CN115340964B - Bacillus strain with swollenin activity and application thereof - Google Patents
Bacillus strain with swollenin activity and application thereof Download PDFInfo
- Publication number
- CN115340964B CN115340964B CN202210731664.XA CN202210731664A CN115340964B CN 115340964 B CN115340964 B CN 115340964B CN 202210731664 A CN202210731664 A CN 202210731664A CN 115340964 B CN115340964 B CN 115340964B
- Authority
- CN
- China
- Prior art keywords
- strain
- bacillus strain
- swollenin
- cffsh
- bacillus
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 241000193830 Bacillus <bacterium> Species 0.000 title claims abstract description 35
- 230000000694 effects Effects 0.000 title claims abstract description 12
- 230000004151 fermentation Effects 0.000 claims abstract description 26
- 238000000855 fermentation Methods 0.000 claims abstract description 26
- 238000006731 degradation reaction Methods 0.000 claims description 12
- 239000000843 powder Substances 0.000 claims description 9
- 238000004321 preservation Methods 0.000 claims description 7
- 240000008892 Helianthus tuberosus Species 0.000 claims description 6
- 235000003230 Helianthus tuberosus Nutrition 0.000 claims description 6
- 239000002068 microbial inoculum Substances 0.000 claims description 6
- 230000001580 bacterial effect Effects 0.000 claims description 3
- 238000000034 method Methods 0.000 claims description 3
- 239000003795 chemical substances by application Substances 0.000 claims 1
- 108010005131 levanase Proteins 0.000 claims 1
- 235000002637 Nicotiana tabacum Nutrition 0.000 abstract description 3
- 230000004952 protein activity Effects 0.000 abstract description 3
- 241000208125 Nicotiana Species 0.000 abstract 1
- 239000002028 Biomass Substances 0.000 description 21
- 108090000623 proteins and genes Proteins 0.000 description 15
- 241000196324 Embryophyta Species 0.000 description 13
- 108090000790 Enzymes Proteins 0.000 description 13
- 102000004190 Enzymes Human genes 0.000 description 13
- 229940088598 enzyme Drugs 0.000 description 13
- 102000004169 proteins and genes Human genes 0.000 description 12
- 239000000047 product Substances 0.000 description 9
- 230000000593 degrading effect Effects 0.000 description 8
- 210000004027 cell Anatomy 0.000 description 7
- 230000015556 catabolic process Effects 0.000 description 6
- 210000002421 cell wall Anatomy 0.000 description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 6
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 5
- 239000001913 cellulose Substances 0.000 description 5
- 229920002678 cellulose Polymers 0.000 description 5
- 239000008103 glucose Substances 0.000 description 5
- 239000006228 supernatant Substances 0.000 description 5
- ZRWPUFFVAOMMNM-UHFFFAOYSA-N Patulin Chemical compound OC1OCC=C2OC(=O)C=C12 ZRWPUFFVAOMMNM-UHFFFAOYSA-N 0.000 description 4
- 238000000105 evaporative light scattering detection Methods 0.000 description 4
- 238000004519 manufacturing process Methods 0.000 description 4
- 244000005700 microbiome Species 0.000 description 4
- 241000209140 Triticum Species 0.000 description 3
- 235000021307 Triticum Nutrition 0.000 description 3
- 230000009471 action Effects 0.000 description 3
- 230000000813 microbial effect Effects 0.000 description 3
- 239000010902 straw Substances 0.000 description 3
- 244000063299 Bacillus subtilis Species 0.000 description 2
- 235000014469 Bacillus subtilis Nutrition 0.000 description 2
- 108010059892 Cellulase Proteins 0.000 description 2
- 108050000194 Expansin Proteins 0.000 description 2
- 229930091371 Fructose Natural products 0.000 description 2
- RFSUNEUAIZKAJO-ARQDHWQXSA-N Fructose Chemical compound OC[C@H]1O[C@](O)(CO)[C@@H](O)[C@@H]1O RFSUNEUAIZKAJO-ARQDHWQXSA-N 0.000 description 2
- 239000005715 Fructose Substances 0.000 description 2
- 229920001202 Inulin Polymers 0.000 description 2
- 108010029541 Laccase Proteins 0.000 description 2
- 244000061176 Nicotiana tabacum Species 0.000 description 2
- 241000320412 Ogataea angusta Species 0.000 description 2
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 2
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 description 2
