CN104560817A - Thermophilic bacillus licheniformis UTM102 for producing phytase and application of thermophilic bacillus licheniformis UTM102 - Google Patents
Thermophilic bacillus licheniformis UTM102 for producing phytase and application of thermophilic bacillus licheniformis UTM102 Download PDFInfo
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Classifications
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- 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
- C12R2001/10—Bacillus licheniformis
-
- 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
- C12N1/205—Bacterial isolates
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
- C05F11/00—Other organic fertilisers
- C05F11/08—Organic fertilisers containing added bacterial cultures, mycelia or the like
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- 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
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- 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
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- 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
- Y02W—CLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
- Y02W30/00—Technologies for solid waste management
- Y02W30/40—Bio-organic fraction processing; Production of fertilisers from the organic fraction of waste or refuse
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- Life Sciences & Earth Sciences (AREA)
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Abstract
The invention provides a thermophilic bacillus licheniformis UTM102 for producing phytase and an application of the thermophilic bacillus licheniformis UTM102. The biological collection number of the thermophilic bacillus licheniformis UTM102 is CGMCC (China General Microbiological Culture Collection Center) No.9508. The thermophilic bacillus licheniformis UTM102 can perform aerobic composting fermentation in organic solid wastes such as sewage sludge, household garbage, animal carcasses, livestock manure, crop straw and the like, can adapt to high-temperature environment and be multiplied massively in the organic solid wastes; a pile is high in heating speed and temperature, can be quickly composted, can degrade organic matter more quickly, minimization and harmlessness are realized more completely, and the organic solid wastes can be converted into biological fertilizers which contain massive bacillus licheniformis, can inhibit plant pathogens, can promote growth of plants and can improve the yield of crops. The bacillus licheniformis UTM102 has good strain performance and remarkable economic efficiency.
Description
Technical field
The present invention relates to fermentation arts, particularly, relate to a kind of carry phytase gene thermophilic lichem bacillus strain UTM102 and the organic solid castoff and prepare the purposes of bio-feritlizer of degrading.
Background technology
A large amount of organic solid castoffs can be produced in the process of exhausting and production.As produced a large amount of mud in town domestic sewage treating processes, it is containing a large amount of harmful levels of pathogens, and organic content enriches in addition, and easily corruption is smelly causes serious secondary pollution.And the waste in agricultural zootechnical production process, as tangerine bar, feces of livestock and poultry and carcase etc. have become serious source of pollution, become first pollution source in certain areas.
Organic solid castoff main composition in town and country is wood fibre metallic substance.The crystal-like structure that lignocellulose is made up of Mierocrystalline cellulose, hemicellulose and xylogen, utilizes During High-Temperature Composting aerobic fermentation to be the effective ways of efficiency utilization organic solid castoff with lignocellulose degradation.Utilize the synergy of multiple-microorganism, make the organic nutrient of various complexity, be converted into solubility, the nutrient easily absorbed and soil ulmin.The high temperature (80 ~ 100 DEG C) that simultaneously compost produces kills germ in material, worm's ovum.Prepare bio-feritlizer by fermentable organic solid castoff, in developing eco-agriculture, there is critical role, not only solve the reluctant problem of waste, change harmful to treasure simultaneously.To have phytase activity, and the Bacillus licheniformis with multiple resistant to elevated temperatures biomass by hydrolyzation enzyme applies to the process of organic solid waste material, and prepare growth-promoting sexual function bio-feritlizer and be not reported.
Summary of the invention
The object of the present invention is to provide a kind of thermophilic Bacillus licheniformis UTM102 bacterial strain and application thereof of phytase generating.
The present invention is from picking up from nature hot spring bed mud, utilize domestication substratum (glucose 10g, extractum carnis 3g, yeast extract 5g, peptone 10g, sodium-chlor 5g, distilled water 1000ml) at 40 DEG C, through many for acclimating, obtain UTM102 bacterial strain after directed screening, it has degraded organic solid castoff characteristic.This bacterial strain (is called for short CGMCC on August 13rd, 2014 at China Committee for Culture Collection of Microorganisms's common micro-organisms center, address: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, Institute of Microorganism, Academia Sinica, postcode 100101) preservation, Classification And Nomenclature is Bacillus licheniformis Bacillus licheniformis, and preserving number is CGMCC No.9508.
