CN109251879A - A kind of Jie meter La series bacillus fermentation process in high density - Google Patents
A kind of Jie meter La series bacillus fermentation process in high density Download PDFInfo
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- CN109251879A CN109251879A CN201811401672.8A CN201811401672A CN109251879A CN 109251879 A CN109251879 A CN 109251879A CN 201811401672 A CN201811401672 A CN 201811401672A CN 109251879 A CN109251879 A CN 109251879A
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- 238000000855 fermentation Methods 0.000 title claims abstract description 82
- 230000004151 fermentation Effects 0.000 title claims abstract description 82
- 241000193830 Bacillus <bacterium> Species 0.000 title claims abstract description 64
- 241000726221 Gemma Species 0.000 claims abstract description 29
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims abstract description 20
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 claims abstract description 18
- 239000007788 liquid Substances 0.000 claims abstract description 18
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- SEGLCEQVOFDUPX-UHFFFAOYSA-N di-(2-ethylhexyl)phosphoric acid Chemical compound CCCCC(CC)COP(O)(=O)OCC(CC)CCCC SEGLCEQVOFDUPX-UHFFFAOYSA-N 0.000 claims 1
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- 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
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Abstract
The invention discloses a kind of Jie meter La series bacillus fermentation process in high density.Fermentation medium of the present invention includes following components: corn flour, bean cake powder, wheat bran, sucrose, calcium carbonate, potassium dihydrogen phosphate and sodium chloride.After the present invention activates Jie meter La series bacillus, through level-one, secondary seed solution preparation and fermented and cultured, when 90% or more thallus is converted into gemma in fermentation liquid, stops fermentation, use two steps fermentative technology in fermentation process, first step purpose is proliferated thallus largely in a short time, second step builds the environment for being unsuitable for thalli growth, promotes transformation of the viable bacteria to gemma, improves gemma production rate, under the conditions of 50L fermentor scale, living bacteria count is up to 7.5 × 109Cfu/mL or more produces gemma rate up to 92.6% or more.Zymotechnique of the present invention is simple, and raw material is cheap and easy to get, at low cost, is suitble to industrialization production.
Description
Technical field
The invention belongs to field of microbial fermentation, and in particular to a kind of Jie meter La series bacillus fermentation process in high density.
Background technique
Plant pest is always the formidable enemy of agricultural production, drastically influences the growth of crops, restricts holding for agricultural
Supervention exhibition causes huge economic loss, therefore the research about control of plant disease is increasingly taken seriously.China at present
Mainly based on chemical pesticide control, long-term a large amount of applications all cause to seriously endanger to ecological environment and our mankind.With life
Object controlling plant diseases research is goed deep into, and this green, environmentally friendly, person poultry harmless can reduce using for chemical pesticide
Control method obtains social more and more concerns and approves, becomes the hot spot of Recent study control of plant disease, wherein short of money
Antibacterium plays an important role in biological control.
Most study is bacillus in antagonistic bacterium at present, and bacillus is many kinds of with its, environmental-friendly, easy
Many advantages, such as being colonized in plant rhizosphere surface, a variety of antibacterial substances such as lipopeptide antibiotic, antibacterial protein generated
Or polypeptide compounds all show very strong bacteriostatic activity, and bacillus endogenous spore to most plants disease fungus, resist
Inverse property is strong, and reproduction speed is fast, is conducive to industrialized production.It is had broad application prospects using microbial control plant disease.
Jie meter La series bacillus (Paenibacillus jamilae) is one kind of bacillus, Jie meter La class bud
Spore bacillus can secrete antibacterial protein, antibiotic, enzyme or polypeptide isoreactivity substance, have well to plant pathogenetic bacteria, fungi
Inhibiting effect significantly improves the disease resistance of plant, while can promote plant growth, improves plant products, is a kind of good life
Anti- Promoting bacteria.Present invention applicant separates from halmeic deposit obtains one plant of Jie meter La series bacillus (Paenibacillus
Jamilae) ZDC-04, deposit number are CGMCC No.16231.Tests prove that the bacterial strain is to the fruit as caused by colletotrichum gloeosporioides Penz
Tree (citrus, apple, grape) anthracnose and the vegetables wilt disease as caused by Fusarium oxysporum all have good control efficiency.
