CN112342163A - Burkholderia and application thereof - Google Patents

Burkholderia and application thereof Download PDF

Info

Publication number
CN112342163A
CN112342163A CN202011269503.0A CN202011269503A CN112342163A CN 112342163 A CN112342163 A CN 112342163A CN 202011269503 A CN202011269503 A CN 202011269503A CN 112342163 A CN112342163 A CN 112342163A
Authority
CN
China
Prior art keywords
strain
nootkatone
valencene
culture medium
bacterial
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
CN202011269503.0A
Other languages
Chinese (zh)
Inventor
任婧楠
彭芷芊
范刚
张璐璐
李晓
潘思轶
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.)
Huazhong Agricultural University
Original Assignee
Huazhong Agricultural 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 Huazhong Agricultural University filed Critical Huazhong Agricultural University
Priority to CN202011269503.0A priority Critical patent/CN112342163A/en
Publication of CN112342163A publication Critical patent/CN112342163A/en
Pending legal-status Critical Current

Links

Images

Classifications

    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12RINDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00Microorganisms ; Processes using microorganisms
    • C12R2001/01Bacteria or Actinomycetales ; using bacteria or Actinomycetales
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00Microorganisms, 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
    • C12N1/20Bacteria; Culture media therefor
    • C12N1/205Bacterial isolates
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00Microorganisms, 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
    • C12N1/20Bacteria; Culture media therefor
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12PFERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P7/00Preparation of oxygen-containing organic compounds
    • C12P7/24Preparation of oxygen-containing organic compounds containing a carbonyl group
    • C12P7/26Ketones

Landscapes

  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Genetics & Genomics (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Biotechnology (AREA)
  • General Engineering & Computer Science (AREA)
  • Microbiology (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Biomedical Technology (AREA)
  • Virology (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Medicinal Chemistry (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

The invention relates to the field of microorganisms, in particular to Burkholderia and application thereof. Firstly, diluting and coating a sample bacterial suspension on a flat plate through primary screening, selecting a single colony, taking valencene as a unique carbon source, carrying out primary screening by drawing a growth curve, and selecting a strain with good growth to carry out the next experiment; secondly, re-screening, analyzing and comparing the yield of the grapefruit ketone generated by bioconversion of valencene by the strains, and selecting strains with high grapefruit ketone yield to perform the next experiment; and (3) carrying out streak separation on the selected strains, and analyzing and measuring the products and the content of the strains in the valencene biotransformation. Compared with the existing physical and chemical synthesis methods, the method has the characteristics of high specificity, mild reaction conditions, high reaction rate, high yield, low cost, few reaction steps and the like, and the produced nootkatone belongs to natural perfume and can be added into foods, cosmetics and other household products.

