolism refers to a process of synthesizing substances which have no definite function on the vital activities of microorganisms by using primary metabolites as precursors during a certain growth period of the microorganisms. The secondary metabolic products are secondary metabolic products, and most of them are compounds with relatively complex molecular structures. They can be classified into antibiotics, hormones, alkaloids, toxins, etc. according to their actions.

In a fourth aspect, the invention claims a bacterial agent.

The microbial inoculum as claimed in the present invention comprises hermiconix cellulolyticus IMB220419 as described in the first aspect above, a culture as described in the second aspect above and/or a metabolite as described in the third aspect above.

The microbial inoculum is a microbial inoculum for hydrolyzing cellulose.

In the above microbial inoculum, the microbial inoculum contains a carrier in addition to the active ingredient. The carrier may be one that is commonly used in the pesticide art and is biologically inert. The carrier can be a solid carrier or a liquid carrier; the solid carrier can be a mineral material, a plant material or a high molecular compound; the mineral material may be at least one of clay, talc, kaolin, montmorillonite, white carbon, zeolite, silica, and diatomaceous earth; the plant material may be at least one of corn flour, bean flour and starch; the high molecular compound can be polyvinyl alcohol and/or polyglycol; the liquid carrier can be an organic solvent, vegetable oil, mineral oil, or water; the organic solvent may be decane and/or dodecane.

In the microbial inoculum, the dosage form of the microbial inoculum can be various dosage forms, such as liquid, emulsion, suspending agent, powder, granule, wettable powder or water dispersible granule.

According to the requirement, the microbial inoculum can also be added with a surfactant (such as Tween 20, Tween 80 and the like), a binder, a stabilizer (such as an antioxidant), a pH regulator and the like.

In a fifth aspect, the invention claims the use of hermiconix cellulolyticus IMB220419 of the first aspect or the culture of the second aspect or the metabolite of the third aspect or the inoculant of the fourth aspect as described above in any one of the following:

(A1) hydrolyzing cellulose;

(A2) preparing a product for hydrolyzing cellulose;

(A3) preparing cellulase;

(A4) preparing the product with the cellulase activity.

In a sixth aspect, the invention claims the use of a cellulolytic female (hermiconix cellulolyticus) IMB220419 as described in the first aspect or a culture as described in the second aspect or a metabolite as described in the third aspect or a bacterial agent as described in the fourth aspect as described in any of the following:

(B1) treating sewage;

(B2) environmental management;

(B3) soil improvement;

(B4) and (5) producing bacterial manure preparation.

In a seventh aspect, the invention claims a product for hydrolyzing cellulose.

The product for hydrolyzing cellulose claimed in the present invention has an active ingredient of hermiconiux cellulolyticus (amb 220419) described in the first aspect or the culture described in the second aspect or the metabolite described in the third aspect or the microbial agent described in the fourth aspect.

In an eighth aspect, the invention claims a product having cellulase activity.

The product having cellulase activity as claimed in the present invention comprises the active ingredient of hermiconiux cellulolyticus (IMB 220419) described in the first aspect or the culture described in the second aspect or the metabolite described in the third aspect or the microbial preparation described in the fourth aspect.

In a ninth aspect, the invention claims a method of hydrolyzing cellulose.

The method for hydrolyzing cellulose claimed by the invention can comprise the following steps: the sample to be treated is treated with herminium cellulolyticus (herminiux cellulolyticus) IMB220419 according to the first aspect or with a culture according to the second aspect or with a metabolite according to the third aspect or with a inoculum according to the fourth aspect.

In a tenth aspect, the present invention claims the use of a cellulolytic female (Herbiconicix cellulolyticus) IMB220419 as described in the first aspect hereinbefore for the preparation of a culture as described in the second aspect hereinbefore or a metabolite as described in the third aspect hereinbefore or a bacterial agent as described in the fourth aspect hereinbefore.

Experiments prove that the cellulolytic female phytophthora (Herbiconiux cellulolyticus) IMB220419 has partially obvious characteristic difference with the conventional female phytophthora, and comprises the aspects of phenotype, physiological biochemistry, cytochemical components and the like. Meanwhile, the phylogenetic analysis based on the gene level further proves that the strain IMB220419 can be different from various effective species of the prior female phytophthora, and fully proves that the strain IMB220419 represents a new species of the female phytophthora and is named as cellulolytic female phytophthora. Meanwhile, the cellulose degradation capability detection also proves that the strain has the cellulose degradation capability and has wide application prospect in the aspect of cellulose degradation. Can be used for sewage treatment, environmental management, soil improvement, bacterial manure preparation production and other aspects.