- 241000499912 Trichoderma reesei Species 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 239000007864 aqueous solution Substances 0.000 description 2
- 230000001413 cellular effect Effects 0.000 description 2
- 229940106157 cellulase Drugs 0.000 description 2
- 238000001816 cooling Methods 0.000 description 2
- 239000006260 foam Substances 0.000 description 2
- 238000004128 high performance liquid chromatography Methods 0.000 description 2
- 230000007062 hydrolysis Effects 0.000 description 2
- 238000006460 hydrolysis reaction Methods 0.000 description 2
- JYJIGFIDKWBXDU-MNNPPOADSA-N inulin Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)OC[C@]1(OC[C@]2(OC[C@]3(OC[C@]4(OC[C@]5(OC[C@]6(OC[C@]7(OC[C@]8(OC[C@]9(OC[C@]%10(OC[C@]%11(OC[C@]%12(OC[C@]%13(OC[C@]%14(OC[C@]%15(OC[C@]%16(OC[C@]%17(OC[C@]%18(OC[C@]%19(OC[C@]%20(OC[C@]%21(OC[C@]%22(OC[C@]%23(OC[C@]%24(OC[C@]%25(OC[C@]%26(OC[C@]%27(OC[C@]%28(OC[C@]%29(OC[C@]%30(OC[C@]%31(OC[C@]%32(OC[C@]%33(OC[C@]%34(OC[C@]%35(OC[C@]%36(O[C@@H]%37[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O%37)O)[C@H]([C@H](O)[C@@H](CO)O%36)O)[C@H]([C@H](O)[C@@H](CO)O%35)O)[C@H]([C@H](O)[C@@H](CO)O%34)O)[C@H]([C@H](O)[C@@H](CO)O%33)O)[C@H]([C@H](O)[C@@H](CO)O%32)O)[C@H]([C@H](O)[C@@H](CO)O%31)O)[C@H]([C@H](O)[C@@H](CO)O%30)O)[C@H]([C@H](O)[C@@H](CO)O%29)O)[C@H]([C@H](O)[C@@H](CO)O%28)O)[C@H]([C@H](O)[C@@H](CO)O%27)O)[C@H]([C@H](O)[C@@H](CO)O%26)O)[C@H]([C@H](O)[C@@H](CO)O%25)O)[C@H]([C@H](O)[C@@H](CO)O%24)O)[C@H]([C@H](O)[C@@H](CO)O%23)O)[C@H]([C@H](O)[C@@H](CO)O%22)O)[C@H]([C@H](O)[C@@H](CO)O%21)O)[C@H]([C@H](O)[C@@H](CO)O%20)O)[C@H]([C@H](O)[C@@H](CO)O%19)O)[C@H]([C@H](O)[C@@H](CO)O%18)O)[C@H]([C@H](O)[C@@H](CO)O%17)O)[C@H]([C@H](O)[C@@H](CO)O%16)O)[C@H]([C@H](O)[C@@H](CO)O%15)O)[C@H]([C@H](O)[C@@H](CO)O%14)O)[C@H]([C@H](O)[C@@H](CO)O%13)O)[C@H]([C@H](O)[C@@H](CO)O%12)O)[C@H]([C@H](O)[C@@H](CO)O%11)O)[C@H]([C@H](O)[C@@H](CO)O%10)O)[C@H]([C@H](O)[C@@H](CO)O9)O)[C@H]([C@H](O)[C@@H](CO)O8)O)[C@H]([C@H](O)[C@@H](CO)O7)O)[C@H]([C@H](O)[C@@H](CO)O6)O)[C@H]([C@H](O)[C@@H](CO)O5)O)[C@H]([C@H](O)[C@@H](CO)O4)O)[C@H]([C@H](O)[C@@H](CO)O3)O)[C@H]([C@H](O)[C@@H](CO)O2)O)[C@@H](O)[C@H](O)[C@@H](CO)O1 JYJIGFIDKWBXDU-MNNPPOADSA-N 0.000 description 2
- 229940029339 inulin Drugs 0.000 description 2
- 238000011031 large-scale manufacturing process Methods 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 239000011159 matrix material Substances 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 239000006041 probiotic Substances 0.000 description 2
- 230000000529 probiotic effect Effects 0.000 description 2
- 235000018291 probiotics Nutrition 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 230000002195 synergetic effect Effects 0.000 description 2
- 241000219194 Arabidopsis Species 0.000 description 1
- 241000194110 Bacillus sp. (in: Bacteria) Species 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 241000221198 Basidiomycota Species 0.000 description 1
- 101710130006 Beta-glucanase Proteins 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- 240000008067 Cucumis sativus Species 0.000 description 1
- 235000010799 Cucumis sativus var sativus Nutrition 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- 244000068988 Glycine max Species 0.000 description 1
- 235000010469 Glycine max Nutrition 0.000 description 1
- 241000045411 Hahella chejuensis Species 0.000 description 1
- 241000235058 Komagataella pastoris Species 0.000 description 1
- 239000012880 LB liquid culture medium Substances 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 240000007594 Oryza sativa Species 0.000 description 1
- 235000007164 Oryza sativa Nutrition 0.000 description 1
- 241000588701 Pectobacterium carotovorum Species 0.000 description 1
- 241000208181 Pelargonium Species 0.000 description 1
- 108091005804 Peptidases Proteins 0.000 description 1
- 239000001888 Peptone Substances 0.000 description 1
- 108010080698 Peptones Proteins 0.000 description 1
- 241000235648 Pichia Species 0.000 description 1
- 108010059820 Polygalacturonase Proteins 0.000 description 1
- 239000004365 Protease Substances 0.000 description 1
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- 241001484137 Talaromyces leycettanus Species 0.000 description 1