The microbial bacterial agent overwhelming majority is acted on viable bacteria, and effective viable count is its important quality index, must in microbial inoculum
Effect must be can be only achieved containing a certain amount of viable count.There is specific living bacteria count in China to the microbial inoculum of various approval productions
The regulation of amount and dosage.Viable count and gemma yield are to measure the important indicator of Jie meter La series bacillus microbial inoculum, but current state
The inside and outside research seldom to the optimization of biological and ecological methods to prevent plant disease, pests, and erosion Jie meter La series bacillus high density fermentation, and it is most in high cell density fermentation optimization
Viable count is only improved, and sporulation number is not high, the low activity and unstable quality that will lead to microbial inoculum of gemma number, the holding time
It is greatly reduced, is not suitable for actual use diseases prevention.Therefore, developing one kind not only has the function of biological and ecological methods to prevent plant disease, pests, and erosion, but also the microbial inoculum product of high gemma number
There is very big meaning for actual application.
Summary of the invention
In view of the above-mentioned problems, the present invention provides a kind of Jie meter La series bacillus fermentation process in high density.The present invention makes
It uses agricultural and sideline product cheap and easy to get to produce Jie meter La series bacillus as carbon and nitrogen sources liquid fermentation, reduces its production cost,
The viable count and gemma yield in unit volume fermentation liquid are improved, is the industrialized production and popularization of Jie meter La series bacillus
Reference is provided.
Present invention firstly provides a kind of fermentation mediums of Jie meter La series bacillus, including following components: corn flour,
Bean cake powder, wheat bran, sucrose, calcium carbonate, potassium dihydrogen phosphate and sodium chloride.Jie meter La series bacillus used in the present invention is outstanding person
Rice draws series bacillus (Paenibacillus jamilae) ZDC-04.
Preferably, fermentation medium of the invention composition are as follows: corn flour 12-30g/L, bean cake powder 5-17g/L, wheat bran 5-
14g/L, sucrose 6-12g/L, calcium carbonate 1-5g/L, potassium dihydrogen phosphate 1-6g/L, sodium chloride 0-5g/L, initial pH value 6.4-
7.4。
It is furthermore preferred that fermentation medium of the invention forms are as follows: corn flour 15g/L, bean cake powder 10g/L, wheat bran 8g/L, sugarcane
Sugared 6g/L, calcium carbonate 5g/L, potassium dihydrogen phosphate 5g/L, sodium chloride 2g/L, initial pH value 6.4-7.4.
The bean cake powder is smashed dregs of beans, and the wheat bran is the wheat bran after wheat crushing, the sodium chloride
For industrial chlorinations sodium, wherein sucrose is level-one white granulated sugar.
The present invention provides application of the above-mentioned fermentation medium in fermentation Jie meter La series bacillus.
The present invention also provides a kind of fermentation process in high density of Jie meter La series bacillus, characterized in that by Jie meter La
After series bacillus activation, through level-one, secondary seed solution preparation and fermented and cultured, when 90% or more thallus is converted into fermentation liquid
When gemma, stop fermentation；
Specifically includes the following steps:
(1) prepared by shaking flask primary seed solution: the Jie meter La series bacillus after activation is transferred to shake-flask seed culture medium,
Temperature is 28-37 DEG C, shaking speed cultivates 16-30h under being 160-200r/min；
The shake-flask seed culture medium composition are as follows: 10 ± 2g/L of peptone, 10 ± 2g/L of yeast extract, sucrose 30 ±
5g/L, pH6.5 ± 0.1.
(2) prepared by seeding tank secondary seed solution: cultured shake-flask seed liquid is transferred by inoculum concentration 1-5% (volume ratio)
Enter in secondary seed medium, cultivates 14-18h at 30 ± 2 DEG C of temperature；
Secondary seed medium composition are as follows: 20 ± 5g/L of corn flour, 15 ± 3g/L of bean cake powder, 30 ± 5g/L of sucrose,
Potassium dihydrogen phosphate 5 ± 1g/L, pH 6.5 ± 0.1.
(3) seed liquor obtained in step (2) is trained by the inoculum concentration access of 3-10% (volume ratio) equipped with above-mentioned fermentation
In the fermentor for supporting base, in 30 ± 2 DEG C of fermented and cultureds, when 90% or more thallus is converted into gemma in fermentation liquid, stop fermentation.
Preferably, speed of agitator 300-400r/min, the liquid amount 50-70% of the step (2), ventilatory capacity 0.4-1m3/
H, tank press 0.03-0.05Mpa.
It is described, when step (3) fermented and cultured, 0-10h, fermentation condition are as follows: speed of agitator 300-400r/min, logical
Tolerance 2-4m3/ h, tank press 0.03-0.05Mpa；Fermentation culture conditions after 11h are as follows: speed of agitator 200-300r/min, lead to
Tolerance 1-2.5m3/ h, tank press 0.03-0.05Mpa.
