CN116083256B - Kazakhstan yeast FJY-3 strain and application thereof - Google Patents
Kazakhstan yeast FJY-3 strain and application thereof Download PDFInfo
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- CN116083256B CN116083256B CN202310084025.3A CN202310084025A CN116083256B CN 116083256 B CN116083256 B CN 116083256B CN 202310084025 A CN202310084025 A CN 202310084025A CN 116083256 B CN116083256 B CN 116083256B
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- gray mold
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Classifications
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- 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
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
- A01N63/30—Microbial fungi; Substances produced thereby or obtained therefrom
- A01N63/32—Yeast
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01P—BIOCIDAL, PEST REPELLANT, PEST ATTRACTANT OR PLANT GROWTH REGULATORY ACTIVITY OF CHEMICAL COMPOUNDS OR PREPARATIONS
- A01P3/00—Fungicides
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23F—COFFEE; TEA; THEIR SUBSTITUTES; MANUFACTURE, PREPARATION, OR INFUSION THEREOF
- A23F5/00—Coffee; Coffee substitutes; Preparations thereof
- A23F5/02—Treating green coffee; Preparations produced thereby
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- 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
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- 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
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/88—Lyases (4.)
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- 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
- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/02—Preparation of oxygen-containing organic compounds containing a hydroxy group
- C12P7/04—Preparation of oxygen-containing organic compounds containing a hydroxy group acyclic
- C12P7/06—Ethanol, i.e. non-beverage
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y402/00—Carbon-oxygen lyases (4.2)
- C12Y402/02—Carbon-oxygen lyases (4.2) acting on polysaccharides (4.2.2)
- C12Y402/02002—Pectate lyase (4.2.2.2)
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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Abstract
The invention discloses a Kazakhstan yeast FJY-3 strain and application thereof. The Kazakhstan yeast FJY-3 strain is preserved in the Guangdong province microorganism strain collection at 2022, 12 months and 4 days, and the preservation number is GDMCC No:63034 the preservation address is building No. 100 and building No. 59 in the Mitsui of Guangzhou City of Guangdong. The Kazakhstan yeast FJY-3 strain has wide application, not only can be used for biological degumming of coffee, but also can be used for degrading coffee peel to ferment and generate ethanol, and is beneficial to recycling of the coffee peel. In addition, the Kazakhstan yeast FJY-3 strain can inhibit the growth of gray mold, italian penicillium, banana fusarium wilt, fusarium graminearum and colletotrichum gloeosporium, and can be used for biological control of plant diseases caused by the pathogenic bacteria.
Description
Technical Field
The invention belongs to the technical field of microorganisms. More particularly, relates to a Kazakhstan yeast FJY-3 strain and application thereof.
Background
The fresh coffee fruit mainly comprises embryo, endosperm, coffee bean silver skin, endocarp, fruit gum layer (mucilage layer), mesocarp and epicarp from inside to outside. The fermentation degumming link is the most important link in the wet processing of coffee, and aims to remove the pectin layer of fresh coffee, improve the cleanliness of coffee beans, avoid microbial contamination and have close relation with the quality of coffee commodity beans.
At present, two methods of natural fermentation and mechanical fermentation are mainly adopted for solving the problem of fermentation degumming in the coffee processing process. The natural fermentation is to soak the peeled coffee into water by taking water as a medium after the coffee is peeled, and to carry out fermentation degumming by virtue of enzymes contained in pectin. However, the natural fermentation degumming period is long, the influence factors are many, the degumming process is not easy to control, the degumming is easy to be incomplete or the fermentation time is too long, so that the sour taste of the coffee beans is too heavy, the quality is reduced, microorganisms in the natural fermentation process are not screened, the variety is various, and the pollution substances generated in the microorganism metabolism process also easily influence the quality of the coffee. The mechanical fermentation is to use machinery to directly remove pectin from the peeled coffee beans. However, mechanical degumming is easy to cause incomplete degumming, and coffee beans are easy to undergo secondary fermentation, so that the quality and the storage period of the commercial coffee beans are affected. Biological degumming is to inoculate the selected degumming strain onto dehulled coffee beans, and decompose colloid by utilizing enzymes secreted by the strain. Compared with natural fermentation, the biological degumming process is easy to control, no toxic or harmful substances are produced in the process, and the method has the advantages of being green, environment-friendly, high in efficiency and the like. However, the prior report about the coffee degumming strain is less, the practical application is also in the experimental demonstration stage, and continuous excavation of the strain which can be used for the coffee biological degumming is of great significance to promote the development of the coffee biological degumming technology.
In addition to the problem of coffee fermentation degumming, the reuse of by-products generated during the processing of coffee is also a concern. The coffee pericarp is one of main byproducts generated in the coffee processing process, and is rich in polysaccharide, crude fiber and other substances. At present, the coffee pericarp is mainly used for preparing drinks, feeds and fertilizers at home and abroad, but the utilization rate is low. Therefore, the industrial utilization value of the coffee peel is necessary to be mined, the utilization method of the coffee peel is provided, the utilization rate of the coffee peel is improved, the economic benefit of enterprises can be increased, and the problem of environmental pollution caused by discarding the coffee peel can be reduced.
Disclosure of Invention
The invention aims to overcome the defects and the shortcomings of the prior art and provide a Kazakhstan yeast FJY-3 strain and application thereof.
The first object of the present invention is to provide a strain of Kazakhstan yeast FJY-3.
The second object of the present invention is to provide the use of the FJY-3 strain in preparing pectase.
A third object of the present invention is to provide the use of the FJY-3 strain in the biological degumming of coffee or in the preparation of a biological degumming formulation for coffee.
The fourth object of the present invention is to provide the use of the FJY-3 strain in preparing ethanol by using coffee pericarp.
The fifth object of the invention is to provide the application of the FJY-3 strain or the bacterial liquid of the FJY-3 strain in inhibiting plant pathogenic bacteria.
The sixth object of the present invention is to provide an application of the FJY-3 strain or the bacterial liquid of the FJY-3 strain in preparing a preparation for inhibiting plant pathogenic bacteria.
The seventh object of the invention is to provide the application of the FJY-3 strain or the bacterial liquid of the FJY-3 strain in preventing and treating plant diseases.
The eighth object of the invention is to provide the application of the FJY-3 strain or the bacterial liquid of the FJY-3 strain in preparing the preparation for preventing and treating plant diseases.
The ninth object of the invention is to provide a plant disease biocontrol agent.
