CN114921350B - Flocculation yeast with antibacterial property and application thereof - Google Patents
Flocculation yeast with antibacterial property and application thereof Download PDFInfo
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- CN114921350B CN114921350B CN202210056137.3A CN202210056137A CN114921350B CN 114921350 B CN114921350 B CN 114921350B CN 202210056137 A CN202210056137 A CN 202210056137A CN 114921350 B CN114921350 B CN 114921350B
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- hanseniaspora
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- flocculation
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- 238000005189 flocculation Methods 0.000 title abstract description 14
- 230000016615 flocculation Effects 0.000 title abstract description 11
- 230000000844 anti-bacterial effect Effects 0.000 title abstract description 8
- 240000004808 Saccharomyces cerevisiae Species 0.000 title abstract description 6
- 230000001580 bacterial effect Effects 0.000 claims abstract description 15
- 241000191967 Staphylococcus aureus Species 0.000 claims description 5
- 244000063299 Bacillus subtilis Species 0.000 claims description 4
- 235000014469 Bacillus subtilis Nutrition 0.000 claims description 4
- 241001149669 Hanseniaspora Species 0.000 claims description 2
- 241000186781 Listeria Species 0.000 claims description 2
- 241000320412 Ogataea angusta Species 0.000 claims 3
- 241001646719 Escherichia coli O157:H7 Species 0.000 claims 1
- 239000003814 drug Substances 0.000 claims 1
- 238000000034 method Methods 0.000 abstract description 3
- 238000012408 PCR amplification Methods 0.000 abstract description 2
- 230000003311 flocculating effect Effects 0.000 abstract description 2
- 238000012163 sequencing technique Methods 0.000 abstract description 2
- 235000009917 Crataegus X brevipes Nutrition 0.000 abstract 1
- 235000013204 Crataegus X haemacarpa Nutrition 0.000 abstract 1
- 235000009685 Crataegus X maligna Nutrition 0.000 abstract 1
- 235000009444 Crataegus X rubrocarnea Nutrition 0.000 abstract 1
- 235000009486 Crataegus bullatus Nutrition 0.000 abstract 1
- 235000017181 Crataegus chrysocarpa Nutrition 0.000 abstract 1
- 235000009682 Crataegus limnophila Nutrition 0.000 abstract 1
- 235000004423 Crataegus monogyna Nutrition 0.000 abstract 1
- 240000000171 Crataegus monogyna Species 0.000 abstract 1
- 235000002313 Crataegus paludosa Nutrition 0.000 abstract 1
- 235000009840 Crataegus x incaedua Nutrition 0.000 abstract 1
- 239000007788 liquid Substances 0.000 description 8
- 241000894006 Bacteria Species 0.000 description 5
- 241000588722 Escherichia Species 0.000 description 5
- 239000002207 metabolite Substances 0.000 description 5
- RJQXTJLFIWVMTO-TYNCELHUSA-N Methicillin Chemical compound COC1=CC=CC(OC)=C1C(=O)N[C@@H]1C(=O)N2[C@@H](C(O)=O)C(C)(C)S[C@@H]21 RJQXTJLFIWVMTO-TYNCELHUSA-N 0.000 description 4
- 239000001963 growth medium Substances 0.000 description 4
- 229960003085 meticillin Drugs 0.000 description 4
- 229920001817 Agar Polymers 0.000 description 3
- 241000588724 Escherichia coli Species 0.000 description 3
- 241000186779 Listeria monocytogenes Species 0.000 description 3
- 241001138501 Salmonella enterica Species 0.000 description 3
- 239000008272 agar Substances 0.000 description 3
- 238000004140 cleaning Methods 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 244000005700 microbiome Species 0.000 description 3
- 235000014493 Crataegus Nutrition 0.000 description 2
- 241001092040 Crataegus Species 0.000 description 2
- 241001052560 Thallis Species 0.000 description 2
- 230000002411 adverse Effects 0.000 description 2
- 238000000855 fermentation Methods 0.000 description 2
- 230000004151 fermentation Effects 0.000 description 2
- 230000005764 inhibitory process Effects 0.000 description 2
- 238000002955 isolation Methods 0.000 description 2
- 238000009630 liquid culture Methods 0.000 description 2
- 239000003755 preservative agent Substances 0.000 description 2
- 238000001228 spectrum Methods 0.000 description 2
- 239000011550 stock solution Substances 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 241000193830 Bacillus <bacterium> Species 0.000 description 1
- 108020004414 DNA Proteins 0.000 description 1
- 108091023242 Internal transcribed spacer Proteins 0.000 description 1
- 108090000526 Papain Proteins 0.000 description 1
- 241000235648 Pichia Species 0.000 description 1
- 239000004365 Protease Substances 0.000 description 1