Further, the activation Jie meter La series bacillus strain method: the Jie meter La class gemma that sandy soil pipe is saved
Bacillus streak inoculation is cultivated on LB culture medium flat plate, and 16-48h is cultivated at 28-37 DEG C, and then picking single colonie scribing line turns
It is connected to LB slant medium, obtains activated spawn after 16-48h is cultivated at 28-37 DEG C；The LB plating medium and the inclined-plane LB is trained
Support the composition of base are as follows: peptone 100g/L, sodium chloride 100g/L, yeast extract 50g/L, agar 15-20g/L, pH 6.5 ±
0.1。
The present invention in Jie meter La series bacillus high cell density fermentation, according to shaking flask first order seed, secondary seed and
The difference of ferment tank situation has prepared different culture mediums respectively to improve seed vitality and promote high density fermentation.In addition,
Two steps fermentative technology is used during fermented and cultured, first step purpose is proliferated thallus largely in a short time, and second
Step, which is built, is unsuitable for the environment of thalli growth, promotes transformation of the viable bacteria to gemma, improves gemma production rate, thus make viable count with
Gemma yield can be improved.
Compared with prior art, the invention has the following advantages:
1) present invention has studied the high cell density fermentation of Jie meter La series bacillus for the first time, is Jie meter La series bacillus
Industrialized production and promote provide reference；
2) raw material of fermentation medium of the present invention is mainly from agricultural and sideline product corn flour cheap and easy to get, bean cake powder, wheat bran,
It is at low cost, it is easily enlarged production；
3) zymotechnique of the present invention is simple, and fermentation period is short (15h), and living bacteria count and gemma yield are high, ferments in 50L
Under the conditions of tank scale, living bacteria count is up to 7.5 × 109Cfu/mL or more produces gemma rate up to 92.6% or more.
Specific embodiment
The separation and screening of 1: Jie meter La series bacillus ZDC-04 of embodiment
(1) separation method of Jie meter La series bacillus ZDC-04
Jie meter La series bacillus ZDC-04 of the invention is located away from Floor of The East China Sea deposit (120.6 ° of E, 34 ° of N), the depth of water
It 17 meters, is separated and is obtained using method of dilution butteron on plate, method particularly includes: it weighs 10g bottom sediment sample and is dissolved in 90mL sterile water,
It is placed in shaking table and vibrates 30min, be sufficiently mixed sample with water, that is, obtain 10-1Dilution.Take the above-mentioned dilution of 1ml to 9ml without
In bacterium water, shakes up and obtain 10-2Dilution.Identical method obtains 10-3、10-4、10-5、10-6Dilution.10 are taken respectively-4、
10-5、10-6100 μ L of dilution in good LB plate, dilution is coated with uniformly with spreader, each concentration repeats 3
It is secondary.It is placed in 30 DEG C of insulating box and cultivates 48h, the different single colonie of picking cultural characteristic carries out plate scribing line purifying, and respectively
Number saves.
(2) screening of colletotrichum gloeosporioides Penz and Fusarium oxysporum antagonistic Bacillus
1. primary dcreening operation: using face-off flat band method, using colletotrichum gloeosporioides Penz and Fusarium oxysporum as target, the scribing line of PDA plate center
Inoculated bacteria, the both ends away from bacterium 2.5cm are inoculated with pathogen bacteria cake, 25 DEG C of constant temperature incubations, and control is only inoculated with pathogen bacteria cake.Often
A processing sets 3 repetitions, when covering with 3/4 culture dish wait compare, measures antibacterial bandwidth, filtering out has obviously above-mentioned pathogen
The bacterium bacterial strain of antagonism.
2. secondary screening: the bacterium bacterial strain after primary dcreening operation prepares seed liquor, and 2mL seed liquor is taken to be inoculated in 100mL LB culture medium,
180r/min, 30 DEG C of constant-temperature shaking culture 48h.Using mycelial growth rate method, by fermentation liquid and after melting, cooling PDA is cultivated
(45~50 DEG C) of base mixings, inverted plate, the PDA culture medium of isometric LB liquid medium is added as compareing, in plate
The pathogen bacteria cake that centre inoculation diameter is 5mm.Each processing sets 3 repetitions, when covering with 3/4 culture dish wait compare, right-angled intersection
Method measures control group and processing group colony diameter, calculates bacteriostasis rate, filters out to colletotrichum gloeosporioides Penz and Fusarium oxysporum antagonism effect
The optimal bacterium bacterial strain of fruit.
