CN102352320B - Novel myceliophthora thermophila strain and application thereof - Google Patents
Novel myceliophthora thermophila strain and application thereof Download PDFInfo
- Publication number
- CN102352320B CN102352320B CN2011102909513A CN201110290951A CN102352320B CN 102352320 B CN102352320 B CN 102352320B CN 2011102909513 A CN2011102909513 A CN 2011102909513A CN 201110290951 A CN201110290951 A CN 201110290951A CN 102352320 B CN102352320 B CN 102352320B
- Authority
- CN
- China
- Prior art keywords
- vmt
- climax
- mould
- silk
- thermophilic
- 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
Abstract
The invention discloses a novel myceliophthora thermophila strain (CLIMAX-VMT). The preservation number of the novel myceliophthora thermophila strain is CGMCC No.5134, the preservation date is August 12th, 2011, and the preservation unit is General Microbiology Center of China Microbial Strain Preservation Management Committee. The invention also discloses a microbial inoculum containing the strain. The novel myceliophthora thermophila strain (CLIMAX-VMT) provided by the invention has very obvious fast fermentation temperature rise and cellulose decomposition performance and is innocuous and harmless to people and livestock. When the strain is used for fermenting organic solid waste materials, the fast temperature rise in 24h can be reached, and the temperature is raised to a value higher than 60 DEG C in 48h and can be maintained for more than 5 days. The strain does not have the obvious antagonism action on most low-temperature germs, i.e. the universality and the matching with other strains of the microbial inoculum are very good.
Description
Technical field
The invention belongs to and be used for the soil and fertilizer field, be specifically related to a kind of agricultural microorganism preparation and application thereof.
Background technology
Agriculture prodn needs soil that the nutritive substance and the living environment of plant growth are provided.For many years, using of chemical fertilizer played keying action to getting agricultural profit.Yet because long-term application chemical fertilizer, fertilizer is under-supply, and all kinds of nutrients ratio imbalances cause farmland ecological environment, soil physical and chemical property and soil microflora to receive destruction in various degree, have influenced the quality of agricultural-food to a certain extent.Therefore, chemical fertilizer pollution has become the world today's one big public hazards, and the present soil resource present situation of China presses for through the people to measure replenishes the soil organism, guarantees agricultural planting level and crop products quality.
In ten big countermeasures of China's ecological environmental protection, clearly advocate and to promote " ecological agriculture "; Preserve the ecological environment and agricultural land soil; Promote the production of pollution-free healthy food; Assurance has the sufficient amount organic matter to return soil, to promote biological cycle in the ecosystem, reaches maintenance and strengthens soil fertility and biological activity purpose.Along with the fast development of China's modern agriculture, particularly the green organic agricultural cultivated area constantly enlarges, and will play the part of more and more important role to the mikrobe that plays an important role in the ecological agriculture cycle art.And the pyroprocessing technology of wherein carrying out organic substance through thermophile bacteria is one of emphasis of numerous agricultural microorganisms technology.
Thermophile bacteria is one type of mikrobe that lives in the hot environment; Its relevant bacterial classification and research and development of products progressively begin to obtain scientist's attention and concern; The high temperature of thermophile bacteria and grow rapidly that characteristic makes it significantly reduce the living contaminants probability than mesophilic bacteria kind is fit to adopt specific suitability for industrialized production technology to make and be applied to the agriculture circular treatment field of the biological high-temperature technology of needs.
Summary of the invention
The present invention at first provides a kind of thermophilic new bacterial strain CLIMAX-VMT of silk mould (Myceliophthora thermophila) that ruins.
The present invention also provides and contains the thermophilic microbial inoculum of ruining the mould CLIMAX-VMT of silk.