- 241000607142 Salmonella Species 0.000 description 1
- 241000191940 Staphylococcus Species 0.000 description 1
- 239000003242 anti bacterial agent Substances 0.000 description 1
- 229940088710 antibiotic agent Drugs 0.000 description 1
- 244000052616 bacterial pathogen Species 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000000443 biocontrol Effects 0.000 description 1
- 229960005091 chloramphenicol Drugs 0.000 description 1
- WIIZWVCIJKGZOK-RKDXNWHRSA-N chloramphenicol Chemical compound ClC(Cl)C(=O)N[C@H](CO)[C@H](O)C1=CC=C([N+]([O-])=O)C=C1 WIIZWVCIJKGZOK-RKDXNWHRSA-N 0.000 description 1
- 239000013078 crystal Substances 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 238000003912 environmental pollution Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 230000002906 microbiologic effect Effects 0.000 description 1
- 238000009629 microbiological culture Methods 0.000 description 1
- 229940055729 papain Drugs 0.000 description 1
- 235000019834 papain Nutrition 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 238000004064 recycling Methods 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
Classifications
-
- 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
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Abstract
The invention discloses a flocculating yeast with antibacterial property and application thereofHanseniaspora spStrain CC-ZZ6, accession number: CGMCC No.23572; the said processHanseniaspora spApplication of bacterial strain CC-ZZ6 in preparing flocculant. The strain is separated from fresh hawthorn, and after PCR amplification and sequencing of ITS and 26S D1/D2 regions, the 26S D1/D2 sequence is shown to be identical with that of BLAST comparison resultHanseniaspora vineaeThe similarity of the ITS region sequence is 99.35%, and the ITS region sequence is shown to be identical to that of the ITS region sequence through BLAST comparison resultHanseniaspora spSimilarity of 99.27%; has certain flocculation capacity (flocculation rate is 51.00%), and has antibacterial effect.
Description
Technical Field
The invention belongs to the technical field of microorganisms, and particularly relates to flocculation yeast with antibacterial property and application thereof.
Background
Antibiotics and chemical preservatives can well achieve the effect of inhibiting pathogenic bacteria or spoilage bacteria, but have adverse effects such as bacterial strain resistance and environmental pollution caused by chemical preservatives. With the enhancement of health consciousness of people, the safe and environment-friendly biological prevention and control technology is attracting more and more attention. Biocontrol is a common practice with microorganisms or metabolites of microorganisms.
Self-flocculating bacteria are generally more tolerant to adverse environments due to their flocculating properties. In the fermentation process, the high-concentration metabolite can be tolerated due to stronger tolerance, so that the high-concentration metabolite can be used for fermenting and producing the yield of target metabolites required by more people, and the high-concentration metabolite is beneficial to the separation of fermentation liquor and thalli and the recycling of thalli. Therefore, the strain with certain self-flocculation capacity has better application value.
Disclosure of Invention
The invention aims to provide flocculation yeast with antibacterial property and application thereof.
Hanseniaspora spStrain CC-ZZ6, deposited with the accession number: CGMCC No.23572.
The said processHanseniaspora spStrain CC-ZZ6 in inhibitionRiemerella anatipestiferATCC11845、Escherichia coliO157:H7、methicillin-resistantStaphylococcus aureusATCC43300、Bacillus subtilisCICC 10732、 Salmonella choleraesuisATCC 50019、Staphylococcus aureusATCC29213、Listeria monocytogenesATCC19111、Escherichia coliApplication in CICC 10899.
The invention providesHanseniaspora spStrain CC-ZZ6, deposited with the accession number: CGMCC No.23572; the said processHanseniaspora spApplication of bacterial strain CC-ZZ6 in preparing flocculant. The strain has a certain flocculation capacity (flocculation rate 51.00%) and has good flocculation effect onRiemerella anatipestifer(ATCC 11845)、Escherichia coliO157:H7、methicillin-resistantStaphylococcus aureus(ATCC43300)、Bacillus subtilis(CICC 10732)、Salmonella choleraesuis(ATCC 50019)、Staphylococcus aureus(ATCC 29213)、Listeria monocytogenes(ATCC 19111)、Escherichia coli(CICC 10899) has antibacterial effect.