The above object of the present invention is achieved by the following technical scheme:
the invention obtains a Kazakhstan yeast (Kazachstania humilis) which can degrade coffee pectin from the fermentation broth of the small-grain coffee and is named as Kazakhstan yeast FJY-3 strain. Besides secreting pectase and degrading citrus pectin and coffee pectin, the FJY-3 strain can also utilize the product of coffee peel pretreated by dilute sulfuric acid and cellulase to generate ethanol, thereby being beneficial to recycling of the byproduct coffee peel generated in the coffee processing process. In addition, the further experiments of the invention find that the FJY-3 strain has an inhibiting effect on gray mold, italian penicillium, banana fusarium, fusarium graminearum and colletotrichum gloeosporium and has a good biocontrol effect on strawberry gray mold. Therefore, the invention claims the Kazakhstan yeast FJY-3 strain and the application thereof.
The invention provides a Kazakhstan yeast FJY-3 strain which is preserved in the Guangdong province microorganism strain collection at the 12 th month 4 of 2022, wherein the preservation number is GDMCC No:63034 the preservation address is building No. 100 and building No. 59 in the Mitsui of Guangzhou City of Guangdong.
The FJY-3 strain can secrete pectase to degrade citrus pectin and coffee pectin. Therefore, the invention claims the application of FJY-3 strain in preparing pectase.
The invention also claims the application of the FJY-3 strain in biological degumming of coffee or preparation of biological degumming preparation of coffee.
The invention also provides a coffee biological degumming fermenting agent, which contains the FJY-3 strain.
The invention also claims the application of the FJY-3 strain in preparing ethanol by using coffee peel.
The invention also provides a method for preparing ethanol by degrading coffee pericarp by using the FJY-3 strain, which comprises the following steps: pretreating coffee peel by dilute sulfuric acid, adding 1.8% (w/v) cellulase, treating at 50deg.C to obtain coffee peel hydrolysate, taking the obtained coffee peel hydrolysate as carbon source, adding nitrogen source, and fermenting and culturing with FJY-3 strain.
Specifically, the dilute sulfuric acid pretreatment conditions are that 0.02M dilute sulfuric acid is used for 30min at 121 ℃.
Specifically, the treatment time of the cellulase is 72h.
The invention also claims the application of the FJY-3 strain or the bacterial liquid of the FJY-3 strain in inhibiting plant pathogenic bacteria.
The invention also claims the application of the FJY-3 strain or the bacterial liquid of the FJY-3 strain in preparing the preparation for inhibiting plant pathogenic bacteria.
Specifically, the plant pathogenic bacteria are one or more of gray mold, italian penicillium, fusarium graminearum, colletotrichum gloeosporium and banana fusarium wilt.
The invention also claims the application of the FJY-3 strain or the bacterial liquid of the FJY-3 strain in preventing and controlling plant diseases.
The invention also claims application of the FJY-3 strain or the bacterial liquid of the FJY-3 strain in preparation of a preparation for preventing and treating plant diseases.
Specifically, the plant disease is one or more of gray mold, blue mold, scab, anthracnose and banana vascular wilt.
More specifically, the plant disease is one or more of gray mold caused by gray mold, penicillium caused by penicillium italicum, gibberellic disease caused by fusarium graminearum, anthracnose caused by colletotrichum gloeosporioides, banana vascular wilt caused by banana vascular wilt.
The invention also provides a plant disease biocontrol preparation, which contains the FJY-3 strain or the bacterial liquid of the FJY-3 strain.
Specifically, the plant disease is one or more of gray mold, blue mold, scab, anthracnose and banana vascular wilt.
The invention also claims application of the FJY-3 strain or the bacterial liquid of the FJY-3 strain in preventing and treating strawberry gray mold or preparing a preparation for preventing and treating strawberry gray mold.
The invention also provides a biocontrol preparation for the strawberry gray mold, which contains the FJY-3 strain or the bacterial liquid of the FJY-3 strain.
Specifically, the concentration of the bacterial liquid is 10 5 ～10 7 cells/mL。
The invention also provides a method for preventing and controlling the gray mold of the strawberries, which comprises the following steps: soaking for 30s with the bacterial liquid of the FJY-3 strain or uniformly spraying the bacterial liquid of the FJY-3 strain on the surface of the strawberry.
Specifically, the concentration of the bacterial liquid is 10 5 ～10 7 cells/mL。
Preferably, the concentration of the bacterial liquid is 10 6 cells/mL, see example 5.
The invention has the following beneficial effects:
the invention provides a Kazakhstan yeast FJY-3 strain which is preserved in the Guangdong province microorganism strain collection at the 12 th month 4 of 2022, wherein the preservation number is GDMCC No:63034 the preservation address is building No. 100 and building No. 59 in the Mitsui of Guangzhou City of Guangdong. The FJY-3 strain can secrete pectase, degrade citrus pectin and coffee pectin, can be used for fermentation degumming links in coffee wet processing, is used for biological degumming of coffee or is used for preparing a coffee biological degumming preparation, has no pectase activity, does not generate methanol harmful to human by catalyzing pectin to remove methyl ester groups, is safe and effective, and is environment-friendly.
Besides being used for biological degumming of coffee, the FJY-3 strain has good tolerance to phenolic substances in coffee peel, can be used for degrading the coffee peel to ferment and generate ethanol, has high concentration of ethanol obtained by fermentation, reaches 62.8g/L, and is the highest yield of ethanol fermentation using the coffee peel as a substrate reported at present. The invention is not only beneficial to recycling the coffee peel and reducing the problem of environmental pollution caused by discarding the coffee peel, but also enriches the sources of raw materials for producing the ethanol, is beneficial to producing the ethanol and increases the economic benefit of enterprises.
In addition, the FJY-3 strain can inhibit the growth of gray mold, italian penicillium, banana fusarium, fusarium graminearum and colletotrichum gloeosporioides, and can be used for biological control of gray mold, blue mold, banana fusarium wilt, scab and anthracnose. The FJY-3 strain has the best effect of inhibiting the gray mold, and the bacterial liquid can effectively inhibit the growth of the gray mold disease spots of the strawberries without affecting the quality of the strawberries.
The invention not only enriches the fungus library for biological degumming of coffee and preventing and controlling the gray mold of the strawberry, but also provides the method for producing ethanol by fermenting the coffee peel and the method for preventing and controlling the gray mold of the strawberry, which is beneficial to recycling the byproducts of coffee processing, namely the coffee peel and producing the ethanol, has convenient operation, safety and environmental protection, and has remarkable environmental benefit, economic benefit and social benefit.