UTM102 provided by the invention is Gram-positive, cell 0.8 × (2.0 ~ 3.5), direct rod shape, Dan Sheng, the nearly middle raw oval gemma of generation, and sporangiocyst expands; (glucose 10g, peptone 10g, yeast extract paste 5g, sodium-chlor 5g, agar 15g, distilled water 1000ml on the substratum containing glucose proteins peptone, pH7.0) the smooth positive garden of bacterium colony, slightly prominent, eggshell is yellow, growth is rapid, cultivate colony diameter about 2 ~ 3mm after 24 hours, optimum growth temperature 40 ~ 60 DEG C for 30 ~ 40 DEG C.The catalase positive, oxidase positive, VP test the positive.
Primer is utilized to be (27f): 5 '-AGA GTT TGA TCC TGG CTC AG-3 ' and (1492r): 5 '-GGT TAC CTT GTT ACG ACT T-3 ' is through the 16S rRNA gene of this bacterial strain of pcr amplification, and the sequence obtained after order-checking is shown in sequence table SEQ ID No:1.Acquisition sequence is submitted to EzTaxon database compare, this gene of result display UTM102 bacterial strain is the highest with the similarity of Bacillus licheniformis, is 99.3%, and this sequence identifies the principal character foundation of this bacterial strain.
Utilize universal primer UP-1S:5 '-GAA GTC ATC ATG ACC GTT CTG CA-3 ' and UP-2Sr:5 '-AGC AGG GTA CGG ATG TGC GAG CC-3 ' through the gyrB gene of this bacterial strain of pcr amplification, the gene order after order-checking is shown in sequence table SEQ ID No:2.The sequence obtained is submitted to NCBI GeneBank database and uses Blast program to carry out sequence alignment, gyrB gene order and the Bacillus licheniformis similarity of result display UTM102 bacterial strain are the highest, and the physio-biochemical characteristics in conjunction with described bacterial strain are differentiate the accidental quality of this bacterial strain.Determine that this bacterial strain is Bacillus licheniformis Bacillus licheniformis.
The invention provides a kind of Bacillus licheniformis (Bacillus licheniformis) bacterial strain UTM102, its deposit number is CGMCC No.9508.
Bacillus licheniformis UTM102 provided by the invention has beta-glucan restriction endonuclease, cellobiase, zytase and phytase activity, and its encoding gene is shown in sequence table SEQ IDNo:3 ~ SEQ ID No:6.
The invention provides the microbial inoculum containing Bacillus licheniformis UTM102.
The invention provides Bacillus licheniformis UTM102 or the application of its microbial inoculum in the biodegradation process of organic solid castoff.
The invention provides a kind of method preparing compost Inoculant containing Bacillus licheniformis UTM102, comprise the following steps:
(1) fermented liquid is prepared: be inoculated in the liquid nutrient medium of glucose proteins peptone by lichem bacillus strain UTM102 activated spawn, carry out fermentative production;
(2) fermented liquid packing becomes liquid bacterial agent, obtains bacteria powder through dehydrating.
In aforesaid method, step (1) lawn that takes a morsel from the bacterial strain UTM102 inclined-plane of the present invention that 4 DEG C are preserved is seeded to the 250ml triangular flask of substratum (glucose 10g, peptone 10g, yeast extract paste 5g, sodium-chlor 5g, distilled water 1000ml, pH7.0) of 40ml containing glucose proteins peptone is housed.Culture temperature 35 ~ 40 DEG C, shaking speed 160-220 rev/min, cultivates 1 ~ 2 day, OD
600stop cultivating when being greater than 1.8, this is shake-flask seed liquid.