Thermophilic ruining provided by the invention extracted in the straw powder heap that the mould CLIMAX-VMT of silk is high temperature heating more than 60 ℃ and obtained production performance and the stable bacterial strain of metabolic activity through long-term screening, purifying, domestication.This bacterial strain has good decomposition of cellulose ability, and through measuring, the cellulase activity is higher in its fermented product.This bacterial strain carries out preservation at China Committee for Culture Collection of Microorganisms common micro-organisms center, and the preservation accession designation number is CGMCC No.5134, and preservation date is on August 12nd, 2011.
The researchist piles at 60 ℃ corn straw; Get the remarkable bacterium piece in high-temperature zone through selecting culture medium culturing; Get meet thermophilic ruin the silk mould characteristic bacterial classification and at test under microscope; Cultivate screening through separating repeatedly after proving single CLIMAX-VMT thalline, obtain the CLIMAX-VMT bacterial strain with a large amount of bacterial strain repeatability.The CLIMAX-VMT bacterial strain is confirmed through colonial morphology, microscopic morphology characteristic etc. that this bacterial strain meets and is thermophilicly ruined the mould characteristic of silk, and certainly hides through China Committee for Culture Collection of Microorganisms's common micro-organisms nucleus.
The thermophilic mould CLIMAX-VMT of silk that ruins of the present invention can be prepared into microbial inoculum, and wherein, the thermophilic viable count of ruining the mould CLIMAX-VMT of silk can be more than 1,500,000,000/gram, is preferably 3,000,000,000/gram.
Of the present invention contain thermophilic ruin the silk mould CLIMAX-VMT microbial inoculum, can prepare as follows:
1) the thermophilic mould CLIMAX-VMT seed liquor of silk of ruining of preparation;
2) the thermophilic solid multiplication of ruining the mould CLIMAX-VMT of silk is cultivated: the seed liquor of step 1) is linked in the solid multiplication substratum 42 ± 2 ℃ aseptic aerobic conditions and cultivated 5～10 days down; Described solid multiplication substratum contains 50～100 parts in wheat bran, 10～40 parts of corn cob meals, 1～10 part in brown sugar, 5～20 parts of soya-bean cakes, 1～3 part in calcium chloride, 1～3 part in sodium-chlor, pH7.0～7.5;
3) substratum behind the step 3) multiplication culture is carried out Air drying, promptly get the thermophilic former bacteria agent of the mould CLIMAX-VMT of silk of ruining.
The present invention also provides and contains the thermophilic preparation method who ruins the mould CLIMAX-VMT microbial inoculum of silk, and it comprises:
1) the thermophilic mould CLIMAX-VMT seed liquor of silk of ruining of preparation;
2) the thermophilic solid multiplication of ruining the mould CLIMAX-VMT of silk is cultivated: the seed liquor of step 1) is linked in the solid multiplication substratum 42 ± 2 ℃ aseptic aerobic conditions and cultivated 7～10 days down; Described solid multiplication substratum contains 50～70 parts in wheat bran, 3～6 parts in brown sugar, 20～40 parts of cottonseed cakes (or soya-bean cake), 1～3 part in sal epsom, 1～3 part in sodium-chlor, 1～4 part in lime carbonate, pH7.0～7.5;
3) substratum behind the step 3) multiplication culture is carried out Air drying, promptly get the thermophilic former bacteria agent of the mould CLIMAX-VMT of silk of ruining.
Wherein, thermophilic preparation of ruining the mould CLIMAX-VMT seed liquor of silk preferably includes following steps:
On the aseptic technique platform, the CLIMAX-VMT in the freeze-drying pipe is linked in the inclined-plane of PDA substratum earlier and cultivated 3-7 days at 42 ℃; Behind the bacterium colony that secures good health, it is inserted in nutrient agar 42 ℃ cultivated 3-7 days, select then grow healthy bacterial classification be linked into through sterilization by sucrose 5～15; Preferred 10 parts, sodium-chlor 3～10 minutes; Preferred 5 parts, Carnis Bovis seu Bubali cream 5～15,3～8 parts of preferred 10 parts, calcium chloride, 0.2～1.0 part of preferred 5 part, potassium primary phosphate; Preferred 0.5 part, 0.01 part of iron trichloride, 1～4 part of yeast extract paste; In preferred 2 parts, the liquid nutrient medium of 1000 parts of compositions of water, and cultivated 3 days, obtains a thermophilic mould seed liquor of ruining 150r/min, 42 ℃.
