CN115011509A - Bacterial strain for degrading cellulose in kitchen waste at high temperature and screening and application thereof - Google Patents
Bacterial strain for degrading cellulose in kitchen waste at high temperature and screening and application thereof Download PDFInfo
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- CN115011509A CN115011509A CN202210483248.2A CN202210483248A CN115011509A CN 115011509 A CN115011509 A CN 115011509A CN 202210483248 A CN202210483248 A CN 202210483248A CN 115011509 A CN115011509 A CN 115011509A
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- 230000001580 bacterial effect Effects 0.000 title claims description 21
- 238000012216 screening Methods 0.000 title abstract description 9
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Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B15/00—Preparation of other cellulose derivatives or modified cellulose, e.g. complexes
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/10—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in agriculture
- Y02A40/20—Fertilizers of biological origin, e.g. guano or fertilizers made from animal corpses
Abstract
The invention relates to the technical field of microbial degradation, in particular to a strain for degrading kitchen waste cellulose at high temperature and screening and application thereof. The kitchen waste strain has the characteristic of degrading cellulose, and experiments on degrading cellulose by kitchen waste show that compared with non-inoculation, the kitchen waste strain inoculated with the strain can obviously improve the degradation efficiency of the kitchen waste, can grow at 45-82 ℃, can obviously degrade the cellulose in the kitchen waste, greatly improve the conversion efficiency of the kitchen waste, shorten the treatment period, improve the utilization level of the kitchen waste, has a great promotion effect on degrading the kitchen waste, and can be used for large-scale actual production.
Description
Technical Field
The invention relates to the technical field of microbial degradation, in particular to a strain for degrading cellulose in kitchen waste at a high temperature and screening and application thereof.
Background
Cellulose is one of substances with a large proportion of kitchen waste, and has a large promotion effect on degradation of the kitchen waste aiming at research on the cellulose in the kitchen waste. The method has the advantages that the method gradually takes the microbial degradation of grease, cellulose, starch, protein and the like in the kitchen waste as a novel method for treating the kitchen waste, can greatly improve the conversion efficiency of the kitchen waste, shortens the treatment period and improves the utilization level of the kitchen waste.
With the increasing importance of China on the treatment of kitchen wastes, more and more enterprises are put into the field of recycling of the kitchen wastes, but the kitchen waste treatment mode basically has the purposes of landfill, feed, fertilizer and the like after crushing, but the utilization rate of the kitchen wastes is low, the cycle period is long, and the kitchen waste treatment mode is not beneficial to environmental protection.
Disclosure of Invention
In view of the above, the technical problem to be solved by the present invention is to provide a strain for high temperature degradation of cellulose in kitchen waste, and screening and application thereof.
The invention provides bacillus with the preservation number of CGMCC No. 24544.
The bacillus strain provided by the invention comprises the following components: the vegetative cells are in a rod-shaped, round-ended, mostly single, rarely paired or chain arrangement, and the cell wall is a gram-positive structure. Aerobic or facultative anaerobic. The colony on the nutrient agar culture medium is transparent, has smooth and moist surface and neat edge, and is picked to be sticky or frozen.
The invention relates to a method for identifying the phylogenetic position of the strain, which is used for sequencing the 16S rDNA sequence of the separated strain. Obtaining 17 reference strain sequences from NCBI (GenBank) database, analyzing the 16S rDNA sequences of the separated strain and the reference strain by using software BioEdit and MEGA11, and constructing a phylogenetic tree of the separated strain and the reference strain. Thus, the Bacillus strain was identified as a strain of Bacillus licheniformis (FIG. 3) and named BGB-85F.
The growth conditions of the strain comprise: the pH value is 6-8, the temperature is 45-82 ℃, the rotating speed is 120-200 r/min, and wide carbon sources and nitrogen sources can be utilized.
In some embodiments, the optimal growth conditions for the strains of the invention include: the pH value is 7.0, the temperature is 55-60 ℃, the rotating speed is 160r/min, and wide carbon sources and nitrogen sources can be utilized.