Shake-flask seed liquid is seeded in the seeding tank containing the substratum (glucose 10g, peptone 10g, yeast extract paste 5g, sodium-chlor 5g, distilled water 1000ml, pH7.0) of glucose proteins peptone and cultivates.Culture temperature 35 ~ 40 DEG C, air flow is 1:0.1 ~ 0.5 (v/v), and stirring velocity is 100 ~ 400 revs/min, incubation time 30 ~ 48 hours.Work as OD
600cultivate for being greater than 1.8 stoppings, this is seeding tank seed liquor.
By seeding tank seed liquor by 0.1 ~ 5% inoculum size, inoculation carries out fermentation culture containing in the fermentor tank of above-mentioned substratum.Culture temperature 35 ~ 40 DEG C, air flow is 1:0.2 ~ 0.6 (v/v), and stirring velocity is 60 ~ 200 revs/min, incubation time 20 ~ 40 hours.Work as OD
600for stopping cultivating when being greater than 1.8, this is the fermented liquid of bacterial strain.
The invention provides lichem bacillus strain UTM102 or the application of microbial inoculum in organic solid waste compost fermentation containing it.
Described organic solid castoff is one or more in downflow sludge, domestic refuse, crop material, carcase or feces of livestock and poultry.
By lichem bacillus strain UTM102 or the method for containing its microbial inoculum, organic solid castoff being carried out to compost, comprise the following steps:
(1) be inoculated in organic solid castoff by the UTM102 or its microbial inoculum that are equivalent to total material weight in wet base 0.1% ~ 0.5%, laggard row aerobic composting fermentation is mixed in mixing thoroughly;
(2) with moisture amendment, the moisture of material being adjusted to water ratio is 45% ~ 65%, and the carbon-nitrogen ratio of this material is 10 ~ 60:1, and heap body keeps oxygen content to be about 8% ~ 15%;
(3) pile body fermentation 12-20 days, fermentation stops; Leftover materials or landfill or burning or packing of sieving obtain bio-feritlizer.
In above-mentioned compost method, in the course of fermentation of step (2), the highest temperature reaches 103 ~ 105 DEG C; Step (3) is down to 30 ~ 35% through the aerobic fermentation material moisture of 12-20 days, and the most of organism in waste is decomposed, and stops fermentation.
The invention provides lichem bacillus strain UTM102 or prepare the application in organic fertilizer containing its microbial inoculum.
The invention provides lichem bacillus strain UTM102 or the application of microbial inoculum in Promoting plant growth containing it.
The present invention is separated to the bacterial strain that a strain has phytase, cellobiase, beta-glucan restriction endonuclease, zytase isoreactivity from hot spring bed mud, prepares bio-feritlizer provide and a kind ofly have farm crop growth-promoting effect concurrently and degraded organic solid castoff makes the microorganism of the innoxious characteristic of its minimizing for utilizing organic solid castoff.UTM102 contains phytase, phytic acid (salt) can be degraded to inositol and inorganic phosphorus by the phytase of its secretion, discharge in the energy metabolism and substance metabolism process that other material of combining with phytic acid (salt) participates in plant simultaneously, thus Promoting plant growth.UTM102 bacterial strain can carry out aerobic composting fermentation in the organic solid wastes such as downflow sludge, domestic refuse, carcase, feces of livestock and poultry, agricultural crop straw, the stable bacterium colony ecosystem is formed in organic materials, a large amount of Fast-propagation, the organism of effectively degrading in organic solid castoff, makes it reach minimizing, innoxious fast; Organic solid castoff can be changed into bio-feritlizer by the microbiobacterial agent of application prepared by the present invention simultaneously, and a large amount of Bacillus licheniformis contained in this fertilizer can Suppressing phytopathogens, can play promoter action, can improve the output of crop to plant-growth.
Accompanying drawing explanation
Fig. 1 is bacterial strain UTM102 systematic evolution tree of the present invention.
Fig. 2 is the microphotograph of bacterial strain UTM102 of the present invention.
Embodiment
Following examples for illustration of the present invention, but are not used for limiting the scope of the invention.Without departing from the spirit and substance of the case in the present invention, the amendment do the inventive method, step or condition or replacement, all belong to scope of the present invention.