- 240000008042 Zea mays Species 0.000 description 1
- 235000016383 Zea mays subsp huehuetenangensis Nutrition 0.000 description 1
- 235000002017 Zea mays subsp mays Nutrition 0.000 description 1
- 230000006578 abscission Effects 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- 244000000005 bacterial plant pathogen Species 0.000 description 1
- 239000003899 bactericide agent Substances 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- GUBGYTABKSRVRQ-QRZGKKJRSA-N beta-cellobiose Chemical compound OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)[C@H](O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@@H]1O GUBGYTABKSRVRQ-QRZGKKJRSA-N 0.000 description 1
- 239000002551 biofuel Substances 0.000 description 1
- 229940041514 candida albicans extract Drugs 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 230000010261 cell growth Effects 0.000 description 1
- 239000013592 cell lysate Substances 0.000 description 1
- 230000008162 cell wall modification Effects 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 238000004891 communication Methods 0.000 description 1
- 230000006854 communication Effects 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 230000004069 differentiation Effects 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 108010093305 exopolygalacturonase Proteins 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 238000002386 leaching Methods 0.000 description 1
- 229920005610 lignin Polymers 0.000 description 1
- 239000008176 lyophilized powder Substances 0.000 description 1
- 235000009973 maize Nutrition 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 230000010534 mechanism of action Effects 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 230000005305 organ development Effects 0.000 description 1
- 235000019319 peptone Nutrition 0.000 description 1
- 230000029553 photosynthesis Effects 0.000 description 1
- 238000010672 photosynthesis Methods 0.000 description 1
- 230000008635 plant growth Effects 0.000 description 1
- 244000000003 plant pathogen Species 0.000 description 1
- 239000010908 plant waste Substances 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 235000009566 rice Nutrition 0.000 description 1
- 230000009758 senescence Effects 0.000 description 1
- 238000005507 spraying Methods 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 230000002459 sustained effect Effects 0.000 description 1
- 230000025366 tissue development Effects 0.000 description 1
- 230000032258 transport Effects 0.000 description 1
- 230000035899 viability Effects 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
- 239000002916 wood waste Substances 0.000 description 1
- 239000012138 yeast extract Substances 0.000 description 1
Classifications
-
- 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
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/02—Monosaccharides
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/14—Preparation of compounds containing saccharide radicals produced by the action of a carbohydrase (EC 3.2.x), e.g. by alpha-amylase, e.g. by cellulase, hemicellulase
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P2201/00—Pretreatment of cellulosic or lignocellulosic material for subsequent enzymatic treatment or hydrolysis
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/07—Bacillus
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Biotechnology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- Microbiology (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Biomedical Technology (AREA)
- Virology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Medicinal Chemistry (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Enzymes And Modification Thereof (AREA)
Abstract
The invention discloses a bacillus strain with swollenin activity and application thereof, and fermentation liquor of the bacillus strain has extremely high swollenin protein activity. The strain is a bacillus strain CFFSH and 013, the original strain is separated from the surface of tobacco leaves, and is identified as the bacillus strain through 16srDNA, and the strain is preserved in China Center for Type Culture Collection (CCTCCNO): m2020635.