The main functions of the strain comprise rapid propagation at high temperature and good degradation effect on cellulose in kitchen waste. The thallus is released into soil, is harmless to human, animals and plants, does not pollute the environment, can enrich the structure of natural flora under certain conditions, and improves the diversity of the natural flora.
Therefore, the invention also provides the application of the bacillus with the preservation number of CGMCC No.24544 in degrading cellulose.
The application of the invention is that the cellulose comprises cellulose in kitchen waste.
The application of the invention is that the pH value of the kitchen waste is 6-8.
The invention provides a cellulose degradation microbial inoculum which comprises bacillus with the preservation number of CGMCC No. 24544.
The cellulose degradation microbial inoculum of the invention has the bacterial content of 2 multiplied by 10 8 cfu/g～9×10 8 cfu/g。
The preparation method of the cellulose degradation microbial inoculum comprises the steps of culturing bacillus with the preservation number of CGMCC No.24544 and collecting thalli.
The preparation method of the invention comprises the following steps of after the thallus is collected, re-suspending the thallus to prepare a bacterial suspension; or mixing the thallus with a protective agent, and preparing powder by a spray drying machine.
The invention also provides a method for degrading cellulose, which comprises the step of inoculating the cellulose degrading microbial inoculum into a substance to be degraded.
The method comprises the step of preparing the degradation product, wherein the degradation product comprises kitchen waste.
The method of the invention, wherein the degradation temperature comprises 45-82 ℃.
The cellulose provided by the invention comprises cellulose in kitchen waste.
In some embodiments, the cellulose of the present invention comprises cellulose in kitchen waste, wherein the cellulose accounts for 10% to 20%, and in other embodiments, the cellulose accounts for 14.8%.
The kitchen waste strain has the characteristic of degrading cellulose, and experiments on the degradation of the cellulose by the kitchen waste show that compared with the strain without inoculation, the strain inoculated with the strain can obviously improve the degradation efficiency of the kitchen waste, can grow at 45-82 ℃, can obviously degrade the cellulose in the kitchen waste, greatly improve the conversion efficiency of the kitchen waste, shorten the treatment period, improve the utilization level of the kitchen waste, has a great promotion effect on the degradation of the kitchen waste, and can be used for large-scale actual production.
Biological preservation Instructions
Biological material: BGB-85F, taxonomic nomenclature: bacillus licheniformis, deposited in China general microbiological culture Collection center at 18.03.2022, with the addresses of: no. 3 of Xilu No. 1 of Beijing, Chaoyang, Beijing, the institute of microbiology, Chinese academy of sciences, with the collection number of CGMCC No. 24544.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, and it is obvious that the drawings in the following description are some embodiments of the present invention, and other drawings can be obtained by those skilled in the art without creative efforts according to the drawings:
FIG. 1 shows a photograph of Bacillus licheniformis BGB-85F, taken under an optical microscope; wherein A in FIG. 1 shows light microscope × 400 Bacillus licheniformis BGB-85F; FIG. 1B shows optical microscope × 1000 (under oil microscope) Bacillus licheniformis BGB-85F;
FIG. 2 shows the colony morphology of Bacillus licheniformis BGB-85F cultured on nutrient agar medium at 60 ℃; wherein A in FIG. 2 shows a colony streaking line and B in FIG. 2 shows a single colony growth morphology;
FIG. 3 shows a phylogenetic tree constructed based on the 16S rDNA sequence;
FIG. 4 shows a photograph of Bacillus licheniformis BGB-85F growth on Congo red medium; wherein A in FIG. 4 represents the treatment group (60 ℃ C.); b in FIG. 4 shows a control group (60 ℃ C.); c in FIG. 4 represents a nutrient agar medium (60 ℃ C.); d in FIG. 4 shows the treatment group (30 ℃ C.);
FIG. 5 shows the effect of the strain Bacillus licheniformis BGB-85F on degrading cellulose in kitchen waste.
