CN105316415A - Gene rapid screening method for synchronously identifying four types of pollution saccharomycetes in beer brewing - Google Patents
Gene rapid screening method for synchronously identifying four types of pollution saccharomycetes in beer brewing Download PDFInfo
- Publication number
- CN105316415A CN105316415A CN201510809739.1A CN201510809739A CN105316415A CN 105316415 A CN105316415 A CN 105316415A CN 201510809739 A CN201510809739 A CN 201510809739A CN 105316415 A CN105316415 A CN 105316415A
- Authority
- CN
- China
- Prior art keywords
- beer
- primer
- yeast bacterium
- black yeast
- seqidno
- 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.)
- Granted
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6844—Nucleic acid amplification reactions
- C12Q1/686—Polymerase chain reaction [PCR]
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biophysics (AREA)
- Immunology (AREA)
- Microbiology (AREA)
- Molecular Biology (AREA)
- Biotechnology (AREA)
- Analytical Chemistry (AREA)
- Physics & Mathematics (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention discloses a gene rapid screening method for synchronously identifying four types of pollution saccharomycetes in beer brewing. According to the method, a primer system composed of four pairs of primers is utilized, a mode of combining a high-flux multi-PCR technology with a high-resolution capillary electrophoresis separation technology is adopted, so that whether pollution saccharomycetes exist in the beer yeast and the fermentation process or not is rapidly determined in the beer production process. Compared with other existing molecular biology detection means based on the PCR technology, four types of beer pollution saccharomycetes, namely Pichia fermentans, Candida medenterica, Hansenula polymorpha and Phodotorula glutinis, can be integrally detected in a reaction, the detection cost is saved, and detection time is saved and shortened to 8 hours from the original 7-14 days.
Description
Technical field
The invention belongs to beer technical field, relate to a kind of gene quick screening method simultaneously identifying four kinds of black yeast bacterium in beer brewing, in particular to one based on multiple PCR technique and capillary electrophoresis separation technology, for the method for black yeast bacterium rapid screening several in beer brewing.
Background technology
Beer can be divided into two large classes according to the difference of zymotechnique and bacterial classification used, i.e. Ai Er (ale) beer and glug (lager) beer.The former uses S. cervisiae (Saccharomycescerevisiae) to ferment at a relatively high temperature, and yeast often floats over upper strata, therefore also known as high fermentation beer.The latter is at low temperature (about 10 DEG C) condition bottom fermentation, and yeast is often sunken to bottom, therefore also known as lower floor's fermentation beer.Glug beer fermentation bacterial classification is called saccharomyces pastorianus (Saccharomycespastorianus).
Brewage is based upon the fermentation that utilizes cereuisiae fermentum and controls harmful microbe to pollute on basis.We are it is desirable that the pure-blood ferment that only has S. cervisiae to participate in, but in whole Process of Beer Brewing, the factors such as preservation, brewing equipment, air, brewing water of yeast saccharomyces cerevisiae bacterial classification all likely causes harmful microorganism to enter in wheat juice or fermented liquid.From wine brewing process upstream, the high-quality of cereuisiae fermentum and purity are the most key, if the yeast itself at first for beer fermentation contains contaminated bacteria (being generally other black yeast bacterium), can to the beer quality of whole even several production batch and the overall special favor of beer, produce grievous injury, such as beer becomes sour, foreign odor.In addition, other black yeast bacterium enter wheat juice or fermented liquid during the fermentation, also can pollute wheat juice and beer.Black yeast bacterium can be the same with S. cervisiae in liquor-making environment, and amount reproduction increases.Therefore, brewageing initially, and in brewing process, the purity of S. cervisiae is being monitored, seeming most important.