Drawings
FIG. 1 is a schematic view ofHanseniaspora sp.The strain CC-ZZ6 is in a form after being dyed by crystal violet;
FIG. 2 is a schematic view ofHanseniaspora sp.The self-flocculation condition of the strain CC-ZZ6 in 5ml YPD liquid culture medium is shown as a state after vortex for 1 min;
FIG. 3 is a schematic view ofHanseniaspora sp.Bacterial inhibition spectrum of strain CC-ZZ6.
Detailed Description
Example 1Hanseniaspora sp.Isolation and identification of Strain CC-ZZ6
1. And (3) separating and purifying: after fresh hawthorns are purchased from a fruit store, the hawthorns are transported back to a laboratory within half an hour; placing fresh fructus crataegi in sterile bag, adding appropriate amount of sterile PBS, and cleaning under shaking to obtain stock solution. 1mL of the stock solution was taken out for gradient dilution, and 100. Mu.L of 10 was taken out, respectively -1 、10 -2 、10 -3 And 10 -4 Is plated on YPD agar plates containing 25. Mu.g/mL chloramphenicol. After the plates were incubated at 30℃for 3-7 days, suspicious colonies on the plates were picked for isolation and purification.
2. Molecular identification: DNA from the purified yeast was extracted as template and the 26S D1/D2 and ITS regions were sequenced by PCR amplification using primers NL1 (5'-GCA TAT CAA TAAGCG GAG GAA AAG-3')/NL 4 (5'-GGT CCG TGT TTC AAG ACGG-3') and ITS1 (5'-TCC GTA GGT GAA CCT GCG G-3')/ITS 4 (5'-TCC TCC GCT TAT TGA TAT GC-3'). After sequencing the PCR product, the 26S D1/D2 sequence was shown to be identical to that of the BLAST alignmentHanseniasporavineaeThe similarity of the ITS region sequence is 99.35%, and the ITS region sequence is shown to be identical to that of the ITS region sequence through BLAST comparison resultHanseniasporasp.The similarity of (2) was 99.27%, so that the strain was tentatively determined to belong toHanseniasporaGenus, named Hansenula polymorphaHanseniasporasp.Bacterial strain CC-ZZ6. The strain is preserved in China general microbiological culture collection center (CGMCC) of 10-11 days 2021, and the preservation number is CGMCC No.23572, wherein the address is CGMCC No. 3 of North west road No. 1, the university of China, and the microbiological institute of China, which are the regions of Korea of Beijing, respectively.
Example 2 determination of self-flocculation Capacity of Strain
Picking upHanseniaspora sp.The strain CC-ZZ6 is cultured in 5ml YPD liquid culture medium at 30 ℃ and 120rpm for 48 hours, and then vortexed for 10sStanding for 10min, carefully sucking 200 mu L of bacterial liquid on the upper layer, measuring an OD600 value, and marking as OD initial; centrifuging the bacterial liquid at 3000rpm for 5min, collecting bacterial liquid, cleaning with PBS, re-suspending in PBS, adding proper amount of papain to decolour the bacterial liquid, centrifuging at 3000rpm for 5min after complete decolouring, collecting bacterial liquid, cleaning with PBS, re-suspending the bacterial liquid in 5mLYPD culture medium, absorbing 200 mu L of upper layer bacterial liquid, measuring OD600 value, and recording as OD after; the flocculation rate (%) = (1-ODinitial/ODafter) ×100. The flocculation rate of the bacteria was calculated to be 51.00%.