Description

Burkholderia and application thereof
Technical Field
The invention relates to the field of microorganisms, in particular to Burkholderia and application thereof.
Background
Nootkatone is one of important substances in pomelo fragrance components, has a persistent strong citrus-flavor, and is mainly added as a perfume in foods and cosmetics. A certain amount of nootkatone is added into cigarettes, so that the fragrance of the cigarettes can be increased, the fragrance of the cigarettes can be enriched, and the stimulation can be reduced; the nootkatone has sweet pericarp and costus scent, so the nootkatone can also be used for preparing edible essences such as pomelo, orange, tropical fruit and the like.
Natural nootkatone is mainly present in essential oils such as bergamot oil, grapefruit oil, lemon oil, citrus oil, lime oil, and tangerine oil. However, the natural content of the grapefruit seed oil is generally low, so that the grapefruit seed oil used in actual production is mainly derived from chemical synthesis. The most common synthetic methods are oxidation using valencene as a starting material, oxidation of valencene using dichromate, or oxidation using a catalyst in the presence of t-butyl hydroperoxide (TBHP). The chemical synthesis methods use a large amount of heavy metal catalysts, which seriously damage the environment, and other available non-heavy metal catalysts have great potential safety hazards although the environment cannot be damaged, and the production efficiency is too low to meet the production requirements.
European food legislation has specifically established that the nootkatone product obtained by chemical synthesis cannot be sold and used as a natural flavour which can only be prepared by physical processes (extraction from natural raw materials) or by enzymatic/microbiological processes. Microbial conversion is the structural modification of exogenous substrates by microbial cells or enzymes, namely the catalytic reaction of a certain enzyme or a series of enzymes produced in the microbial metabolic process on the substrates, so that the perfumes produced by microbial conversion also belong to natural perfumes. The microbial conversion has the characteristics of high specificity, mild reaction conditions, high reaction rate, high yield, low cost, few reaction steps and the like. At present, the method is widely applied to aspects of organic synthesis, new drug development, in-vitro metabolite prediction, spice production and the like.
Disclosure of Invention
The invention aims to efficiently and accurately screen out strains capable of transforming nootkatone by adopting a screening method taking valencene as a unique carbon source, and transform the valencene which is rich in natural content and low in application value into natural spice nootkatone with stronger fragrance, wider application and higher commercial value.
In order to realize the aim, the invention provides burkholderia, the preservation number is CCTCC NO: m2020510, and the screening method comprises the following steps:
(1) taking 10g of soil sample from research center of orange engineering technology in Hubei province, adding the soil sample into 90g of sterilized normal saline, and mixing uniformly.
(2) 2mL of the mixed liquid is put into 100mL of minimum culture medium, and 1mL of valencene (0.22 μm filter membrane filtration sterilization, 4 ℃ storage) is added, and shake culture is carried out at 37 ℃ and 121r/min for 2-7 days.
(3) Diluting the bacterial liquid obtained in the step (2) to 10 degrees with sterilized normal saline-1、10-2、10-3、10-4、 10-5And 10-6And 6 dilution gradients in total, coating the gradient bacterial liquid on a plate to separate strains, and culturing at 37 ℃ until bacterial colonies completely grow.
(4) After the bacterial colony grows out in the step (3), selecting a plate with proper concentration to pick out a single bacterial colony, and inoculating the single bacterial colony in an LB culture medium for activated culture.