Black yeast bacterium mainly contains Pichiafermentans, Candidamesenterica, Hansenulapolymorpha, Rhodotorulaglutinis.Detection mode for beer black yeast bacterium is mainly divided into two classes, and the first kind is the culture-based method based on substratum, and domestic and international most of brew-house adopts this class methods, but detection time is long; Make waste mash dense, wherein the black yeast bacterium of trace is cultivated by substratum simultaneously, is often difficult to find, therefore cannot Instructing manufacture technician monitors beer production in time; Equations of The Second Kind is the molecular biology for detection of PCR-based technology, round pcr drastically increases detection efficiency and sensitivity from nucleic acid, gene level, can be qualitative to planting, belonging to or a class object bacteria, detect quick and qualitative ability is strong, therefore no matter for the monitoring of finished beer quality, or the investigation of tracing to the source of process problems, the application of round pcr all has broad prospects and profound significance, but primary first-order equation can only detect a quasi-microorganism, and existence detects the saccharomycetic sensitivity problem of micropollution from a large amount of wine waste mash.
Great many of experiments confirms, black yeast bacterium Pichiafermentans, Candidamesenterica, Hansenulapolymorpha, Rhodotorulaglutinis, discriminating is a more complicated job, these black yeast bacterium are usually with comparatively micro-form, make wine uses yeast fungus with high density to mix, testing process is complicated and easily undetected, is thus necessary to carry out detection method more responsive more fast.In order to monitor the microbial status of beer production more in time, be necessary to develop a kind of detection method of, better effects if faster for common beer black yeast bacterium speed.
Summary of the invention
The object of the invention is the defect for prior art, overcome black yeast bacterium Pichiafermentans in conventional beer, Candidamesenterica, Hansenulapolymorpha, Rhodotorulaglutinis detection technique complex operation, the deficiencies such as time-consuming effort, a kind of gene quick screening method simultaneously identifying four kinds of black yeast bacterium in beer brewing is provided, utilize the primer system be made up of 4 pairs of primers, by the mode adopting high-throughout multiple PCR technique to combine with high-resolution capillary electrophoresis separation technology, thus realize determining to beer production the detection that whether there is black yeast bacterium in cereuisiae fermentum and fermenting process fast.
Technical scheme of the present invention is:
The present invention analyzes sample pin 4 kinds of black yeast bacterium, if wherein any one contaminated bacteria can be detected, then in interpret sample containing the brewery sludge with beer pollution capacity.
A quick screening method for beer black yeast bacterium, adopts 4 heavy PCR to increase the gene of 4 kinds of contaminated bacterias simultaneously, then by 4 heavy pcr amplification products described in capillary electrophoresis separation, applies primer SEQIDNO.1 ~ 8 in described 4 heavy PCR reactions.
The inventive method concrete steps comprise as follows:
Step (1), beer black yeast bacterium DNA profiling extract:
Adopt general DNA extraction method and purification process, from wine brewing sample thief high yeast concentration bacterium, or from wine brewing process sample thief in wheat juice or fermented liquid, extract to obtain DNA.
Step (2), multiplexed PCR amplification:
After multiplexed PCR amplification reaction system vibration mixing, carry out multiplexed PCR amplification, obtain amplified production;
Described multiplexed PCR amplification reaction system is the MgCl of 20ul by 100nM forward mix primer 2ul, 100nM reverse mix primer 2ul, 25mM
24ul, the Taq polysaccharase 0.7ul of 5 × PCRBuffer4ul, 5U/ul, DNA profiling 1ul and surplus distilled water; Wherein in forward mix primer, the concentration ratio of SEQIDNO.1 primer, SEQIDNO.3 primer, SEQIDNO.5, SEQIDNO.7 primer is 1:1:1:1, and in reverse mix primer, the concentration ratio of SEQIDNO.2 primer, SEQIDNO.4 primer, SEQIDNO.6, SEQIDNO.8 primer is 1:1:1:1;
The condition of described multiplexed PCR amplification reaction is 95 DEG C of denaturation 10min; With 94 DEG C of 30s; 58 DEG C of 30s; 70 DEG C of 1min are 1 circulation, carry out 35 circulations altogether; 4 DEG C of insulations;
The each primer sequence of 4 heavy primer system of the gene test for black yeast bacterium of table 1 the present invention design
Step (3), capillary electrophoresis separation condition:
Separation system: amplified production and the volume content of the middle gained of step described in 1ul (2) are 95% deionized formamide 38.5ul, and 400bpDNAMarker0.5ul mixes; Separation condition: 50 DEG C, under 6.0KV voltage, be separated 35min;
Step (4), Testing and appraisal:
By the amplified production of gained in described step (2) through capillary electrophoresis separation, automatically retrieved by multiple gene expression genetic analysis systems, collect fluorescent signal in sample and judge whether in gene fragment length to be that characteristic peak appears in 142bp, 147bp, 335bp, 359bp corresponding position; If there is characteristic peak in 142bp corresponding position, be then judged as in this detection sample containing black yeast bacterium Pichiafermentans; If there is characteristic peak in 147bp corresponding position, be then judged as in this detection sample containing black yeast bacterium Candidamesenterica; If there is characteristic peak in 335bp corresponding position, be then judged as in this detection sample containing black yeast bacterium Hansenulapolymorpha; If there is characteristic peak in 359bp corresponding position, be then judged as in this detection sample containing black yeast bacterium Rhodotorulaglutinis.
