CN112094746A - Isolated culture method of caproic acid bacteria in pit mud - Google Patents
Isolated culture method of caproic acid bacteria in pit mud Download PDFInfo
- Publication number
- CN112094746A CN112094746A CN202010950504.5A CN202010950504A CN112094746A CN 112094746 A CN112094746 A CN 112094746A CN 202010950504 A CN202010950504 A CN 202010950504A CN 112094746 A CN112094746 A CN 112094746A
- Authority
- CN
- China
- Prior art keywords
- caproic acid
- pit mud
- acid bacteria
- electric field
- culture
- 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
Images
Classifications
-
- 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
- C12N1/02—Separating microorganisms from their culture media
-
- 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
- C12N13/00—Treatment of microorganisms or enzymes with electrical or wave energy, e.g. magnetism, sonic waves
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Zoology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Organic Chemistry (AREA)
- Genetics & Genomics (AREA)
- Wood Science & Technology (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Biomedical Technology (AREA)
- Medicinal Chemistry (AREA)
- Tropical Medicine & Parasitology (AREA)
- Virology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
Abstract
The invention provides a separation culture method of caproic acid bacteria in pit mud, which comprises the steps of enrichment culture by using an RCM liquid culture medium, and then applying an external electric field to separate and culture the caproic acid bacteria in the pit mud of strong aromatic white spirit, shortens the acquisition time of the caproic acid bacteria, strengthens the caproic acid production capacity of the caproic acid bacteria, can meet the separation culture requirements of microorganisms in the research directions of white spirit brewing, vinegar brewing, fermented food and the like, and has the characteristic of simplicity, rapidness and high efficiency, the separation culture time of the caproic acid bacteria is greatly shortened, the production efficiency is improved, the aging of the pit mud is accelerated, and the high-quality wine yield of a manufacturer is improved.
Description
Technical Field
The invention relates to a separation culture method of microorganisms for wine making, in particular to a separation culture method of caproic acid bacteria in pit mud.
Background
China has a long wine culture, white spirit dominates in wine and beverage, strong aromatic white spirit accounts for over 70% of the white spirit market, and the quality of the strong aromatic white spirit is closely related to the quality of pit mud. A large amount of pit mud is needed for building a new pit and maintaining an old pit. The quality of the pit mud seriously influences the quality of the strong aromatic Chinese spirits. Through the research on pit mud microorganisms for many years, the artificial pit mud technology has developed to a certain extent. In the present stage, the pit mud manufacturing technology with a good application effect is to add various enhanced functional bacteria into the pit mud in the pit mud manufacturing process, so that the aims of accumulation of aroma substances produced in the pit mud and enrichment of beneficial floras are achieved, and simultaneously, the aging of the pit mud is accelerated. The caproic acid bacteria in the pit mud are microorganisms for producing flavor substance ethyl caproate in the strong aromatic white spirit, so the caproic acid bacteria are the most commonly used added bacterial strains in the pit mud manufacturing process. In order to obtain high-quality addable caproic acid bacteria, the best method is to screen from pit mud. Most of microorganisms in pit mud are non-culturable microorganisms, so that the screening of high-quality caproic acid bacteria is very difficult, and the traditional caproic acid bacteria obtaining method is long in time.
The traditional caproic acid bacteria are obtained by the following method:
1. preparation of pit mud suspension
Weighing pit mud, adding the pit mud into a triangular flask containing sterile normal saline and sterile glass beads, and oscillating at 37 ℃ and 220r/min for 30-60min to obtain pit mud suspension;
2. isolation culture of pit mud caproic acid bacteria
Killing the nutritive somatic cells by pit mud suspension in water bath at 80 ℃ for 10min, then inoculating the pit mud suspension into an RCM liquid culture medium, and carrying out enrichment culture in an anaerobic culture box at 37 ℃ for 4-6 days;
respectively sucking 10 liquid culture mediums after enrichment culture-3～10-5Coating 100 mu L of the diluted bacterial suspension on a corresponding RCM solid culture medium plate, coating three plates on each dilution, placing the inoculated plate in an anaerobic incubator, and culturing for 4 days at 37 ℃;
the colonies with various characteristics are subjected to plate streaking purification for many times, and pure caproic acid bacteria can be obtained after three rounds of purification for about 30 days.
The time required for obtaining the caproic acid bacteria by adopting the traditional method is about 1-2 months, and the caproic acid yield of the screened caproic acid bacteria is about 11-13 mg/mL.
With the development of economic society of China, the demand of high-end white spirit is more and more, and further, the demand of high-quality pit mud is increased. The method for accelerating the aging of pit mud mainly comprises the step of adding artificially cultured caproic acid bacteria into new pit mud, so that the screening of high-quality caproic acid bacteria is the key for manufacturing high-quality pit mud. How to quickly obtain the pit mud becomes urgent in the field of brewing the strong aromatic Chinese spirits by using caproic acid bacteria.
Disclosure of Invention
The invention aims to solve the technical problem of providing a method for separating and culturing caproic acid bacteria in pit mud.
In order to solve the technical problems, the technical scheme of the invention is as follows:
a separation culture method of caproic acid bacteria in pit mud utilizes an electric field auxiliary screening technology to separate and culture the pit mud caproic acid bacteria, and comprises the following specific steps:
(1) preparation of pit mud suspension
Weighing a proper amount of pit mud, adding the pit mud into a triangular flask containing 50mL of sterile normal saline and 20 sterile glass beads (phi is 3mm), and oscillating the mixture for 30min at 37 ℃ at 220 r/min;
(2) enrichment culture of pit mud caproic acid bacteria
Inoculating pit mud suspension with volume fraction of 3-5% into 300mL of RCM liquid culture medium, and performing enrichment culture in an anaerobic incubator at 37 ℃ for 4 days;
(3) electric field auxiliary primary screen
Pouring 200mL of liquid culture medium after enrichment culture into an electric field auxiliary screening device, loading 5-15V voltage between two electrode plates, observing that a small amount of fine foam is generated at a negative plate after 1-4h, and observing obvious substance adhesion on the negative plate;
(4) electric field auxiliary re-screening: eluting the colony gathered on the electrode plate into a fresh 200mL RCM liquid culture medium by using a sterile cotton swab, carrying out anaerobic culture for 4 days, and then screening again;
(5) repeating the steps (3) and (4) until obtaining pure caproic acid bacterial strain, and preserving the bacterial strain in RCM solid culture medium for standby.
The caproic acid bacteria obtained by the separation culture method of the caproic acid bacteria in the pit mud takes about 12 to 30 days, and the caproic acid bacteria obtained by screening has caproic acid yield of about 15 to 18 mg/mL.
Preferably, in the method for isolating and culturing caproic acid bacteria in pit mud, the Reinforced Clostridium (RCM) liquid medium: 10g/L of peptone, 10g/L of beef extract, 3g/L of yeast extract, 5g/L of glucose, 1g/L of soluble starch, 5g/L of sodium chloride, 3g/L of sodium acetate and 0.5g/L of cysteine hydrochloride, sterilizing at 121 ℃ for 20min, and naturally adjusting pH; RCM solid medium: to the RCM liquid medium was added 2% agar.
Preferably, in the isolated culture method of caproic acid bacteria in pit mud, the electric field auxiliary screening device is a microorganism culture device with platinum electrode plates at two ends, and is externally connected with a direct current power supply, so that the voltage of the direct current power supply can be changed, and the change of experimental parameters within the range of 3-15V is realized.
Preferably, the isolated culture method of caproic acid bacteria in the above-mentioned cellar mud, the auxiliary screening device of electric field is in application number 201721435531.9 the microorganism sieving mechanism, the auxiliary screening device of electric field includes negative pole groove, anode tank, connects the platinum plate electrode of liquid bridge and a pair of external DC power supply, negative pole groove, anode tank and connection liquid bridge divide into the triplex with this sieving mechanism, connect the liquid bridge and arrange negative pole groove and anode tank middle part in, the waters degree of depth that should connect the liquid bridge top is less than the waters degree of depth of negative pole groove and anode tank, the negative plate of a pair of platinum plate electrode is arranged in the negative pole inslot, the anode plate is arranged in the anode tank.
Preferably, in the method for isolated culture of caproic acid bacteria in pit mud, the specific operation process of the electric field auxiliary screening device is as follows:
(1) inoculating the enriched culture solution into an electric field auxiliary screening device (cathode tank, anode tank, connecting liquid bridge) containing 200ml RCM liquid culture medium at an inoculation amount of 10%;
(2) placing the electric field auxiliary screening tank in an anaerobic incubator at 37 ℃, switching on power supplies at two ends of the electric field auxiliary screening tank to generate a stable electric field between the cathode platinum electrode plate and the anode platinum electrode plate, and adjusting the electric field intensity by changing the external voltage;
(3) the boosting procedure is as follows: keeping the initial 5-10V for 0.5-1h, adjusting to 10-12V for 0.5-1h, then increasing to 12-15V until thallus appears on the electrode plate, and the culture solution around the electrode plate becomes turbid.
Has the advantages that:
the isolation culture method of the caproic acid bacteria in the pit mud is characterized in that RCM liquid culture medium is used for enrichment culture, then an external electric field is applied, the caproic acid bacteria in the pit mud of the strong aromatic white spirit can be isolated and cultured, the method shortens the acquisition time of the caproic acid bacteria, strengthens the caproic acid production capacity of the caproic acid bacteria, can meet the culture requirements of microorganisms in the research directions of white spirit brewing, vinegar brewing, fermented food and the like, and the caproic acid bacteria caproic acid yield obtained by the method can reach 15-18 mg/mL.
Drawings
FIG. 1 is a schematic structural diagram of a microorganism screening device in the pit mud caproic acid bacterium separation and culture method.
In the figure: 1-container cover 2-cathode tank 3-anode tank 4-connecting liquid bridge 5-1-cathode plate 5-2-anode plate
Detailed Description
An electric field auxiliary screening device used in the following embodiment is shown in fig. 1, and comprises a container cover 1, a cathode tank 2 (cathode microorganism culture area), an anode tank 3 (anode microorganism culture area), a connecting liquid bridge 4 and a pair of platinum electrode plates externally connected with a direct current power supply, wherein the direct current power supply can linearly adjust voltage and current (ensure that the voltage between the two electrodes of the electrode plates is adjustable), the screening device is divided into three parts by the cathode tank, the anode tank and the connecting liquid bridge, the connecting liquid bridge is arranged in the middle of the cathode tank and the anode tank (the connecting liquid bridge is a shallow water area connecting the cathode tank and the anode tank), the depth of the water area above the connecting liquid bridge is smaller than that of the cathode tank and the anode tank, the container cover covers the cathode tank, the anode tank and the connecting liquid bridge, through holes are arranged on the container cover, and a pull rod is connected on the platinum electrode plates, be provided with the sealed pad that has locate function on the pull rod, the pull rod is worn out in by the through-hole of container lid, sealed pad can follow the pull rod and slide from top to bottom and with the through-hole overlap joint is fixed fixes and fixes a position, and then adjusts the electrode plate and insert the degree of depth in cathode trough and/or anode tank, the cathode plate 5-1 of a pair of platinum electrode plate is arranged in the cathode trough in, the anode plate 5-2 is arranged in the anode tank in, the platinum electrode plate is detachable structure, just the degree of depth that the platinum electrode plate deepened the culture solution is linear adjustable, and the microorganism of screening is at the cathodic region enrichment growth, finally collects on the cathode plate and obtains the target bacterial strain, cathode trough and anode tank can be used for cultivateing the microorganism simultaneously.
Example 1
Screening caproic acid bacteria from 5-age pit mud of Sichuan certain winery:
(1) preparation of pit mud suspension
Weighing 2.5g of pit mud, adding into a triangular flask containing 50mL of sterile physiological saline and 20 particles of sterile glass beads (phi is 3mm), and shaking at 37 ℃ and 220r/min for 30 min;
(2) enrichment culture of pit mud caproic acid bacteria
Inoculating pit mud suspension with volume fraction of 3% into RCM liquid culture medium of 300mL, and performing enrichment culture in an anaerobic incubator at 37 ℃ for 4 days.
(3) Electric field auxiliary primary screen
Pouring 200mL of liquid culture medium after enrichment culture into an electric field auxiliary screening device, loading 8V voltage between two electrode plates, changing the loading voltage to 12V after 0.5h of culture, changing the loading voltage to 14V after 1h of culture, and observing that obvious substances are attached to a negative plate to stop loading voltage;
(4) electric field auxiliary re-screening: eluting the colony gathered on the electrode plate into a fresh 200mL RCM liquid culture medium by using a sterile cotton swab, placing the colony in an anaerobic incubator for 4 days at 37 ℃, and operating according to the step (3);
(5) repeating the steps (3) and (4) for 3 times, performing microscopic examination on the strains washed off from the negative electrode plate to obtain pure strains, and preserving the strains for later use. The screening was carried out for a total of 20 days.
(6) Identification and investigation of strains: and identifying the separated strain by adopting a 16S rDNA sequencing technology, wherein the identification result is Clostridium sp. Under the optimized fermentation condition, the caproic acid content in the fermentation liquid of the strain reaches 16.24 mg/mL.
(7) The method for measuring the caproic acid in the fermentation liquor comprises the following steps: and (3) adopting GC-FID for analysis, centrifuging the fermentation liquor at 12000r/min for 10min to remove thalli, filtering the supernatant through a 0.22 mu m filter membrane, and collecting the supernatant into a sample injection bottle. The gas phase method comprises the following steps: DB-WAX capillary column (30m × 250 μm × 0.25 μm), sample size of 1 μ L, split ratio of 6: 1, carrying nitrogen gas at a rate of 1mL/min, maintaining the temperature of the column oven at 50 ℃ for 2min, and then increasing the temperature to 230 ℃ at a rate of 15 ℃/min for 10 min.
Example 2
Screening caproic acid bacteria from 15-age pit mud of Sichuan certain winery:
(1) preparation of pit mud suspension
Weighing 2.5g of pit mud, adding into a triangular flask containing 50mL of sterile physiological saline and 20 particles of sterile glass beads (phi is 3mm), and shaking at 37 ℃ and 220r/min for 30 min;
(2) enrichment culture of pit mud caproic acid bacteria
Inoculating pit mud suspension with volume fraction of 3% into RCM liquid culture medium of 300mL, and performing enrichment culture in an anaerobic incubator at 37 ℃ for 4 days.
(3) Electric field auxiliary primary screen
Pouring 200mL of liquid culture medium after enrichment culture into an electric field auxiliary screening device, loading 6V voltage between two electrode plates, changing the loading voltage to 10V after 0.5h of culture, changing the loading voltage to 13V after 0.5h of culture, and observing obvious substance adhesion on a negative plate to stop loading voltage;
(4) electric field auxiliary re-screening: eluting the colony gathered on the electrode plate into a fresh 200mL RCM liquid culture medium by using a sterile cotton swab, placing the colony in an anaerobic incubator for 6 days at 37 ℃, and operating according to the step (3);
(5) repeating the steps (3) and (4) for 2 times, performing microscopic examination on the strains washed off from the negative electrode plate to obtain pure strains, and preserving the strains for later use. The screening takes 12 days.
(6) Identification and investigation of strains: and identifying the separated strain by adopting a 16S rDNA sequencing technology, wherein the identification result is Clostridium sp. Under the optimized fermentation condition, the caproic acid content in the fermentation liquid of the strain reaches 18.33 mg/mL.
(7) The method for measuring hexanoic acid in the fermentation broth was the same as described in example 1.
The electric field auxiliary screening principle of the electric field auxiliary screening device is as follows: the low redox potential of the growth environment is maintained by the electron transfer process in the respiratory chain in the growth process of anaerobic caproic acid bacteria, in the process, the organisms of the caproic acid bacteria need electrons, and a large amount of electrons are provided on the cathode electrode plate for the utilization of the electrons, so that sufficient nutrition is provided for the growth of the caproic acid bacteria, the caproic acid bacteria near the cathode can grow rapidly, the growth of anaerobic strains is facilitated under the low redox potential, and the caproic acid bacteria can be obtained by the method. And the caproic acid bacteria are enriched through repeated rounds of caproic acid bacteria, and finally the high-activity caproic acid bacteria are obtained.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.
Claims (5)
1. A separation culture method of caproic acid bacteria in pit mud is characterized in that: the pit mud caproic acid bacteria is isolated and cultured by using an electric field auxiliary screening technology, and the method comprises the following specific steps:
(1) preparation of pit mud suspension
Weighing a proper amount of pit mud, adding the pit mud into a triangular flask filled with sterile normal saline and sterile glass beads, and oscillating the mixture for 30min at the temperature of 37 ℃ at the speed of 220 r/min;
(2) enrichment culture of pit mud caproic acid bacteria
Inoculating pit mud suspension with volume fraction of 3% into 300mL of RCM liquid culture medium, and performing enrichment culture in an anaerobic incubator at 37 ℃ for 4 days;
(3) electric field auxiliary primary screen
Pouring 200mL of liquid culture medium after enrichment culture into an electric field auxiliary screening device, loading 5-15V voltage between two electrode plates, observing that a small amount of fine foam is generated at a negative plate after 1-4h, and observing obvious substance adhesion on the negative plate;
(4) electric field auxiliary re-screening: eluting the colony gathered on the electrode plate into a fresh 200mL RCM liquid culture medium by using a sterile cotton swab, carrying out anaerobic culture for 4 days, and then screening again;
(5) repeating the steps (3) and (4) until obtaining pure caproic acid bacterial strain, and preserving the bacterial strain in RCM solid culture medium for standby.
2. The method for separating and culturing caproic acid bacteria in pit mud according to claim 1, characterized in that: the RCM liquid culture medium: 10g/L of peptone, 10g/L of beef extract, 3g/L of yeast extract, 5g/L of glucose, 1g/L of soluble starch, 5g/L of sodium chloride, 3g/L of sodium acetate and 0.5g/L of cysteine hydrochloride, sterilizing at 121 ℃ for 20min, and naturally adjusting pH; RCM solid medium: to the RCM liquid medium was added 2% agar.
3. The method for separating and culturing caproic acid bacteria in pit mud according to claim 1, characterized in that: the electric field auxiliary screening device is a microorganism culture device with platinum electrode plates at two ends, is externally connected with a direct current power supply, can change the voltage of the direct current power supply, and realizes the change of experimental parameters within the range of 3-15V.
4. The method for separating and culturing caproic acid bacteria in pit mud according to claim 3, characterized in that: the electric field auxiliary screening device comprises a cathode tank, an anode tank, a connecting liquid bridge and a pair of platinum electrode plates externally connected with a direct current power supply, wherein the cathode tank, the anode tank and the connecting liquid bridge divide the screening device into three parts, the connecting liquid bridge is arranged in the middle of the cathode tank and the anode tank, the water area depth above the connecting liquid bridge is smaller than that of the cathode tank and the anode tank, and the cathode plate of the pair of platinum electrode plates is arranged in the cathode tank and the anode plate of the pair of platinum electrode plates is arranged in the anode tank.
5. The method for separating and culturing caproic acid bacteria in pit mud according to claim 1, characterized in that: the specific operation process of the electric field auxiliary screening device is as follows:
(1) inoculating the enriched culture solution into an electric field auxiliary screening device containing 200ml RCM liquid culture medium in an inoculation amount of 10%;
(2) placing the electric field auxiliary screening tank in an anaerobic incubator at 37 ℃, switching on power supplies at two ends of the electric field auxiliary screening tank to generate a stable electric field between the cathode platinum electrode plate and the anode platinum electrode plate, and adjusting the electric field intensity by changing the external voltage;
(3) the boosting procedure is as follows: keeping the initial 5-10V for 0.5-1h, adjusting to 10-12V for 0.5-1h, then increasing to 12-15V until thallus appears on the electrode plate, and the culture solution around the electrode plate becomes turbid.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2020109261619 | 2020-09-07 | ||
| CN202010926161 | 2020-09-07 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN112094746A true CN112094746A (en) | 2020-12-18 |
Family
ID=73750846
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010950504.5A Pending CN112094746A (en) | 2020-09-07 | 2020-09-11 | Isolated culture method of caproic acid bacteria in pit mud |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN112094746A (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112812997A (en) * | 2021-01-12 | 2021-05-18 | 江南大学 | Method for quickly enriching glucose-utilizing caproic acid-producing flora from pit mud and application |
| CN113106032A (en) * | 2021-03-17 | 2021-07-13 | 湖北工业大学 | Culture medium for screening glucose-inhibited acetohexogen and application thereof |
| CN113151061A (en) * | 2021-03-24 | 2021-07-23 | 湖北工业大学 | Glucose-inhibited oxytoca |
| CN113186121A (en) * | 2021-04-06 | 2021-07-30 | 江南大学 | Caproic acid bacteria capable of utilizing various substrates and application thereof |
| CN113502219A (en) * | 2021-05-22 | 2021-10-15 | 山东景阳冈酒厂有限公司 | Screening and identifying device for high-quality pit mud high-yield caproic acid bacteria |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104651440A (en) * | 2015-01-14 | 2015-05-27 | 同济大学 | Biological extension method for electrically promoted carbon chain and biological extension device for electrically promoted carbon chain |
| CN107619826A (en) * | 2017-11-01 | 2018-01-23 | 天津科技大学 | A kind of biodegradable screening technique for using microorganism |
| CN207552340U (en) * | 2017-11-01 | 2018-06-29 | 天津科技大学 | A kind of microbe to screen device |
| CN109971686A (en) * | 2019-04-29 | 2019-07-05 | 四川轻化工大学 | A kind of caproic acid bacteria proliferated culture medium, caproic acid fermentation and caproic acid bacterial screening method |
| CN111321082A (en) * | 2020-03-21 | 2020-06-23 | 湖北古襄阳酒业有限公司 | Pit mud microorganism separation method |
-
2020
- 2020-09-11 CN CN202010950504.5A patent/CN112094746A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104651440A (en) * | 2015-01-14 | 2015-05-27 | 同济大学 | Biological extension method for electrically promoted carbon chain and biological extension device for electrically promoted carbon chain |
| CN107619826A (en) * | 2017-11-01 | 2018-01-23 | 天津科技大学 | A kind of biodegradable screening technique for using microorganism |
| CN207552340U (en) * | 2017-11-01 | 2018-06-29 | 天津科技大学 | A kind of microbe to screen device |
| CN109971686A (en) * | 2019-04-29 | 2019-07-05 | 四川轻化工大学 | A kind of caproic acid bacteria proliferated culture medium, caproic acid fermentation and caproic acid bacterial screening method |
| CN111321082A (en) * | 2020-03-21 | 2020-06-23 | 湖北古襄阳酒业有限公司 | Pit mud microorganism separation method |
Non-Patent Citations (2)
| Title |
|---|
| 姚继承，等: "影响己酸菌生长代谢的因素与培养应用", 《酿酒科技》 * |
| 张鹏程，等: "阴极大小对微生物电合成系统还原二氧化碳产有机物的影响", 《环境工程学报》 * |
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112812997A (en) * | 2021-01-12 | 2021-05-18 | 江南大学 | Method for quickly enriching glucose-utilizing caproic acid-producing flora from pit mud and application |
| CN112812997B (en) * | 2021-01-12 | 2023-04-28 | 江南大学 | Method for rapidly enriching glucose utilization type caproic acid-producing flora from pit mud and application of method |
| CN113106032A (en) * | 2021-03-17 | 2021-07-13 | 湖北工业大学 | Culture medium for screening glucose-inhibited acetohexogen and application thereof |
| CN113151061A (en) * | 2021-03-24 | 2021-07-23 | 湖北工业大学 | Glucose-inhibited oxytoca |
| CN113151061B (en) * | 2021-03-24 | 2022-06-17 | 湖北工业大学 | Glucose-inhibited oxytoca |
| CN113186121A (en) * | 2021-04-06 | 2021-07-30 | 江南大学 | Caproic acid bacteria capable of utilizing various substrates and application thereof |
| WO2022213898A1 (en) * | 2021-04-06 | 2022-10-13 | 江南大学 | Clostridium butyricum capable of utilizing various substrates and use thereof |
| CN113502219A (en) * | 2021-05-22 | 2021-10-15 | 山东景阳冈酒厂有限公司 | Screening and identifying device for high-quality pit mud high-yield caproic acid bacteria |
| CN113502219B (en) * | 2021-05-22 | 2022-09-06 | 山东景阳冈酒厂有限公司 | Screening and identifying device for high-quality pit mud high-yield caproic acid bacteria |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN112094746A (en) | Isolated culture method of caproic acid bacteria in pit mud | |
| CN106939288B (en) | Application of lactobacillus plantarum SG5 in production of gamma-aminobutyric acid | |
| CN111378605B (en) | Lactobacillus plantarum for biological deacidification of high-yield volatile ester compounds and application of lactobacillus plantarum in fruit wine | |
| CN103087871A (en) | Brewing method of aged perry, and brewing method of pear brandy | |
| CN110093285B (en) | Acid-resistant lactobacillus fermentum and application thereof | |
| CN110564580B (en) | Method for producing vinegar containing pyrroloquinoline quinone through microbial co-culture fermentation | |
| CN108179119B (en) | Fermentation process for improving aroma quality of ice wine by using non-saccharomyces cerevisiae | |
| CN113607793A (en) | Method for constructing biological membrane catalytic electrode with high activity | |
| CN107858292B (en) | Antrodia camphorata strain and preservation method thereof | |
| CN114250157B (en) | Saccharomycopsis fibuligera cx-3 strain for high yield of aroma substances and application thereof | |
| CN113957016B (en) | Bacillus subtilis and method for preparing milk-flavored cordyceps sinensis fermentation liquor by using same | |
| CN112812997B (en) | Method for rapidly enriching glucose utilization type caproic acid-producing flora from pit mud and application of method | |
| JPWO2017217116A1 (en) | New microalgae with aggregation ability | |
| CN113930368A (en) | Bacillus vallismortis and application thereof in vinegar brewing | |
| Maicas et al. | Malolactic fermentation in wine with high densities of non-proliferating Oenococcus oeni | |
| CN110591852B (en) | Application and method of eurotium scherzeri in microbial rejuvenation of pit mud of white spirit cellar | |
| RU2069229C1 (en) | Method of $$$-iron preparing | |
| CN113502233B (en) | Mei Ji Yeast and application thereof in wine brewing | |
| CN113151061B (en) | Glucose-inhibited oxytoca | |
| CN117431189B (en) | Lactobacillus paracasei subspecies paracasei strain QH-20029 and application thereof | |
| CN110846242A (en) | Lactobacillus hilgardii strain and application thereof | |
| CN112522122B (en) | Fruit wine deacidification strain and application thereof | |
| CN113773977B (en) | Yeast strain with low ethanol yield and high aroma yield and application thereof | |
| CN115820470B (en) | Bacillus amyloliquefaciens ZH804 and application thereof | |
| CN112625963B (en) | Clostridium sarmentosum capable of producing 4-methylphenol and application thereof |
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 | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20201218 |
|
| WD01 | Invention patent application deemed withdrawn after publication |