Deposit description

And (3) classification and naming: cellulolytic female (Herbiconiux cellulolyticus);

according to the biological material: IMB 220419;

the preservation organization: china general microbiological culture Collection center;

the preservation organization is abbreviated as: CGMCC;

address: xilu No.1 Hospital No. 3, Beijing, Chaoyang, Beicheng;

the preservation date is as follows: 20/2022/4/month;

the registration number of the collection center: CGMCC No. 24729.

Drawings

FIG. 1 shows the results of the measurement of polar lipid components of the strain IMB 220419.

FIG. 2 is a screening picture of the cellulose degrading ability of the strain IMB 220419.

FIG. 3 shows the construction of a phylogenetic tree based on the gene sequences of strain IMB220419 and 16S rRNA of related species of the family Microbacteriaceae (phylogenetic tree is Brevibacterium metalllius NM2E3^T(GenBank access No. km874400) as an exo-group).

Detailed Description

The present invention is described in further detail below with reference to specific embodiments, which are given for the purpose of illustration only and are not intended to limit the scope of the invention. The examples provided below serve as a guide for further modifications by a person skilled in the art and do not constitute a limitation of the invention in any way.

The experimental procedures in the following examples, unless otherwise indicated, are conventional and are carried out according to the techniques or conditions described in the literature in the field or according to the instructions of the products. Materials, reagents and the like used in the following examples are commercially available unless otherwise specified.

Example 1 isolation, screening and characterization of Cellulosimicrobium cellulolyticus (Herbiconiux cellulolyticus) IMB220419

First, separation and screening of strain IMB220419

The strain IMB220419 is obtained by separating stems of orange daphne plants collected from Yunnan Zhongdian. The specific separation operation is as follows: the separation culture medium used is R2A culture medium, and the specific components are as follows: 0.5g of glucose, 0.5g of soluble starch,

Peptone 0.5g, yeast extract 0.5g, acid hydrolyzed casein 0.5g, sodium pyruvate 0.3g, dipotassium hydrogen phosphate 0.3g, heptahydrateMagnesium sulfate 0.05g, agar 15g, ddH₂O1L, pH value 7.2. And (3) strain separation operation procedures: firstly, using sodium hypochlorite (the available chlorine content is 4.5%, v/v) and alcohol (75%, v/v) to disinfect the surface of the plant; then washing away the surface disinfectant by using sterile water; transferring the plant tissue with the sterilized surface into a sterile culture dish, and airing the moisture on the surface of the plant tissue in a super clean bench; then cutting off the tail end of the plant tissue, and crushing the middle part of the rest plant tissue; spreading appropriate amount of pulverized plant tissue on R2A separation culture medium, and incubating at 28 deg.C for 2-3 weeks. Selecting a plate with better microbial diversity, inoculating single colonies with complete colonies and different morphological characteristics on an R2A plate, and streaking for culture to obtain a pure culture for subsequent research. Meanwhile, the obtained pure culture takes 20% (v/v) glycerol as a protective agent and is respectively preserved in liquid nitrogen at ultralow temperature and preserved at minus 80 ℃. Obtaining a strain of Herbiconiux flava DSM 26474^TThe most closely related (98.34% similarity of 16S rRNA genes) new species, strain number IMB 220419.

II, identification of a strain IMB220419

The strain is subjected to morphological, physiological and biochemical, cytochemical and gene level researches by using an R2A culture medium at the temperature of 28 ℃, and other special conditions are described.

1. Cell morphology observation and physiological and biochemical characteristic detection of strain IMB220419

The growth temperature of the strain IMB220419 is detected in the range of 4, 10, 28, 30, 32, 37, 42 and 45 ℃; 6 (0, 1, 3, 5,7, 10) concentration gradients with a salt tolerance concentration (NaCl) detection range of 0-10% (0-10g/100 ml); growth pH was measured as 10 (4, 5, 6, 7, 8, 9, 10, 11, 12, 13) gradients between pH 4-13. The physiological and biochemical characteristics of the strain are detected by using detection kits such as API 50CH, API ZYM, BiOLOG GEN III and the like. Other physiological characteristics of the strain, including gram stain profile, oxygen demand, contact enzyme activity, oxidase activity, gelatin hydrolysis activity, starch hydrolysis activity and cellulose hydrolysis activity, were mainly described in "handbook of actinomycete systems identification" (Xu L H. Actinomycetes systems: principles, methods and practices. Beijing: Science Press,2007, 93-108.).

