CN108865938A - A kind of bacillus stearothermophilus of high yield gemma - Google Patents
A kind of bacillus stearothermophilus of high yield gemma Download PDFInfo
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- CN108865938A CN108865938A CN201810759522.8A CN201810759522A CN108865938A CN 108865938 A CN108865938 A CN 108865938A CN 201810759522 A CN201810759522 A CN 201810759522A CN 108865938 A CN108865938 A CN 108865938A
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- gemma
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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
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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Abstract
The object of the present invention is to provide a kind of bacillus stearothermophiluses of high yield gemma, for the YB1-1 strain of Geobacillus stearothermophilus Geobacillus stearothermophilus, in the China Committee for Culture Collection of Microorganisms's common micro-organisms center being deposited in positioned at Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3 on April 8th, 2018, deposit number is CGMCC No：15559；Provided YB1-1 plants for producing gemma.The gemma yield for the YB1-1 bacterial strain that the present invention is screened reaches 80% or more.Have compared to other bacillus stearothermophiluses (gemma yield 30%) and significantly improve, can be used for producing gemma, meet the needs of large-scale commercial production.
Description
Technical field
The invention belongs to microbe to screen technical fields, and in particular to a kind of bacillus stearothermophilus of high yield gemma.
Background technique
Generated hypopus when gemma stops growing for bacterium, gemma have very the poor environments such as low temperature, high temperature, arid
Strong resistance, and the sprouting of bacillus stearothermophilus gemma produces acid but do not produce gas, Bromocresol purple in acid condition can
Yellow is become from purple, can be used to quickly detect antibiotic residue situation in dairy products using this principle.If people's long-term consumption is anti-
The exceeded milk of raw cellulose content will lead to human body and generate drug resistance, makes internal flora imbalance, causes serious adverse consequences.Using thermophilic
Hot Bacillus stearothermophilus gemma detection Residue of Antibiotics in Milk is the mode of rapid and convenient economy the most so far.
But the gemma yield of current existing bacillus stearothermophilus is lower, therefore obtains the high bacterial strain pair of gemma yield
Actual production is of great significance.
Summary of the invention
The object of the present invention is to provide a kind of bacillus stearothermophiluses of high yield gemma, to make up the prior art not
Foot.
The bacillus stearothermophilus of high yield gemma provided by the present invention is Geobacillus stearothermophilus
YB1-1 plants of (Geobacillus stearothermophilus) is deposited on April 8th, 2018 positioned at Chaoyang District, Beijing City
The China Committee for Culture Collection of Microorganisms's common micro-organisms center of the institute 3 of North Star West Road 1, deposit number CGMCC
No：15559；
Provided YB1-1 plants for producing gemma.
The gemma yield for the YB1-1 bacterial strain that the present invention is screened reaches 80% or more.Compared to other stearothermophilus gemma
Bacillus (gemma yield 30%), which has, to be significantly improved, and can be used for producing gemma, meets the needs of large-scale commercial production.
Specific embodiment
Applicant screened during preparing gemma obtain one plant can efficiently sporiferous stearothermophilus gemma bar
Bacterial strain, and a kind of method for preparing gemma is established, to make up the deficiencies in the prior art.
The present invention is described in detail below with reference to embodiment
The screening of 1 bacillus stearothermophilus of embodiment
Applicant has found certain a batch of bud during carrying out gemma fermenting and producing using bacillus stearothermophilus
The sporiferous efficiency of spore bacillus fermentation significantly improves, and in culture medium culture is more than 72h by the bacillus stearothermophilus of the batch
Afterwards, discovery colonial morphology changes, and it is the opaque shape of milky that part bacterium colony changes from light yellow clear shape, by two kinds of forms
Bacterium colony purify respectively, two kinds of purifying bacterial strains are obtained, wherein transparent bacterial strain is in activation medium (nutrient agar 3%, CaCl2
0.01%, FeSO4·7H2O 0.001%, MnCl2·4H2O 0.3%, MgSO40.01%) gemma yield is about after culture for 24 hours
10%, milky bacterial strain gemma yield after activation medium culture for 24 hours is 80%.And reference culture stearothermophilus ground gemma bar
It is 30% that bacterium (CICC10267) cultivates gemma yield for 24 hours under above-mentioned similarity condition.The bacterial strain of the high yield gemma of above-mentioned screening passes through
It is accredited as Geobacillus stearothermophilus, therefore is named as YB1-1 plants of Geobacillus stearothermophilus.
YB1-1 plants of the Geobacillus stearothermophilus (Geobacillus tearothermophilus) obtained is screened, in
It is deposited in the Chinese microorganism strain preservation management committee positioned at Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3 on April 8th, 2018
Member's meeting common micro-organisms center, deposit number are CGMCC No：15559.
The property that YB1-1 plants of Geobacillus stearothermophilus is as follows：Gram-positive, aerobic amphimicrobian, hydrogen peroxide
Enzyme is negative, contact enzyme positive, and rod-shaped under microscope, gemma is at ellipticity.
2YB1-1 plants of production gemma effect detections of embodiment
Respectively by YB1-1 plants of the Geobacillus stearothermophilus of screening and reference standard bacterial strain Geobacillus stearothermophilus
10267 plants of CICC are cultivated as steps described below：
The activation of step 1) gemma：
By strain inoculated in strain activation and culture base (nutrient agar 3%, CaCl20.01%,
FeSO4·7H2O 0.001%, MnCl2·4H2O 0.3%, MgSO40.01%) 65 DEG C of incubator cultures, are placed in
8h。
The amplification of step 2) gemma
Take the bacillus stearothermophilus after gemma activates of 1g, the gemma amplification culture medium (nutrient meat of inoculation and 1L
Soup 1.5%, CaCl20.05%, FeSO4·7H2O 0.001%, MnCl2·4H2O 0.5%, MgSO40.01%, carboxymethyl
Sodium cellulosate 0.1%), 60 DEG C are placed in, 120r/min shaking bath culture 32h.