If do not specialize, the conventional means that technique means used in embodiment is well known to those skilled in the art.
The Isolation and ldentification of embodiment 1 Bacillus licheniformis UTM102
Take 1 gram of hot spring bed mud sample to be placed in the 250ml triangular flask that many little granulated glass spherees and 50ml sterilized water are housed, 40 DEG C of constant-temperature tables shake 1 hour, leave standstill 30 minutes.Aseptically Aspirate supernatant 1ml, access is equipped with in the 250ml triangular flask of the sterilized domestication substratum of 50ml (glucose 10g, extractum carnis 3g, yeast extract 5g, peptone 10g, sodium-chlor 5g, 1000ml distilled water), cultivates 3 days (rotating speed 160 revs/min) for 40 DEG C.With same method, again draw 1ml pregnant solution, Secondary Culture 3 generation.
Get the bacteria suspension 1ml after cultivation 3 generation, join in 9ml sterilized water, be mixed with 10 by 10 times of dilution methods
-1, 10
-2, 10
-3, 10
-4, 10
-5, 10
-6, 10
-7extent of dilution.Each extent of dilution is respectively got 0.1ml and is coated on the flat board of 3 qualification substratum containing phytic acid ca.The sterilized water getting 0.1ml operates by the same method, compares.Cultivate 3 days for 40 DEG C.Select bacterium colony dispersion dull and stereotyped preferably, from single bacterium colony that upper picking degraded circle is large.And on qualification substratum, line separation and purification 3 times repeatedly, be seeded in (glucose 10g, peptone 10g, yeast extract paste 5g, sodium-chlor 5g, agar 15g, distilled water 1000ml on the substratum containing proof agar, pH7.0) slant culture is to abundant, preserve at 4 DEG C, for subsequent use.
Above-mentionedly identify that the formula of substratum is: phytic acid ca 0.1% ~ 0.5%, glucose 3 ~ 5%, NH
4nO
30.5 ~ 0.8%, MnSO
44H
2o 0.002 ~ 0.005%, FeSO
47H
2o0.003 ~ 0.005%, MgSO
47H
2o 0.03 ~ 0.06%, KC1 0.03 ~ 0.07%, 0.04% tetrabromo-mcresolsulfonphthalein solution 1.5 ~ 2.0% (v/v), agar 1.5 ~ 2%, pH5.0 ~ 6.0.
With the DNA of bacterial strain of the present invention for template, its 16S rRNA gene order of pcr amplification, primer is (27f): 5 '-AGA GTT TGA TCC TGG CTC AG-3 ' and (1492r): 5 '-GGT TAC CTT GTT ACG ACT T-3 '.PCR response procedures is: 95 DEG C of denaturations 5 minutes, 95 DEG C of sex change 30 seconds, 53 DEG C of annealing 45 seconds, and 72 DEG C extend 90 seconds, 30 circulations, 72 DEG C of extensions 10 minutes.Amplified production checks order after its purity of electrophoresis detection, and sequencing result is shown in shown in sequence table SEQ ID No:1.The 16S rRNA gene order obtained is compared by EzTaxon database (http://www.ezbiocloud.net/), and the highest with the similarity of bacterial strain Bacilluslicheniformis, homology is 99.3%.Fig. 1 is based on UTM102 bacterial strain and close bacterial strain 16S rRNA gene order thereof, utilizes the systematic evolution tree (maximum likelihood method) of Mega5.0 phyletic evolution software building, illustrate its Phylogenetic and Bacillus licheniformis nearest.
Utilize universal primer UP-1S:5 '-GAA GTC ATC ATG ACC GTT CTG CA-3 ' and UP-2Sr:5 '-AGC AGG GTA CGG ATG TGC GAG CC-3 ' through the gyrB gene of this bacterial strain of pcr amplification, sequencing result is shown in that sequence that sequence table SEQ ID No:2 obtains is submitted to NCBI GeneBank database and uses Blast program to carry out sequence alignment, and gyrB gene order and the Bacillus licheniformis similarity of result display UTM102 bacterial strain are the highest.