Drawings
FIG. 1 is a colony morphology of FJY-3 strain.
FIG. 2 is a diagram showing the bacterial morphology of FJY-3 strain.
FIG. 3 is a sequence phylogenetic tree of FJY-3 strain based on the D1/D2 region of 26S rDNA.
FIG. 4 is a sequence phylogenetic tree of FJY-3 strain based on ITS.
FIG. 5 is a standard curve of BSA.
FIG. 6 shows the standard curve of D-galacturonic acid.
FIG. 7 is a standard curve for ethanol vapor phase.
FIG. 8 shows statistical results of the bacterial inhibition rate of FJY-3 strain against 5 different plant pathogens, different letters representing significant differences (p < 0.05).
FIG. 9 shows the growth inhibitory effect of the bacterial liquid of FJY-3 strain on gray mold lesions of strawberry.
FIG. 10 is a graph showing the statistical result of growth inhibition of bacterial liquid of FJY-3 strain on gray mold lesions of strawberry; CK: only inoculating gray mold; tr: by 1X 10 6 cell/mL FJY-3 strain bacterial liquidInoculating gray mold after soaking; ** indicating that the difference was extremely remarkable (p<0.01)。
FIG. 11 shows the growth inhibitory effect of bacterial liquids of FJY-3 strains at different concentrations on gray mold lesions of strawberry.
FIG. 12 is a graph showing the statistical results of growth inhibition of bacterial solutions of FJY-3 strains at different concentrations on gray mold lesions of strawberry; CK: only inoculating gray mold; the different letters represent significant differences (p < 0.05).
Detailed Description
The invention is further illustrated in the following drawings and specific examples, which are not intended to limit the invention in any way. Unless specifically stated otherwise, the reagents, methods and apparatus employed in the present invention are those conventional in the art.
Reagents and materials used in the following examples are commercially available unless otherwise specified.
The FJY-3 strain in the embodiment of the invention is Kazakhstan yeast (Kazachstania humilis) FJY-3 strain which is preserved in the microorganism strain preservation center of Guangdong province in the year 12 and 4 of 2022, and the preservation number is GDMCC No:63034 the preservation address is building No. 100 and building No. 59 in the Mitsui of Guangzhou City of Guangdong.
PDA medium: 200g of potato, and adding a proper amount of ddH 2 O, boiling for 20min, filtering with four layers of gauze, adding glucose 20g, agar 20g, ddH 2 O is fixed to 1000mL, and the pH is natural; sterilizing at 121deg.C for 20min.
YPD medium: peptone 20g, yeast extract 10g, glucose 20g, agar 20g, ddH 2 O is fixed to 1000mL, and the pH is natural; sterilizing at 121deg.C for 20min.
Bradford color development solution: 35mg of Coomassie brilliant blue G-250 was weighed and dissolved in 25mL of a mixture of 95% ethanol and 50mL of 88% phosphoric acid, and ddH was added 2 O is fixed to 500mL, and the qualitative filter paper is stored in a brown bottle for standby after being filtered.
Fermentation medium: 5g/L of citrus pectin or coffee peel, 0.3g/L of KCl and K 2 HPO 4 0.4g/L, yeast powder 10g/L, ddH 2 And (3) sterilizing for 20min at 121 ℃ with the constant volume of O to 1000 mL.
EXAMPLE 1 isolation, purification and characterization of FJY-3 Strain
1. Isolation and purification of FJY-3 Strain
The FJY-3 strain is separated from fermentation liquor of small-grain coffee (Coffea arabica L.) of south-screen Zhennan island river in Pu' er city of Yunnan province, and the strain is separated by adopting a flat plate dilution method, and the separation and purification processes are as follows:
the fermentation broth was first filtered with six layers of gauze and the filtrate was diluted in a gradient (100, 10) -1 、10 -2 、10 -3 、10 -4 And 10 -5 ) 100. Mu.L of dilutions of different concentrations were pipetted separately into PDA medium (containing 100mg/mL kanamycin) and 3 replicates were set for each gradient; culturing for 4-5 d at 28 ℃ until colonies grow out, picking single colonies, streaking and inoculating the single colonies into YPD culture medium, purifying and culturing for 2-3 d, purifying for 3 times continuously, and numbering and preserving strains by a glycerol method.
2. Identification of FJY-3 Strain
(1) Morphological identification
The FJY-3 strain was streaked onto YPD solid medium and after 48h the colony morphology was observed. In addition, single colonies were picked up and placed in 50. Mu.L of sterile water, 5. Mu.L was pipetted onto a slide glass and observed for bacterial status under an optical microscope at 100-fold magnification.
As shown in FIG. 1, the colony morphology of FJY-3 strain is circular, milky white, smooth in edge, moist in surface, opaque and convex in middle as shown in FIG. 1. As shown in FIG. 2, the form of the cells of the FJY-3 strain was shown in FIG. 2, and it was found that FJY-3 was a single-cell fungus, and the cells were round or oval, and the cell size was 0.6-1.0. Mu.m.times.1.8-3.0. Mu.m.
(2) Molecular biological identification
(1) Cell culture
The FJY-3 strain is activated in YPD solid culture medium, cultured for 48 hours at 28 ℃ and inoculated in YPD liquid culture medium, placed in a shaking table at 28 ℃ and 180r/min and cultured for 12 hours.
(2) PCR amplification
The method for extracting the genomic DNA of the FJY-3 strain is carried out according to the specification of an OMEGA fungus DNA extraction kit. PCR amplification primers used for molecular identification of strains are shown below:
ITS rRNA primer:
ITS3：5’-GATGAAGAACGYAGYRAA-3’；
ITS4：5’-TCCTCCGCTTATTGATATGC-3’；
26S rDNA D1/D2 region sequence primer:
NL-1：5’-GCATATCAATAAGCGGAGGAAAAG-3’；
NL-4：5’-GGTCCGTGTTTCAAGACGG-3’；
the PCR reaction system (25. Mu.L total) was: 2 XPCR Buffer 12.5. Mu.L, 2mM dNTPs 5. Mu.L, 10pmoL/mL upstream primer 0.75. Mu.L, 10pmoL/mL downstream primer 0.75. Mu.L, KOD FX (1.0U/. Mu.L) 0.5. Mu. L, DNA 1.0 and 1.0. Mu. L, ddH 2 O4.5. Mu.L. The PCR amplification procedure was: pre-denaturation at 94℃for 5min, denaturation at 98℃for 10s, annealing at 58℃for 30s, extension at 68℃for 40s, total of 35 cycles, and extension at 68℃for 5min.