Detailed Description
The invention provides a bacterial strain for degrading kitchen waste cellulose at high temperature and screening and application thereof, and a person skilled in the art can appropriately improve process parameters by referring to the content. It is expressly intended that all such similar substitutes and modifications which would be obvious to one skilled in the art are deemed to be included in the invention. While the methods and applications of this invention have been described in terms of preferred embodiments, it will be apparent to those of ordinary skill in the art that variations and modifications in the methods and applications described herein, as well as other suitable variations and combinations, may be made to implement and use the techniques of this invention without departing from the spirit and scope of the invention.
1 separation and purification of kitchen waste degrading strains
1) Collecting a kitchen sample: the method is characterized by collecting kitchen waste treatment plants from great village of Beijing, and registering date and other information.
2) Primary separation of target bacteria: weighing 10g of the collected sample, adding into a sterilized triangular flask containing 100ml of normal saline under aseptic condition, sufficiently oscillating, at 28 deg.C for 30min, diluting to 10% -7 The selected dilution concentration was 10 -5 、10 -6 、10 -7 0.1ml to 0.2ml of the dilution is respectively absorbed, a plate is coated on nutrient agar (the formula is shown in table 1), and the liquid is reversely cultured for 24h to 48h at the temperature of 60 ℃ after being absorbed completely. Picking single clone on the plate, picking different colony streaks according to the shape of microorganism on the plateAnd (4) performing line separation and purification (culturing according to the culture conditions) for 2-3 times until the bacterial colony is single, numbering, recording, and refrigerating for storage.
3) Preliminary identification: the separated and purified strain is examined and observed under microscope, as shown in figure 1.
TABLE 1 nutrient agar medium formulation
| Name of reagent | Dosage of | Name of reagent | Dosage of |
| Peptone | 10g/L | NaCl | 5g/L |
| Beef extract | 3g/L | Agar-agar | 25g/L |
| Distilled water | To 1L |
2 Properties of the Individual Strain
1) Morphological characteristics
The strain is as follows: the vegetative cells are rod-shaped and round-ended, most of the vegetative cells are arranged in a single, a few pairs or a chain, and the cell walls are gram-positive structures. The optimal growth temperature is 55-60 ℃, the growth temperature is 45-82 ℃, the minimum growth temperature is 45 ℃, and the growth temperature is aerobic or facultative anaerobic. The colony cultured in nutrient agar medium at 55 deg.C for 16h is transparent, has smooth and moist surface and neat edge, and is selected to be viscous or frozen (figure 2).
2) Characteristics of culture
Experiments (shake culture) and temperatures with different pH values were performed, and the results are shown in Table 2 below:
TABLE 2 influence of different pH values and temperature treatments on the amount of live bacteria cultured
Therefore, the optimal growth conditions of the strain are as follows: the pH value is 7.0, the temperature is 55-60 ℃, the rotating speed is 160r/min, and wide carbon sources and nitrogen sources can be utilized.
3) Functional characteristics
In the kitchen waste compost, the main function is that the compost can be rapidly propagated at high temperature, and has good degradation effect on cellulose in the kitchen waste. The thallus is released into soil, is harmless to human, animals and plants, does not pollute the environment, can enrich the structure of natural flora under certain conditions, and improves the diversity of the natural flora.
16S rDNA sequencing of 3 Strain
To identify the phylogenetic position of the present strain, the 16S rDNA sequence of the isolated strain was sequenced.
Obtaining 17 reference strain sequences from NCBI (GenBank) database, analyzing the 16S rDNA sequences of the separated strain and the reference strain by using software BioEdit and MEGA11, and constructing a phylogenetic tree of the separated strain and the reference strain. Thus, the Bacillus strain was identified as a strain of Bacillus licheniformis (FIG. 3) and named BGB-85F.