In conjunction with strain morphology feature (Fig. 2) and physiological and biochemical property, be Bacillus licheniformis this identification of strains, called after UTM102, and on August 13rd, 2014 in (address: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, China Committee for Culture Collection of Microorganisms's common micro-organisms center, Institute of Microorganism, Academia Sinica, postcode 100101) preservation, Classification And Nomenclature is Bacillus licheniformis Bacillus licheniformis, and preserving number is CGMCC No.9508.
Embodiment 2 bacterial strain UTM102 cellulase activity detects and related gene sequence
By the dibbling of bacillus UTM102 dibbling method at Xylo-Mucine substratum (Xylo-Mucine 5g, KH
2pO
4lg, agar 17g, NaNO
33g, KCL 0.5g, MgSO
40.5g, FeSO
40.01g, distilled water 1000ml, pH 5.5 ~ 6.0) on, cultivate 48 hours for 40 DEG C.Adopt 0.2% congo red staining 30 minutes, wash away dye liquor with distilled water, then soak 1 hour with the NaC1 that concentration is 1mol/L, finally use the acetate solution constant color of 5%.Form water white transparency circle in periphery of bacterial colonies and show this bacterium eccrine fiber element enzyme.Cellulase is multienzyme mixture, and it is made up of cellobiase, beta-glucan restriction endonuclease etc.
According to beta-glucan incision enzyme gene design primer betaglu-f:5 '-CGA TGT TGTTCA TGC CGG CT-3 ' and betaglu-r:5 '-TTG CCA GCG TGT GTG ACAGC-3 ', with bacterial strain UTM102 DNA of the present invention for template carries out PCR reaction, PCR reaction conditions is 95 DEG C of denaturations 5 minutes, 95 DEG C of sex change 30 seconds, anneal 45 seconds for 55 DEG C, 72 DEG C extend 90 seconds, 30 circulations, 72 DEG C extend 10 minutes, obtain the fragment of about 2.0kb, order-checking also will use Blast program to carry out sequence alignment at NCBI GeneBank database, this gene segment encodes beta-glucan restriction endonuclease, its gene order is shown in sequence table SEQ ID No:3.
According to degenerated primer Cellobiase-f:5 '-GAA GGCATT CCT TAT CAT TC-3 ' and the Cellobiase-r:5 '-ACG GTC ATA CTC AGCGTA AG-3 ' of cellobiase genes design, and with bacterial strain UTM102 DNA of the present invention for template, carry out pcr amplification reaction, PCR response procedures is: 95 DEG C of denaturations 5 minutes, 95 DEG C of sex change 30 seconds, anneal 45 seconds for 53 DEG C, 72 DEG C extend 90 seconds, 35 circulations, 72 DEG C extend 10 minutes, the nucleotide fragments of about 2kb size is obtained by electrophoresis detection, order-checking also will use Blast program to carry out sequence alignment at NCBI GenBank database, this sequence fragment encoding fiber disaccharidase gene, its gene order is shown in sequence table SEQ ID No:4.
Embodiment 3 bacterial strain UTM102 xylanase activity detects and gene order
Accurately take beech xylan (Sigma) (being accurate to 0.001 gram), the beech xylan solution (best matching while using, glycan easily decomposes in acid condition) of 1% is made into 50mmol/LNaAc-HAc (pH 5.0) damping fluid; Get the supernatant liquor (fermention medium: glucose 5g, peptone 15g, yeast extract paste 5g, sodium-chlor 5g, distilled water 1000ml of 1ml UTM102 culture in addition, pH7.0), become the enzyme liquid of proper concn with 50mmol/L NaAc-HAc (pH 5.0) buffer, ensure that photoabsorption is between 0.2 ~ 0.6.Then get 0.9ml 1% beech xylan solution, add the enzyme liquid that 0.1ml UTM102 prepares, be incubated 30 minutes in 55 DEG C of water-baths after, add 1ml distilled water and 2ml DNS reagent.Boiling water bath 5 minutes after mixing, be cooled to room temperature, adding distil water constant volume is to 25ml.Mix rear spectrophotometer and measure its OD value at wavelength 540nm place.The enzyme liquid simultaneously using the supernatant liquor of inactivator (100 DEG C are boiled 10 minutes) to prepare makes blank.UTM102 bacterial strain can be hydrolyzed beech xylan after measured, illustrates that it has the activity of zytase.