(5) Primary screening: taking appropriate amount of activated bacteria liquid, adding 1% valencene in minimum culture medium (0.22 μm filter membrane filtration sterilization, 4 deg.C storage), culturing at 37 deg.C under 210r/min, and measuring bacteria liquid OD every 12h600Values, growth curves of the strains were plotted.
(6) And (5) selecting strains with good growth according to the growth curve drawn in the step (5), and transferring the strains to an LB culture medium for activated culture.
(7) Taking 1mL of bacterial liquid to be cultured in 100mLLB culture medium at 37 ℃ at 210r/min, and obtaining the OD of bacterial liquid600When the value reaches 0.6-0.7, 0.1% valencene is added (0.22 μm filter membrane filtration sterilization,stored at 4 ℃), and cultured for further 72 h.
(8) Re-screening: and respectively taking fermentation liquor cultured for 24h, 48h and 72h for SPME-GC-MS detection analysis, and detecting the yield of the nootkatone generated by strain conversion.
(9) Selecting strains with high yield of nootkatone, performing plate streaking separation, and continuously separating for 3 times. And picking single colonies on the plate to perform activated culture in an LB culture medium.
(10) Transferring 1mL of bacterial solution into 100mLLB culture medium, culturing at 37 deg.C and 210r/min until bacterial solution OD600When the value reaches 0.6-0.7, 0.1% valencene is added (0.22 μm filter membrane filtration sterilization, 4 deg.C storage), and culture is continued for 72 h.
(11) SPME-GC-MS detection is carried out on fermentation liquor cultured for 24h, 48h and 72h respectively, determination and analysis are carried out on transformation products, and the analysis results of the obtained bacterial strain biotransformation products are shown in Table 1:
Figure RE-GDA0002883298160000031
Figure RE-GDA0002883298160000032
TABLE 1
(12) The colony morphology of the strain on a solid culture medium is observed morphologically according to a manual of identifying common bacteria systems, and a plurality of physiological and biochemical indexes of the strain are measured at the same time, and the analysis of the physiological and biochemical characteristics of the obtained strain is shown in table 2:
Figure RE-GDA0002883298160000041
TABLE 2
(13) Extracting DNA of the strain, carrying out PCR amplification and sequencing of 16SrDNA, and carrying out strain homology analysis.
Preferably, the minimum medium components in step (2) are as follows: 6g/LNa2HPO4、0.50 g/LNaCl、3g/LKH2PO4、1g/LNH4Cl, dissolved in 0.85% physiological saline.
Preferably, the composition of the LB medium in step (4) is as follows: 10g/L peptone, 5g/L yeast extract, 10g/L NaCl, dissolved in distilled water.
Preferably, all of the operations described are performed in a sterile operating station, maintaining a sterile environment.
The invention also provides a spice, wherein the nootkatone in the spice is prepared by the following steps of preserving in the formula of CCTCC NO: m2020510 by bioconversion of Burkholderia.
Biological preservation Instructions
And (3) classification and naming: burkholderia plantarii PZQ14, burkholderia sp. Is preserved in China center for type culture Collection (CMGWU university, Wuhan, China) in 2020 on 9/17 th month with the preservation number of CCTCCNO: M2020510.
The invention has the beneficial effects that:
(1) the nootkatone produced by the present invention belongs to natural flavors. Compared with the existing production technology of nootkatone synthesized by a chemical synthesis method, the method for producing nootkatone by a microbiological method has the characteristics of high specificity, mild reaction conditions, high reaction rate, high yield, low cost, few reaction steps and the like.
(2) The invention provides a screening method of a bacterial strain which can use valencene as a unique carbon source and has high conversion rate of nootkatone. The screening method is efficient and accurate, and has the advantages of reducing workload, improving efficiency and the like.
Drawings
FIG. 1 is a graph showing the growth profile of the strain obtained in step (5);
FIG. 2 is a graph showing a comparative analysis of the yield of nootkatone obtained in step (8);
FIG. 3 is a cross-sectional drawing of a plate of four strains obtained by the final screening;