The identification result shows that: the strain IMB220419 is gram-positive bacteria and is aerobic. The strain can form wet, smooth and milky colony after being cultured on R2A culture medium at 28 ℃ for 72 hours. The growth tolerance range of the strain is 4-37 ℃, 0-5% NaCl and pH 6.0-11.0, and the optimal growth conditions are 28 ℃, 0-1% NaCl and pH 8.0-9.0.

The strain IMB220419 has positive gelatin liquefaction reaction, positive cellulose hydrolysis and negative nitrate reduction reaction, and the strain and a closely related strain represent a strain H^T、H.solani DSM 19813^T、H.flava DSM 26474^T、H.moechotypicolaDSM 25800^TThe difference in the characteristic is shown in table 1.

TABLE 1 Difference characteristics of the strain IMB220419 with other efficiently described species of the genus Eatoria

Note: positive, negative, w, weak positive, ND, no relevant data was searched.

As can be seen from the results shown in Table 1, the strain IMB220419 of the present invention is significantly different from the published related strains of the female Physarum in the aspects of partial physiological and biochemical characteristics, cytochemical characteristics, genetic compositions, etc.

2. Detection of the cytochemical characteristics of Strain IMB220419

The strain IMB220419 is tested for its cytochemical components of fatty acids, quinone type, polar lipids, etc., by GC-gas chromatography, HPLC-liquid chromatography and TLC-thin layer chromatography (Sasser M. identification of bacteria by gas chromatography of cellular lipids, MIDI Technical Note 101.Newark, DE: MIDI inc., 1990.Minnikin DE, O' Donnell AG, Goodfellow M, Alderson G, Atharye M et al. an integrated process for the extraction of bacteria specific peptides, J microorganisms Methods 1984; 2: 233. Asaru 241).

The experimental result shows that the main fatty acid component of the strain IMB220419 is ai-C_15:0(51.0％),i-C_16:0(16.2％),ai-C_15:1A (10.6%), the major polar lipid components include Diphosphatidylglycerol (DPG), Phosphatidylglycerol (PG), Glycolipids (GL) and small amounts of other polar lipid components (PL), as shown in fig. 1.

3. Detection of cellulase Activity of Strain

Plate screening was performed using sodium carboxymethylcellulose CMC-Na as the sole source of C in the plate to detect cellulase activity of strain IMB220419 (Reinhold-Hurek, B., Hurek. T., Claeyssens, M., van Montagu, M. (1993) Cloning, expression in Escherichia coli, and characterization of cellular enzymes of Azoarcus sp., a root-contracting diazol. J Bacteriol 175, 7056. 7065.). The method comprises the following specific steps: CMC-Na screening medium ((NH) is prepared₄)₂SO₄ 4g，NaCl 0.1g，MgSO₄·7H₂0 0.1g，CaCl₂0.1g, yeast extract 0.5g, Fe (III) EDTA 0.033g, CMC-Na 2g, agar 15g, 1L ddH₂O, pH 7.0, selecting a single colony of a strain IMB220419 in the logarithmic growth phase, inoculating the single colony on a CMC-Na screening culture medium, culturing for 3 days, and detecting the cellulase activity of the strain by using Congo red dye liquor. As shown in FIG. 2, a clear circle was formed around strain IMB220419, indicating that strain IMB220419 has a better cellulolytic activity. And the same experimental method was used to perform the secondary screening verification to find the same experimental results as the primary screening (Teather, R.M., Wood, P.J (1982) Use of Congo red-polysaccharide interactions in the evaluation and characterization of cellular bacteria from the said boron road, applied Environ Microbiol 43, 777-780).

4. Determination of phylogenetic position of strain

Genomic DNA of strain IMB220419 was extracted for sequencing and the 16S rRNA gene sequences (SEQ ID No.1) were aligned online in the International authoritative bacteriological taxonomic analysis database (http:// www.ezbiocloud.net /) (Kim, O.S., Cho, Y.J., Lee, K., Yoon SH, Kim M, Na H, Park SC, Jeon YS, Lee JH, Yi H, Won S, Chun J. (2012), Introducing EzTaxon-e: a proryotic 16S rRNA gene sequence data with a phylotypes that is present in expressed uncultured spectra. int J Evost, 62: 716. 721.). The results show that the strain IMB220419 of the present invention has the highest similarity to the species of the genus Oestramus. 16S rRNA gene sequence of the strain IMB220419 of the invention and known bacterial species (H.flava DSM 26474)^T) The highest similarity of (c) was 98.34%, which is below 98.75% (threshold for dividing bacterial species). According to the general reference "Stackelbrandt E, Ebers J.2006, Tarsonomic parameters accessed: Tarnised gold standards. Microbiol Today,33: 152-. The strain IMB220419 is 1 new species of the genus estetrum.