(3) Sequence determination and construction of phylogenetic tree
The PCR product was sent to the Option (ITS) sequence of FJY-3 strain obtained by sequencing by the same method as follows:
TTTAGGGTGACTGCGGAGGTCATTAAAGAAATGATTGGGGGAGCCCGCCTGCGCTTAGCTGCGCGGTGGCGCTCCGCCTCTTACACACAGTGGAGTAATTACTTATTCTTGGGGTGGACGAAAGTCCATCTCGCCAAACACAAACAACTATTTTCTATTATACAACGTCATCAAATCTGCTTACGCAGTAACCAAAATATTCAAAACTTTCAACAACGGATCTCTTGGTTCTCGCATCGATGAAGAACGCAGCGAAATGCGATACGTAATGTGAATTGCAGAATTCCGTGAATCATCGAATCTTTGAACGCACATTGCGCCCCTTGGTATTCCAGGGGGCATGCCTGTTTGAGCGTCATTTCCTTCTCAAGCACCTGTGCTTGGTTGTGGGTGACACTCTCTCGAGTTAGCTTGAAATTGCTGGCCGCACTGCGGTGGAGCAGTTGGCTTGTCTGTCGTGCGCGGTGCCTCGGCGCCGGGCGTGGCTGGCATGCGATTGTCGTACTAGGTTTTACCAATTCGGCAGGAGCATGGCAGACAGAGAGACAATACAACCGCCCT CCCCCTGGCTAACAGTACTCTTTAAGTTTGACCTCAAATCAGGTAGGAATACCCGCTGAACTTAAGATAAAAAAAAGGGGGG
the sequence of the region 26S rDNA D1/D2 of FJY-3 strain is as follows:
CGGGGGGGATGCTCGTACGGCGAGTGAGCGGCAAAAGCTCAAATTTGAAATCTGGTACCTTCGGTGCCCGAGTTGTAATTTGTAGAGGGCGACTTTGGGGCGGCTCCTTGTCTATGTTCCTTGGAACAGGACGTCATAGAGGGTGAGAATCCCGTGTGGCGAGGAGTGCGGTTCCGTGTAAAGCGCTCTCGAAGAGTCGAGTTGTTTGGGAATGCAGCTCTAAGTGGGTGGTAAATTCCATCTAAAGCTAAATATTGGCGAGAGACCGATAGCGAACAAGTACAGTGATGGAAAGATGAAAAGAACTTTGAAAAGAGAGTGAAAAAGTACGTGAAATTGTTGAAAGGGAAGGGCATTTGATCAGACATGGTGTTTTGCGCCCCCCGCTCCTTGTGGGTGGGGGACTCTCGCAGCTCACTGGGCCAGCATCAGTTTTGGCGGCCGGACAAAACTGCAGGAACGTAGCTTGCTTCGGGAAGTGTTACAGCCTGCAGGAATACGGCCAGCCGGGACTGAGGAATGCGATTCGTCAAGGATGCTGGCATAATGGTTATATGCCGCCCGTCTAAAACCCCGGGACCA
BLAST similarity comparison and analysis are carried out on the sequencing result in NCBI database and the ITS sequence and 26S rDNA D1/D2 region sequence of the known strain, and the similarity between the FJY-3 strain and the ITS sequences and 26S rDNA D1/D2 region sequences of a plurality of Kazakhstan yeast strains is found to be more than 98%. The invention uses MEGA 11 software to construct a sequence evolution tree by a neighbor-joining method, wherein the sequence evolution tree of the FJY-3 strain based on the 26S rDNA D1/D2 region is shown in figure 3, and the sequence evolution tree of the FJY-3 strain based on ITS is shown in figure 4. As can be seen from FIGS. 3 and 4, the FJY-3 strain was closest to the reported affinity for Kazakhstan yeast. Combining morphological and molecular identification results, the FJY-3 strain was identified as Kazakhstan yeast (Kazachstania humilis).
EXAMPLE 2 determination of pectase Activity of FJY-3 Strain
1. Experimental method
1. Crude enzyme protein concentration determination
(1) BSA standard curve formulation
A BSA standard solution was prepared in accordance with Table 1, 100. Mu.L of the mixture was aspirated from each of the 1-6 tubes, and 5mL of Bradford color development solution was added. Fully and uniformly mixing, standing for 5min, taking a tube 1 as a blank control, and measuring the light absorption value at 595 nm; with BSA standard concentration as X-axis, OD 595 nm And the absorbance value is the Y axis to make a standard curve.
Table 1 BSA standard solution formulation
(2) Protein concentration determination
The FJY-3 strain is inoculated on YPD culture medium, cultured for 1-2 d at 28 ℃ until single colony grows, picked up and placed in a 50mL conical flask containing 25mL YPD liquid culture medium, and shake-cultured for 24h at 28 ℃ under 150r/min to obtain seed liquid. 100 mu L of seed liquid is inoculated into 50mL of fermentation culture medium (two fermentation culture media are used in the embodiment, citrus pectin and coffee peel powder are respectively used as substrates) for fermentation culture, after 48h of shaking culture at 28 ℃ and 150r/min, centrifugation is carried out at 4 ℃ and 8000r/min for 15min, the obtained supernatant is crude enzyme liquid, and the protein concentration of the crude enzyme liquid is measured.
Taking 100 mu L of crude enzyme liquid sample, adding 5mL of Bradford color development liquid, fully mixing uniformly, standing for 5min, taking a tube No. 1 in Table 1 as a blank control, measuring the light absorption value at 595nm, and calculating the protein concentration of the measured sample by using a standard curve.
2. Determination of Polygalacturonase (PG) Activity
(1) D-galacturonic acid standard curve formulation
Preparing 1mg/mL of D-galacturonic acid standard solution; taking 9 test tubes, adding 0, 20, 40, 60, 80, 100, 120, 140 and 160 μl standard solution respectively, adding 0.5mL DNS reagent, mixing thoroughly, boiling water bath for 5min, taking out, cooling to room temperature with cold water, and adding ddH 2 O, respectively fixing the volumes of the tubes to 4mL, uniformly mixing, taking a test tube No. 1 without the D-galacturonic acid standard solution as a blank control, and measuring the light absorption value at 520 nm; the content of D-galacturonic acid (μg) is taken as X axis, OD 520 nm And the absorbance value is the Y axis to make a standard curve.