Inhibiting rate (%)=(control colony radius (mm)-processing colony radius (mm))/control colony radius (mm) ×
100
(3) result and analysis: utilizing dilution-plate method isolated 20 plants of bacteriums from bottom sediment, and stand facing each other flat band method
7 plants of antagonistic strains for having obvious inhibitory effect to above-mentioned pathogen are filtered out, account for the 35% of separated bacterial strain, wherein bacterial strain ZDC-
04 antagonistic effect is best, and the antibacterial bandwidth to colletotrichum gloeosporioides Penz and Fusarium oxysporum is respectively 17.00mm and 15.50mm.Choosing
The fermentation liquid of the bacterial strain is taken to dilute 10 times of progress mycelial growth rate method tests in PDA culture medium, the results showed that the bacterial strain
The inhibiting rate that fermentation liquid grows colletotrichum gloeosporioides Penz and Fusarium oxysporum mycelia is respectively 100% and 96.5%.
The present invention is also proved by field trial: the fermentation liquid of Jie meter La series bacillus ZDC-04 is to citrus anthracnose
The preventive effect of blade and fruit is respectively 81.53% and 81.82%.
The identification of 2: Jie meter La series bacillus ZDC-04 of embodiment
(1) strain morphology: thallus elongated rod shape, size are (0.5~1.1) μ m (3.2~5.2) μm, produce gemma, gram
Stained positive.30 DEG C of culture 48h on LB culture medium, bacterium colony is white, round, edge protuberance.
(2) Molecular Identification result
The 16S rRNA result of gene sequence determination (SEQ-1) of the bacterial strain is as follows:
AAGCTTGCTTCTAATTAACCTAGCGGCGGACGGGTGAGTAACACGTAGGCAACCTGCCCACAAGACAGG
GATAACTACCGGAAACGGTAGCTAATACCCGATACATCCTTTTCCTGCATGGGAGAAGGAGGAAAGGCGGAGCAATC
TGTCACTTGTGGATGGGCCTGCGGCGCATTAGCTAGTTGGTGGGGTAAAGGCCTACCAAGGCGACGATGCGTAGCCG
ACCTGAGAGGGTGATCGGCCACACTGGGACTGAGACACGGCCCAGACTCCTACGGGAGGCAGCAGTAGGGAATCTTC
CGCAATGGGCGAAAGCCTGACGGAGCAACGCCGCGTGAGTGATGAAGGTTTTCGGATCGTAAAGCTCTGTTGCCAGG
GAAGAACGCTTGATAGAGTAACTGCTCTTGAAGTGACGGTACCTGAGAAGAAAGCCCCGGCTAACTACGTGCCAGCA
GCCGCGGTAATACGTAGGGGGCAAGCGTTGTCCGGAATTATTGGGCGTAAAGCGCGCGCAGGCGGCTCTTTAAGTCT
GGTGTTTAATCCCGAGGCTCAACTTCGGGTCGCACTGGAAACTGGGGAGCTTGAGTGCAGAAGAGGAGAGTGGAATT
CCACGTGTAGCGGTGAAATGCGTAGAGATGTGGAGGAACACCAGTGGCGAAGGCGACTCTCTGGGCTGTAACTGACG
CTGAGGCGCGAAAGCGTGGGGAGCAAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGATGAATGCTAGGTG
TTAGGGGTTTCGATACCCTTGGTGCCGAAGTTAACACATTAAGCATTCCGCCTGGGGAGTACGGTCGCAAGACTGAA
ACTCAAAGGAATTGACGGGGACCCGCACAAGCAGTGGAGTATGTGGTTTAATTCGAAGCAACGCGAAGAACCTTACC
AGGTCTTGACATCCCTCTGACCGGTCTAGAGATAGATCTTTCCTTCGGGACAGAGGAGACAGGTGGTGCATGGTTGT
CGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTATGCTTAGTTGCCAGCAGGTCAA
GCTGGGCACTCTAAGCAGACTGCCGGTGACAAACCGGAGGAAGGTGGGGATGACGTCAAATCATCATGCCCCTTATG
ACCTGGGCTACACACGTACTACAATGGCCGGTACAACGGGAAGCGAAGGAGCGATCTGGAGCCAATCCTAGAAAAGC
CGGTCTCAGTTCGGATTGTAGGCTGCAACTCGCCTACATGAAGTCGGAATTGCTAGTAATCGCGGATCAGCATGCCG
CGGTGAATACGTTCCCGGGTCTTGTACACACCGCCCGTCACACCACGAGAGTTTACAACACCCGAAGTCGGTGAGGT
AACC
Compared according to Gen-Bank sequence homology, bacterial strain ZDC-04 is similar to Paenibacillus jamilae sequence
Property is 100%, therefore bacterial strain ZDC-04 is accredited as Jie meter La series bacillus (Paenibacillus jamilae).The bacterial strain
China Committee for Culture Collection of Microorganisms's common micro-organisms center (abbreviation CGMCC, ground were preserved on August 8th, 2018
Location is Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, Institute of Microorganism, Academia Sinica), deposit number CGMCC
No.16231。
Embodiment 3: the screening of fermentation medium
One, the activation of strain
The Jie meter La series bacillus ZDC-04 streak inoculation that sandy soil pipe saves is cultivated on LB culture medium flat plate,
48h is cultivated at 30 DEG C, then picking microscopy, the scribing line switching of typical single colonie is trained at the test tube slant LB culture medium, 30 DEG C
Activated spawn is obtained after supporting 48h, it is spare；
The composition of LB culture medium are as follows: peptone 100g/L, sodium chloride 100g/L, yeast extract 50g/L, agar 15-
6.5 ± 0.1,121 DEG C of high pressure steam sterilization 30min of 20g/L, pH.