According to degenerated primer Xylanase-f:5 '-GCG CTG ACCTAT AAC G-3 ' and the Xylanase-r:5 '-CGC TCA CAG TGG ATT C-3 ' of xylanase gene design, and with bacterial strain UTM102DNA of the present invention for template, carry out pcr amplification reaction, PCR response procedures is: 95 DEG C of denaturations 5 minutes, 95 DEG C of sex change 30 seconds, anneal 45 seconds for 56 DEG C, 72 DEG C extend 90 seconds, 35 circulations, 72 DEG C extend 10 minutes, the nucleotide fragments of about 1.3kb size is obtained by electrophoresis detection, order-checking also will use Blast program to carry out sequence alignment at NCBI GeneBank database, obtain this sequence fragment encoding xylanase gene, its gene order is shown in sequence table SEQ ID No:5.
Embodiment 4 bacterial strain UTM102 phytase activity detects and gene order
By the streak inoculation of UTM102 bacterial strain on Phytate Ca medium solid plate, filling a prescription is: phytic acid ca 0.5%, glucose 5%, NH
4nO
30.8%, MnSO
44H
2o 0.005%, FeSO
47H
2o 0.005%, MgSO
47H
2o 0.06%, KC1 0.07%, 0.04% tetrabromo-mcresolsulfonphthalein solution 2.0% (v/v), agar 1.5 ~ 2%, pH5.0 ~ 6.0.Phytic acid ca degraded is produced transparent hydrolysis circle by the phytase that UTM102 produces.
According to degenerated primer Phytase-f:5 '-CGC AGC ATCCTT ATG G-3 ' and the Phytase-r:5 '-TCT GAT TGG CTG GTT G-3 ' of phytase gene design, and with bacterial strain UTM102DNA of the present invention for template, carry out pcr amplification reaction, PCR response procedures is: 95 DEG C of denaturations 5 minutes, 95 DEG C of sex change 30 seconds, anneal 45 seconds for 55 DEG C, 72 DEG C extend 90 seconds, 30 circulations, 72 DEG C extend 10 minutes, the nucleotide fragments of about 1kb size is obtained by electrophoresis detection, order-checking also will use Blast program to carry out sequence alignment at NCBI GenBank database, obtain this sequence fragment coding phytase gene, its gene order is shown in sequence table SEQ ID No:6.
The preparation of embodiment 5 bacterial strain UTM102 liquid bacterial agent and bacteria powder
The lawn that takes a morsel from 4 DEG C of bacterial strain UTM102 inclined-planes of the present invention preserved is seeded to and the 250ml triangular flask of 40ml containing the substratum (glucose 10g, peptone 10g, yeast extract paste 5g, sodium-chlor 5g, distilled water 1000ml, pH7.0) of glucose proteins peptone is housed.Culture temperature 37 DEG C, shaking speed 180 revs/min, cultivates 2 days, OD
600stop cultivating when being greater than 1.8, this is shake-flask seed liquid.
Shake-flask seed liquid is seeded in the seeding tank containing the substratum (glucose 10g, peptone 10g, yeast extract paste 5g, sodium-chlor 5g, distilled water 1000ml, pH7.0) of glucose proteins peptone and cultivates.Culture temperature 37 DEG C, air flow is 1:0.3 (v/v), and stirring velocity is 200 revs/min, incubation time 38 hours.Work as OD
600cultivate for being greater than 1.8 stoppings, this is seeding tank seed liquor.
By seeding tank seed liquor by 0.1 ~ 5% inoculum size, inoculation carries out fermentation culture containing in the fermentor tank of above-mentioned substratum.Culture temperature 37 DEG C, air flow is 1:0.4 (v/v), and stirring velocity is 100 revs/min, incubation time 30 hours.Work as OD
600for stopping cultivating when being greater than 1.8, this is the fermented liquid of bacterial strain.