FIG. 4 is a graph showing the growth of the strain in LB medium;
Detailed Description
Example one
The screening steps of the strain which takes valencene as a unique carbon source and has high conversion rate of nootkatone are as follows:
(1) 10g of soil sample is taken from research center of orange engineering technology in Hubei province. The soil sample was added to 90g of sterilized normal saline and mixed well. Taking 2mL of uniformly mixed liquid into 100mL of minimum culture medium, adding 1mL of valencene (0.22 mu m filter membrane filtration sterilization, preservation at 4 ℃), performing shake culture at 37 ℃ at 121r/min for 2-7 days, separating strains by using a gradient dilution coating method, performing culture at 37 ℃ until the strains completely grow, and picking out single colonies.
(2) Primary screening: and inoculating the picked single colony in LB culture medium for activating culture. Taking appropriate amount of bacterial liquid, adding 1% valencene (0.22 μm filter membrane filtration sterilization, 4 deg.C preservation), culturing at 37 deg.C under 210r/min, and measuring bacterial liquid OD every 12h600Values, growth curves of the bacteria were plotted. And selecting a strain with good growth, and transferring the strain to an LB culture medium for enrichment culture.
(3) Re-screening: taking 1mL of bacterial liquid to be cultured in 100mLLB culture medium at 37 ℃ at 210r/min, and obtaining the OD of bacterial liquid600And when the value reaches 0.6-0.7, adding 0.1% valencene in valencene (0.22 mu m filter membrane filtration sterilization, preservation at 4 ℃), continuously culturing for 72h, respectively taking fermentation liquor cultured for 24h, 48h and 72h, carrying out SPME-GC-MS detection analysis, detecting the yield of the grapefruit ketone generated by strain conversion, and selecting a strain with high yield of the grapefruit ketone for the next experiment.
(4) Analytical determination of the bioconversion products of the strains: the selected strains were subjected to plate streaking separation and separated 3 times in succession. Selecting single colony on the plate, performing activated culture in LB culture medium, transferring to 100mLLB culture medium, culturing at 37 deg.C under 210r/min, and culturing as bacterial liquid OD600When the value reaches 0.6-0.7, adding 0.1% valencene myrcene (0.22 mu m filter membrane filtration sterilization, preservation at 4 ℃), continuously culturing for 72h, respectively taking fermentation liquor cultured for 24h, 48h and 72h, carrying out SPME-GC-MS detection, and carrying out determination analysis on the conversion product.
(5) GC-MS analysis of the conversion products:
accurately weighing 5mL of the centrifuged fermentation liquid, transferring the centrifuged fermentation liquid into a 20mL jaw sample bottle, adding 1.8g of NaCl to promote volatilization of volatile components, sealing the fermentation liquid by using a polytetrafluoroethylene spacer, balancing the fermentation liquid on a magnetic stirrer at a constant temperature of 40 ℃ for 15min, inserting an activated 50/30 mu m DVB/CAR/PDMS extraction head (activated at 270 ℃ for 1h) through the spacer, pushing out a fiber head, adsorbing the fermentation liquid in a headspace for 40min, and then inserting a GC-MS sample inlet for desorption for 5 min.
Gas chromatography conditions: 6890N type gas chromatograph has capillary column HP-5(30m × 250 μm × 0.25 μm), temperature programming, initial temperature of 40 deg.C, holding for 3min, heating to 160 deg.C at 3 deg.C/min, holding for 2min, heating to 220 deg.C at 8 deg.C/min, and holding for 3 min. The injection port temperature was 250 ℃. Mass spectrum conditions: 5975B mass spectrometer with ion source temperature 230 deg.C, quadrupole rod temperature 150 deg.C, ionization mode EI, electron energy 70eV, and mass range of 45-550 AMU/sec16
And (3) qualitative analysis: analyzing and identifying by GC-MS combined instrument and utilizing C6~C30Retention time of normal paraffins the retention index of each chromatographic peak was calculated. The analysis result is subjected to preliminary retrieval and data analysis by using a computer spectrum (NIST05/WILEY7.0), and then is compared and subjected to manual spectrogram analysis by combining with retention indexes of documents to confirm each chemical composition of the volatile substances.