Provided by the inventionly thermophilicly ruin that silk is mould to have a good decomposition of cellulose ability, through measuring, the cellulase activity is higher in its fermented product, can be used for handling agricultural organic solid refuse.Wherein, it is 10～40 that the organic solid refuse of accessible agricultural is preferably C/N, initial moisture 35～70%, the organic solid refuse of pH5～9.
The invention provides a kind of thermophilic new bacterial strain of a mould of ruining, it has very significant quick fermentation intensification and decomposition of cellulose performance, and nontoxic to people and animals.This bacterium adopts the liquid+solid combined fermentation method to carry out value added by manufacture; The mode of adsorbing into solid preparation with traditional liquid fermenting is compared; It is better that this method is carried out the efficient biological activity higher, its product of bacterial classification propagation, and preservation period is longer, can preserve at normal temperatures to reach more than 24 months.And in the microbial inoculum, the activity of cellulase can reach more than the 300U/g.With the organic solid refuse of this strain fermentation, can reach 24 hours and be rapidly heated, more than the temperature rise to 60 in 48 hours ℃ and continue more than 5 days.Bacterial strain of the present invention does not have tangible antagonism property to most of psychrophiles, and promptly this microbial inoculum is general and fine with the compatibleness of other bacterial strains.
Embodiment
Following examples are used to explain the present invention, but are not used for limiting scope of the present invention.
Do not specify that " part " described in the application is meant " weight part " if having.
Embodiment 1 thermophilic evaluation of ruining the mould CLIMAX-VMT of silk
The thermophilic mould CLIMAX-VMT of silk that ruins of the present invention has following colonial morphology and form:
It is fast to go up colony growth at wort agar substratum (MEA), and 40 ℃ of dark conditions were cultivated colony diameter 40-50mm 5 days down; Bacterium colony initial stage white, bacteria colony white becomes grey pink colour to beige afterwards, and quality is cotton-shaped; Back side beige does not have water-soluble pigment.
Hyphae colorless, many separations, wide 2-5 μ m.Conidiophore does not have specialization, on mycelia top or lateral short handle and urniform protuberance, produces spore.Give birth to or adnation on the conidium top, and Dan Sheng sees that seldom chain is living, oval or near clavate, and base portion is truncate, and is colourless to light brown, smooth or coarse, 4.5-7.5 * 2.8-4.5 μ m.
Above-mentioned characteristic all meets the thermophilic mould characteristic of silk of ruining.
Measured the thermophilic sequence of ruining the ITS4 of the mould CLIMAX-VMT of silk of the present invention, its sequence is shown in SEQ ID No.1.This sequence and the known thermophilic homology of sequence of ruining the mould ITS4 of silk are higher.
Really to ruin silk mould for thermophilic can to confirm as the bacterial strain that is separated to of the present invention through above-mentioned characteristic.