All effective species strains of the female species and 16S rRNA gene sequences of some species strains of the family Microbacteriaceae that are adjacent to the female species were selected to construct phylogenetic trees (FIG. 3).

5. Whole genome information of strain

The whole genome sequence of strain IMB220419 contains 5.3Mbp, with a genome G + C content of 68.8%. Strain IMB220419 genome and its closest relatives h.flava DSM 26474^TThe genome of (A) has an average nucleotide similarity value (ANI) of 80.3%, which is well below the limit for distinguishing bacterial species of 95% (Kim, M., Oh, H.S., Park, S.C., and Chun, J. (2014.) Towards a taxonomic gene family average nucleotide identity and 16S rRNA gene sequence identity for protocols of prokaryotes. int J. Syst alcohol 64,346-351.doi: 10.1099/ijs.0.059774-0). Thus, the whole genome sequence information of strain IMB220419 supports the taxonomic status of its new species.

In conclusion, the strain IMB220419 of the invention has partially obvious characteristic differences with the existing female Phyllostachys strains, including the aspects of phenotype, physiology, biochemistry, cytochemical components and the like. Meanwhile, the phylogenetic analysis based on the gene level further proves that the strain IMB220419 can be distinguished from various effective species of the conventional female phytophthora, and fully proves that the strain IMB220419 represents a new species of the female phytophthora, namely the cellulolytic female phytophthora. Meanwhile, the cellulose degradation capability detection also proves that the strain has the cellulose degradation capability and has wide application prospect in the aspect of cellulose degradation. Can be used for sewage treatment, environmental management, soil improvement, bacterial manure preparation production and other aspects.

The strain IMB220419 has been deposited in China general microbiological culture Collection center (CGMCC) with the preservation number of CGMCC No.24729 in 20 days at 4 months at 2022, and the suggested classification is named as cellulose-degrading female bacteria (Herbiconiux cellulolyticus).

The present invention has been described in detail above. It will be apparent to those skilled in the art that the invention can be practiced in a wide range of equivalent parameters, concentrations, and conditions without departing from the spirit and scope of the invention and without undue experimentation. While the invention has been described with reference to specific examples, it will be appreciated that the invention may be further modified. In general, this application is intended to cover any variations, uses, or adaptations of the invention following, in general, the principles of the invention and including such departures from the present disclosure as come within known or customary practice within the art to which the invention pertains. The use of some of the essential features is made possible within the scope of the claims attached below.

<110> institute of medical and Biotechnology of Chinese academy of medical sciences