Table 2 four kinds of black yeast bacterium
Compared with prior art the present invention has following features:
One, the present invention can carry out in same PCR pipe, simple to operate, can determine whether there is beer black yeast bacterium fast, and detected result is accurate;
They are two years old, by analyzing gene determination beer black yeast bacterium Pichiaermentans, Candidamesenterica, Hansenulapolymorpha, Rhodotorulaglutinis exists at the trace of brewage initially and in process, harmful yeast in Timeliness coverage beer, realizes effective monitoring and prophylactic effect;
They are three years old, the present invention relative to current other molecular Biological Detection means based on round pcr, can in primary first-order equation integrated detection 4 kinds of beer black yeast bacterium, saved testing cost, save detection time, detection time shortened to 8h by former 7 ~ 14 days.
Accompanying drawing explanation
Fig. 1 is the multiplex PCR-capillary electrophoresis spectrogram of the specific embodiment of the invention 1;
Fig. 2 is the multiplex PCR-capillary electrophoresis spectrogram of the specific embodiment of the invention 2.
Embodiment
Below in conjunction with specific embodiment, the present invention is further analyzed.
Embodiment 1:
(1) cultivation of beer black yeast bacterium and DNA profiling extract:
The beer black yeast bacterium Pichiaermentans that picking plate culture medium grows, Candidamesenterica, Hansenulapolymorpha, the mono-bacterium colony of Rhodotorulaglutinis, the DNA profiling of beer black yeast bacterium is extracted according to the operation instruction of the GenomicDNAPurificationKit test kit of Qiagen company, and using ultraviolet spectrophotometer under 260nm wavelength condition, measure the DNA quality of extraction, DNA solution is diluted to 250ng/ul, and-20 DEG C save backup.
(2) Quadruple-PCR amplification
The primer system is the SEQIDNO.1-8 described in table 1,20ulPCR reaction system: the MgCl of 100nM forward mix primer 2ul, 100nM reverse mix primer 2ul, 25mM
24ul, the Taq polysaccharase 0.7ul of 5 × PCRBuffer4ul, 5U/ul, DNA profiling 1ul, supply distilled water and make cumulative volume be 20ul; 95 DEG C of denaturation 10min; With 94 DEG C of 30s; 58 DEG C of 30s; 70 DEG C of 1min are 1 circulation, carry out 35 circulations altogether; 4 DEG C of insulations;
(3) capillary electrophoresis separation condition
Separation system: the amplified production of the middle gained of step described in 1ul (2) and 95% deionized formamide 38.5ul, 400bpDNAMarker0.5ul mixes;
Separation condition: 50 DEG C, under 6.0KV voltage, be separated 35min;
(4) data analysis
The GenomeLabGeXPGeneticAnalysisSystem analytical system of Beckman Ku Erte is used automatically to analyze separating resulting, shown in result Fig. 1, the gene of 4 kinds of black yeast bacterium is all detected, use the present invention in primary first-order equation, the characteristic peak of 4 kinds of black yeast bacterium genes in sample can be captured.