Embodiment 2 thermophilic preparations of ruining the former bacteria agent of the mould CLIMAX-VMT of silk
The present invention adopts liquid-solid combined ferment culture process to carry out thermophilic cultivation of ruining the mould microbial inoculum of silk, is linked into the solid culture multiplication culture again through first liquid multiplication culture, and formed microbial inoculum has very strong bioenzyme activity, and concrete steps comprise:
1) seed preparation and liquid multiplication culture: will preserve the thermophilic freeze-drying pipe of ruining the mould CLIMAX-VMT of silk earlier and carry out activation, concrete grammar is on the aseptic technique platform, the bacterial classification in the freeze-drying pipe to be linked in the inclined-plane of PDA substratum earlier to cultivate 5 days at 42 ℃; Behind the bacterium colony that secures good health; It is inserted in nutrient agar 42 ℃ cultivated 5 days; Select then grow healthy bacterial classification be linked into through sterilization by in 10 parts of sucrose, 5 parts in sodium-chlor, 10 parts of Carnis Bovis seu Bubali creams, 5 parts in calcium chloride, 0.5 part of potassium primary phosphate, 0.01 part of iron trichloride, 2 parts of yeast extract pastes, 1000 parts of liquid nutrient mediums of forming of water; And cultivated 3 days 150r/min, 42 ℃, obtains a thermophilic mould seed liquor of ruining.
2) solid multiplication is cultivated: the thermophilic mould seed liquor of silk of ruining of step 1) is linked into by in 52 parts in wheat bran, 30 parts of corn cob meals, 5 parts in brown sugar, 10 parts of soya-bean cakes, 1.5 parts in calcium chloride, 1.5 parts of solid culture mediums (pH7.0) through high-temperature sterilization more than 120 ℃ of forming of sodium-chlor, under 42 ℃ aseptic aerobic conditions, cultivated 7 days.
(3) drying treatment: after solid multiplication is cultivated and finished it is put in kiln and carries out Air drying.Driedly thermophilicly ruin silk mould to detect a living bacteria count through a colony counting method be 3,000,000,000/gram, preserve as the thermophilic mould former bacteria preparation packing warehouse-in of silk of ruining.
Measured the cellulase activity in the substratum after the above-mentioned propagation through the method for routine, be 350U/g, shown that the thermophilic mould CLIMAX-VMT of silk that ruins of the present invention has higher cellulolytic activity.
The embodiment 2 thermophilic mould CLIMAX-VMT of silk that ruin handle agricultural organic solid refuse
The C/N of cow dung is adjusted to 30, initial moisture 50%, pH7.0.The microbial inoculum of embodiment 2 preparations is mixed in the above-mentioned cow dung, and input amount is 0.3% of a cow dung weight, adds the superphosphate of lime of said mixture weight 10% again.When temperature of charge reaches 60～70 ℃ and be reduced to 20～40 ℃ of end that can be judged as fermentative processing.
After the fermentation ends, through detecting, the cow dung after the treated of the present invention, its colibacillus of excrement crowd＜100/gram, roundworm mortality ratio 100%.The temperature of charge index can reach 24 hours and is rapidly heated, more than the temperature rise to 60 in 48 hours ℃ and continue more than 5 days.
Cow dung before becoming thoroughly decomposed is compared with the cow dung of utilizing the CLIMAX-VMT high-temperature maturing; The latter has naked eyes can distinguish a large amount of mycelia; The higher materials of content of cellulose such as the weeds stalk in the cow dung have been decomposed rotten, show that the thermophilic mould effectively Mierocrystalline cellulose of silk of ruining of CLIMAX-VMT decomposes utilization.
Embodiment 3 thermophilic compatiblenesies of ruining the mould CLIMAX-VMT of silk and other psychrophile
With the thermophilic mould CLIMAX-VMT microbial inoculum of silk of ruining of the present invention: Bacillus subtillis microbial inoculum: yeast saccharomyces cerevisiae microbial inoculum: zeolite powder=1: 1: 1: 1 ratio is mixed, and viable count such as following table are said separately for they.Respectively according to the data of joining at 1: 3 behind the zeolite powder, the bacterium number average that draws each bacterium test in the 30th day does not have significantly sacrificing through the various bacterium of comparison, explain thermophilicly to ruin that silk is mould to possess the good many bacterial classifications flexibility (table 1) of ten minutes.