Two, the determination of fermentation medium
Fermentation medium of the invention is mainly obtained by following steps:
1, the screening of Jie meter La series bacillus fermentation medium
The present invention studies Jie meter La series bacillus for the first time, and the present invention is consulted using NCBI to middle National IP Network related both at home and abroad
Document discovery, has no the report of Jie meter La series bacillus fermentation medium, therefore can only use for reference and Jie meter La series bacillus
The fermentation medium of the nearest bacterial strain of affiliation, has found out 11 kinds, then in conjunction with growth condition, the principle of fermentation and production
Spore condition etc. screens 11 kinds of fermentation mediums, and 3 kinds of fermentation of bacillus culture mediums have finally been determined, have carried out subsequent reality
It tests.
2, the determination of Jie meter La series bacillus ZDC-04 fermentation medium components
The principle and Sporulation condition of fermentation medium are analyzed, and combines practical raw material to buy approach and cost etc., and combine
Single factor experiment method medium component is screened, and evaluation of result uses dilution plate counting method.Finally determine main medium
Ingredient is corn flour, bean cake powder, wheat bran, sucrose.
3, the determination of Jie meter La series bacillus ZDC-04 fermentation medium components content
Fermentation medium nutritional ingredient finally determines medium component and content: corn flour 15g/ by orthogonal experiment
L, bean cake powder 10g/L, wheat bran 8g/L, sucrose 6g/L, calcium carbonate 5g/L, potassium dihydrogen phosphate 5g/L and sodium chloride 2g/L.
4, the determination of fermentation temperature
After fermentation medium inoculation, setting fermentation temperature is respectively 25,28,30,35,37 DEG C, shaking speed 200r/min,
Viable bacteria amount and gemma rate are measured, test is in triplicate.Finally determine that its optimum culturing temperature is 30 DEG C.
5, the determination of fermentation pH
Setting initial pH value of medium is respectively as follows: 6.0,6.2,6.4,6.6,6.8,7.0,7.2,7.4, after inoculation, culture
Temperature is 37 DEG C, shaking speed 200r/min, measures viable bacteria amount and gemma rate, test is in triplicate.Finally determine its Optimal pH
Value is 6.5 ± 0.1.
Embodiment 4:50L fermentor high density fermentation Jie meter La series bacillus ZDC-04
Shake-flask seed culture medium: peptone 10g/L, yeast extract 10g/L, sucrose 30g/L, pH 6.5 ± 0.1；
Secondary seed medium: corn flour 20g/L, bean cake powder 15g/L, sucrose 30g/L, potassium dihydrogen phosphate 5g/L, pH
6.5±0.1；
Fermentation medium: corn flour 15g/L, bean cake powder 10g/L, wheat bran 8g/L, sucrose 6g/L, calcium carbonate 5g/L, phosphoric acid
Potassium dihydrogen 5g/L, sodium chloride 2g/L, initial pH6.5 ± 0.1.
(1) prepared by shaking flask primary seed solution: shake-flask seed culture medium, 121 DEG C of high pressures are packed into 1 500mL triangular flask
Jie meter La series bacillus ZDC-04 after activating in embodiment 1 is used oese by steam sterilizing 30min in superclean bench
It accesses in shake-flask seed culture medium, condition of culture are as follows: 30 DEG C of temperature, shaking speed 200r/min, incubation time are for 24 hours.