Embodiment 2:
Two large classes of beer, i.e. Ai Er (ale) beer and glug (lager) beer, the former uses S. cervisiae (Saccharomycescerevisiae), glug beer fermentation bacterial classification is saccharomyces pastorianus (Saccharomycespastorianus), in the upstream of wine brewing process, exist with high density.This example can prove that the present invention is to black yeast bacterium Pichiafermentans, Candidamesenterica, Hansenulapolymorpha, the detection specificity of Rhodotorulaglutinis, namely detection method of the present invention is under only there is situation in S. cervisiae (Saccharomycescerevisiae) existence or saccharomyces pastorianus (Saccharomycespastorianus), non-indicating characteristic peak, 2 kinds of uncontaminations are commonly used yeast (Saccharomycescerevisiae and Saccharomycespastorianus) and are existed and can not produce interference to detection.
Operation steps of the present invention as described in Example 1.
(1) cultivation of brewery sludge and DNA profiling extract:
The S. cervisiae (Saccharomycescerevisiae) that picking plate culture medium grows exists and saccharomyces pastorianus (Saccharomycespastorianus) single bacterium colony, the DNA profiling of S. cervisiae and saccharomyces pastorianus is extracted according to the operation instruction of the GenomicDNAPurificationKit test kit of Qiagen company, and use ultraviolet spectrophotometer under 260nm wavelength condition, measure the DNA quality of extraction, DNA solution is diluted to 250ng/ul, and-20 DEG C save backup.
(2) Quadruple-PCR amplification
The primer system is the SEQIDNO.1-8 described in table 1,20ulPCR reaction system: the MgCl of 100nM forward mix primer 2ul, 100nM reverse mix primer 2ul, 25mM
24ul, the Taq polysaccharase 0.7ul of 5 × PCRBuffer4ul, 5U/ul, DNA profiling 1ul, supply distilled water and make cumulative volume be 20ul; 95 DEG C of denaturation 10min; With 94 DEG C of 30s; 58 DEG C of 30s; 70 DEG C of 1min are 1 circulation, carry out 35 circulations altogether; 4 DEG C of insulations;
(3) capillary electrophoresis separation condition
Separation system: the amplified production of the middle gained of step described in 1ul (2) and 95% deionized formamide 38.5ul, 400bpDNAMarker0.5ul mixes;
Separation condition: 50 DEG C, under 6.0KV voltage, be separated 35min;
(4) data analysis
The GenomeLabGeXPGeneticAnalysisSystem analytical system of Beckman Ku Erte is used automatically to analyze separating resulting, result shows, characteristic peak is not had to occur, namely the existence of S. cervisiae (Saccharomycescerevisiae) or saccharomyces pastorianus (Saccharomycespastorianus) does not affect detected result, there will not be false positive results.
Embodiment 3:
In wine brewing process, uncontamination fermented yeast bacterium (Saccharomycescerevisiae and Saccharomycespastorianus) fermented bacterium exists with high density, black yeast bacterium Pichiafermentans, Candidamesenterica, Hansenulapolymorpha, Rhodotorulaglutinis exist with lower concentration.When this example can prove that the present invention coexists to the black yeast bacterium of trace and high gravity fermentation bacterial classification, the present invention is to the saccharomycetic susceptibility of detection micropollution.
This example adopts 2 kinds of uncontaminations of brewage to commonly use yeast (Saccharomycescerevisiae and Saccharomycespastorianus), concentration ratio with 100,000 times: 1 and black yeast bacterium are mixed into sample, to detect susceptibility of the present invention.
The S. cervisiae (Saccharomycescerevisiae) respectively picking plate culture medium grown, add in 100mL substratum with saccharomyces pastorianus (Saccharomycespastorianus) single bacterium colony, be cultured in aerobic environment and make bacteria suspension, spectrophotometric about OD=0.2.Black yeast bacterium Pichiafermentans, the mono-bacterium colony of Candidamesenterica, Hansenulapolymorpha, Rhodotorulaglutinis adds in 5mL substratum, is cultured to and makes bacteria suspension in aerobic environment, spectrophotometric about OD=0.2.Bacteria suspension concentration is calculated according to OD value, with the concentration of 100,000 times: 1, by S. cervisiae or saccharomyces pastorianus, and the mixing of black yeast bacterium.Getting this bacteria suspension of 1mL adds in 1.5ml centrifuge tube, 13,000 × g centrifugal 2min, and collecting cell precipitates, and removes supernatant liquor; Add the thorough re-suspended cell of 50mMEDTA of 480ul; Add the 10mg/ml lyase bacterium of 120ul, hatch 60min, 13 for 37 DEG C, the centrifugal 2min of 000 × g, removes supernatant liquor.