Fermented liquid packing becomes liquid bacterial agent, and fermentation liquor dehydrates and obtains bacteria powder.
The microbial inoculum compost fermentation process domestic sludge that embodiment 6 utilizes bacterial strain UTM102 to prepare
By domestic sludge: moisture amendment (1: 0.8) preparation material (scale of construction ratio), if do not add and add obtained UTM102 microbial inoculum 2 experimental group of 1% embodiment 5.Compost starting condition is moisture 63.1%, C/N value is 35:1, adopts aerobic composting fermentation method.The compost fermentation time is 20 days, and every day period is in heap body difference meter record temperature.Compost at 0 day, 5 days, 10 days, 15 days to fall the mode turning of groove sampling and measuring sample water ratio.Both temperature and change of soil water content the results are shown in Table 1.
Table 1 compost temperature and change of soil water content situation
Add bacterium liquid group moisture lower than 40% during compost fermentation 15 days, reached national Sludge landfill standard, and do not add bacterium liquid group and take 20 days and just can reach standard of landfill; The former loss of weight about 60%, the latter's loss of weight about 50%.
Embodiment 7 utilizes UTM102 microbial inoculum compost fermentation to prepare bio-feritlizer
With the maize straw after feces of livestock and poultry and pulverizing for main material, the UTM102 microbial inoculum pulvis that 400kg swine excrement and 600kg maize straw and 1kg embodiment 5 obtain mixes to be mixed thoroughly, regulate moisture to be about 55% to water ratio, be placed in fermenter and carry out aerobic composting fermentation.Simultaneously not add the test of microbial inoculum of the present invention for contrast.Period every 3 ~ 5 days with the mode of falling groove turning once.When the moisture content of material is lower than 30%, and stop fermentation when heap temperature no longer rises.This material is through the bio-feritlizer that sieves to obtain.To carbon-nitrogen ratio and the germination index analysis in table 2 of prepared bio-feritlizer.
The impact of UTM102 microbial inoculum on compost maturity inoculated by table 2
The above is only the preferred embodiment of the present invention.It should be pointed out that for those skilled in the art, under the prerequisite not departing from the technology of the present invention principle, can also make some improvements and modifications, these improvements and modifications also should be considered as protection scope of the present invention.
Claims (10)
1. Bacillus licheniformis (Bacillus licheniformis) bacterial strain UTM102, its deposit number is CGMCC No.9508.
2. lichem bacillus strain UTM102 as claimed in claim 1, it is characterized in that, it can produce beta-glucan restriction endonuclease, cellobiase, zytase, phytase.
3. lichem bacillus strain UTM102 as claimed in claim 2, is characterized in that, described beta-glucan restriction endonuclease coding nucleotide sequence contains nucleotide sequence described in SEQ ID NO:3 or its complementary nucleotide sequence; Cellobiase coding nucleotide sequence contains nucleotide sequence described in SEQ ID NO:4 or its complementary nucleotide sequence; Zytase coding nucleotide sequence contains nucleotide sequence described in SEQ ID NO:5 or its complementary nucleotide sequence; Phytase coding nucleotide sequence contains nucleotide sequence described in SEQ ID NO:6 or its complementary nucleotide sequence.
4. the microbial inoculum containing lichem bacillus strain UTM102 described in claim 1.
5. prepare a method for the compost Inoculant containing Bacillus licheniformis UTM102, it is characterized in that, comprise the following steps:
(1) fermented liquid is prepared: be inoculated in the liquid nutrient medium of glucose proteins peptone by lichem bacillus strain UTM102 activated spawn, carry out fermentative production;
(2) fermented liquid packing becomes liquid bacterial agent or obtains pulvis through dehydrating.
6. the arbitrary described lichem bacillus strain UTM102 of claim 1-3 or the application of microbial inoculum according to claim 4 in organic solid waste compost fermentation.
7. apply as claimed in claim 6, it is characterized in that, described organic solid castoff is one or more in downflow sludge, domestic refuse, crop material, carcase or feces of livestock and poultry.