(2) prepared by seeding tank secondary seed solution: by seed liquor obtained in step (1) after microscopy is without living contaminants, pressing
In 2% inoculum concentration access 10L secondary seed tank, condition of culture: 30 DEG C of temperature, speed of agitator 300r/min, ventilatory capacity
0.6m3/ h, tank press 0.04Mpa, cultivate 14h.
(3) 50L fermentor high density fermentation culture: by seed liquor obtained in step (2) after microscopy is without living contaminants
It is accessed in 50L fermentor by 5% inoculum concentration, fermentation tank capacity 30L.Ferment 0-10h, condition of culture are as follows: temperature 30 ± 0.1
DEG C, speed of agitator 400r/min, ventilatory capacity 3m3/ h, tank press 0.03-0.05Mpa；Fermentation culture conditions after 11h: temperature 30
± 0.1 DEG C, speed of agitator 300r/min, ventilatory capacity 1m3/ h, tank press 0.05Mpa.Fermentation liquid microscopy is found 90% or more in the visual field
When thalli morphology is gemma, fermentation stops, fermentation period 15h, and fermentation liquid is spare.
(4) thalline quantity and gemma yield calculate
1) measurement of thallus sum: dilution plate rubbing method counts, digit cfu/mL；
2) measurement of total spore content: bacterium solution kills nutrition body cell, using gradient dilution in 80 DEG C of heating water bath 15min
Spread plate；
3) gemma rate calculation formula: referring to sample, measurement viable bacteria is total respectively for gemma calculating according to total viable count calculating and after inactivating
Several and gemma number, calculation formula are as follows:
Gemma rate=(gemma number/total viable count) × 100%
(5) experimental result:
Under the conditions of 50L fermentor scale, fermentation period 15h, living bacteria count is up to 7.5 × 109Cfu/mL or more is produced
Gemma rate is up to 92.6% or more.
The foregoing is merely illustrative of the preferred embodiments of the present invention, is not intended to limit the invention, all in essence of the invention
Within mind and principle, any modification, equivalent replacement, improvement and so on be should all be included in the protection scope of the present invention.
SEQUENCE LISTING
<110>agricultural science and technology Co., Ltd is reached in Qingdao
<120>a kind of Jie meter La series bacillus fermentation process in high density
<130> 0
<160> 1
<170> PatentIn version 3.3
<210> 1
<211> 1382
<212> DNA
<213>the 16S rRNA gene order of Jie meter La series bacillus ZDC-04
<400> 1
aagcttgctt ctaattaacc tagcggcgga cgggtgagta acacgtaggc aacctgccca 60
caagacaggg ataactaccg gaaacggtag ctaatacccg atacatcctt ttcctgcatg 120
ggagaaggag gaaaggcgga gcaatctgtc acttgtggat gggcctgcgg cgcattagct 180
agttggtggg gtaaaggcct accaaggcga cgatgcgtag ccgacctgag agggtgatcg 240
gccacactgg gactgagaca cggcccagac tcctacggga ggcagcagta gggaatcttc 300
cgcaatgggc gaaagcctga cggagcaacg ccgcgtgagt gatgaaggtt ttcggatcgt 360
aaagctctgt tgccagggaa gaacgcttga tagagtaact gctcttgaag tgacggtacc 420
tgagaagaaa gccccggcta actacgtgcc agcagccgcg gtaatacgta gggggcaagc 480
gttgtccgga attattgggc gtaaagcgcg cgcaggcggc tctttaagtc tggtgtttaa 540
tcccgaggct caacttcggg tcgcactgga aactggggag cttgagtgca gaagaggaga 600
gtggaattcc acgtgtagcg gtgaaatgcg tagagatgtg gaggaacacc agtggcgaag 660
gcgactctct gggctgtaac tgacgctgag gcgcgaaagc gtggggagca aacaggatta 720
gataccctgg tagtccacgc cgtaaacgat gaatgctagg tgttaggggt ttcgataccc 780
ttggtgccga agttaacaca ttaagcattc cgcctgggga gtacggtcgc aagactgaaa 840
ctcaaaggaa ttgacgggga cccgcacaag cagtggagta tgtggtttaa ttcgaagcaa 900
cgcgaagaac cttaccaggt cttgacatcc ctctgaccgg tctagagata gatctttcct 960
tcgggacaga ggagacaggt ggtgcatggt tgtcgtcagc tcgtgtcgtg agatgttggg 1020
ttaagtcccg caacgagcgc aacccttatg cttagttgcc agcaggtcaa gctgggcact 1080
ctaagcagac tgccggtgac aaaccggagg aaggtgggga tgacgtcaaa tcatcatgcc 1140
ccttatgacc tgggctacac acgtactaca atggccggta caacgggaag cgaaggagcg 1200
atctggagcc aatcctagaa aagccggtct cagttcggat tgtaggctgc aactcgccta 1260
catgaagtcg gaattgctag taatcgcgga tcagcatgcc gcggtgaata cgttcccggg 1320
tcttgtacac accgcccgtc acaccacgag agtttacaac acccgaagtc ggtgaggtaa 1380
cc 1382
Claims (10)
1. a kind of fermentation medium of Jie meter La series bacillus, including following components: corn flour, bean cake powder, wheat bran, sucrose,
Calcium carbonate and potassium dihydrogen phosphate.