(2) PG Activity assay
Mixing 400 μl crude enzyme solution, 400 μl 1% PGA (polygalacturonic acid) solution (prepared with 0.05mol/L acetic acid-sodium acetate buffer, pH 5.0) and 400 μl 50mmol/L acetic acid-sodium acetate buffer thoroughly, mixing, standing in 45deg.C water bath for 30min, adding 0.6mL DNS reagent to terminate the reaction, boiling for 5min, immediately cooling to room temperature with cold water, adding 2.2mL ddH to each tube 2 O, shake well, measure its OD 520 nm Numerical value, blank control is added with equivalent amount of crude enzyme inactivated by boiling water bath for 15 min. After measurement, in the standardThe corresponding D-galacturonic acid content was found on the curve.
The enzyme activity units of PG are defined as: at 45℃1mg of crude protein was used to decompose pectin to 1mg of D-galacturonic acid in 1min with an enzyme amount of 1 enzyme activity unit (U.mg -1 ·min -1 )。
PG enzyme Activity= (A) 1 ×N×K 2 )/(A 2 ×0.4×K 1 ×T)
A 1 : OD of experimental group 520 A value; a is that 2 : OD of experimental group 595 A value; 0.4: volume of enzyme solution (mL) used in enzyme activity measurement; k (K) 1 : slope of D-galacturonic acid standard curve; k (K) 2 : slope of BSA standard curve; t: the reaction time (min) was 30min; n: supernatant dilution fold.
3. Determination of Pectin Lyase (PL) Activity
The reaction system was 1mL, and contained 200. Mu.L of 1% PGA solution, 200. Mu.L of 20mmol/L buffer (Tris HCl+CaCl) 2 pH 8.5), 200. Mu.L of the crude enzyme solution was reacted at 50℃for 20 minutes, 400. Mu.L of 0.1mol/L HCl was added to stop the reaction, and then absorbance at 235nm was measured. The control group is firstly added with buffer solution for reaction for 20min, and then crude enzyme solution is added.
A standard enzyme activity unit (mu mol. Mg) -1 ·min -1 ) The definition is as follows: the polygalacturonic acid was cleaved per minute to yield an enzyme amount of 1. Mu. Mol of unsaturated galacturonic acid.
PL enzyme activity= (a) 3 X reaction volume x 1000 x dilution x K 2 )/(T×0.2×A 2 ×4600)
A 3 : OD of experimental group 235 A value; a is that 2 : OD of experimental group 595 A value; 1000:1mg = 1000 μg; k (K) 2 : slope of BSA standard curve; t: the reaction time (min) was 20min;0.2: crude enzyme liquid volume (mL) used in enzyme activity measurement; 4600: molar absorption coefficient of unsaturated galacturonic acid.
4. Pectin methylesterase (pectin methylesterase, PME) Activity assay
18mL ddH was taken 2 O,2mL of 0.1mol/L NaCl solution, 10mL of 1% (w/v) pectin solutionMixing, adding 1mL of crude enzyme solution at a constant temperature of 35 ℃, regulating the pH of the solution to 7.5 by using 0.05mol/L NaOH solution and HCl, starting timing, and continuously adding 0.01mol/L NaOH solution by using a burette to keep the pH of a reaction system at 7.5 all the time. After the start of the titration, the total volume of NaOH solution consumed was recorded every 2min, plotted as the total volume of NaOH solution consumed versus the reaction time, and pectin methylesterase activity was calculated from the initial slope of the resulting line. The enzyme activity is expressed in terms of micromoles of acid produced per minute, and the unit of enzyme activity is mu mol.min -1 ·mL -1 。
2. Experimental results
In the invention, the BSA standard solution concentration is taken as an abscissa, the light absorption value at 595nm is taken as an ordinate, linear regression is performed, a standard curve is drawn, the standard curve of the obtained BSA is shown in FIG. 5, and the equation of the standard curve of the protein content of the obtained BSA is as follows: y=0.8123x+0.0169 (R 2 =0.9965). According to the measurement, when citrus pectin is used as a substrate, the protein concentration of the fermentation broth of the FJY-3 strain is 0.099+/-0.028 mg/mL, and when coffee peel powder is used as a substrate, the protein concentration of the fermentation broth of the FJY-3 strain is 0.158+/-0.007 mg/mL.
Linear regression is performed by taking the D-galacturonic acid sugar content as an abscissa and the light absorption value at 520nm as an ordinate, and a standard curve is drawn, wherein the obtained standard curve of the D-galacturonic acid is shown in figure 6, and the equation of the obtained standard curve of the D-galacturonic acid is as follows: y=0.0044 x-0.0116 (R 2 =0.9900). Through measurement, the PG enzyme activities of FJY-3 strain taking citrus pectin and coffee peel powder as substrates are respectively 14.46 U.mg -1 ·min -1 And 30.89 U.mg -1 ·min -1 The method comprises the steps of carrying out a first treatment on the surface of the The PL enzyme activities of FJY-3 strain taking orange pectin and coffee peel powder as substrates are respectively 0.022 mu mol.mg -1 ·min -1 And 0.037. Mu. Mol.mg -1 ·min -1 The method comprises the steps of carrying out a first treatment on the surface of the The PME activity of the FJY-3 strain was 0, i.e. the FJY-3 strain had no PME activity.
The result shows that the FJY-3 strain can secrete pectase to degrade citrus pectin and coffee pectin. Meanwhile, the method has no pectin methylesterase activity, so that methanol harmful to human is not generated by catalyzing pectin to remove methyl ester groups, and the method can be used for biological degumming in the coffee fermentation process and for preparing the coffee biological degumming starter.
EXAMPLE 3 determination of ethanol production by FJY-3 Strain
1. Experimental method
1. Ethanol gas phase standard curve
Ethanol standard solutions of 0, 0.5, 1, 2.5, 5, 10 and 20mg/mL are respectively prepared, n-propanol is selected as an internal standard, and chromatographic grade n-propanol is used for preparing an internal standard solution of 10 mg/mL.
500. Mu.L of each of the prepared ethanol standard solution and the prepared internal standard solution was mixed in a 2mL centrifuge tube, filtered through a 0.22 μm needle filter into a gas phase bottle using a 1mL syringe, and the cap was sealed and placed in a gas chromatograph for measurement.
The ethanol concentration was analyzed and detected by gas chromatography. The temperature of the injector of the gas chromatograph is 250 ℃; the chromatographic column has the specification that: 19091J-413 (30 m. Times.0.32 mm. Times.0.25 μm); the column temperature box adopts the programming temperature: the initial temperature is 40 ℃, and the temperature is kept for 3min; heating to 140 ℃ at 10 ℃/min; then heating to 230 ℃ at 50 ℃/min; finally, keeping at 230 ℃ for 2min; the temperature of the detector is controlled at 300 ℃, wherein the carrier gas is nitrogen, and the flow rate is 30mL/min; the flow rate of hydrogen is 40mL/min; the air flow rate was 350mL/min. The sample loading per sample was 0.5. Mu.L.
2. Sample ethanol yield determination
The nitrogen source required for the ethanol fermentation culture is added according to the components of the YPD medium: 10g/L yeast powder, 20g/L peptone and 180g/L glucose and 160g/L coffee peel hydrolysate as carbon sources. Filtering the supernatant obtained by fermentation through a 0.22 mu m needle filter membrane to obtain a sample liquid; the method for measuring the ethanol concentration of the sample liquid is the same as that of the standard liquid.
2. Experimental results
Taking the ethanol concentration as an abscissa, taking the ratio of the ethanol peak area to the internal standard peak area as an ordinate, performing linear regression, drawing a standard curve, wherein the standard curve of the obtained ethanol gas phase is shown in fig. 7, and the obtained standard curve equation is as follows: y= 0.0658x-0.0031 (R 2 = 0.9986). Performing fermentation on FJY-3 by taking 180g/L glucose as a carbon source, and stopping fermentation after 54 hoursAll the fermentation broths were centrifuged separately and the supernatants were diluted 2-fold and mixed with 500uL of each internal standard, filtered through a 0.22 μm needle filter, and analyzed in the gas phase. The yield of ethanol from FJY-3 strain was 72.5g/L and 0.47g/g.
Pretreating coffee pericarp with dilute sulfuric acid (0.02M, 121 ℃) for 30min, adding 1.8% (w/v) cellulase, treating at 50deg.C for 72 hr, concentrating the obtained hydrolysate mainly composed of fructose (22.6 g/L) and glucose (20.4 g/L) to total sugar concentration of 160g/L for ethanol fermentation. After 54h fermentation, the ethanol concentration reaches 62.8g/L, which is the highest yield of ethanol fermentation using coffee pericarp as a substrate reported at present, and shows that the FJY-3 strain has good tolerance to phenolic substances in the coffee pericarp.
EXAMPLE 4 determination of bacteriostatic Capacity of FJY-3 Strain
The invention uses a plate counter test to find that FJY-3 strain has inhibition effect on Botrytis cinerea, italian penicillium (Penicillium italicum), fusarium graminearum (Fusarium graminearum), colletotrichum gloeosporioides (Colletotrichum gloeosporioides) and banana vascular wilt (F.oxysporum f.sp.cube).
The plate confrontation test process comprises the following steps: inoculating fresh mycelium blocks of Botrytis cinerea, penicillium Italipes, fusarium graminearum, fusarium oxysporum and colletotrichum gloeosporium at the center of PDA plate, streaking and inoculating fresh bacterial liquid (OD) of FJY-3 strain on both sides of the mycelium blocks 600 nm =1.0), the distance between the inoculation lines on both sides is 5cm; in the PDA plate as a control, only pathogenic bacterial pieces were inoculated. PDA plates with different treatments are placed at 28 ℃ for culture, the size of pathogenic bacteria is observed after 7d of culture, each treatment is repeated 3 times, the diameters of control bacterial colonies and the diameters of bacterial colonies of a treatment group are measured, and the inhibition rate of FJY-3 strain on the growth of pathogenic bacteria hyphae is calculated. Pathogenic bacterial hypha growth antibacterial rate = [ (control group colony diameter-treatment group colony diameter)/control group colony diameter]×100％。
The statistical results of the bacterial inhibition rate of the FJY-3 strain on 5 different plant pathogens are shown in figure 8, and as can be seen from figure 8, the FJY-3 strain has inhibition effects on Botrytis cinerea, penicillium italicum, fusarium oxysporum, fusarium graminearum and colletotrichum gloeosporium, and the hypha inhibition rates are 83.61%, 60.56%, 53.78%, 54.55% and 47.95% respectively.
Example 5 control Effect of FJY-3 Strain on strawberry gray mold
As shown by the experimental result of example 4, the FJY-3 strain has the best inhibition effect on Botrytis cinerea (Botrytis cinerea) causing mildew of fruits and vegetables, so that the control effect of the FJY-3 strain on the gray mold is further measured by taking the gray mold of strawberries as an example.
1. Inhibition effect of FJY-3 strain on strawberry gray mold
Inoculating FJY-3 strain into 250mL triangular flask containing sterilized 50mL YPD liquid medium, shaking culturing at 28deg.C for 12 hr/min, diluting with sterile water to 10 times, counting with blood cell counting plate, and diluting bacterial solution of FJY-3 strain to 1×10 6 cells/mL。
Preparation of gray mold fungus blocks: inoculating fresh mycelium blocks of Botrytis cinerea in the center of PDA culture medium, attaching sterilized 6mm filter paper sheets on the culture medium around the mycelium blocks, culturing at 28deg.C for 7 days, and spreading mycelium on the filter paper sheets for use.
The strawberry fruits to be tested are selected to have uniform size and maturity, the surfaces of the fruits are free of mechanical injuries such as scratch and the like and diseases and insect injuries, the fruits are washed by sterile water, soaked and sterilized for 2min by 70% ethanol, washed by the sterile water for 2 times to wash out residual ethanol, and placed in a sterile operation table to be naturally dried for standby. The strawberries were then subjected to the following treatments: (1) control group 1 (CK 1): soaking in sterile water for 30s; (2) control group 2 (CK 2): by 1X 10 6 Soaking the bacterial liquid of the cells/mL FJY-3 strain for 30s; (3) control group 3 (CK 3): inoculating gray mold; (4) treatment group (Tr): by 1X 10 6 Soaking the cells/mL of the bacterial liquid of the FJY-3 strain for 30s, culturing at 23 ℃ for 24 hours, and inoculating the gray mold fungus blocks. After naturally airing the surface of the strawberry, putting the strawberry into a plastic tray lined with wet gauze, sealing with a preservative film, and culturing at 23 ℃; each treatment was repeated for 9 fruits and the test was repeated 2 times. After 5d culture, the antibacterial rate and the lesion growth size are recorded, so that the biocontrol effect of the Kazakhstan yeast on the gray mold of the strawberry is evaluated.
The growth inhibition effect of the bacterial liquid of the FJY-3 strain on the strawberry gray mold disease spots is shown in figure 9, and the statistical result of the growth inhibition of the bacterial liquid of the FJY-3 strain on the strawberry gray mold disease spots is shown in figure 10. As can be seen from FIGS. 9 and 10, the growth of the gray mold-inoculated strawberry fruit was 15mm in size, and 1X 10 was used 6 After the bacterial liquid of the cells/mL FJY-3 strain is soaked, strawberry fruits of gray mold are inoculated, the growth size of the disease spots is 1mm, and the bacteriostasis rate reaches 93.33%; the FJY-3 strain has obvious biocontrol effect on the gray mold of strawberry fruits.
2. Inhibition effect of different concentrations of FJY-3 strain on strawberry gray mold
Inoculating FJY-3 strain into 250mL triangular flask containing sterilized 50mL YPD liquid medium, shaking culturing at 28deg.C for 12 hr/min, diluting with sterile water to 10 times, counting with blood cell counting plate, and diluting bacterial solution of FJY-3 strain to concentration of 10 4 、10 5 、10 6 、10 7 cells/mL. The following treatments were performed on the strawberries: (1) control group (CK): inoculating gray mold; (2) treatment group (Tr): respectively using 10 4 、10 5 、10 6 、10 7 Soaking the cells/mL of the bacterial liquid of the FJY-3 strain for 30s, culturing at 23 ℃ for 24 hours, and inoculating the gray mold. After naturally airing the surface of the strawberry, putting the strawberry into a plastic tray lined with wet gauze, sealing with a preservative film, and culturing at 23 ℃; each treatment was repeated for 9 fruits and the test was repeated 2 times. After 5d culture, the growth of the fruit spots is recorded, so that the biocontrol effect of the FJY-3 strains with different inoculation concentrations on the gray mold of the strawberry is evaluated.
The growth inhibition effect of bacterial solutions of different concentrations of FJY-3 strains on the gray mold lesions of the strawberries is shown in figure 11, and the statistical result of the growth inhibition of bacterial solutions of different concentrations of FJY-3 strains on the gray mold lesions of the strawberries is shown in figure 12. As can be seen from fig. 11 and 12, strawberry fruit warp 10 4 、10 5 、10 6 、10 7 The growth length of the bacterial liquid of the cells/mL FJY-3 strain and the bacterial spot of the inoculated gray mold after soaking are 5mm, 1.5mm, 1mm and 1mm respectively, and the bacteriostasis rates are 68.75%, 90.63%, 93.75% and 93.75%, 10% respectively 5 Bacterial solution pair of FJY-3 strain with concentration above cells/mLThe biocontrol effect of gray mold reaches more than 90 percent, 10 6 cells/mL is the optimal treatment concentration to inhibit the growth of Botrytis cinerea.
Example 6 Effect of FJY-3 Strain on strawberry postharvest storage quality
Pretreatment of strawberry fruits to be tested the following treatments were performed on strawberries as in example 5: (1) control group (CK): soaking in sterile water for 2min; (2) treatment group (Tr): by 1X 10 6 The bacterial solution of the cells/mL FJY-3 strain is soaked for 2min. After naturally airing the surface of the strawberry, putting the strawberry into a plastic tray lined with wet gauze, sealing with a preservative film, and culturing at 20 ℃; 5 storage quality indexes of the strawberries are recorded every day for 7 days; each treatment was repeated for 9 fruits and the test was repeated 3 times.
(1) Weight loss rate
Weight loss ratio at initial strawberry weight M 0 And each time the weight M of the strawberries is measured 1 The difference is expressed as a percentage of the original sample weight, and is calculated by weighing as follows: weight loss ratio = [ (M) 0 -M 1 )/M 0 ]X 100%; the measurements were performed 3 times in parallel and averaged.
(2) Hardness of
The hardness change of the strawberries in Kg/cm during storage is measured by adopting a GY-3 pointer fruit durometer 2 Or Pa; the measurements were performed 3 times in parallel and averaged.
(3) Soluble solids content (TSS)
10g of strawberries were ground into homogenate by a mortar, the homogenate was centrifuged by a centrifuge (8000 r/min,10 min), and the supernatant was collected, and the content of soluble solids in each group of strawberries was measured by an NK-55T high-precision digital display glycometer, and the measurement was performed in parallel for 3 times, and an average value was obtained.
(4) Vitamin C content (Vc content)
The measurement of vitamin C content of strawberry was carried out using a vitamin C test kit (phosphomolybdic acid microplate method) produced by Shanghai source leaf biotechnology Co.
(5) Titratable Acidity (TA)
An acid-base titration method is adopted. 10g of strawberry is weighed, ground into homogenate in a mortar, washed into a 50mL centrifuge tube with a small amount of distilled water, and the volume is fixed to 50mL to be used as a sample extracting solution, and the extracting solution is centrifuged for 10min at 8000 r/min. Respectively taking 10mL of centrifugal sample solution in 3 triangular flasks, adding 2 drops of 1% phenolphthalein reagent, titrating to reddish color by using 0.1mol/L NaOH standard solution, and parallelly measuring for 3 times at the end point of 1min, wherein the formula is as follows:
the measurement results of the 5 storage quality indexes of the strawberries are shown in table 2, and as shown in table 2, the weight loss rate, hardness, soluble solids, titratable acid and vitamin C content of the strawberry fruits treated by the FJY-3 strain are not significantly different from those of the control group, so that the FJY-3 strain has no influence on the quality of the strawberries, and can be used for preventing and treating gray mold of the strawberries.
TABLE 2 Effect of Kazakhstan Yeast FJY-3 on the post-harvest storage quality of strawberry
Note that: CK is a control group without FJY-3 strain; tr is 1×10 6 Treatment group of cells/mL FJY-3 strain. The same letter in the same column represents the difference insignificant (p > 0.05).
The above examples are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above examples, and any other changes, modifications, substitutions, combinations, and simplifications that do not depart from the spirit and principle of the present invention should be made in the equivalent manner, and the embodiments are included in the protection scope of the present invention.
Claims (10)
1. Kazakhstan yeast strainKazachstania humilis) A fJY-3 strain, characterized in that said strain was deposited at the collection of microbiological bacterial strains in the cantonese province at month 4 of 2022 under the accession number gdMCC No:63034.
2. use of the FJY-3 strain of claim 1 for the preparation of pectase.
3. Use of the FJY-3 strain of claim 1 for biological degumming of coffee or for preparing a biological degumming formulation for coffee.
4. Use of the FJY-3 strain of claim 1 for the preparation of ethanol from coffee cherry husks.
5. The use of the FJY-3 strain of claim 1 or the bacterial liquid of the FJY-3 strain of claim 1 in inhibiting plant pathogenic bacteria, characterized in that the plant pathogenic bacteria is Botrytis cinerea @Botrytis cinerea) Penicillium Italian mouldPenicillium italicum) Fusarium graminearumFusarium graminearum) Anthrax of gum sporeColletotrichum gloeosporioides) Banana wilt germF.oxysporum f.sp.cubense) One or more of the following.
6. Use of the FJY-3 strain of claim 1 or the bacterial liquid of the FJY-3 strain of claim 1 for the preparation of a preparation for inhibiting plant pathogenic bacteria, wherein the plant pathogenic bacteria are one or more of gray mold, penicillium italicum, fusarium graminearum, colletotrichum gloeosporium, banana fusarium wilt.
7. Use of the FJY-3 strain of claim 1 or the bacterial liquid of the FJY-3 strain of claim 1 for controlling plant diseases, wherein the plant diseases are one or more of gray mold, blue mold, scab, anthracnose and banana vascular wilt.
8. Use of the FJY-3 strain of claim 1 or the bacterial liquid of the FJY-3 strain of claim 1 for preparing a preparation for controlling plant diseases, wherein the plant diseases are one or more of gray mold, blue mold, scab, anthracnose and banana vascular wilt.
9. The use according to claim 7 or 8, wherein the plant disease is one or more of gray mold caused by gray mold, blue mold caused by penicillium italicum, scab caused by fusarium graminearum, anthracnose caused by colletotrichum gloeosporioides, banana vascular wilt caused by banana vascular wilt.
10. A plant disease biocontrol preparation, which is characterized in that the biocontrol preparation contains the FJY-3 strain of claim 1 or the bacterial liquid of the FJY-3 strain of claim 1; the plant diseases are one or more of gray mold, blue mold, scab, anthracnose and banana vascular wilt.
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Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109321479A (en) * | 2018-11-27 | 2019-02-12 | 宜宾五粮液股份有限公司 | Acidproof saccharomyces cerevisiae and application thereof |
| CN111961600A (en) * | 2020-08-27 | 2020-11-20 | 宜宾五粮液股份有限公司 | Solid-state fermentation aroma-producing yeast and application thereof |
| CN113201478A (en) * | 2021-05-27 | 2021-08-03 | 广西民族大学 | Siamese bacillus Gxun-36 secreting iron carrier and application thereof |
| CN114410489A (en) * | 2021-12-31 | 2022-04-29 | 云南大学 | Abnormal yeast Weikehan yeast CAP5 strain and application thereof |
| CN114806762A (en) * | 2022-03-30 | 2022-07-29 | 江南大学 | Method for reducing and controlling generation of isoamyl alcohol in white spirit fermentation process by regulating and controlling sugar consumption rate |
| CN115777921A (en) * | 2023-01-29 | 2023-03-14 | 贵州南山婆食品加工有限公司 | Preparation method of functional fruit and vegetable pulp and solid powder fermented by composite strain |
| CN116138429A (en) * | 2022-09-16 | 2023-05-23 | 新疆康美瑞农业发展有限公司 | Short Kazakhstan yeast XJ-65 and application thereof in pepper fermentation |
| CN116590164A (en) * | 2023-06-28 | 2023-08-15 | 云南大学 | Kluyveromyces pichia pastoris FJY-4 strain and application thereof |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106604647A (en) * | 2014-08-29 | 2017-04-26 | 杜邦营养生物科学有限公司 | Method of fermenting cocoa beans |
-
2023
- 2023-01-17 CN CN202310084025.3A patent/CN116083256B/en active Active
Patent Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109321479A (en) * | 2018-11-27 | 2019-02-12 | 宜宾五粮液股份有限公司 | Acidproof saccharomyces cerevisiae and application thereof |
| CN111961600A (en) * | 2020-08-27 | 2020-11-20 | 宜宾五粮液股份有限公司 | Solid-state fermentation aroma-producing yeast and application thereof |
| CN113201478A (en) * | 2021-05-27 | 2021-08-03 | 广西民族大学 | Siamese bacillus Gxun-36 secreting iron carrier and application thereof |
| CN114410489A (en) * | 2021-12-31 | 2022-04-29 | 云南大学 | Abnormal yeast Weikehan yeast CAP5 strain and application thereof |
| CN114806762A (en) * | 2022-03-30 | 2022-07-29 | 江南大学 | Method for reducing and controlling generation of isoamyl alcohol in white spirit fermentation process by regulating and controlling sugar consumption rate |
| CN116138429A (en) * | 2022-09-16 | 2023-05-23 | 新疆康美瑞农业发展有限公司 | Short Kazakhstan yeast XJ-65 and application thereof in pepper fermentation |
| CN115777921A (en) * | 2023-01-29 | 2023-03-14 | 贵州南山婆食品加工有限公司 | Preparation method of functional fruit and vegetable pulp and solid powder fermented by composite strain |
| CN116590164A (en) * | 2023-06-28 | 2023-08-15 | 云南大学 | Kluyveromyces pichia pastoris FJY-4 strain and application thereof |
Non-Patent Citations (3)
| Title |
|---|
| Interactions between Kazachstania humilis Yeast Species and Lactic Acid Bacteria in Sourdough;Belén Carbonetto等;Microorganisms;20200226;第1-10页 * |
| 哈萨克斯坦酵母FJY-3菌株对草莓灰霉病的生防效果;孔德婷等;中国植物病理学会2023年学术年会论文集;20230815;第621页 * |
| 甘蔗尾中酵母菌的分离与鉴定;郭艳霞;杨承剑;唐庆凤;彭开屏;唐振华;李孟伟;李丽莉;梁辛;谢芳;;中国酿造;20181025(第10期);第77-80页 * |
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