Example 3 bacteriostasis Spectrum experiment
Picking upHanseniaspora sp.Bacterial strain 32CC-ZZ6 was centrifuged at 3000rpm for 5min at 30℃in 5ml YPD liquid medium to collect the cells, and the cells were deflocculated as described in example 2, after which 10. Mu.L of the cells were aspirated and streaked at the center of YPD agar plates and cultured at 30℃for 72h. The indicator bacteria cultivated by the TSB culture mediumRiemerella anatipestifer(ATCC 11845)、Escherichia coliO157:H7、methicillin-resistantStaphylococcus aureus(ATCC43300)、Bacillus subtilis(CICC 10732)、 Salmonella choleraesuis(ATCC 50019)、Staphylococcus aureus(ATCC 29213)、 Listeria monocytogenes(ATCC 19111)、Escherichia coli(CICC 10899)) to a colony concentration of about 1X 10 6 cfu/mL, 10. Mu.L of each was taken out and was perpendicular to YPD agar plates after 72h of incubationHanseniaspora sp.The strain CC-ZZ6 is streaked, after streaking is completed, the plate is cultivated for 20 to 24 hours at 37 ℃, and observation is carried outHanseniaspora sp.Bacterial inhibition of indicator bacteria by strain CC-ZZ6. As can be seen from FIG. 3, theHanseniaspora sp.Strain CC-ZZ6 pairRiemerella anatipestifer(ATCC 11845)、Escherichia coliO157:H7、 methicillin-resistantStaphylococcus aureus(ATCC43300)、Bacillus subtilis(CICC 10732)、 Salmonella choleraesuis(ATCC 50019)、Staphylococcus aureus(ATCC 29213)、 Listeria monocytogenes(ATCC 19111)、Escherichia coli(CICC 10899) has antibacterial effect.
Claims (2)
1. Hansenula polymorpha (Hansenula polymorpha)Hanseniaspora sp) Strain CC-ZZ6, deposited with the accession number: CGMCC No.23572.
2. The Hansenula polymorpha of claim 1Hanseniaspora sp) Preparation of bacterial strain CC-ZZ6 for inhibitionRiemerella anatipestiferATCC 11845、Escherichia coli O157:H7、Methicillin- resistant Staphylococcus aureus ATCC43300、Bacillus subtilis CICC 10732、Staphylococcus aureus ATCC 29213、Listeria monocytogenesATCC 19111Escherichia coliApplication of CICC 10899 in medicine is provided.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210056137.3A CN114921350B (en) | 2022-01-18 | 2022-01-18 | Flocculation yeast with antibacterial property and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210056137.3A CN114921350B (en) | 2022-01-18 | 2022-01-18 | Flocculation yeast with antibacterial property and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN114921350A CN114921350A (en) | 2022-08-19 |
| CN114921350B true CN114921350B (en) | 2024-03-22 |
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| CN202210056137.3A Active CN114921350B (en) | 2022-01-18 | 2022-01-18 | Flocculation yeast with antibacterial property and application thereof |
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103740604A (en) * | 2014-01-26 | 2014-04-23 | 北京绿环国际科技有限公司 | Hanseniaspora uvarum and application thereof |
| CN112574929A (en) * | 2021-01-11 | 2021-03-30 | 吉林大学 | Paenibacillus gibilinus YPG26 and medical application thereof |
| CN112940961A (en) * | 2021-01-04 | 2021-06-11 | 中冶华天南京工程技术有限公司 | Microbial flocculant and preparation method and application thereof |
| CN113403211A (en) * | 2021-06-16 | 2021-09-17 | 青岛农业大学 | Hansenula polymorpha MP1861 with bacteriostatic effect on grape juice and application thereof |
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- 2022-01-18 CN CN202210056137.3A patent/CN114921350B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103740604A (en) * | 2014-01-26 | 2014-04-23 | 北京绿环国际科技有限公司 | Hanseniaspora uvarum and application thereof |
| CN112940961A (en) * | 2021-01-04 | 2021-06-11 | 中冶华天南京工程技术有限公司 | Microbial flocculant and preparation method and application thereof |
| CN112574929A (en) * | 2021-01-11 | 2021-03-30 | 吉林大学 | Paenibacillus gibilinus YPG26 and medical application thereof |
| CN113403211A (en) * | 2021-06-16 | 2021-09-17 | 青岛农业大学 | Hansenula polymorpha MP1861 with bacteriostatic effect on grape juice and application thereof |
Non-Patent Citations (3)
| Title |
|---|
| Flocculation and coflocculation of bacteria by yeasts;X Peng等;《Appl Microbiol Biotechnol . 》;第55卷(第6期);777-781 * |
| 桃褐腐病菌拮抗酵母的筛选、鉴定及其抑菌活性初探;王晨;《中国酿造》;第36卷(第05期);63-66 * |
| 耐甲氧西林金黄色葡萄球菌生物防治酵母菌的筛选、鉴定及评价;沈勇;《中国博士学位论文全文数据库基础科学辑》(第2期);A006-453 * |
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