Description
Technical Field
The invention relates to a strain, in particular to a bacillus strain with swollenin activity, and belongs to the technical field of strains.
Background of the invention:
Swollenin (Expansin, also known as swollenin or expansin) is a protein that causes relaxation of the plant cell wall and plays a key role in plant cell expansion and a series of vital events involving cell wall modification. Swollenins are encoded by multiple gene families, and studies have shown that the swollenin superfamily consists of four gene subfamilies.
Cell walls play a vital role in various cellular activities such as differentiation, transport and communication, senescence, abscission, plant-pathogen interactions, and ultimately plant growth. It provides both the mechanical strength required for plants and the plasticity required for plant tissue and organ development. Although the biochemical mechanism of action of swollenins is not completely understood, it is believed that the action of swollenins on the cell wall brings about this urgent plasticity, which itself, although not an enzyme, can relax the cell wall, causing a sustained decrease in the tensile strength of the cell wall, which in turn causes the cell to expand.
Swollenin was first discovered in cucumber by S McQueen-Mason (DOI: 10.1105/tpc.4.11.1425), after which it was found to be ubiquitous in a wide variety of plants, including Arabidopsis, tobacco, rice, maize, soybean, and the like. However, plant-derived swollenins have the problems of low expression yield and difficulty in mass production.
In addition to plant sources, the prior art has found that microorganisms, which are partially colonized on plant surfaces, also have the ability to secrete and produce microbial swollenin proteins. Including expansin-like proteins from Pelargonium dish (Kawata et al, 2015), bsExlx1 from Bacillus subtilis (Kerff et al, 2008), hcExlx2 from marine bacteria Hahella chejuensis (Lee et al, 2010), pcExlx fine bacteria Pectobacterium carotovorum from plant pathogenic bacteria (Olarte-Lozano et al, 2014) and ScExlx1 from Basidiomycetes Schizophylum (Tovar-Herrera et al, 2015). These microbial expansins have demonstrated a variety of capabilities for attaching and colonizing plants with microorganisms. In addition, some researchers are evaluating the potential use of these microbial expansins in the conversion of cellulosic biomass for biofuel production as a means of decomposing the cellulosic structure (Georgelis et al, 2015).
The swollenin does not hydrolyze glycosidic bonds in cellulose molecules, but has a CBD binding domain similar to cellulose enzyme and a sequence similar to plant swollenin, can have expansion and disintegration effects on cellulose and cell walls, and improves the action efficiency of cellulose. The swollenin can effectively convert lignocellulose biomass substrates through the synergistic effect with biomass degrading enzymes such as cellulase and the like, so that the economic benefit of fermentation production is improved.
The patent application fungus-derived swollenin protein, and its gene and use (CN 201710269769.7) discloses a swollenin protein derived from Talaromyces leycettanus bacteria. However, the production of this protein still requires expression by recombinant Pichia (Pichia pastoris) cells, saccharomyces cerevisiae (Saccharomyces cerevisiae) cells or Hansenula polymorpha (Hansenula polymorpha) cells.
The low-cost and rapid large-scale production of the swollenin protein is an important condition for the industrial application. The bacillus strain grows rapidly, has high protein expression, is an important probiotic, has high safety, and is very suitable for large-scale production of swollenin protein. Meanwhile, the bacillus is a probiotic of many plants, and can be used for producing the bacterial swollenin protein BsExlx in bacillus subtilis in the earliest period in 2008. Finding strains that produce high levels of cellular bacterial swollenin protein is a key technology lacking in the prior art.
Disclosure of Invention
The invention aims to provide a bacillus strain with swollenin activity, and fermentation liquor of the bacillus strain has extremely high swollenin protein activity. The strain is a bacillus strain CFFSH013, the original strain is separated on the surface of tobacco leaves by the inventor, and is identified as a bacillus strain through 16srDNA, and the strain is preserved in China center for type culture collection, and addresses: university of martial arts in chinese; classification naming: bacillus CFFSH013 (Bacillus sp.cffsh 013); the preservation center number is CCTCC NO: m2020635; the preservation date is: 10 months and 26 days 2020. Viability of the cultures the present collection was tested at 10 days 11 in 2020 and was found to be viable.