2. a kind of fermentation medium of Jie meter La series bacillus as described in claim 1, characterized in that the fermented and cultured
Base are as follows: corn flour 12-30g/L, bean cake powder 5-17g/L, wheat bran 5-14g/L, sucrose 6-12g/L, calcium carbonate 1-5g/L, di(2-ethylhexyl)phosphate
Hydrogen potassium 1-6g/L, sodium chloride 0-5g/L, initial pH value 6.4-7.4.
3. a kind of fermentation medium of Jie meter La series bacillus as claimed in claim 2, characterized in that the fermented and cultured
Base are as follows: corn flour 15g/L, bean cake powder 10g/L, wheat bran 8g/L, sucrose 6g/L, calcium carbonate 5g/L, potassium dihydrogen phosphate 5g/L, chlorination
Sodium 2g/L, initial pH value 6.4-7.4.
4. a kind of fermentation medium of Jie meter La series bacillus as described in any one of claim 1-3, characterized in that
The Jie meter La series bacillus is Jie meter La series bacillus ZDC-04, which is CGMCC No.16231.
5. a kind of Jie meter La series bacillus fermentation process in high density, characterized in that after the activation of Jie meter La series bacillus, warp
Level-one, secondary seed solution preparation and fermented and cultured stop fermentation when 90% or more thallus is converted into gemma in fermentation liquid；Institute
Fermented and cultured is stated using fermentation medium described in any one of claim 1-3.
6. a kind of Jie meter La series bacillus fermentation process in high density as claimed in claim 5, characterized in that including following step
It is rapid:
(1) prepared by shaking flask primary seed solution: the Jie meter La series bacillus after activation being transferred to shake-flask seed culture medium, in temperature
It is shaking table culture 16-30h at 28-37 DEG C；
(2) prepared by seeding tank secondary seed solution: cultured shake-flask seed liquid being transferred by inoculum concentration 1-5% and is trained into secondary seed
It supports in base, cultivates 14-18h at 30 ± 2 DEG C of temperature；
(3) secondary seed solution obtained in step (2) is equipped with to the fermentor of fermentation medium by the inoculum concentration access of 3-10%
In, in 30 ± 2 DEG C of fermented and cultureds, when 90% or more thallus is converted into gemma in fermentation liquid, stop fermentation.
7. a kind of Jie meter La series bacillus fermentation process in high density as claimed in claim 6, characterized in that the step
(3) when fermented and cultured, 0-10h, fermentation condition are as follows: speed of agitator 300-400r/min, ventilatory capacity 2-4m3/ h, tank press 0.03-
0.05Mpa；Fermentation culture conditions after 11h are as follows: speed of agitator 200-300r/min, ventilatory capacity 1-2.5m3/ h, tank pressure
0.03-0.05Mpa。
8. a kind of Jie meter La series bacillus fermentation process in high density as claimed in claims 6 or 7, characterized in that the outstanding person
It is Jie meter La series bacillus ZDC-04 that rice, which draws series bacillus, which is CGMCC No.16231.
9. a kind of Jie meter La series bacillus fermentation process in high density as claimed in claims 6 or 7, characterized in that
The shake-flask seed culture medium are as follows: 10 ± 2g/L of peptone, 10 ± 2g/L of yeast extract, sucrose 30 ± 5g/L, pH
6.5±0.1；
The secondary seed medium are as follows: 20 ± 5g/L of corn flour, 15 ± 3g/L of bean cake powder, 30 ± 5g/L of sucrose, biphosphate
Potassium 5 ± 1g/L, pH 6.5 ± 0.1.