Compared with the prior art, the kitchen waste strain has the following effects that the kitchen waste strain has the characteristic of degrading cellulose, and the experiment of degrading the cellulose by the kitchen waste shows that compared with the non-inoculation method, the kitchen waste strain inoculated with the strain can obviously improve the degradation efficiency of the kitchen waste, does not grow at the temperature of below 45 ℃, can grow at the temperature of between 45 and 82 ℃, can obviously degrade the cellulose in the kitchen waste, and can be used for actual production.
Embodiments of the present invention will be described in detail below with reference to examples, but it will be understood by those skilled in the art that the following examples are only illustrative of the present invention and should not be construed as limiting the scope of the present invention. The examples, in which specific conditions are not specified, were conducted under conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products available commercially.
EXAMPLE 1 isolation and purification of the Strain
1 isolation and selection of strains
After the kitchen waste is crushed, collecting samples in three periods of initial stage, temperature rise and later stage of natural fermentation, sending the samples to a laboratory, respectively diluting the three samples by 100 times by using sterile water into a triangular flask, oscillating at 25 ℃, 30min and 160rpm, and standing for 15 min. Respectively culturing in nutrient medium, Gao's I medium and PDA medium by plating, and setting two culture temperatures of 28 deg.C and 55 deg.C. And (3) placing the strains at the temperature of 28 ℃ and 55 ℃ at 60 ℃ for screening and culturing, and screening strains which are suitable for high-temperature growth of the degradation of the kitchen waste.
2 purification of the Strain
After the bacteria grow out of the plate, scraping a small amount of bacterial colonies from the plate by using a sterilized toothpick, diluting the bacterial colonies with sterile water, and then carrying out streak culture on the plate again. If no monoclonal colony appears, streaking and purifying on a plate repeatedly until the monoclonal colony appears, purifying for many times, and keeping the colony morphology uniform until no other colony with different morphology appears.
The strain is as follows: the vegetative cells are rod-shaped and round-ended, most of the vegetative cells are arranged in a single, a few pairs or a chain, and the cell walls are gram-positive structures. The optimal growth temperature is 55-60 ℃, the minimum growth temperature is 45 ℃, the growth can be carried out at 45-82 ℃, and the growth is aerobic or facultative anaerobic. The colony cultured in nutrient agar medium at 60 deg.C for 16h is creamy yellow, has wrinkles on surface, is irregular round, and is picked into viscous or frozen state (figure 2).
By applying the separation and purification method, repeated purification is carried out for a plurality of generations, streaking culture is carried out in a nutrient solid culture medium (shown in table 1), a single bacterial strain is obtained, the bacterial strain has the capacity of degrading cellulose by virtue of plate culture medium culture, and the bacterial strain with stronger kitchen waste degradation function is obtained by separation and purification in the embodiment.
Example 2 sequencing of 16S rDNA sequence of Individual strains of Strain
PCR specific amplification is carried out on the strain monoclonal bacteria liquid, primers are 27F (SEQ ID NO: 2) and 1492R (SEQ ID NO: 3) respectively, and enzyme is 2 XStar Mix. And detecting the PCR amplification product by an electrophoretic imaging technology, observing whether the PCR amplification product has a band, and using the residual PCR amplification product for sequence determination. The sequencing result is shown in SEQ ID NO: 1 is shown. The PCR reaction system is as follows:
table 3: 16SrDNA 2 XStartmix enzyme reaction system
Multiple reference strain sequences were obtained from the ncbi (genbank) database, and the 16S rDNA sequences of the isolates and the reference strains were analyzed for full length using software BioEdit and MEGA11 to construct phylogenetic trees of the isolates and the reference strains (fig. 3). Thus, the bacterial relationship of the strain is determined to be Bacillus licheniformis and is named BGB-85F (Jiabovin No. 85).
Example 3 test of degrading cellulose by kitchen garbage
1 purpose of the test: and (3) detecting the cellulose degrading capability of the strain.