The aim of the invention is achieved by the following technical scheme.
A bacillus strain with swollenin activity, said strain being of the type having a accession number cctccc NO: strain of M2020635.
A product having swollenin activity, which is any one of a fermentation broth containing bacillus strain CFFSH, bacillus strain CFFSH013, a fermentation broth supernatant of bacillus strain CFFSH013, a fermentation extract of bacillus strain CFFSH013, and a protein product of bacillus strain CFFSH 013.
A method for accelerating biomass degradation process comprises using Bacillus strain CFFSH (preservation number is CCTCC NO: M2020635) in combination with other biomass degrading enzyme.
Further, the mode of using the bacillus strain CFFSH (with the preservation number of CCTCC NO: M2020635) together with other biomass degrading enzymes comprises using fermentation products of the bacillus strain CFFSH013 together with other biomass degrading enzymes.
A microbial inoculum for accelerating biomass degradation process comprises Bacillus strain CFFSH (preservation number is CCTCC NO: M2020635).
Furthermore, the microbial inoculum for accelerating the biomass degradation process also contains other strains capable of producing biomass degradation enzymes.
The term "biomass" in the present application refers to various organisms produced by photosynthesis using the atmosphere, water, land, etc., that is, all living and growing organic substances are generally called biomass, and include all plants, microorganisms, and animals and wastes produced by the plants, microorganisms, such as crops, crop wastes, wood wastes, and biological source components such as biological source cellulose, starch, lignin, etc.
The concept of "biomass degrading enzyme" in the present application refers to degrading enzymes such as cellose, protease, pectinase, laccase and the like capable of decomposing various biomasses and components thereof.
Detailed Description
The technical features of the present invention are further described below in conjunction with specific embodiments.
Example 1: culture of Bacillus strain CFFSH013
Preparing LB liquid culture medium (yeast extract 5g/L, peptone 10 g/L), sterilizing at 115deg.C for 20min, cooling to room temperature, inoculating 1% (v/v) selected Bacillus strain CFFSH, culturing at 37deg.C for 24 hr, and storing at 4deg.C.
In order to facilitate the determination of protein activity, different fermentation products are obtained, and fermentation liquid of bacillus strain CFFSH013,013 is treated as follows:
(1) CFFSH013 fermentation broth: preserving at 4deg.C for standby after fermentation, and shaking uniformly before use;
(2) CFFSH013 inactivated fermentation broth: heating the fermentation liquor to 100 ℃ after fermentation, cooling to 4 ℃ and preserving for later use;
(3) CFFSH013 fermenting supernatant, namely centrifuging the fermentation liquor by using a refrigerated centrifuge (4 ℃ C., 8000 rpm) after fermentation, discarding the lower layer thallus, and collecting supernatant for storage at 4 ℃ for later use.
(4) CFFSH013 fermenting the supernatant to obtain lyophilized powder, centrifuging the fermentation broth with a refrigerated centrifuge (4 deg.C, 8000 rpm) after fermentation, discarding the lower layer thallus, collecting supernatant, standing at-80deg.C for 12 hr to pre-freeze to ice, lyophilizing the fermentation broth to obtain powder with a vacuum freeze dryer, and collecting the powder.
(5) CFFSH013 cell lysate of 013 cells, after fermentation, the fermentation broth is centrifuged by a refrigerated centrifuge (4 ℃ C., 8000 rpm), the lower layer of cells are resuspended by distilled water, and then crushed by an ultrasonic cell crusher, and collected and stored at 4 ℃ for standby.
Example 2: biomass degradation ability (Jerusalem artichoke powder)
The jerusalem artichoke slices are ground into powder to serve as a biomass degradation matrix, beta-fructosan enzyme (produced by Henan Yi Biotechnology Co., ltd.) is used for measuring the content of glucose and fruit carbon released from hydrolysis liquid after hydrolysis by adopting a high performance liquid chromatography evaporative light scattering detection analysis method (HPLC-ELSD, chromatographic column: fei Roman MARS MCa). The experimental Blank (BK) was prepared by adding inulin directly to water to prepare an aqueous solution having a final concentration of 5% (w/w), and the glucose-based fructose content was directly measured. The experimental blank was prepared by adding inulin to water to prepare an aqueous solution with a final concentration of 5% (w/w), and then reacting for 30min in a 40℃water bath shaker, after which the glucose and fructose contents were determined. The measurement conditions and treatment conditions of the other experimental groups are listed in table 1.
