CN107699610A - For detecting two pairs of specific primers of acidproof Bacillus acidi lactici content and its application during liquor fermentation - Google Patents
For detecting two pairs of specific primers of acidproof Bacillus acidi lactici content and its application during liquor fermentation Download PDFInfo
- Publication number
- CN107699610A CN107699610A CN201710952009.6A CN201710952009A CN107699610A CN 107699610 A CN107699610 A CN 107699610A CN 201710952009 A CN201710952009 A CN 201710952009A CN 107699610 A CN107699610 A CN 107699610A
- Authority
- CN
- China
- Prior art keywords
- pairs
- pit mud
- sequence
- specific primers
- primer
- 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.)
- Pending
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
- C12Q1/6851—Quantitative amplification
-
- 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
- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
- C12Q1/6888—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms
- C12Q1/689—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms for bacteria
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Analytical Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biophysics (AREA)
- Immunology (AREA)
- Microbiology (AREA)
- Molecular Biology (AREA)
- Biotechnology (AREA)
- Physics & Mathematics (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
It is used to detect two pairs of specific primers of acidproof Bacillus acidi lactici content and its application during liquor fermentation the invention discloses a kind of, the forward primer that described two pairs of specific primers include LA2 is shown in sequence 1, and LA2 reverse primer is shown in sequence 2；LA3 forward primer is shown in sequence 3, and LA3 reverse primer is shown in sequence 4, and for quantitative determining the application of drinks corruption bacterial content and application for distinguishing quality pit mud and common pit mud and pit mud inferior.The invention discloses 2 pairs of specific primers for detecting drinks corruption bacterial content.By the measure to microorganism drinks corruption bacterial content in the pit mud in the high-quality pit of white wine and common pit, fermented grain, it is found that the content percentage of drinks spoilage organisms can be used for distinguishing different grades of pit.
Description
Technical field
The invention belongs to biological technical field, and specifically the present invention relates to one kind to detect Lactobacillus
Acetotolerans 2 pairs of Species-specific primers and its application in quality of white spirit control.
Background technology
White wine is the traditional fermented food in China, there is long history, is constantly subjected to liking for people from ancient times to the present.In
State's traditional liquor be using rich in amylaceous Cereals class as raw material, using Chinese distiller's yeast as saccharifying agent, using solid-state, semisolid or liquid
State is fermented, the alcoholic drink through distilling, storing and hook tune forms.
Bacillus acidi lactici is the bacterium for referring to make carbohydrate fermentation produce lactic acid, is a category lactic acid maximum in lactic acid bacteria
Bacillus belongs to Lctobacteriaceae, in Daqu, Chinese yeast and its fermented grain be present in liquor production, lactic acid bacteria is in white wine taste
And also played a very important role on fragrance.Lactic acid bacteria energy fermenting carbohydrate generates lactic acid, and breast is produced by esterification in fermented grain
It is one of microorganism main in pit mud again that acetoacetic ester ethyl lactates, which make white wine have unique fragrance Bacillus acidi lacticis, often with
The mushrooms such as butyric, caproic acid bacteria, methane backeria, sulfate reducing bacteria, nitrate reduction bacterium coexist " symbiosis ".Some lactic acid bacterias are also same
The materials such as a small amount of Shi Shengcheng acetic acid, ethanol, propionic acid, butyric acid, pyruvic acid, it is common to adjust wine body flavor.In wine fermentation process
In the growth restriction of microorganism made due to environmental pressures such as alcohol contents, the bacterium of a few species keeps living under these conditions
Power, its content can cause fermented quality to reduce or ferment failure, be identified as drinks putrefactive microorganisms after exceeding certain limit.Make
It is more difficult that a kind of culture is proved to be for important drinks spoilage organisms Lactobacillus acetotolerans, and is often gone out
A kind of microorganism in present southern china winery produces lactic acid in addition, Lactobacillus acetotolerans have
Ability；It is the end-product of acetic acid and acetyl as carbohydrate fermentation to influence one of key of drinks mouthfeel.Liquor production
Appropriate lactic acid is needed, otherwise, no lactic acid and ethyl lactate, liquor flavor extreme difference.But lactic acid will excessively make fermented grain become sour, not only
Distillation yield is low, and vinosity can also be deteriorated.