2 test materials
2.1 test materials: kitchen waste (crushed and undegraded) Congo red culture medium
2.2 test strains: BGB-85F, BGB-86F
2.3 main test instruments and equipment: an ultra-clean workbench, an autoclave, a constant-temperature shaking incubator, a 500mL conical flask, tweezers, a culture dish, a glass rod, an inoculating ring, a 1.5mL centrifuge tube, a puncher, a coating rod, sterile filter paper and the like.
2.4 test drugs and reagents
(1) Test drugs: NaCl, peptone, yeast powder, beef extract, Congo red reagent, sodium nitrate, disodium hydrogen phosphate, potassium dihydrogen phosphate, magnesium sulfate, potassium chloride, acid hydrolyzed casein, sodium carboxymethylcellulose, agar and the like.
(2) Test reagents:
1) nutrient liquid culture medium: weighing 10g of peptone, 5g of beef extract and 10g of NaCl, dissolving in 1L of pure water, and carrying out pH: 6.8-7.5, 121 ℃, 30min and sterilizing.
2) Nutrient solid culture medium: weighing 10g of peptone, 5g of beef extract, 10g of NaCl and 20g of agar, dissolving in 1L of pure water, and carrying out pH: 6.8-7.5, 121 ℃, 30min and sterilizing.
3) Congo red medium: weighing 0.5g of yeast powder, 0.2g of Congo red reagent, 1.0g of sodium nitrate, 1.2g of disodium hydrogen phosphate, 0.9g of monopotassium phosphate, 0.5g of magnesium sulfate, 0.5g of potassium chloride, 0.5g of acid hydrolyzed casein, 5.0g of cellulose powder and 25g of agar in 1L of pure water, wherein the pH value is as follows: 6.9-7.1, at 121 ℃, for 30min, sterilizing, and pouring the plate on a super clean bench for later use.
4) Kitchen waste: and (3) putting the crushed kitchen waste into a 500ml conical flask, placing 200g of the crushed kitchen waste into each flask, sterilizing at 121 ℃ for 30min, and detecting that the initial cellulose content is 14.8%.
3 test procedure
3.1 Strain activation culture and preparation of microbial inoculum
Activating test strains stored at-80 ℃ for 3 times, transferring the activated test strains to a nutrient liquid culture medium for culture, culturing to a logarithmic phase, detecting the growth condition of the strains by using a nucleic acid protein tester, estimating the bacterial content by using an OD value, and counting by using a dilution coating flat plate for use.
Centrifuging the above bacteria fermentation liquid at 5000r/min for 5min with a centrifuge, removing supernatant, and leaving bacteria suspension. The bacterial suspension is uniformly mixed with a protective agent (the protective agent is proportioned, the mass concentration is 20-50% of dextrin, 20-95% of anhydrous sodium sulfate and 1-10% of sodium metabisulfite) according to the mass ratio of 19: 1, and the mixture is completely and uniformly mixed with the fermentation centrifugal supernatant according to the mass ratio of 1: 1. Spray drying is carried out by utilizing a spray dryer, the air inlet temperature of the spray dryer is set to be 165 ℃, and the air outlet temperature is controlled to be 90 ℃. And (3) detecting the amount of the live bacteria of the spray-dried microbial inoculum by using a dilution coating plate method for later use.
3.2 Congo red staining identification experiment of Strain
And (3) selecting colonies of the activated strains by utilizing an inoculating loop, culturing on a Congo red culture medium under the culture condition of 60 ℃ for 36 hours, and performing blank control, low-temperature (30 ℃) treatment control and BGB-86F treatment.
The experimental results are shown in fig. 4, in the treatment groups, no transparent circles appear in the blank control, the low temperature (30 ℃) control and the BGB-86F treatment, but more significant transparent circles appear in the treatment groups. Compared with the control and other treatments, the treatment group can utilize sodium carboxymethyl cellulose in Congo red culture medium as a carbon source to decompose cellulose.