The results (Table 1) show that the fermentation product of CFFSH strain alone does not have obvious effect of degrading biomass, but can greatly improve the action efficiency of beta-fructosan and beta-glucanase when the fermentation product is combined with other biomass degrading enzymes, and show that the fermentation product has synergistic effect similar to that of swollenin.
TABLE 1 Biomass degradation of Jerusalem artichoke powder
Example 3: biomass degradation (wheat straw foam)
The treatment process of the treatment matrix of the embodiment comprises the steps of taking 200g of wheat straw powder, spraying 200g of water, adding CFFSH g of fermentation product and other enzymes or bactericides according to the treatment mode listed in table 2, uniformly mixing, storing and reacting for 7 days at 30 ℃ in a incubator with 60% humidity, adding 200g of 0.1% HCL solution, shaking for 12 hours at 25 ℃ to leach out glucose, taking the leaching solution, and measuring the glucose content by adopting a high performance liquid chromatography evaporative light scattering detection analysis method (HPLC-ELSD, chromatographic column: fimbrane MARS M Ca). The cellulase used in this example was purchased from the biological engineering (Shanghai) stock company and the laccase was purchased from Shandong Xuanyang Biotech Co. Trichoderma reesei is derived from purified strain of Trichoderma reesei preparation Kang Fengyuan of Shandong Bobi Yuan kang biotechnology Co.
As shown in Table 2, CFFSH A013 fermented product has a greatly improved efficiency in dissociating biomass with an additional enzyme preparation/microbial inoculum.
TABLE 2 Biomass degradation treatment conditions for wheat straw foam
Claims (5)
1. A strain of bacillus having swollenin activity, characterized by: the strain is bacillus strain CFFSH and 013, and the preservation number is CCTCC NO: strain of M2020635.
2. A product comprising the bacillus strain having swollenin activity of claim 1, characterized in that: the product is any one of fermentation liquor containing bacillus strain CFFSH and bacillus strain CFFSH 013.
3. A method for accelerating the degradation process of jerusalem artichoke powder is characterized by comprising the following steps: the bacillus strain CFFSH is taken as a bacillus strain CFFSH, and the preservation number is CCTCC NO: m2020635; is used in combination with glucanase or levanse.
4. A microbial inoculum for accelerating the degradation process of jerusalem artichoke powder is characterized in that: the microbial inoculum contains bacillus strain CFFSH and 013 with the preservation number of CCTCC NO: m2020635.
5. The microbial inoculum for accelerating the degradation process of jerusalem artichoke powder according to claim 4, which is characterized in that: the bacterial agent also contains other strains capable of producing glucanase or levanase.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210731664.XA CN115340964B (en) | 2022-06-26 | 2022-06-26 | Bacillus strain with swollenin activity and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210731664.XA CN115340964B (en) | 2022-06-26 | 2022-06-26 | Bacillus strain with swollenin activity and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN115340964A CN115340964A (en) | 2022-11-15 |
| CN115340964B true CN115340964B (en) | 2024-05-14 |
Family
ID=83947722
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210731664.XA Active CN115340964B (en) | 2022-06-26 | 2022-06-26 | Bacillus strain with swollenin activity and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN115340964B (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115404180A (en) * | 2022-06-26 | 2022-11-29 | 上海龙殷生物科技有限公司 | Bacillus fragrans and application thereof in tobacco |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102712893A (en) * | 2009-07-03 | 2012-10-03 | 帝斯曼知识产权资产管理有限公司 | Talaromyces strains and enzyme compositions |
| CN107345212A (en) * | 2016-12-30 | 2017-11-14 | 广西中烟工业有限责任公司 | The bacillus and its application in reconstituted tobacoo of one plant of cellulase-producing and laccase |
| CN107488221A (en) * | 2017-04-24 | 2017-12-19 | 中国农业科学院饲料研究所 | The expansion fibroin and its gene of originated from fungus and application |