Due to the difference for brewageing plant area's building time and region location of white wine, in different pits acid composition into
Point and content have a larger difference, the content difference of the composition of certain micro-organisms in high-quality pit and common pit is also apparent from
These differences are closely related with brewageing quality, are to brewage an important factor for quality control process needs quantitative determination wherein
Larger difference be present between common pit and high-quality pit, it is necessary to have in Lactobacillus acetotolerans content
The very strong method of specificity is measured the bacterium to the bacterium and is found in the fermentation process of a variety of white wine to be belonged in fermented grain
Still, this category of Bacillus acidi lactici Lactobacillus is currently known hundreds of different kind, white wine hair to absolute predominance strain
Relate generally to nearly 50 kinds different Bacillus acidi lactici during ferment and most Bacillus acidi lacticis not yet obtain careful research, its
Genomic information is not also very comprehensive, so the specific primer design currently for Lactobacillus acetotolerans
It is rarely reported.
The content of the invention
The present invention is by designing specific primer, so as to quantitative determine Lactobacillus acetotolerans
This important industrial microorganism, by judging liquor fermentation to the measure of Lactobacillus acetotolerans contents
The quality of pond pit mud quality.
To achieve these goals, the present invention uses following technical step：
A kind of two pairs of specific primers for being used to detect acidproof Bacillus acidi lactici content during liquor fermentation, its feature exist
In：The forward primer that two pairs of described specific primers include LA2 is shown in sequence 1, and LA2 reverse primer is shown in sequence 2；LA3 is just
See sequence 3 to primer, LA3 reverse primer is shown in sequence 4.
Moreover, the step of measure drinks spoilage organisms Lactobacillus acetotolerans contents, is as follows：
(1) the grand genome of sample is extracted, using genome as template, is expanded using two pairs of above-mentioned primers, then carried out
Agarose gel electrophoresis is tested, and is tentatively judged according to the position of electrophoretic band and light levels；
(2) PCR (Polymerase Chain Reaction) product is sequenced, analysis result, verified simultaneously with this
Specific amplification of the above-mentioned primer of quantitative measurement in specific sample；
(3) QPCR can be used after specific amplification confirms to drinks spoilage organisms Lactobacillus acetotolerans
Quantified.
Two pairs of specific primers for detecting acidproof Bacillus acidi lactici content during liquor fermentation are used to quantitative determine wine
The application of class spoilage organisms Lactobacillus acetotolerans contents.
Moreover, the dynamic changing process of the content of microorganisms in pit mud and fermented grain is measured and tracked.
Two pairs of specific primers for detecting acidproof Bacillus acidi lactici content during liquor fermentation are used to distinguish high-quality cellar for storing things
The application of mud and common pit mud and pit mud inferior.
Moreover, content of the pit mud inferior than Lactobacillus acetotolerans in quality pit mud typically will
More than 10 times, the content that the common pit mud compares Lactobacillus acetotolerans in quality pit mud is also high, but
It is no greater than 10 times.
The present invention compared with prior art, possesses advantages below and beneficial effect：
The advantage of the present invention:(1) design method is advanced, be can be designed that for limited flora background of brewageing come 100%
Specific primer quantifies for Lactobacillus acetotolerans'；(2) other complicated flora backgrounds can be directed to
Specific primer corresponding to Lactobacillus acetotolerans is designed, and with background strain full-length genome information
Constantly improve updates the specificity of primer；(3) can be established using specific primer with fast and low-cost
Lactobacillus acetotolerans quantitative approach；(4) by Lactobacillus in pit mud
It the measure of acetotolerans bacterial contents, can quickly judge the quality of pit mud quality, be advantageous to the quality control of fermentation process
System.
This patent existing full-length genome information that this belongs to by Lactobacillus, with reference to PRIMER-BLAST primers
Design tool, the microorganism species under liquor fermentation background are carried out with full-length genome information and as the excellent of primer specificity
First excluded ranges, two couples of specific primer are devised using the pit mud of liquor fermentation and the grand genome of fermented grain as template amplification target
Product, amplified production confirm the specific amplification of these two pair primer after series of validation, can be used for liquor fermentation process
The measure tracking of middle Lactobacillus acetotolerans contents.
Embodiment
Below by specific embodiment, the invention will be further described, and following examples are descriptive, is not limit
Qualitatively, it is impossible to which protection scope of the present invention is limited with this.
Present invention design specific primer, it is this so as to quantitative determine Lactobacillus acetotolerans
Important industrial microorganism, by judging liquor fermentation Chi Jiao to the measure of Lactobacillus acetotolerans contents
The quality of shale amount.
(1) design of primers：Lactobacillus acetotolerans full-length genome information is collected, collects Bacillus acidi lactici
Belong to full-length genome information known to the whole of other strains of Lactobacillus, one group is designed using Primer-Blast
Lactobacillus acetotolerans specific primers, primer sequence carry out BLAST (www.ncbi.nlm.nih.gov)
Bioinformatic analysis, the sequence-specific of primer is primarily determined that, if it find that any one sequence of primer pair can be with sample
The genome sequence of other species shows higher matching (more than 70%) in present context flora, then collects the full-length genome of the species
Information, the information (species name and accession number) is placed in weight in the exclusion background of Primer-Blast systems
New design primer, until the BLAST output results of any one sequence of designed primer and the matching degree of other species are not
Higher than 70%.
(2) specificity verification：Extract the grand genome sample of flora in fermentation process；It is utilized respectively above-mentioned specific primer
Expand the grand genome sample of flora；Amplified production carries out TA cloning experimentations, and TA corresponding to each pair primer clones positive bacterium colony immediately
15 progress DNA sequencings of picking；Sequencing result carries out BLAST bioinformatic analysis, judges what species is sequence belong to；It is if complete
The insetion sequence of 15, portion clone is all Lactobacillus acetotolerans genome sequence, then the primer pair
Specificity is 100%, is just specific primer.
(3) method for quantitatively determining：The Lactobacillus acetotolerans in arbitrary sample are carried out using QPCR
It is quantitative, wherein required quantitative criterion material is exactly the PCR primer of the target sequence come out using primer amplified, to this
Target sequence in PCR primer, which is quantified to copy/uL, can be used to QPCR standards.
Embodiment 1
The extraction of pit mud or the grand genome of fermented grain
Extract grand genome using Shanghai life work B518233-0100 kits, weigh 100~300mg pit mud (if
It is that fermented grain then needs to crush in advance, 100~300mg is weighed after crushing), the Buffer SCL of 65 DEG C of preheatings of 400ul are added, are shaken
Swing and shake up, be placed in 65 DEG C of water-bath 5min.12000rpm room temperatures centrifuge 3min, draw centrifugation of the supernatant to a clean 1.5ml
Guan Zhong.Isometric Buffer SP are added, overturns and mixes, ice bath 10min, it is white or light to add solution after Buffer SP are mixed
Yellow muddiness shape.12000rpm room temperatures centrifuge 3min, draw supernatant into clean 1.5ml centrifuge tube.Add 200ul
Chloroform, fully mix, 12000rpm centrifugation 5min, draw upper strata aqueous phase into clean 1.5ml centrifuge tube.Add
The Buffer SB of 1.5 times of volumes, its whole is added in adsorption column with pipettor after fully mixing, is stored at room temperature 2min,
12000rpm centrifuges 30s, outwells waste liquid in collecting pipe.Adsorption column is put back in collecting pipe, adds 700ul Wash
Solution, 12000rpm centrifuge 30s, outwell waste liquid in collecting pipe.Adsorption column is put back in collecting pipe, adds 300ul Wash
Solution, 12000rpm centrifuge 1min, outwell waste liquid in collecting pipe.Adsorption column is put back in collecting pipe, 12000rpm centrifugations
2min.Adsorption column is taken out, is put into a new 1.5ml centrifuge tube, 100ul TE Buffer are added in adsorbed film center, it is quiet
3min, 12000rpm centrifugation 2min are put, obtained DNA solution is placed in -20 DEG C and preserves or be directly used in follow-up test.
Embodiment 2
The amplification of design of primers and target dna
Species specificity is carried out to Lactobacillus acetotolerans full-length genome using Primer-Blast
The design of primer, the purpose for designing the specific primer of certain species are generally basede on two, first, identifying the species, and detect certain
Whether the species are included in one biocoene, and another purpose is in it is determined that abundance of the species in whole microbiologic population.
Table 1Lactobacillus acetotolerans Species-specific primer
The extension increasing sequence of 2 two pairs of primers of table
PCR reaction systems:NPK02buffer (2 ×) 6ul, Species-specific primer LA2F LA2R (or LA3F LA3R)
1.5ul, template 0.15ul (the use of the grand genome biased sample of fermented grain being template), Taq enzyme 0.25ul, ddH2O 5.1ul.94
DEG C -4min, (94 DEG C of -30s, 57 DEG C of -40s, 72 DEG C of -40s) × 35 are circulated, 72 DEG C of -2min, 37 circulations.By PCR after purification
Product does TA clones and blue hickie screening experiment.
Embodiment 3
The specificity of checking primer is cloned using TA
The product 0.5ul of PMD19-T carriers 0.3ul, LA2 (or LA3) PCR after purification, sterilizing ddH2O 4.2ul
Solution I 5ul, mix, add 30ul competent escherichia coli cell DH5 α ice baths 40 minutes, then at 37 DEG C of isothermal vibration devices
Middle culture activation 1h.X-gal 40ul, IPTG 40ul, 160ul dH2O are mixed, are coated on LB ammonia benzyl (100ug/ml) solid
Culture medium (LBA), treats its drying, and the bacterium solution 200ul of activation is continued to be coated on the dried LBA solid cultures of X-gal+IPTG
On base.37 DEG C of constant temperature are incubated overnight.Random picking single bacterium colony 15 delivers to the total length sequencing that sequencing company carries out insetion sequence.
The result of sequencing is subjected to BLAST analyses, RDP analyses (http://rdp.cme.msu.edu/), whether amplified production is belonged to
Target Bacillus acidi lactici carries out qualitative and quantitative analysis.Whole 15 cloned sequences are target sequence, and person could be surveyed by specificity
Examination.
Embodiment 4
QPCR quantitative determines Lactobacillus acetotolerans contents
Purpose band in mixing pit mud sample is expanded using LA2 primers and is purified, and obtains QPCR standard DNA 30ng/ul,
The DNA solution is by 10 times of doubling dilution as master sample.
DNA (copy/L)=6.02*1023(copy/mol)*DNA concentration(μg/μL)/[DNA length
(bp)*660(daltons/bp)]。
QPCR reaction systems are：NPK626ul, Primer 1.5ul, Taq enzyme 0.25ul, templet gene group to be measured
4.25ul；QPCR conditions：94 DEG C of -4min, (94 DEG C of -30s, 57 DEG C of -40s, 72 DEG C of -40s) × 45 are circulated, and 72 DEG C of -2min, are used
Instrument be ABI Step-One systems, last quantitative result is shown in Table lattice 3-4.
As a result show：Quality pit mud and the Lactobacillus acetotolerans average contents of pit mud inferior are present
Notable difference, and the Lactobacillus acetotolerans average contents of common pit mud fall between.
Table 3 determines Lactobacillus acetotolerans contents in 11 pit mud samples by primers of LA2
Table 4 is with LA2Lactobacillus acetotolerans contents in 24 pit mud samples are determined for primers
Table 1 is according to Lactobacillus acetotolerans whole genome sequence and background flora full-length genome sequence
Two pairs of Species-specific primers of the best results that row are designed and filtered out.
Target sequence corresponding to two pairs of primers of the best results of table 2.
Table 3 quantitative determines Lactobacillus in 11 pit mud samples using LA2 as primer using QPCR
Acetotolerans contents, as a result show that quality pit mud contains with Lactobacillus acetotolerans in pit mud inferior
Amount has notable difference.
Table 4 quantitative determines Lactobacillus in 24 pit samples using LA2 as primer using QPCR
Acetotolerans contents, show the average content of Lactobacillus acetotolerans in fermented grain apparently higher than cellar for storing things
Mud, but the assay of indivedual samples also brings along relatively large deviation due to the inhomogeneities of sampling.
Sequence table
<110>Harbin Institute of Technology（Weihai）
Anhui Rui Si Weirs Science and Technology Ltd.
<120>For detecting two pairs of specific primers of acidproof Bacillus acidi lactici content and its application during liquor fermentation
<160> 6
<170> SIPOSequenceListing 1.0
<210> 1
<211> 20
<212> DNA
<213>LA2 forward primer (2 Ambystoma laterale x Ambystoma jeffersonianum)
<400> 1
gtgcagctta agggcttcct 20
<210> 2
<211> 21
<212> DNA
<213>LA2 reverse primer (2 Ambystoma laterale x Ambystoma jeffersonianum)
<400> 2
atctgtgcag attgttccct t 21
<210> 3
<211> 20
<212> DNA
<213>LA3 forward primer (2 Ambystoma laterale x Ambystoma jeffersonianum)
<400> 3
agagtgtccc gagtagtccc 20
<210> 4
<211> 20
<212> DNA
<213>LA3 reverse primer (2 Ambystoma laterale x Ambystoma jeffersonianum)
<400> 4
aactgctcaa gaagggaccg 20
<210> 5
<211> 140
<212> DNA
<213>The extension increasing sequence (2 Ambystoma laterale x Ambystoma jeffersonianum) of LA2 primers
<400> 5
ctgtgcagct taagggcttc ctttactaaa cctttattca ttaatttagt cttgttttaa 60
attttctatc accagcgatt gacacagagc cacaataatt ccatcctgta aagttgttga 120
attaaccatt acgtcagaat 140
<210> 6
<211> 109
<212> DNA
<213>The extension increasing sequence (2 Ambystoma laterale x Ambystoma jeffersonianum) of LA3 primers
<400> 6
cttatcactc ttagtcaaat aattaagcag ggatgatttc ccaacattag gacgacccac 60
gattgcggtg gctaaaccat tacgtaagac ggtcccttct tgagcagtt 109
Claims (6)
- A kind of 1. two pairs of specific primers for being used to detect acidproof Bacillus acidi lactici content during liquor fermentation, it is characterised in that： The forward primer that two pairs of described specific primers include LA2 is shown in sequence 1, and LA2 reverse primer is shown in sequence 2；LA3 forward direction is drawn Thing is shown in sequence 3, and LA3 reverse primer is shown in sequence 4.
- 2. two pairs of specific primers for being used to detect acidproof Bacillus acidi lactici content during liquor fermentation described in right 1, its It is characterised by：The step of determining drinks spoilage organisms Lactobacillus acetotolerans contents is as follows：(1) the grand genome of sample is extracted, using genome as template, is expanded using two pairs of above-mentioned primers, then carries out agar Sugared gel electrophoresis experiment, is tentatively judged according to the position of electrophoretic band and light levels；(2) PCR (Polymerase Chain Reaction) product is sequenced, analysis result, verified and quantitative with this Weigh specific amplification of the above-mentioned primer in specific sample；(3) QPCR can be used to carry out drinks spoilage organisms Lactobacillus acetotolerans after specific amplification confirms It is quantitative.
- 3. two pairs of specific primers for detecting acidproof Bacillus acidi lactici content during liquor fermentation are used to quantitative determine drinks The application of spoilage organisms Lactobacillus acetotolerans contents.
- 4. application according to claim 3, it is characterised in that：To the dynamic change of the content of microorganisms in pit mud and fermented grain Process is measured and tracked.
- 5. two pairs of specific primers for detecting acidproof Bacillus acidi lactici content during liquor fermentation are used to distinguish quality pit mud With the application of common pit mud and pit mud inferior.
- 6. application according to claim 5, it is characterised in that：The pit mud inferior is than in quality pit mud Lactobacillus acetotolerans content is typically greater than 10 times, and the common pit mud is compared in quality pit mud Lactobacillus acetotolerans content is also high, but is no greater than 10 times.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710952009.6A CN107699610A (en) | 2017-10-13 | 2017-10-13 | For detecting two pairs of specific primers of acidproof Bacillus acidi lactici content and its application during liquor fermentation |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710952009.6A CN107699610A (en) | 2017-10-13 | 2017-10-13 | For detecting two pairs of specific primers of acidproof Bacillus acidi lactici content and its application during liquor fermentation |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN107699610A true CN107699610A (en) | 2018-02-16 |
Family
ID=61184103
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710952009.6A Pending CN107699610A (en) | 2017-10-13 | 2017-10-13 | For detecting two pairs of specific primers of acidproof Bacillus acidi lactici content and its application during liquor fermentation |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107699610A (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110499379A (en) * | 2019-09-18 | 2019-11-26 | 武汉轻工大学 | Primer, the method and kit for detecting Lactobacillus sp. |
| CN112226523A (en) * | 2020-06-01 | 2021-01-15 | 广州南沙珠江啤酒有限公司 | Specific detection probe and kit for acetic acid-resistant lactobacillus and application of specific detection probe and kit |
| CN112831546A (en) * | 2019-11-25 | 2021-05-25 | 中国食品发酵工业研究院有限公司 | Quantitative detection method for key functional microorganisms in yeast for making hard liquor |
| CN114292929A (en) * | 2021-11-30 | 2022-04-08 | 绍兴文理学院 | Molecular marker for quantitative lactobacillus acidophilus resistance and method for absolutely quantifying bacterial community composition in yellow wine fermentation process |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105331702A (en) * | 2015-11-05 | 2016-02-17 | 安徽瑞思威尔科技有限公司 | Method for fast and quantitatively measuring microbial community composition in fermentation process of baijiu |
| CN106222249A (en) * | 2016-07-14 | 2016-12-14 | 哈尔滨工业大学（威海） | The method for designing measuring the species-specific primer of known group information species in microbiologic population and the method measuring strain content |
| CN106906302A (en) * | 2017-04-27 | 2017-06-30 | 江南大学 | A kind of quick screening, identification and the method for quantifying lactobacillus acetotolerans in fermented food |
-
2017
- 2017-10-13 CN CN201710952009.6A patent/CN107699610A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105331702A (en) * | 2015-11-05 | 2016-02-17 | 安徽瑞思威尔科技有限公司 | Method for fast and quantitatively measuring microbial community composition in fermentation process of baijiu |
| CN106222249A (en) * | 2016-07-14 | 2016-12-14 | 哈尔滨工业大学（威海） | The method for designing measuring the species-specific primer of known group information species in microbiologic population and the method measuring strain content |
| CN106906302A (en) * | 2017-04-27 | 2017-06-30 | 江南大学 | A kind of quick screening, identification and the method for quantifying lactobacillus acetotolerans in fermented food |
Non-Patent Citations (5)
| Title |
|---|
| 于春涛: "不同产区浓香型白酒窖泥中细菌多样性分析", 《食品研究与开发》 * |
| 周恒刚编著: "《白酒生产指南》", 31 January 2000, 中国轻工业出版社 * |
| 张敬慧主编: "《酿酒微生物》", 31 January 2015, 中国轻工业出版社 * |
| 张文学等: "PCR技术对浓香型白酒糟醅细菌菌群的解析", 《四川大学学报（工程科学版）》 * |
| 黄丹等: "荧光定量PCR在研究白酒发酵糟醅优势菌中的应用", 《酿酒科技》 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110499379A (en) * | 2019-09-18 | 2019-11-26 | 武汉轻工大学 | Primer, the method and kit for detecting Lactobacillus sp. |
| CN112831546A (en) * | 2019-11-25 | 2021-05-25 | 中国食品发酵工业研究院有限公司 | Quantitative detection method for key functional microorganisms in yeast for making hard liquor |
| CN112226523A (en) * | 2020-06-01 | 2021-01-15 | 广州南沙珠江啤酒有限公司 | Specific detection probe and kit for acetic acid-resistant lactobacillus and application of specific detection probe and kit |
| CN114292929A (en) * | 2021-11-30 | 2022-04-08 | 绍兴文理学院 | Molecular marker for quantitative lactobacillus acidophilus resistance and method for absolutely quantifying bacterial community composition in yellow wine fermentation process |
| CN114292929B (en) * | 2021-11-30 | 2024-01-02 | 绍兴文理学院 | Molecular marker for quantifying lactobacillus-resistant bacteria and method for absolutely quantifying bacterial colony composition in yellow wine fermentation process |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Andorrà et al. | Effect of oenological practices on microbial populations using culture-independent techniques | |
| Song et al. | Analysis of microflora profile in Korean traditional nuruk | |
| Jansen et al. | Prolonged selection in aerobic, glucose-limited chemostat cultures of Saccharomyces cerevisiae causes a partial loss of glycolytic capacity | |
| Cocolin et al. | Direct profiling of the yeast dynamics in wine fermentations | |
| CN107699610A (en) | For detecting two pairs of specific primers of acidproof Bacillus acidi lactici content and its application during liquor fermentation | |
| Vegas et al. | Evaluation of representativity of the acetic acid bacteria species identified by culture-dependent method during a traditional wine vinegar production | |
| Choi et al. | Effect of low pH start-up on continuous mixed-culture lactic acid fermentation of dairy effluent | |
| CN111471794A (en) | Primer, probe, kit and method for RT-QPCR (reverse transcription-quantitative polymerase chain reaction) detection of fusarium graminearum | |
| Spano et al. | In vivo PCR-DGGE analysis of Lactobacillus plantarum and Oenococcus oeni populations in red wine | |
| CN105349690B (en) | method for monitoring fermentation process of solid-state brewed vinegar based on acid production index of microbial community | |
| CN111378774A (en) | Primer group, kit and method for rapidly detecting Listeria monocytogenes | |
| CN112226523B (en) | Specific detection probe and kit for acetic acid-resistant lactobacillus and application of specific detection probe and kit | |
| Yong et al. | Development of a specific real-time PCR assay targeting the poly-γ-glutamic acid synthesis gene, pgsB, for the quantification of Bacillus amyloliquefaciens in solid-state fermentation | |
| CN112779349A (en) | Probe, kit and application for absolute quantification of schizosaccharomyces pombe | |
| JP2007505638A (en) | Specific rapid detection method for beverage spoilage microorganisms | |
| KR101767744B1 (en) | A primer and probe set for specific and rapid quantitative detection of Lactobacillus kefiri and a real-time PCR method using the same | |
| Khullar et al. | Contamination issues in a continuous ethanol production corn wet milling facility | |
| CN112899382B (en) | Detection method for identifying amycolatopsis | |
| JP3338924B2 (en) | Oligonucleotide for detecting lactic acid bacteria and determination method using the same | |
| Delaherche et al. | Intraspecific diversity of Oenococcus oeni strains determined by sequence analysis of target genes | |
| CN114317808A (en) | Quantitative detection kit, detection method and application of thermophilic fungi | |
| CN112608985A (en) | Primer and method for quickly identifying and quantifying schizosaccharomyces pombe | |
| CN102433385B (en) | Primers for identifying leuconostoc mesenteroides and application thereof | |
| Pribadhi et al. | Identification of Gaba-Producing Lactic Acid Bacteria From Cincalok Fermentation Based on the 16S Rrna and Gad Genes | |
| CN110578014A (en) | Kit and method for detecting candida parapsilosis nucleic acid |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20180216 |
|
| RJ01 | Rejection of invention patent application after publication |