3.3 experiment for degrading cellulose in kitchen wastes by using bacterial strain
Inoculating sterilized conical flask and kitchen waste into the microbial inoculum obtained in the step 1, dissolving the microbial inoculum by using sterile water to dilute the microbial inoculum to obtain the microbial inoculum with the number of (2 multiplied by 10) 8 cfu/ml)～(9×10 8 cfu/ml), inoculating 20ml of the waste mixture into each bottle, and ensuring that the bacteria amount of the kitchen waste mixture is (2X 10) 8 cfu/g)～(9×10 8 cfu/g). Sealing the conical bottle mouth with a sealing film, culturing in a constant-temperature incubator at 60 ℃ for 72h, observing once every 24h, uniformly sampling on an ultra-clean workbench, preventing pollution, detecting the cellulose content in the kitchen waste, setting 3 times of repetition, setting blank control, and adding equivalent sterile water.
The results are shown in table 3, and the 72h test results show that the degradable cellulose content in the treatment groups is 28.6%, 60.8% and 79.5% at 24h, 48h and 72h respectively compared with the control group. Compared with the control, the strain can significantly degrade the cellulose in the material, see figure 5.
Table 4: cellulose degrading effect of bacterial strain
3.4 method for determining cellulose content: content determination of crude fiber in GB/T6434-2006 feed
TABLE 520 h-72 h degradation effect
Therefore, the bacillus licheniformis BGB-85F obtained by separation and identification can normally grow at high temperature, the Congo red identification culture medium is used for identifying the property of degrading cellulose, the microbial inoculum prepared from the zymocyte liquid is used for remarkably degrading the cellulose in the kitchen waste, the application effect is better in the scene of recycling the actual kitchen waste, the conversion efficiency of the kitchen waste can be greatly improved, and the application significance is good.
Finally, it should be noted that: the above embodiments are only used to illustrate the technical solution of the present invention, and not to limit the same; while the invention has been described in detail and with reference to the foregoing embodiments, it will be understood by those skilled in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some or all of the technical features may be equivalently replaced; and the modifications or the substitutions do not make the essence of the corresponding technical solutions depart from the scope of the technical solutions of the embodiments of the present invention.
Sequence listing
<110> Beijing Jia Bo Wen Biotech Co., Ltd
<120> bacterial strain for high-temperature degradation of cellulose in kitchen waste and screening and application thereof
<130> MP22006943
<160> 3
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<213> Bacillus licheniformis (Bacillus licheniformis)
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gttacctcac cgacttcggg tgttacaaac tctcgtggtg tgacgggcgg tgtgtacaag 60
gcccgggaac gtattcaccg cggcatgctg atccgcgatt actagcgatt ccagcttcac 120
gcagtcgagt tgcagactgc gatccgaact gagaacagat ttgtgggatt ggcttagcct 180
cgcggcttcg ctgccctttg ttctgcccat tgtagcacgt gtgtagccca ggtcataagg 240
ggcatgatga tttgacgtca tccccacctt cctccggttt gtcaccggca gtcaccttag 300
agtgcccaac tgaatgctgg caactaagat caagggttgc gctcgttgcg ggacttaacc 360
caacatctca cgacacgagc tgacgacaac catgcaccac ctgtcactct gcccccgaag 420
gggaagccct atctctaggg ttgtcagagg atgtcaagac ctggtaaggt tcttcgcgtt 480
gcttcgaatt aaaccacatg ctccaccgct tgtgcgggcc cccgtcaatt cctttgagtt 540
tcagtcttgc gaccgtactc cccaggcgga gtgcttaatg cgtttgctgc agcactaaag 600
ggcggaaacc ctctaacact tagcactcat cgtttacggc gtggactacc agggtatcta 660
atcctgttcg ctccccacgc tttcgcgcct cagcgtcagt tacagaccag agagtcgcct 720
tcgccactgg tgttcctcca catctctacg catttcaccg ctacacgtgg aattccactc 780
tcctcttctg cactcaagtt ccccagtttc caatgaccct ccccggttga gccgggggct 840
ttcacatcag acttaagaaa ccgcctgcgc gcgctttacg cccaataatt ccggacaacg 900
cttgccacct acgtattacc gcggctgctg ccacgtagtt agccgtggct ttctggttag 960
gtaccgtcaa ggtaccgccc tattcgaccg gtactggttc ttccctacca ccagagtttt 1020
acgatccgaa aaccttcatc actcacgcgg cgttgctccg tcagactttc gtccattgcg 1080
gaagattccc tactgctgcc tcccgtagga gtctgggccg tgtttcagtc ccagtgtggc 1140
cgatcaccct ctcaggtcgg ttacgcatcg tcgccttggt gagccgttac ctcaccaact 1200
agttaatgcg ccgcgggtcc atcggtaagt ggtagctaaa agccaccttt tatgattgaa 1260
ccatgcggtt caatcaagca tccggtatta gccccggttt cccggagtta tcccagtctt 1320
acaggcaggt tacccacgtg ttactcaccc gtccgccgct gacctaaggg agcaagctcc 1380
cgt 1383
<210> 2
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<213> Artificial Sequence (Artificial Sequence)
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<210> 3
<211> 22
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
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tacggctacc ttgttacgac tt 22
Claims (10)