Quantitative analysis: and quantifying the nootkatone by adopting an external standard method. Other products are quantified by using cyclohexanone as an internal standard and comparing and calculating the peak area of the cyclohexanone with the peak area of the aroma substances. The aroma concentration and conversion are calculated according to the following formula:
Figure RE-GDA0002883298160000071
Figure RE-GDA0002883298160000072
example two
Strain identification
(1) Morphological and physiological biochemical assays: the colony morphology of the strain on a solid culture medium is observed morphologically according to a 'handbook for identifying common bacteria systems', and a plurality of physiological and biochemical indexes of the strain are measured at the same time.
(2) Extracting bacterial DNA by using a DNA extraction kit, and amplifying 16SrDNA by using a PCR technology, wherein the PCR reaction conditions are as follows: pre-denaturation at 95 ℃ for 5 min; denaturation at 95 ℃, 30sec, annealing at 58 ℃, 30sec, extension at 72 ℃, 45sec, 35 cycles; terminal extension at 72 ℃ for 5 min. And (3) detecting the PCR amplification product by agarose gel electrophoresis, sending the PCR amplification product to Wuhantianyihuiyuan biotechnology limited company for sequencing, and performing homology analysis and comparison on a sequencing result and a known 16SrDNA sequence in GenBank.
EXAMPLE III
The growth curve of the strain in LB medium is drawn:
inoculating strain seed solution into LB culture medium at 1%, culturing at 37 deg.C under 210r/min, and measuring OD every four hours with sterile LB culture medium as blank control600Value, triplicate, in OD600The average is the ordinate and the time is the abscissa, and the growth curve of the strain is plotted.
The above description is only a preferred embodiment of the present invention, and does not limit the present invention, and those skilled in the art can make various modifications and variations. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Sequence listing
<110> university of agriculture in Huazhong
<120> Burkholderia and application thereof
<141> 2020-11-13
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1426
<212> DNA
<213> Burkholderia plantarii sp. PZQ14)
<400> 1
tgtgcggctg ccttaccatg caagtcgaac ggcagcacgg gtgcttgcac ctggtggcga 60
gtggcgaacg ggtgagtaat acatcggaac atgtcctgta gtgggggata gcccggcgaa 120
agccggatta ataccgcata cgatctacgg atgaaagcgg gggaccttcg ggcctcgcgc 180
tatagggttg gccgatggct gattagctag ttggtggggt aaaggcctac caaggcgacg 240
atcagtagct ggtctgagag gacgaccagc cacactggga ctgagacacg gcccagactc 300
ctacgggagg cagcagtggg gaattttgga caatgggcga aagcctgatc cagcaatgcc 360
gcgtgtgtga agaaggcctt cgggttgtaa agcacttttg tccggaaaga aatccttggt 420
tctaatatag ccgggggatg acggtaccgg aagaataagc accggctaac tacgtgccag 480
cagccgcggt aatacgtagg gtgcaagcgt taatcggaat tactgggcgt aaagcgtgcg 540
caggcggttt gttaagaccg atgtgaaatc cccgggctca acctgggaac tgcattggtg 600
actggcaagc tagagtatgg cagagggggg tagaattcca cgtgtagcag tgaaatgcgt 660
agagatgtgg aggaataccg atggcgaagg cagccccctg ggccaatact gacgctcatg 720
cacgaaagcg tggggagcaa acaggattag ataccctggt agtccacgcc ctaaacgatg 780
tcaactagtt gttggggatt catttcctta gtaacgtagc taacgcgtga agttgaccgc 840
ctggggagta cggtcgcaag attaaaactc aaaggaattg acggggaccc gcacaagcgg 900
tggatgatgt ggattaattc gatgcaacgc gaaaaacctt acctaccctt gacatggtcg 960
gaatcccgct gagaggtggg agtgctcgaa agagaaccgg cgcacaggtg ctgcatggct 1020
gtcgtcagct cgtgtcgtga gatgttgggt taagtcccgc aacgagcgca acccttgtcc 1080
ttagttgcta cgcaagagca ctctaaggag actgccggtg acaaaccgga ggaaggtggg 1140
gatgacgtca agtcctcatg gcccttatgg gtagggcttc acacgtcata caatggtcgg 1200
aacagagggt tgccaacccg cgagggggag ctaatcccag aaaaccgatc gtagtccgga 1260
ttgcactctg caactcgagt gcatgaagct ggaatcgcta gtaatcgcgg atcagcatgc 1320
cgcggtgaat acgttcccgg gtcttgtaca caccgcccgt cacaccatgg gagtgggttt 1380
taccagaagt ggctagtcta accgcaagga ggacggtcac cacggt 1426