The compatibleness that table 1 is thermophilic ruins the mould CLIMAX-VMT of silk and other psychrophile
The above only is a preferred implementation of the present invention; Should be pointed out that for those skilled in the art, under the prerequisite that does not break away from know-why of the present invention; Can also make some improvement and retouching, these improvement and retouching also should be regarded as protection scope of the present invention.
Claims (6)
1. thermophilic new bacterial strain CLIMAX-VMT of silk mould (Myceliophthora thermophila) that ruins, its preserving number is CGMCC No.5134.
2. the microbial inoculum that contains the said bacterial strain of claim 1.
3. microbial inoculum according to claim 2, the wherein thermophilic viable count of ruining the mould CLIMAX-VMT of silk is more than 1,500,000,000/gram.
4. according to claim 2 or 3 described microbial inoculums, it is characterized in that, prepare as follows:
1) the thermophilic mould CLIMAX-VMT seed liquor of silk of ruining of preparation;
2) the thermophilic solid multiplication of ruining the mould CLIMAX-VMT of silk is cultivated: the seed liquor of step 1) is linked in the solid multiplication substratum 42 ± 2 ℃ aseptic aerobic conditions and cultivated 5～10 days down; Described solid multiplication substratum contains 50～100 parts in wheat bran, 10～40 parts of corn cob meals, 1～10 part in brown sugar, 5～20 parts of soya-bean cakes, 1～3 part in calcium chloride, 1～3 part in sodium-chlor, pH7.0～7.5;
3) with step 2) substratum behind the multiplication culture carries out Air drying, promptly gets the thermophilic former bacteria agent of the mould CLIMAX-VMT of silk of ruining.
5. the application of the said bacterial strain of claim 1 in the organic solid waste treatment of agricultural.
6. application according to claim 5 is characterized in that, the C/N of the organic solid refuse of described agricultural is 10～40, initial moisture 35～70%, pH5～9.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011102909513A CN102352320B (en) | 2011-09-29 | 2011-09-29 | Novel myceliophthora thermophila strain and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011102909513A CN102352320B (en) | 2011-09-29 | 2011-09-29 | Novel myceliophthora thermophila strain and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN102352320A CN102352320A (en) | 2012-02-15 |
| CN102352320B true CN102352320B (en) | 2012-10-31 |
Family
ID=45575960
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2011102909513A Active CN102352320B (en) | 2011-09-29 | 2011-09-29 | Novel myceliophthora thermophila strain and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102352320B (en) |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105087415A (en) * | 2014-05-19 | 2015-11-25 | 段一皋 | Efficient and safe crop straw feed fermentation inoculant, solid additive and preparation method thereof |
| CN111690702A (en) * | 2020-06-23 | 2020-09-22 | 中国科学院过程工程研究所 | Method for degrading plant biomass by cooperation of hydrothermal pretreatment and high-temperature mixed enzyme |
| CN114591849A (en) * | 2022-05-10 | 2022-06-07 | 中国农业科学院北京畜牧兽医研究所 | Method for constructing myceliophthora thermophila strain with high homologous recombination efficiency and application thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101717727A (en) * | 2009-12-23 | 2010-06-02 | 贵州大学 | Myceliophthora thermophilia strain and application thereof in aspect of producing keratinase |
| CN101786911A (en) * | 2010-02-04 | 2010-07-28 | 北京微邦生物工程有限公司 | Microorganism and enzyme powder composite biological agent for microbial organic fertilizer |
-
2011
- 2011-09-29 CN CN2011102909513A patent/CN102352320B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101717727A (en) * | 2009-12-23 | 2010-06-02 | 贵州大学 | Myceliophthora thermophilia strain and application thereof in aspect of producing keratinase |
| CN101786911A (en) * | 2010-02-04 | 2010-07-28 | 北京微邦生物工程有限公司 | Microorganism and enzyme powder composite biological agent for microbial organic fertilizer |