8. apply as claimed in claim 6, it is characterized in that, comprise the following steps:
(1) be inoculated in organic solid castoff by the UTM102 or its microbial inoculum that are equivalent to total material weight in wet base 0.1% ~ 0.5%, laggard row aerobic composting fermentation is mixed in mixing thoroughly;
(2) with moisture amendment, the moisture of material being adjusted to water ratio is 45% ~ 65%, and the carbon-nitrogen ratio of this material is 10 ~ 60:1, and heap body keeps oxygen content to be about 8% ~ 15%;
(3) pile body fermentation 12-20 days, fermentation stops; Leftover materials or landfill or burning or packing of sieving obtain bio-feritlizer.
9. the application in organic fertilizer prepared by the arbitrary described lichem bacillus strain UTM102 of claim 1-3 or microbial inoculum according to claim 4.
10. the arbitrary described lichem bacillus strain UTM102 of claim 1-3 or the application of microbial inoculum according to claim 4 in Promoting plant growth.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105695354A (en) * | 2016-02-19 | 2016-06-22 | 大地绿源环保科技(北京)有限公司 | Process and application for treating municipal sewage sludge by means of ultrahigh-temperature aerobic composting fermentation |
| CN110951649A (en) * | 2019-12-26 | 2020-04-03 | 江苏大道生物环境科技有限公司 | Bacillus for treating industrial excess sludge and application method thereof |
| CN112175875A (en) * | 2020-10-13 | 2021-01-05 | 蛋壳城矿环保科技发展(广州)有限公司 | Preparation method and application of ultrahigh-temperature aerobic composite biological agent |
| CN114717125A (en) * | 2021-01-04 | 2022-07-08 | 山东农业大学 | Thermophilic bacillus licheniformis AMCC101380 and application thereof in high-temperature composting of tailstocks |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101892182A (en) * | 2010-06-07 | 2010-11-24 | 中国农业大学 | A strain of Bacillus licheniformis and its application in promoting cellulose degradation |
-
2014
- 2014-12-30 CN CN201410842678.4A patent/CN104560817B/en not_active Expired - Fee Related
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101892182A (en) * | 2010-06-07 | 2010-11-24 | 中国农业大学 | A strain of Bacillus licheniformis and its application in promoting cellulose degradation |
Non-Patent Citations (2)
| Title |
|---|
| 李秋鹏等: "一株嗜热地衣芽孢杆菌及其产高温淀粉酶的初步研究", 《工业微生物》 * |
| 王菊等: "地衣芽孢杆菌的研究进展", 《中国饲料》 * |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105695354A (en) * | 2016-02-19 | 2016-06-22 | 大地绿源环保科技(北京)有限公司 | Process and application for treating municipal sewage sludge by means of ultrahigh-temperature aerobic composting fermentation |
| CN105695354B (en) * | 2016-02-19 | 2019-09-13 | 大地绿源环保科技(北京)有限公司 | The technique and application of superhigh temperature aerobic composting fermentation processing municipal sludge |
| CN110951649A (en) * | 2019-12-26 | 2020-04-03 | 江苏大道生物环境科技有限公司 | Bacillus for treating industrial excess sludge and application method thereof |
| CN110951649B (en) * | 2019-12-26 | 2021-03-26 | 江苏大道生物环境科技有限公司 | Bacillus for treating industrial excess sludge and application method thereof |
| CN112175875A (en) * | 2020-10-13 | 2021-01-05 | 蛋壳城矿环保科技发展(广州)有限公司 | Preparation method and application of ultrahigh-temperature aerobic composite biological agent |
| CN114717125A (en) * | 2021-01-04 | 2022-07-08 | 山东农业大学 | Thermophilic bacillus licheniformis AMCC101380 and application thereof in high-temperature composting of tailstocks |
| CN114717125B (en) * | 2021-01-04 | 2023-06-20 | 山东农业大学 | A thermophilic Bacillus licheniformis AMCC101380 and its application in high-temperature composting of tail vegetables |
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