DNA profiling is extracted afterwards according to the operation instruction of the GenomicDNAPurificationKit test kit of Qiagen company, multi-PRC reaction system uses Thermo-DNAPolymeraseDNA test kit, use 4 heavy PCR reaction systems, the primer system is the SEQIDNO.1-8 described in table 1,20ulPCR reaction system: 100nM forward mix primer 2ul, the reverse mix primer 2ul of 100nM, the MgCl24ul of 25mM, 5 × PCR damping fluid 4ul, the Taq polysaccharase 0.7ul of 5U/ul, DNA profiling 1ul, supplies distilled water and makes cumulative volume be 20ul, 95 DEG C of denaturation 10min, with 94 DEG C of 30s, 58 DEG C of 30s, 70 DEG C of 1min are 1 circulation, carry out 35 circulations altogether, 4 DEG C of insulations, capillary electrophoresis separation condition: adopt the above-mentioned multiplexed PCR amplification product of separation system: 1ul and 95% deionized formamide 38.5ul, 400bpDNAMarker0.5ul mixes, separation condition: at 50 DEG C, under 6.0KV voltage, be separated 35min, the GenomeLabGeXPGeneticAnalysisSystem analytical system of Beckman Ku Erte is used automatically to analyze separating resulting, obtain multiplex PCR-capillary electrophoresis spectrogram, the S. cervisiae (Saccharomycescerevisiae) with 100,000 times of concentration, or under saccharomyces pastorianus (Saccharomycespastorianus) Coexistence Situation, black yeast bacterium Pichiafermentans can be detected, Candidamesenterica, Hansenulapolymorpha, the characteristic peak of Rhodotorulaglutinis gene, result shows, the S. cervisiae (Saccharomycescerevisiae) of 100000 times of concentration mixes with Pichiafermentans, the gene expression characteristics peak of Pichiafermentans can be detected, containing a large amount of uncontamination yeast in shown interpret sample, interference is not caused to the sensitivity detecting black yeast bacterium, and show high sensitivity.
Sequence table
Above-described embodiment is not that the present invention is not limited only to above-described embodiment for restriction of the present invention, as long as meet application claims, all belongs to protection scope of the present invention.
SEQUENCELISTING
<110> Electronic University Of Science & Technology Of Hangzhou
<120> identifies the gene quick screening method of four kinds of black yeast bacterium in beer brewing simultaneously
<130>1
<160>8
<170>PatentInversion3.3
<210>1
<211>37
<212>DNA
<213> synthetic
<400>1
tcgatgacactcagacatactcaccaggtccagacgt37
<210>2
<211>39
<212>DNA
<213> synthetic
<400>2
gtacgactcactatagggagcagacaaatcactccaagg39
<210>3
<211>42
<212>DNA
<213> synthetic
<400>3
tcgatgacactcagacatcatgctacaaaccccgactcacga42
<210>4
<211>40
<212>DNA
<213> synthetic
<400>4
gtacgactcactatagggatgaaccaccgccggcatgaag40
<210>5
<211>39
<212>DNA
<213> synthetic
<400>5
tcgatgacactcagacatgcaaattacccaatcctgaca39
<210>6
<211>38
<212>DNA
<213> synthetic
<400>6
gtacgactcactatagggaggtttgccaacacagagtg38
<210>7
<211>38
<212>DNA
<213> synthetic
<400>7
tcgatgacactcagacattatcccctggcaacactttg38
<210>8
<211>37
<212>DNA
<213> synthetic
<400>8
gtacgactcactatagggatttgcgacgacacccagt37
Claims (5)
1. identify the gene quick screening method of four kinds of black yeast bacterium in beer brewing simultaneously, it is characterized in that the method comprises the following steps:
Step (1), beer black yeast bacterium DNA profiling extract:
Adopt general DNA extraction method and purification process, from wine brewing sample thief high yeast concentration bacterium, or from wine brewing process sample thief in wheat juice or fermented liquid, extract to obtain DNA;
Step (2), multiplexed PCR amplification:
After multiplexed PCR amplification reaction system vibration mixing, carry out multiplexed PCR amplification, obtain amplified production;
Described multiplexed PCR amplification reaction system is the MgCl of 20ul by 100nM forward mix primer 2ul, 100nM reverse mix primer 2ul, 25mM
24ul, the Taq polysaccharase 0.7ul of 5 × PCRBuffer4ul, 5U/ul, DNA profiling 1ul and surplus distilled water;
Step (3), capillary electrophoresis separation condition:
Step (4), Testing and appraisal:
By the amplified production of gained in described step (2) through capillary electrophoresis separation, automatically retrieved by multiple gene expression genetic analysis systems, collect fluorescent signal in sample and judge whether in gene fragment length to be that characteristic peak appears in 142bp, 147bp, 335bp, 359bp corresponding position; If there is characteristic peak in 142bp corresponding position, be then judged as in this detection sample containing black yeast bacterium Pichiafermentans; If there is characteristic peak in 147bp corresponding position, be then judged as in this detection sample containing black yeast bacterium Candidamesenterica; If there is characteristic peak in 335bp corresponding position, be then judged as in this detection sample containing black yeast bacterium Hansenulapolymorpha; If there is characteristic peak in 359bp corresponding position, be then judged as in this detection sample containing black yeast bacterium Rhodotorulaglutinis.
2. the gene quick screening method simultaneously identifying four kinds of black yeast bacterium in beer brewing as claimed in claim 1, it is characterized in that the concentration ratio of SEQIDNO.1 primer, SEQIDNO.3 primer, SEQIDNO.5, SEQIDNO.7 primer in step (2) forward mix primer is 1:1:1:1, in reverse mix primer, the concentration ratio of SEQIDNO.2 primer, SEQIDNO.4 primer, SEQIDNO.6, SEQIDNO.8 primer is 1:1:1:1.
3. the gene quick screening method simultaneously identifying four kinds of black yeast bacterium in beer brewing as claimed in claim 1, is characterized in that the condition of the multiplexed PCR amplification reaction described in step (2) is 95 DEG C of denaturation 10min; With 94 DEG C of 30s; 58 DEG C of 30s; 70 DEG C of 1min are 1 circulation, carry out 35 circulations altogether; 4 DEG C of insulations.
4. the gene quick screening method simultaneously identifying four kinds of black yeast bacterium in beer brewing as claimed in claim 1, it is characterized in that step (3) separation system: amplified production and the volume content of the middle gained of step described in 1ul (2) are 95% deionized formamide 38.5ul, and 400bpDNAMarker0.5ul mixes.
5. the as claimed in claim 1 gene quick screening method simultaneously identifying four kinds of black yeast bacterium in beer brewing, is characterized in that step (3) separation condition: 50 DEG C, under 6.0KV voltage, be separated 35min.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510809739.1A CN105316415B (en) | 2015-11-20 | 2015-11-20 | The gene quick screening method of four kinds of black yeast bacterium in beer brewing is identified simultaneously |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510809739.1A CN105316415B (en) | 2015-11-20 | 2015-11-20 | The gene quick screening method of four kinds of black yeast bacterium in beer brewing is identified simultaneously |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN105316415A true CN105316415A (en) | 2016-02-10 |
| CN105316415B CN105316415B (en) | 2018-11-13 |
Family
ID=55244732
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510809739.1A Active CN105316415B (en) | 2015-11-20 | 2015-11-20 | The gene quick screening method of four kinds of black yeast bacterium in beer brewing is identified simultaneously |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105316415B (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107058578A (en) * | 2017-05-31 | 2017-08-18 | 太原格因生物科技有限公司 | The detection probe of rhodotorula glutinis, genetic chip and method in dairy products |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103361439A (en) * | 2013-07-30 | 2013-10-23 | 青岛啤酒股份有限公司 | Method for integrated detection of nine beer-spoilage bacteria |
| CA2913217A1 (en) * | 2013-06-18 | 2014-12-24 | Anheuser-Busch Inbev Sa | Method for preparing a fermented beverage and beverage thus produced |
-
2015
- 2015-11-20 CN CN201510809739.1A patent/CN105316415B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CA2913217A1 (en) * | 2013-06-18 | 2014-12-24 | Anheuser-Busch Inbev Sa | Method for preparing a fermented beverage and beverage thus produced |
| CN103361439A (en) * | 2013-07-30 | 2013-10-23 | 青岛啤酒股份有限公司 | Method for integrated detection of nine beer-spoilage bacteria |