Non-Patent Citations (4)
| Title |
|---|
| Enzymatic hydrolysis of pretreated cellulosic wastes by the cellulase complex of Myceliophthora thermophila D-14 to produce ethanol;Samir K.Roy等;《Fuel》;19910630;第70卷(第6期);第757-760页 * |
| Samir K.Roy等.Enzymatic hydrolysis of pretreated cellulosic wastes by the cellulase complex of Myceliophthora thermophila D-14 to produce ethanol.《Fuel》.1991,第70卷(第6期),第757-760页. |
| 中国毁丝霉属嗜热型真菌两新记录种;王冬梅等;《菌物研究》;20040630;第2卷(第2期);第47-49页 * |
| 王冬梅等.中国毁丝霉属嗜热型真菌两新记录种.《菌物研究》.2004,第2卷(第2期),第47-49页. |
Also Published As
| Publication number | Publication date |
|---|---|
| CN102352320A (en) | 2012-02-15 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103232272B (en) | A kind of compound organic microbial fertilizer and manufacture method thereof | |
| CN105567612B (en) | A kind of degradation composite bacteria agent preparation of garden waste and application | |
| CN103992184B (en) | A kind of biological organic fertilizer | |
| CN104212746B (en) | A kind of changing food waste of salt tolerant processes composite bacteria agent and its preparation method and application | |
| CN105176881B (en) | A method of producing the engineering microbial inoculum and its production active bio-organic fertilizer of active bio-organic fertilizer | |
| CN107857689A (en) | The technique that a kind of bioanalysis prepares bio-organic fertilizer using chicken manure | |
| CN107365718B (en) | Bacillus megaterium MYB3 and application thereof in straw fermented feed | |
| CN103045511B (en) | Cellulase production bacillus licheniformis with phosphate-solubilizing and potassium-releasing function and application of same | |
| CN101914445B (en) | Indigenous probiotic microorganism solid fungicide and preparation method and application thereof | |
| CN104293694A (en) | Preparation method for sludge aerobic composting composite inoculum | |
| CN101659934A (en) | Antagonistic bacteria preventing and removing continuous cropping banana Panama wilt disease and microbial organic fertilizer thereof | |
| CN103484421B (en) | A kind of chlamydosporic method of liquid fermenting scale up test Gliocladium roseum | |
| CN109402015A (en) | One plant of heat bites bacillus amyloliquefaciens and its application | |
| CN103896646A (en) | Preparation method of microorganism organic fertilizer | |
| CN101914443B (en) | Complex microbial inoculum for swine production by fermentation bed started quickly at low temperature and preparation method thereof | |
| CN105441360A (en) | Organic fertilizer fermentation inoculant | |
| CN103159570A (en) | Solid microbial fertilizer and preparation method thereof | |
| CN108203352A (en) | A kind of production method of kitchen garbage organic fertilizer | |
| CN105543149B (en) | One plant of new bacillus megaterium and its application | |
| CN102703364B (en) | Anoxybacillus mongoliensis UTM501 and applications thereof | |
| CN103387428A (en) | Preparation method for organic material decomposition agent | |
| CN101948780A (en) | Antagonist bacterium for preventing and treating continuous cropping hot pepper epidemic disease and microbial organic fertilizer thereof | |
| CN106916756B (en) | Fungus strain, microbial agent, straw soil remediation agent and application thereof | |
| CN108865927B (en) | Bacterial strain for low-temperature glycolysis of corn straw and fermentation culture method and application thereof | |
| CN102334589B (en) | Method for preparing fermented feed from waste bacillus fermentation liquor |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20170310 Address after: 102200 Zhongguancun science and Technology Park, Changping Hua Hua Road, No. 11, No. Patentee after: Beijing Jinggeng Tianxia Agricultural Technology Co., Ltd. Address before: 102200 Zhongguancun science and Technology Park, Changping Hua Hua Road, No. 11, No. Patentee before: Beijing Climax Biotechnology Co., Ltd. |
|
| TR01 | Transfer of patent right |