10. a kind of Jie meter La series bacillus fermentation process in high density as claimed in claims 6 or 7, characterized in that the step
Suddenly the speed of agitator 300-400r/min of (2), liquid amount 50-70%, ventilatory capacity 0.4-1m3/ h, tank press 0.03-0.05Mpa.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111073837A (en) * | 2019-12-31 | 2020-04-28 | 盐城工学院 | Fermentation method for promoting Paenibacillus polymyxa to produce spores |
| CN112410237A (en) * | 2019-08-21 | 2021-02-26 | 朝阳华星生物工程有限公司 | Industrial production fermentation process of high-yield bacillus mucilaginosus |
| CN112521948A (en) * | 2020-12-01 | 2021-03-19 | 中国农业科学院农业资源与农业区划研究所 | Fertilizing method for improving secondary salinization greenhouse soil |
| CN112760254A (en) * | 2019-09-29 | 2021-05-07 | 淮阴工学院 | Method for preventing and treating tomato bacterial wilt |
| CN114874941A (en) * | 2022-05-11 | 2022-08-09 | 淮阴师范学院 | Paenibacillus phyllophilus with raw starch hydrolysis capacity and application thereof |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105670976A (en) * | 2016-03-18 | 2016-06-15 | 湖北中化东方肥料有限公司 | Fermentation culture medium for bacillus amyloliquefaciens and application of fermentation culture medium |
| CN106396862A (en) * | 2016-08-29 | 2017-02-15 | 佛山市艳晖生物科技有限公司 | Disease controlling microbial fertilizer special for tuberous crops and preparation method thereof |
| CN106701619A (en) * | 2016-12-20 | 2017-05-24 | 湖南泰谷生物科技股份有限公司 | High-density fermentation method for bacillus amyloliquefaciens and preparation method of microbial agent of bacillus amyloliquefaciens |
| CN108617406A (en) * | 2018-08-24 | 2018-10-09 | 湖北欣恺生物科技有限公司 | A kind of preparation method of CNNS's Marasmius solid medium and dry mycelium |
-
2018
- 2018-11-22 CN CN201811401672.8A patent/CN109251879B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105670976A (en) * | 2016-03-18 | 2016-06-15 | 湖北中化东方肥料有限公司 | Fermentation culture medium for bacillus amyloliquefaciens and application of fermentation culture medium |
| CN106396862A (en) * | 2016-08-29 | 2017-02-15 | 佛山市艳晖生物科技有限公司 | Disease controlling microbial fertilizer special for tuberous crops and preparation method thereof |
| CN106701619A (en) * | 2016-12-20 | 2017-05-24 | 湖南泰谷生物科技股份有限公司 | High-density fermentation method for bacillus amyloliquefaciens and preparation method of microbial agent of bacillus amyloliquefaciens |
| CN108617406A (en) * | 2018-08-24 | 2018-10-09 | 湖北欣恺生物科技有限公司 | A kind of preparation method of CNNS's Marasmius solid medium and dry mycelium |
Non-Patent Citations (2)
| Title |
|---|
| JOSE ANTONIO MORILLO等: "Production of a Metal-Binding Exopolysaccharide by Paenibacillus Jamilae Using Two-Phase Olive-Mill Waste as Fermentation Substrate", 《CURRENT MICROBIOLOGY》 * |
| 吴金鹏主编: "《食品微生物学》", 31 October 1992, 农业出版社 * |
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112410237A (en) * | 2019-08-21 | 2021-02-26 | 朝阳华星生物工程有限公司 | Industrial production fermentation process of high-yield bacillus mucilaginosus |
| CN112760254A (en) * | 2019-09-29 | 2021-05-07 | 淮阴工学院 | Method for preventing and treating tomato bacterial wilt |
| CN112760254B (en) * | 2019-09-29 | 2022-06-03 | 淮阴工学院 | Method for preventing and treating tomato bacterial wilt |
| CN111073837A (en) * | 2019-12-31 | 2020-04-28 | 盐城工学院 | Fermentation method for promoting Paenibacillus polymyxa to produce spores |
| CN111073837B (en) * | 2019-12-31 | 2023-08-25 | 盐城工学院 | Fermentation method for promoting Paenibacillus polymyxa to produce spores |
| CN112521948A (en) * | 2020-12-01 | 2021-03-19 | 中国农业科学院农业资源与农业区划研究所 | Fertilizing method for improving secondary salinization greenhouse soil |
| CN112521948B (en) * | 2020-12-01 | 2022-04-15 | 中国农业科学院农业资源与农业区划研究所 | Fertilizing method for improving secondary salinization greenhouse soil |
| CN114874941A (en) * | 2022-05-11 | 2022-08-09 | 淮阴师范学院 | Paenibacillus phyllophilus with raw starch hydrolysis capacity and application thereof |
| CN114874941B (en) * | 2022-05-11 | 2023-08-11 | 淮阴师范学院 | Paenibacillus phylloides with starch hydrolysis capability and application thereof |
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