Non-Patent Citations (2)
| Title |
|---|
| SATOSHI SHIMOTSU ET AL.: "Investigation of beer-spoilage ability of", 《INSTITUTE OF BREWING & DISTILLING》 * |
| 雷剑: "啤酒生产中酵母污染的防治研究", 《中国优秀硕士学位论文全文数据库工程科技辑》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107058578A (en) * | 2017-05-31 | 2017-08-18 | 太原格因生物科技有限公司 | The detection probe of rhodotorula glutinis, genetic chip and method in dairy products |
Also Published As
| Publication number | Publication date |
|---|---|
| CN105316415B (en) | 2018-11-13 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103361439B (en) | Method for integrated detection of nine beer-spoilage bacteria | |
| Ivey et al. | Detection and identification of microorganisms in wine: a review of molecular techniques | |
| CN1402795A (en) | Method of judging flocculating properties of bottom brewer's yeast | |
| Vegas et al. | Evaluation of representativity of the acetic acid bacteria species identified by culture-dependent method during a traditional wine vinegar production | |
| Bokulich et al. | Profiling the yeast communities of wine fermentations using terminal restriction fragment length polymorphism analysis | |
| Lebuhn et al. | Microbiology and molecular biology tools for biogas process analysis, diagnosis and control | |
| Larralde-Corona et al. | Rational selection of mixed yeasts starters for agave must fermentation | |
| CN103882105A (en) | Method for measuring content of saturated hydrocarbon-degrading gene AlkB in petroleum-contaminated soil | |
| CN106497916A (en) | A kind of construction method in the NK cell polygenic variations library for high-flux sequence detection and its application | |
| CN105316415A (en) | Gene rapid screening method for synchronously identifying four types of pollution saccharomycetes in beer brewing | |
| CN103305623A (en) | Multiple rapid detection method of two food-borne pathogenic bacteria and detection primer groups as well as kit | |
| CN112226523A (en) | Specific detection probe and kit for acetic acid-resistant lactobacillus and application of specific detection probe and kit | |
| US20210172004A1 (en) | Universal lactic acid bacteria quantification kit for fermentation monitoring | |
| CN105132561A (en) | Quick detection method capable of simultaneously identifying three beer-spoilage bacteria megasphaera, pectinatus and pediococcus | |
| CN112813142B (en) | MicroRNA capture magnetic bead, preparation method and detection method of MicroRNA | |
| Zara et al. | Detection, quantification, and identification of yeast in winemaking | |
| CN114317808A (en) | Quantitative detection kit, detection method and application of thermophilic fungi | |
| CN103397092B (en) | Method for rapidly screening beer contaminating bacterium | |
| Pozo-Bayón et al. | Molecular approaches for the identification and characterisation of oenological lactic acid bacteria | |
| CN105316414B (en) | The gene quick screening method of a variety of thermoduric bacterias in fruit drink is identified simultaneously | |
| CN105316413B (en) | A kind of gene quick screening method that can detect four kinds of contaminated bacterias in fruit drink simultaneously | |
| CN102277449B (en) | Subtype identification method of corynebacterium crenatum bacteriophage | |
| CN108251513A (en) | A kind of method of quick judgement wine brewing pit mud quality | |
| CN116926218B (en) | Probe combination, gene chip, kit and method for detecting ascophyllum sp | |
| Storgårds et al. | Brewing control systems: microbiological analysis |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| EE01 | Entry into force of recordation of patent licensing contract |
Application publication date: 20160210 Assignee: Shaoxing Keqiao Intelligent Agricultural Research Institute Co Ltd Hangzhou University of Electronic Science and technology Assignor: HANGZHOU DIANZI University Contract record no.: X2023980030245 Denomination of invention: Rapid gene screening method for simultaneous identification of four contaminated yeasts in beer brewing Granted publication date: 20181113 License type: Common License Record date: 20230109 |
|
| EE01 | Entry into force of recordation of patent licensing contract |