The preparation of step 3) gemma
Take that 1mL bacterium solution is measured and microscopy, the microscopy method are：50uL bacteria suspension is taken to be placed in glass slide center,
Heating fixation is carried out on alcolhol burner, carries out spore staining 5min with 6.7% malachite green at 75 DEG C after the completion of fixing, and is dyed
It finishes and rinses malachite green well, redye 5min using husky of common dye, in 100 times of oil microscopic observations after rinsing well.It can be seen that
Thallus is dyed to red, and gemma is green.
The result shows that YB1-1 plants of gemma yields are 80%, and 10267 plants of gemma yields of CICC are only 30%；Meanwhile
YB1-1 plants of bacterium solutions are 0.87 in 600nm light absorption value, illustrate that the bacterial strain can the vigorous growth under above-mentioned condition of culture.It will finally train
Obtained YB1-1 strain bacterium solution room temperature 6000r/min centrifugation 6min is supported, precipitating is taken, presses 50 times of its weight with sterile saline
It redissolves, in 100 DEG C of heating water bath 30min, spore solution is finally made.Spore solution obtained can be used for commodity production.
Embodiment 3：YB1-1 plants of expansion fermentations produce the detection of gemma ability
By after the YB1-1 plants of method cultures according to embodiment 2, then expand be inoculated in 10L fermentor (nutrient broth 1.5%,
CaCl20.05%, FeSO4·7H2O 0.001%, MnCl2·4H2O 0.5%, MgSO40.01%), 55 DEG C of cultivation temperature,
Stirring rate 100r/min, ventilatory capacity 3L/min, gemma yield is produced up to 75% with 1L fermentation system gemma after culture 72 hours
Rate 80% is not much different, and existing bacillus stearothermophilus, and gemma ability is produced in large volume fermentation process cannot keep.
Claims (2)
1. a kind of bacillus stearothermophilus, which is characterized in that the bacillus stearothermophilus is stearothermophilus soil gemma
YB1-1 plants of bacillus (Geobacillus stearothermophilus) is deposited on April 8th, 2018 positioned at court of Beijing
The China Committee for Culture Collection of Microorganisms's common micro-organisms center of the institute 3 of positive area's North Star West Road 1, deposit number are
CGMCC No：15559.
2. application of the bacillus stearothermophilus described in claim 1 in production gemma.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201810759522.8A CN108865938B (en) | 2018-07-11 | 2018-07-11 | Bacillus stearothermophilus with high yield of spores |
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| CN201810759522.8A CN108865938B (en) | 2018-07-11 | 2018-07-11 | Bacillus stearothermophilus with high yield of spores |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110438060A (en) * | 2019-07-22 | 2019-11-12 | 南京巨鲨显示科技有限公司 | A kind of preparation of Geobacillus stearothermophilus gemma, enrichment method |
| CN111943741A (en) * | 2020-08-21 | 2020-11-17 | 四川润格生物科技有限公司 | Technical method for treating animals died of illness by enzyme method |
| CN114181859A (en) * | 2021-12-10 | 2022-03-15 | 中国科学院青岛生物能源与过程研究所 | Geobacillus stearothermophilus and method for producing lactic acid by utilizing lignocellulose |
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| CN105368746A (en) * | 2015-12-01 | 2016-03-02 | 山东新华医疗器械股份有限公司 | Preparation method of Bacillus stearothermophilus spores |
| CN106282068A (en) * | 2016-09-28 | 2017-01-04 | 中国科学院苏州生物医学工程技术研究所 | The preparation method of resistance controllable type bacillus spore |
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2018
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Patent Citations (2)
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| CN105368746A (en) * | 2015-12-01 | 2016-03-02 | 山东新华医疗器械股份有限公司 | Preparation method of Bacillus stearothermophilus spores |
| CN106282068A (en) * | 2016-09-28 | 2017-01-04 | 中国科学院苏州生物医学工程技术研究所 | The preparation method of resistance controllable type bacillus spore |
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| THEREZA CHRISTINA VESSONI PENNA 等: "Effect of Media on Spore Yield and Thermal Resistance of Bacillus stearothermophilus", 《APPL BIOCHEM BIOTECHNOL》 * |
| 吴潇等: "嗜热脂肪芽孢杆菌的特性及应用 ", 《中国消毒学杂志》 * |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110438060A (en) * | 2019-07-22 | 2019-11-12 | 南京巨鲨显示科技有限公司 | A kind of preparation of Geobacillus stearothermophilus gemma, enrichment method |
| CN111943741A (en) * | 2020-08-21 | 2020-11-17 | 四川润格生物科技有限公司 | Technical method for treating animals died of illness by enzyme method |
| CN114181859A (en) * | 2021-12-10 | 2022-03-15 | 中国科学院青岛生物能源与过程研究所 | Geobacillus stearothermophilus and method for producing lactic acid by utilizing lignocellulose |
| CN114181859B (en) * | 2021-12-10 | 2023-09-01 | 中国科学院青岛生物能源与过程研究所 | Geobacillus stearothermophilus and method for producing lactic acid by using lignocellulose |
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| CN108865938B (en) | 2021-05-28 |
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