Claims (8)

1. Burkholderia, the preservation number is CCTCC NO: m2020510.
2. Burkholderia according to claim 1, characterized in that 16S rDNA is as indicated in the SIPO sequence listing.
3. Use of burkholderia according to claim 1 for the production of the natural flavor nootkatone.
4. A preservation number is CCTCC NO: the screening method of M2020510 Burkholderia is characterized by comprising the following steps:
(1) taking 10g of soil sample from research center of orange engineering technology in Hubei province, adding the soil sample into 90g of sterilized normal saline, and mixing uniformly;
(2) taking 2mL of uniformly mixed liquid, adding 1mL of valencene into 100mL of minimum culture medium, and performing shake culture at 37 ℃ for 2-7 days;
(3) diluting the bacterial liquid obtained in the step (2) to 10 degrees with sterilized normal saline-1、10-2、10-3、10-4、10-5And 10-66 dilution gradients in total, coating each gradient bacterial liquid on a plate separation strain, and culturing at 37 ℃ until bacterial colonies completely grow;
(4) after the bacterial colony grows out in the step (3), selecting a flat plate with proper concentration to pick out a single bacterial colony, and inoculating the single bacterial colony in an LB culture medium for activated culture;
(5) primary screening: taking a proper amount of activated and cultured bacterial liquid in a minimum culture medium, adding 1% valencene and myrcene, performing shake culture at 37 ℃, and measuring the OD of the bacterial liquid every 12h600Value, plotting the growth curve of the strain;
(6) Selecting strains with good growth according to the growth curve drawn in the step (5), and transferring the strains to an LB culture medium for activated culture;
(7) taking 1mL of bacterial liquid into 100mL of LB culture medium, performing shake culture at 37 ℃, and obtaining bacterial liquid OD600Adding 0.1% valencene to 0.6-0.7, and culturing for 72 hr;
(8) re-screening: carrying out SPME-GC-MS detection analysis on fermentation liquor cultured for 24h, 48h and 72h respectively, and detecting the yield of the grapefruit ketone generated by strain conversion;
(9) selecting strains with high yield of nootkatone, performing plate streaking separation, and continuously separating for 3 times. Picking single colony on the plate to be activated and cultured in an LB culture medium;
(10) transferring 1mL of the bacterial solution into 100mL of LB culture medium, performing shake culture at 37 ℃, and obtaining OD of the bacterial solution600Adding 0.1% valencene to 0.6-0.7, and culturing for 72 hr;
(11) carrying out SPME-GC-MS detection on fermentation liquor cultured for 24h, 48h and 72h respectively, and determining and analyzing a conversion product;
(12) the colony morphology of the strain on a solid culture medium is observed morphologically according to a 'handbook for identifying common bacteria systems', and a plurality of physiological and biochemical indexes of the strain are measured at the same time.
(13) Extracting DNA of the strain, carrying out PCR amplification and sequencing of 16S rDNA, and carrying out homology analysis of the strain.
5. A flavor containing nootkatone, characterized in that nootkatone is prepared from nootkatone having a preservation number of CCTCC NO: m2020510 by bioconversion of Burkholderia.
6. The method for screening Burkholderia according to claim 4, wherein the minimum medium in the step (2) comprises: 6g/L Na2HPO4、0.50g/L NaCl、3g/L KH2PO4、1g/L NH4Cl, and dissolved in 0.85% physiological saline.
7. The method for producing natural flavor nootkatone by biotransformation according to claim 4, wherein the LB medium composition in step (4) comprises: 10g/L peptone, 5g/L yeast extract, 10g/L NaCl, dissolved in distilled water.
8. The method for producing nootkatone, a natural flavor, by bioconversion of claim 4, wherein all of said steps are performed in a sterile processing station.
CN202011269503.0A 2020-11-13 2020-11-13 Burkholderia and application thereof Pending CN112342163A (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN202011269503.0A CN112342163A (en) 2020-11-13 2020-11-13 Burkholderia and application thereof

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN202011269503.0A CN112342163A (en) 2020-11-13 2020-11-13 Burkholderia and application thereof

Publications (1)

Publication Number Publication Date
CN112342163A true CN112342163A (en) 2021-02-09

Family

ID=74363684

Family Applications (1)

Application Number Title Priority Date Filing Date
CN202011269503.0A Pending CN112342163A (en) 2020-11-13 2020-11-13 Burkholderia and application thereof

Country Status (1)

Country Link
CN (1) CN112342163A (en)

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105189772A (en) * 2013-03-15 2015-12-23 阿迈瑞斯公司 Use of phosphoketolase and phosphotransacetylase for production of acetyl-coenzyme a derived compounds
CN110869500A (en) * 2017-01-26 2020-03-06 马努斯生物合成股份有限公司 Metabolic engineering for microbial production of terpenoid products

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105189772A (en) * 2013-03-15 2015-12-23 阿迈瑞斯公司 Use of phosphoketolase and phosphotransacetylase for production of acetyl-coenzyme a derived compounds
CN110869500A (en) * 2017-01-26 2020-03-06 马努斯生物合成股份有限公司 Metabolic engineering for microbial production of terpenoid products

Non-Patent Citations (6)

* Cited by examiner, † Cited by third party
Title
VLADA B URLACHER等: "Recent advances in oxygenase-catalyzed biotransformations", 《CURRENT OPINION IN CHEMICAL BIOLOGY》 *
XIAO LI等: "Catalytic condition optimization in the conversion of nootkatone from valencene by Yarrowia lipolytica", 《JOURNAL OF FOOD PROCESSING AND PRESERVATION》 *
冯堃: "中国主要柚类果皮成分及活性研究", 《中国优秀博硕士学位论文全文数据库(硕士)工程科技Ⅰ辑》 *
刘均洪等: "加氧酶催化生物转化研究最新进展", 《化工生产与技术》 *
彭芷芊等: "圆柚酮生物转化菌种筛选及培养基组分优化", 《华中农业大学学报》 *
李 晓等: "瓦伦西亚橘烯生物转化生成圆柚酮", 《食品科学》 *

Similar Documents

Publication Publication Date Title
CN113307848A (en) Cyclo-serine-valine-isoleucyl-leucyl peptide with antifungal and free radical scavenging activities and preparation method thereof
CN113264987A (en) Cyclochromo-threo-valine-isoleucin with antifungal and free radical scavenging activity and preparation method thereof
CN112746029B (en) Hansenula polymorpha strain QTX22 for producing aroma substances in grape juice and application of strain
CN114250157B (en) Saccharomycopsis fibuligera cx-3 strain for high yield of aroma substances and application thereof
CA3039479C (en) A strain of yeast saccharomyces bayanus subsp. uvarum dbvpg36p, its use in the fermentative production of foods and a method for the selection of the strain
CN112725235B (en) Novel strain of clostridium and application thereof
CN111235307A (en) Preparation and application method of fermentation type red date wine composite strain
CN112342163A (en) Burkholderia and application thereof
CN108795773B (en) Musk mould strain and perfume prepared from same
CN108795771B (en) Musk mould strain and perfume prepared from same
CN112522123B (en) Acid-resistant saccharomyces cerevisiae and application thereof in high-acidity fruit fermented wine
CN116410874A (en) Pichia pastoris fermentation strain and application thereof in preparation of fermented sour meat
CN113122470A (en) Strain screening and application for high-yield production of methyl benzoate and methyl anthranilate
CN112625963B (en) Clostridium sarmentosum capable of producing 4-methylphenol and application thereof
Calabretti et al. Aglianico and Fiano wines obtained with an autochthonous non-Saccharomyces yeast
CN113046259B (en) Clostridium triquetrum producing 4-methylphenol and application thereof
CN112662597B (en) Bacillus pumilus for high yield of trimethyl pyrazine and tetramethyl pyrazine
CN112940959B (en) Klebsiella capable of degrading limonene and application thereof
CN115353983B (en) ZR21 of zygosaccharomyces rouxii and DH06 of Debaryomyces hansenii and method for preparing fermented soybean by using same
CN110982710B (en) Wine cellar verruca umbilicalis leveling and application thereof in aging of new wine
CN115820432B (en) Coriolus versicolor trametes versicolor producing strain and application thereof
CN112746028B (en) Louisitania red wintergreen spore-locked spore yeast strain QTX26 for high yield of aroma substances and application thereof
CN110305912B (en) Method for extracting spice from jasmine Marasmius pauciflorus strain and application of extracted spice in tobacco
CN116103169A (en) Pichia kudriavzevii and use thereof
CN116656561A (en) Wessella fusion and application thereof

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
WD01 Invention patent application deemed withdrawn after publication
WD01 Invention patent application deemed withdrawn after publication

Application publication date: 20210209