| CN113652379A (en) * | 2021-09-27 | 2021-11-16 | 中国科学院广州能源研究所 | Bacillus cell wall depolymerized GIEC and application thereof |
-
2022
- 2022-06-26 CN CN202210731664.XA patent/CN115340964B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102712893A (en) * | 2009-07-03 | 2012-10-03 | 帝斯曼知识产权资产管理有限公司 | Talaromyces strains and enzyme compositions |
| CN107345212A (en) * | 2016-12-30 | 2017-11-14 | 广西中烟工业有限责任公司 | The bacillus and its application in reconstituted tobacoo of one plant of cellulase-producing and laccase |
| CN107488221A (en) * | 2017-04-24 | 2017-12-19 | 中国农业科学院饲料研究所 | The expansion fibroin and its gene of originated from fungus and application |
| CN113652379A (en) * | 2021-09-27 | 2021-11-16 | 中国科学院广州能源研究所 | Bacillus cell wall depolymerized GIEC and application thereof |
Non-Patent Citations (4)
| Title |
|---|
| A potential cellulose microfibril swelling enzyme isolated from Bacillus sp.AY8 enhances cellulose hydrolysis;Md. Azizul Haque等;Process Biochemistry;第807-815页 * |
| Crystal structure and activity of Bacillus subtilis YoaJ (EXLX1), a bacterial expansin that promotes root colonization;Kerffa等;PNAS;20081104;第105卷(第44期);第16876–16881页 * |
| 扩展蛋白在纤维素降解中作用的研究进展;由林鹭等;中国生物制品学杂;第31卷(第12期);第1409-1416页 * |
| 膨胀素对植物根系生长发育的调控;文文乙豪等;生命科学;20130930;第25卷(第9期);第922-931页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN115340964A (en) | 2022-11-15 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN110331109B (en) | Bacillus subtilis and culture method and application thereof | |
| CN109355227B (en) | Streptomyces violaceus strain and application thereof in cellulose degradation | |
| CN111363684B (en) | Composite microbial inoculum for efficiently degrading wood fibers and application thereof in composting | |
| CN101914450B (en) | Fungus agent for biologically dewatering compost and preparation method thereof | |
| Hamouda et al. | Bioethanol production by various hydrolysis and fermentation processes with micro and macro green algae | |
| CN109161495B (en) | Composite microbial inoculum for efficiently degrading straw cellulose | |
| CN108823102B (en) | Cold region straw rotten fungus Mortierella sarnyensis strain and application thereof in rice straw rotten | |
| Abada et al. | Optimization of cellulase production from Bacillus albus (MN755587) and its involvement in bioethanol production | |
| US20020012980A1 (en) | Method for simultaneous saccharification and fermentation of spent cellulose sausage casings | |
| CN115340964B (en) | Bacillus strain with swollenin activity and application thereof | |
| CN108865927B (en) | Bacterial strain for low-temperature glycolysis of corn straw and fermentation culture method and application thereof | |
| CN115851545A (en) | Korean bacillus and culture medium for improving activity of enzyme produced by same | |
| CN109439585B (en) | Katy bacteria adipate and application thereof in cellulose degradation | |
| CN113652379B (en) | Bacillus cell wall depolymerization GIEC and application thereof | |
| TW201420760A (en) | Bacillus subtilis and application thereof | |
| CN103352016A (en) | Method for preparing biological fertilizer by utilizing Alteromonas colwelliana A321 to ferment enteromorpha | |
| CN112029681B (en) | Preparation of special liquid composite microbial inoculum for decomposing vinasse | |
| CN114480205A (en) | Bacillus amyloliquefaciens and application thereof in brewing of solid-state fermented vinegar | |
| CN108841743B (en) | Cold region straw rotten bacterial strain and preparation method and application thereof | |
| CN112725194B (en) | Fungus Flavodon sp.x10 for high yield of cellulase and application thereof | |
| CN113201460B (en) | Medicinal fungus trichoderma brevicompactum and application thereof | |
| CN1834222A (en) | Fetid aspergillic strain and uses | |
| CN104818220B (en) | One plant is screened the Rhizopus oryzae bacterial strain JHSW01 obtained from rotten stalk | |
| CN111172062B (en) | Sphingobacterium multivorum and application thereof | |
| CN109988729B (en) | Normal-temperature compound strain system and application thereof in degradation of lignocellulose |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |