CN117511755B - Saccharomycetes tectorial membrane yeast and application thereof - Google Patents

Saccharomycetes tectorial membrane yeast and application thereof Download PDF

Info

Publication number
CN117511755B
CN117511755B CN202311385212.1A CN202311385212A CN117511755B CN 117511755 B CN117511755 B CN 117511755B CN 202311385212 A CN202311385212 A CN 202311385212A CN 117511755 B CN117511755 B CN 117511755B
Authority
CN
China
Prior art keywords
yeast
strain
saccule
application
microorganism
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.)
Active
Application number
CN202311385212.1A
Other languages
Chinese (zh)
Other versions
CN117511755A (en
Inventor
罗小叶
李琳玲
刘婉琳
邱树毅
王春晓
王晓丹
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Guizhou University
Original Assignee
Guizhou University
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by Guizhou University filed Critical Guizhou University
Priority to CN202311385212.1A priority Critical patent/CN117511755B/en
Publication of CN117511755A publication Critical patent/CN117511755A/en
Application granted granted Critical
Publication of CN117511755B publication Critical patent/CN117511755B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

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
    • C12N1/14—Fungi; Culture media therefor
    • C12N1/16—Yeasts; Culture media therefor
    • C12N1/165—Yeast isolates
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12G—WINE; PREPARATION THEREOF; ALCOHOLIC BEVERAGES; PREPARATION OF ALCOHOLIC BEVERAGES NOT PROVIDED FOR IN SUBCLASSES C12C OR C12H
    • C12G1/00—Preparation of wine or sparkling wine
    • C12G1/02—Preparation of must from grapes; Must treatment and fermentation
    • C12G1/0203—Preparation of must from grapes; Must treatment and fermentation by microbiological or enzymatic treatment
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P1/00—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes
    • C12P1/02—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes by using fungi
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12G—WINE; PREPARATION THEREOF; ALCOHOLIC BEVERAGES; PREPARATION OF ALCOHOLIC BEVERAGES NOT PROVIDED FOR IN SUBCLASSES C12C OR C12H
    • C12G2200/00—Special features
    • C12G2200/05—Use of particular microorganisms in the preparation of wine
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/645—Fungi ; Processes using fungi

Landscapes

  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Health & Medical Sciences (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Genetics & Genomics (AREA)
  • Mycology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Biotechnology (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Engineering & Computer Science (AREA)
  • Microbiology (AREA)
  • Medicinal Chemistry (AREA)
  • Botany (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Biomedical Technology (AREA)
  • Virology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Molecular Biology (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Abstract

The invention discloses a saccule-buckling compound film yeast (Saccharomycopsis fibuligera) FBKL2.8DCJS1 which is characterized in that the preservation number is CCTCC NO . M2020891; and the application of the saccule-covered yeast in producing acid, ester and alcohol substances. The saccule-covered yeast has wide application range to natural environment conditions such as temperature, pH and the like, has strong tolerance to bad environment, and the microorganism can decompose and structurally change fermentation raw materials on the molecular level through the metabolism of the microorganism to generate a plurality of metabolites favorable for organisms.

Description

Saccharomycetes tectorial membrane yeast and application thereof
Technical Field
The invention belongs to the technical field of biology, and particularly relates to a saccule-covering membrane yeast, and further relates to functional application of substances such as acid production, alcohols, esters and the like.
Technical Field
The saccharomycete, which is the most important microorganism in the wine brewing process, has important influence on the quality and style of the wine, and along with the rapid development of the wine industry in China, the development and utilization of the saccharomyces cerevisiae resources in China are improved. The current use of commercial strains in large numbers results in wines that tend to show a phenomenon of homogeneity and a monotonous flavour. Therefore, in order to fully utilize wild yeast resources, the wild yeast strains with special functions are screened from vineyard soil, grape berries and grape mash, so that the special grape wine is brewed, and the method has great significance to the middle grape wine industry.
The brewing function of yeast mainly produces alcohols and esters, namely, the produced alcohols, except ethanol, produce higher alcohols such as isoamyl alcohol, isobutanol, phenethyl alcohol and the like, and esters such as ethyl acetate, ethyl octanoate, ethyl palmitate and the like, thus endowing various fermented fruit wines with certain charm. According to the prior reports, pichia kudriavzevii, saccharomyces cerevisiae, hansenula debaryomyces, schizosaccharomyces pombe and the like all have the capability of producing esters and alcohols, and currently, yeasts with acid production performance in wine brewing are reported only recently, and especially, the Saccharomyces cerevisiae has only been reported to study the esters and alcohols. Therefore, the separation and screening of yeast strain resources with special acid-producing substances, alcohol substances and ester substances has important significance for brewing fruit wine.
Disclosure of Invention
The invention aims to provide a saccule-buckling compound film yeast.
It is another object of the present invention to provide the use of the yeast in acids, esters and alcohols.
The invention relates to a yeast, which has the name: oocyst composite yeast (Saccharomycopsis fibuligera), strain number FBKL2.8DCJS1, its morphological characteristics: inoculating the strain on YPD agar medium by streaking, culturing for 2d at 28 ℃, wherein the diameter of the bacterial colony is 6-9 mm, the bacterial colony is round , the color is off-white, the texture is dry and is villous, the bacterial colony edge is blanket-shaped, the back of the bacterial colony is yellow, and the strain is preserved in China center for type culture collection (China) at 12 months and 11 days in 2020: university of martial arts, deposit number: cctccc N O M2020891.
The invention discloses separation of a saccule-covered yeast (Saccharomycopsis fibuligera), which comprises the following steps:
separation of the oocyst membrane yeasts (Saccharomycopsis fibuligera):
Taking 10g of broken grape sample from the epidermis of a Guizhou wild grape sample, adding the broken grape sample into a triangular flask filled with 90ml of sterile physiological saline and glass beads, oscillating for 30min at a temperature of 28 ℃ by a shaking table, and uniformly mixing; sucking 1mL of bacterial suspension in an ultra-clean workbench, injecting the bacterial suspension into a test tube filled with 9mL of sterile physiological saline, carrying out gradient dilution, and taking a gradient of 10 -1、10-2、10-3、10-4、10-5 for pouring and coating a flat plate respectively; after inverted culturing at 28 ℃ for 2d, counting, selecting single colony with typical yeast colony morphology, inoculating and streaking on YPD agar medium, repeating for 3-4 times until pure strain is obtained, then numbering each strain respectively, and finally streaking on inclined plane for low-temperature preservation.
(II) culture medium:
Identification medium: bengalia culture medium, peptone 5g, glucose 10g, potassium dihydrogen phosphate 1g, magnesium sulfate (MgSO4.7H2O) 0.5g, agar 20g, chloramphenicol 0.1g,1/3000 Bengalia solution 100mL, distilled water 1000mL. Preservation medium: wort agar medium, malt extract powder 130g/L, chloramphenicol 0.1g/L, agar 15/L, pH5.8-6.2. Seed culture medium: glucose 1g, yeast extract 0.5g, peptone 1g, distilled water 50mL, and sterilization at 121℃for 20min.
Liquid fermentation medium: 10g of glucose, 0.5g of yeast extract, 1g of peptone, 50mL of distilled water and sterilizing at 115 ℃ for 20min.
And (III) seed conservation and passage of the strain:
1. Strain preservation culture medium:
(1) Wort agar medium: 130g/L malt extract powder, 0.1g/L chloramphenicol, 15g/L agar, pH5.8-6.2, and sterilizing at 115deg.C for 20min;
(2) YPD medium: 10g of yeast extract, 20g of peptone, 20g of glucose, 20g of agar, 1000ml of distilled water, pH6.0 and sterilization at 115 ℃ for 20min.
2. The preservation method comprises 4 steps:
(1) The preservation method of the subculture yeast strains comprises the following steps: inoculating yeast on malt wort agar slant culture medium, culturing at 28deg.C for 2d, sealing rubber stopper with sterilized solid paraffin, transferring to 4deg.C refrigerator, and storing in 4deg.C refrigerator for one month, and periodically transplanting.
(2) The preservation method of the sand tube yeast strain comprises the following steps: taking clean sea sand, washing with 1mol/L sodium hydroxide solution, washing with 1mol/L hydrochloric acid solution, washing with running water, subpackaging into test tubes, sterilizing at a depth of about 2-3 cm by dry heat, adding about 1mL of yeast culture solution, uniformly mixing with sand, drying in a vacuum drying tube, and sealing for preservation after complete drying;
(3) Vacuum freeze drying yeast strain preservation method: suspending microbial cells or spores to be preserved in a suitable protective agent, freezing the cells in advance, freezing the water, and then sublimating the ice under vacuum to remove most of the water, some of the remaining non-frozen water, and removing the water from the cells by evaporation;
(4) Liquid paraffin method: the method is also called as mineral oil preservation method, which is a seed preservation method of inoculating yeast seed on a proper slant culture medium, culturing under the optimum condition until robust thalli are obtained, injecting sterilized liquid paraffin to cover the whole slant, and standing at a drying place with the temperature of 4-6 ℃ for preservation, and is an auxiliary method of periodic transplanting preservation method.
Ecological characteristics of strain (IV):
The bacterial colony formed by the strain is cultured for 2 days on YPD agar medium at 28 ℃, the diameter of the bacterial colony is 6-9mm, the bacterial colony is round , the color is gray white, the texture is dry and is villous, the bacterial colony is opaque, the edge of the bacterial colony is blanket-shaped, the bacterial colony is not easy to pick up, the microscopic examination cell is elliptical, the scanning electron microscope examination cell is elliptical and oblong, and the pseudohypha and ascospores can be formed.
(Fifth) culture characteristics of the seed culture
(1) The culture temperature is 20-30 ℃, and the optimum temperature is 28-30 ℃;
(2) The culture pH is 3.0-7.5, and the optimal range is 4.0-6.0.
The cells are elliptic, with candida and ascospores, and budding, and according to the characteristics and identification handbook of saccharomycetes, the detected sequences are subjected to screening, separation, purification, microscopic examination, physiological biochemistry and 16S rDNA sequencing in a sample, the detected sequences are subjected to sequence inversion in DNAMAN software, BLAST comparison is performed in NCBI, a phylogenetic tree of the separated strain is established by using MEGA5 software, and the separated strain is determined to be the sacculus laminating yeast (Saccharomycopsis fibuligera).
The invention discloses application of a saccule-covered yeast (Saccharomycopsis fibuligera) in acid, ester and alcohol production.
Compared with the prior art, the invention has obvious beneficial effects, and the technical scheme can be adopted as follows: the saccule-covered yeast (Saccharomycopsis fibuligera) has wide application range to natural environment conditions such as temperature, pH and the like, has strong tolerance to adverse environment, and the microorganism can decompose and change the structure of fermentation raw materials on the molecular level through the metabolism of the microorganism, so that a plurality of metabolites favorable for organisms are generated.
Drawings
FIG. 1 is a colony morphology of a strain according to the present invention;
FIG. 2 is a diagram showing the morphology of yeast cells observed under a 100-fold microscope after staining with merocyanin of the strain according to the invention;
FIG. 3 is a diagram showing the structural morphology of the candida yeast mycelium observed under a 100-fold microscope after the strain of the invention is subjected to merocyanin staining;
FIG. 4 is a diagram showing the morphology of the strain of the present invention observed under a scanning electron microscope at a magnification of 5000;
FIG. 5 is a diagram showing the morphology of the strain of the present invention observed 2400-fold under a scanning electron microscope;
FIG. 6 is a graph of glucose tolerance of the strain of the invention;
FIG. 7 is a graph showing lactic acid resistance of the strain of the present invention;
FIG. 8 is a graph showing the change of the organic acid produced by the strain of the present invention.
Detailed Description
The invention is further described below with reference to the accompanying drawings.
Referring to fig. 1-8;
Taking 10g of broken grape sample from the epidermis of a Guizhou wild grape sample, adding the broken grape sample into a triangular flask filled with 90ml of sterile physiological saline and glass beads, oscillating for 30min at a temperature of 28 ℃ by a shaking table, and uniformly mixing; sucking 1mL of bacterial suspension in an ultra-clean workbench, injecting the bacterial suspension into a test tube filled with 9mL of sterile physiological saline, carrying out gradient dilution, and taking a gradient of 10 -1、10-2、10-3、10-4、10-5 for pouring and coating a flat plate respectively; after inverted culturing at 28 ℃ for 2d, counting, selecting single colony with typical yeast colony morphology, inoculating and streaking on YPD agar medium, repeating for 3-4 times until pure strain is obtained, then numbering each strain respectively, and finally streaking on inclined plane for low-temperature preservation.
Inoculating the strain on YPD culture medium by streaking, culturing at 28deg.C for 2d, and culturing colony diameter of 6-9mm, wherein the colony is round , has off-white color, dry texture and is villous, opaque, and the edge of the colony is blanket-shaped, so that the colony is not easy to pick up; the microscopic examination cells are elliptical, and the scanning electron microscopic examination cells are elliptical and oblong, so that pseudohypha and ascospores can be formed.
Test example:
Physiological and biochemical feature analysis: according to the results of the physiological and biochemical carbon source utilization experiments, the strain can utilize carbon sources widely and comprises the following steps: glucose, lactose, fructose, sucrose, galactose, starch, xylose, maltose, cellobiose, sorbitol, mannitol, salicin and the like, which shows that the strain has wider adaptability.
TABLE 1 use of saccule-buckling complex film yeast (Saccharomycopsis fibuligera) physiological and biochemical carbon
Carbon source Results
Salicin +
Xylose +
Mannitol (mannitol) +
Maltose +
Cellobiose +
Sorbitol +
Glucose +
Lactose and lactose +
Fructose +
Sucrose +
Galactose +
Starch +
Note that: positive, can be utilized; negative, unavailable.
Alcohol production capacity measurement: the alcohol production capacity of the yeast was measured, and the alcohol content was 18.4.+ -. 0.29% VoL.
Sugar tolerance determination: the glucose tolerance of the yeast is measured, the glucose tolerance of the strain can reach 300Biex, and the strain can still keep weak growth under 400 Biex.
Acid resistance measurement: the lactic acid resistance of the yeast is measured, the strain is suitable for growing at pH 4.0-6.0 and the optimal growing pH is 5.0, and the strain is found to be at least resistant to pH3.0, the strain growth vigor can still reach 3.29 x 10 7 CFU/ml, and weak growth can be maintained at pH 2.0.
Determination of organic acid production ability: the organic acid production capacity of the yeast was measured and the yeast was metabolically produced during fermentation: l-malic acid, D-malic acid, lactic acid, acetic acid, wherein the content of L-malic acid is the highest, up to 0.76mg/ml.
Determination of higher alcohol production Capacity: determining the higher alcohol produced by the yeast, wherein the main higher alcohol is 9.907mg/ml of isoamyl alcohol, 2.334mg/ml of linalool, 2.307mg/ml of beta-phenethyl alcohol and 1.031mg/ml of isobutanol; and (3) measuring the capacity of ester-producing substances: the capacity of the yeast for producing esters is measured, and the yeast fermentation mainly produces 0.452mg/ml of ethyl n-caproate, 1.184mg/ml of isoamyl acetate, 1.704mg/ml of ethyl acetate and 0.204mg/ml of ethyl caprylate.
Table 2 units of volatile component change of the sacculus-covering yeast (Saccharomycopsis fibuligera): mg/mL
The invention is, of course, capable of other and further embodiments and of modification in accordance with the invention, obvious to those skilled in the art, without departing from the spirit and substance of the invention, but these modifications and variations are intended to be within the scope of the appended claims.

Claims (1)

1. An application of a saccule-covered yeast (Saccharomycopsis fibuligera) FBKL2.8DCJS1 in producing L-malic acid, which is characterized in that the preservation number is CCTCC NO. M2020891.
CN202311385212.1A 2023-10-25 2023-10-25 Saccharomycetes tectorial membrane yeast and application thereof Active CN117511755B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN202311385212.1A CN117511755B (en) 2023-10-25 2023-10-25 Saccharomycetes tectorial membrane yeast and application thereof

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN202311385212.1A CN117511755B (en) 2023-10-25 2023-10-25 Saccharomycetes tectorial membrane yeast and application thereof

Publications (2)

Publication Number Publication Date
CN117511755A CN117511755A (en) 2024-02-06
CN117511755B true CN117511755B (en) 2024-08-23

Family

ID=89750403

Family Applications (1)

Application Number Title Priority Date Filing Date
CN202311385212.1A Active CN117511755B (en) 2023-10-25 2023-10-25 Saccharomycetes tectorial membrane yeast and application thereof

Country Status (1)

Country Link
CN (1) CN117511755B (en)

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN117025422B (en) * 2022-12-23 2024-06-04 贵州大学 Yeast and application thereof

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN112998252A (en) * 2021-01-17 2021-06-22 贵州大学 Microbial compound fermentation tartary buckwheat enzyme and preparation method thereof

Family Cites Families (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP3253779A1 (en) * 2015-02-06 2017-12-13 Cargill, Incorporated Modified glucoamylase enzymes and yeast strains having enhanced bioproduct production
CN106085888B (en) * 2016-06-17 2019-04-23 四川伯禹生物工程有限公司 Saccharomycopsis fibuligera new strains and its cultural method and purposes
CN112601818A (en) * 2018-07-25 2021-04-02 诺维信公司 Enzyme-expressing yeast for producing ethanol
CN109971661B (en) * 2019-04-03 2020-07-31 佛山市海天(高明)调味食品有限公司 Ester-producing yeast ZB406 and application thereof
CN114250157B (en) * 2021-11-17 2022-11-15 华南农业大学 Saccharomycopsis fibuligera cx-3 strain for high yield of aroma substances and application thereof
CN115786145A (en) * 2022-11-25 2023-03-14 北京红星股份有限公司 Separated fibula-coated yeast strain, saccharomyces cerevisiae composition and application of composition in production of bran koji white spirit
CN116875472A (en) * 2023-06-13 2023-10-13 贵州大学 Saccharomycetes complex film yeast strain for producing biological film and tolerance and ester producing ability thereof

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN112998252A (en) * 2021-01-17 2021-06-22 贵州大学 Microbial compound fermentation tartary buckwheat enzyme and preparation method thereof

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
扣囊复膜酵母在白酒生产中的应用研究进展;王乃军等;酿酒科技;20230318;摘要、第116页1.1节 *

Also Published As

Publication number Publication date
CN117511755A (en) 2024-02-06

Similar Documents

Publication Publication Date Title
CN105861346B (en) One plant of high-yield urea, the abnormal Brunswick Durham yeast strain for producing wind taste and its application in food production
CN111534439B (en) Penicillium oxalicum SDF-25 strain and application thereof
CN117511755B (en) Saccharomycetes tectorial membrane yeast and application thereof
CN107287127B (en) Lactic acid-resistant pichia pastoris capable of producing ester
CN103525743B (en) Clostridium sporogenes and purpose thereof
WO2023104005A1 (en) Ester-producing yeast strain and use thereof
CN102994398B (en) Urease high-yielding Aspergillus sydowii and application thereof
CN113151123B (en) Bacillus cereus for producing cellulase and xylanase at low temperature
CN106753994B (en) Method for improving alcohol content of alcohol fermentation liquor and reducing isoamyl alcohol content by using high-ester-yield indigenous aroma-producing yeast enhanced yeast
CN104164395A (en) Clostridium beijerinckii for hydrogen generation via fermentation as well as fermentation method and application of clostridium beijerinckii
CN111100825B (en) Bacillus and application thereof in industry
CN110564625B (en) Saline-alkali resistant aspergillus flavus and separation method and application thereof
CN111019849B (en) Pseudocercospora viticola and application thereof
CN103205369B (en) Novel brewing yeast strain with L-apple acid degrading property and application of novel brewing yeast strain
CN108841889B (en) Method for producing griseofulvin serving as major component of tranexamycin by microbial fermentation
CN107164246B (en) High-temperature-resistant yeast and application thereof
CN117286064A (en) Bacillus licheniformis capable of reducing acidity and higher alcohol content of fermented grains, method and application thereof in white spirit production
CN114480222B (en) Paenibacillus kriging spaceflight mutant and application thereof
CN105062904A (en) Saccharomycopsis fibuligera and application thereof
CN113502282A (en) Method for producing pectinase preparation by solid-state fermentation of penicillium
Ueno et al. Fermentation of Molasses by Several Yeasts from Hot Spring Drain and Phylogeny of the Unique Isolate Producing Ethanol at 55^ oC
CN106399212A (en) Streptomyces bangladeshensisstrain FBKL4.004 and application thereof
CN113151014A (en) Simple and rapid monascus separating and storing method
CN117025422B (en) Yeast and application thereof
CN1302864A (en) High-content mycose saccharomycetes and its preparing process

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
GR01 Patent grant
GR01 Patent grant