<120> cellulolytic female phytophthora and application thereof

<130> GNCLN221538

<160> 1

<170> PatentIn version 3.5

<210> 1

<211> 1501

<212> DNA

<213> Artificial sequence

<400> 1

gacgaacgct ggcggcgtgc ttaacacatg caagtcgaac ggtgaacaag gagcttgctt 60

cttgggatca gtggcgaacg ggtgagtaac acgtgagtaa cctgcccttg actctgggat 120

aagcgttgga aacgacgtct aataccggat acgacttccg acggcatcgt ctgggggtgg 180

aaagattttt tggtcaagga tggactcgcg gcctatcagc ttgttggtga ggtaatggct 240

caccaaggcg acgacgggta gccggcctga gagggtgacc ggccacactg ggactgagac 300

acggcccaga ctcctacggg aggcagcagt ggggaatatt gcacaatggg cgcaagcctg 360

atgcagcaac gccgcgtgag ggatgacggc cttcgggttg taaacctctt ttagtaggga 420

agaagggagc ttgctcttga cggtacctgc agaaaaagca ccggctaact acgtgccagc 480

agccgcggta atacgtaggg tgcaagcgtt gtccggaatt attgggcgta aagagctcgt 540

aggcggtttg tcgcgtctgc tgtgaaaact ggaggctcaa cctccagcct gcagtgggta 600

cgggcagact agagtgcggt aggggagatt ggaattcctg gtgtagcggt ggaatgcgca 660

gatatcagga ggaacaccga tggcgaaggc agatctctgg gccgtaactg acgctgagga 720

gcgaaagcgt ggggagcgaa caggattaga taccctggta gtccacgccg taaacgttgg 780

gaactagatg tggggaccat tccacggtct ccgtgtcgca gctaacgcat taagttcccc 840

gcctggggag tacggccgca aggctaaaac tcaaaggaat tgacgggggc ccgcacaagc 900

ggcggagcat gcggattaat tcgatgcaac gcgaagaacc ttaccaaggc ttgacatata 960

cgagaacggg ccagaaatgg tcaactcttt ggacactcgt aaacaggtgg tgcatggttg 1020

tcgtcagctc gtgtcgtgag atgttgggtt aagtcccgca acgagcgcaa ccctcgttct 1080

atgttgccag cacgtcatgg tgggaactca taggagactg ccggggtcaa ctcggaggaa 1140

ggtggggatg acgtcaaatc atcatgcccc ttatgtcttg ggcttcacgc atgctacaat 1200

ggccggtaca aagggctgca ataccgtaag gtggagcgaa tcccaaaaag ccggtctcag 1260

ttcggattga ggtctgcaac tcgacctcat gaagtcggag tcgctagtaa tcgtggatca 1320

gcaacgccac ggtgaatacg ttcccgggcc ttgtacacac cgcccgtcaa gtcatgaaag 1380

tcggtaacac ccaacgccag tggcctaacc ctttttggga gggagctgtc taaggtggga 1440

tcggtgatta ggactaagtc gtaacaaggt agccgtaccg gaaggtgcgg ctggatcacc 1500

t 1501

Claims

1. The accession number of the cellulolytic female bacteria (Herbiconiux cellulolyticus) IMB220419 in China general microbiological culture Collection center is CGMCC No. 24729.

2. The culture of the female cellulolytic bacterium (Herbiconicix cellulolyticus) IMB220419 according to claim 1, which is obtained by culturing the female cellulolytic bacterium (Herbiconicix cellulolyticus) IMB220419 according to claim 1 in a bacterial culture medium.

3. A metabolite of cellulolytic female bacteria (Herbiconiux cellulolyticus) IMB220419 according to claim 1.

4. The microbial inoculum is characterized in that: the microbial preparation comprises a cellulolytic female bacteria (Herbiconicux cellulolyticus) IMB220419 according to claim 1, a culture according to claim 2 and/or a metabolite according to claim 3.

5. The microbial inoculum according to claim 4, which is characterized in that: the microbial inoculum is a microbial inoculum for hydrolyzing cellulose.

6. Use of a cellulolytic female bacteria (Herbiconiux cellulolyticus) IMB220419 according to claim 1 or of a culture according to claim 2 or of a metabolite according to claim 3 or of a bacterial preparation according to claim 4 or 5 in any of the following:

(A1) hydrolyzing cellulose;

(A2) preparing a product for hydrolyzing cellulose;

(A3) preparing cellulase;

(A4) preparing a product with cellulase activity.

7. Use of a cellulolytic female bacteria (Herbiconiux cellulolyticus) IMB220419 according to claim 1 or of a culture according to claim 2 or of a metabolite according to claim 3 or of a bacterial preparation according to claim 4 or 5 in any of the following:

(B1) treating sewage;

(B2) environmental management;

(B3) soil improvement;

(B4) and (5) producing bacterial manure preparation.

8. A product for hydrolyzing cellulose, the active ingredient of which is the cellulolytic female bacteria (hermiconix cellulolyticus) IMB220419 of claim 1 or the culture of claim 2 or the metabolite of claim 3 or the microbial agent of claim 4 or 5;

or

A product having cellulase activity, the active ingredient of which is hermiconiux cellulolyticus (IMB 220419) of claim 1 or the culture of claim 2 or the metabolite of claim 3 or the microbial inoculum of claim 4 or 5.

9. A method of hydrolyzing cellulose comprising the steps of: treating a sample to be treated with a cellulolytic female bacteria (Herbiconiux cellulolyticus) IMB220419 according to claim 1 or with a culture according to claim 2 or with a metabolite according to claim 3 or with a microbial inoculum according to claim 4 or 5.

10. Use of a cellulolytic female bacteria (Herbiconiux cellulolyticus) IMB220419 according to claim 1 for the preparation of a culture according to claim 2 or a metabolite according to claim 3 or a bacterial preparation according to claim 4 or 5.