1. Bacillus with preservation number of CGMCC No. 24544.
2. Application of bacillus with the preservation number of CGMCC No.24544 in degrading cellulose.
3. Use according to claim 2, wherein the cellulose comprises cellulose from kitchen waste.
4. The application of the kitchen waste as claimed in claim 3, wherein the pH of the kitchen waste is 6-8.
5. The cellulose degradation microbial inoculum is characterized by comprising bacillus with the preservation number of CGMCC No. 24544.
6. The cellulose-degrading bacterial agent according to claim 5, wherein the cellulose-degrading bacterial agent is a microorganism belonging to the genus BacillusThe content is 2 x 10 8 cfu/g～9×10 8 cfu/g。
7. The method for producing a cellulose-degrading bacterial agent according to claim 5 or 6, wherein a Bacillus having a preservation number of CGMCC No.24544 is cultured and the bacterial cells are collected.
8. A method for degrading cellulose, comprising inoculating the cellulose-degrading microbial agent of claim 5 or 6 to a substance to be degraded.
9. The method of claim 8, wherein the material to be degraded comprises kitchen waste.
10. The method of claim 8 or 9, wherein the degradation temperature comprises 45 ℃ to 82 ℃.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101892182A (en) * | 2010-06-07 | 2010-11-24 | 中国农业大学 | Bacillus licheniformis and application thereof in promotion of cellulose degradation |
| CN103820361A (en) * | 2014-01-15 | 2014-05-28 | 深圳市铁汉生态环境股份有限公司 | High-temperature cellulose degradation bacterium and application thereof |
| CN105647832A (en) * | 2016-03-05 | 2016-06-08 | 北京林业大学 | High-temperature-resistant garden waste decomposition bacterium FHM1 and application thereof |
| CN106957807A (en) * | 2017-03-24 | 2017-07-18 | 广西大学 | A kind of lichem bacillus strain TA65 and its application in compost maturity is promoted |
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101892182A (en) * | 2010-06-07 | 2010-11-24 | 中国农业大学 | Bacillus licheniformis and application thereof in promotion of cellulose degradation |
| CN103820361A (en) * | 2014-01-15 | 2014-05-28 | 深圳市铁汉生态环境股份有限公司 | High-temperature cellulose degradation bacterium and application thereof |
| CN105647832A (en) * | 2016-03-05 | 2016-06-08 | 北京林业大学 | High-temperature-resistant garden waste decomposition bacterium FHM1 and application thereof |
| CN106957807A (en) * | 2017-03-24 | 2017-07-18 | 广西大学 | A kind of lichem bacillus strain TA65 and its application in compost maturity is promoted |
Non-Patent Citations (1)
| Title |
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| 万文娟;张赟彬;聂志妍;毛彦佳;许静;喻莹: "耐高温餐厨垃圾分解细菌的筛选、鉴定及应用", 食品与机械, vol. 35, no. 006 * |
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