CN106566778B - A kind of tea seeds saccharomycete and its application - Google Patents
A kind of tea seeds saccharomycete and its application Download PDFInfo
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- A23F3/00—Tea; Tea substitutes; Preparations thereof
- A23F3/06—Treating tea before extraction; Preparations produced thereby
- A23F3/08—Oxidation; Fermentation
- A23F3/10—Fermentation with addition of microorganisms or enzymes
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- 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
- C12P7/64—Fats; Fatty oils; Ester-type waxes; Higher fatty acids, i.e. having at least seven carbon atoms in an unbroken chain bound to a carboxyl group; Oxidised oils or fats
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Abstract
The invention discloses a kind of tea seeds saccharomycete and its applications, its Latin literary fame of the tea seeds saccharomycete is known as: Meyerozyma caribbica JJZ11, the tea seeds saccharomycete was deposited in China typical culture collection center (CCTCC) on September 9th, 2016, deposit number is CCTCC M 2016470, preservation address are as follows: Wuhan City, Hubei Province Wuchang District Bayi Road Luo Jia Shan.The present invention utilizes tea seeds yeast fermentation technique, and tea seeds grease body can be extracted first from the mixture of tea seeds constituent, after purification by tea seeds grease body, then extracts tea seed oil from grease body and comes out；Other constituent residual quantities of tea seeds are minimum in the tea seed oil extracted in this way, ensure that the quality of tea seed oil, and enormously simplify the work of tea seed oil following process.
Description
Technical field
The present invention relates to a kind of tea seeds saccharomycete and its applications, belong to microorganisms technical field.
Background technique
Tea seed oil is the edible oil produced using the seed of tea tree (Camellia sinensis) as raw material, Yin Qifu
East olive oil is known as containing various bioactive components.There is the tea seed oil production method formally reported mainly to have at present cold
Squeezing method, hot squeezing method, aqueous enzymatic method, common solvent lixiviation process and carbon dioxide supercritical extraction method, the production of these tea seed oils
Technique there are problems that some being difficult to overcome.To seek to solve the problems, such as by way of Jiang Jinzhong reports a kind of tealeaves not long ago
Seed oil production new technique --- tea seed oil biofermentation production technology, the core of the technique are by biofermentation by tea seeds
Water slurry is divided into three layers, then takes out upper layer heating production tea seed oil.The technique is with production cost is low, yield is high, oil quality
High, the advantages that tea seed oil flavor is dense.
Since biofermentation layering belongs to fermentation scope, generating process is necessarily required to quickly and effectively fermentative microorganism.
However, containing a large amount of Tea Saponin in tea seeds water slurry, Tea Saponin is inhibited for most of saccharomycete, so, greatly
Most saccharomycete are difficult the flourish in tea seeds water slurry, therefore, can find one kind quickly breeding life in tea seeds water slurry
Long saccharomycete at the technique critical issue.It is purified repeatedly, qualification test, we, which have looked for, a kind of can make tea seeds water slurry
The yeast of Rapid Fermentation --- tea seeds yeast (Meyerozyma caribbica JJZ11).Tea seeds yeast and other
The maximum difference of Meyerozyma caribbica bacterial strain is that it can be caused using the tea polysaccharide in tea seeds water slurry as carbon source
Tea seeds water slurry Rapid Fermentation generates CO2And other metabolites, but not all Meyerozyma caribbica bacterial strain is equal
With this ability.
The external Meyerozyma caribbica of report earliest is Kurtzman, and is named by him.Meyerozyma
Although caribbica has the generic name and kind name of oneself, but it is the nothing of fermentation Candida (Candida fermentati)
Property type, so, also known as phorozoon fermentation Candida (anamorph Candida fermentati).Kim was reported
A bacterial strain of Meyerozyma caribbica: Meyerozyma caribbicaMG20W, it is believed that the bacterial strain has thermophilic salt
Characteristic illustrates that Meyerozyma caribbica yeast has and adapts to along with tea seeds yeast has the ability of anti-Tea Saponin
The ability of different growing environment, thus there is wide developing and utilizingpotentiality.
Summary of the invention
The object of the present invention is to provide a kind of tea seeds saccharomycete, Latin literary fame is known as: Meyerozyma
Caribbica JJZ11, the tea seeds saccharomycete were deposited in China typical culture collection center on September 9th, 2016
(CCTCC), deposit number is CCTCC M 2016470, preservation address are as follows: Wuhan City, Hubei Province Wuchang District Bayi Road Luo Jia Shan.
The ITSrDNA sequence of above-mentioned tea seeds yeast is as shown in SEQ ID NO.1,26SrDNA sequence such as SEQ ID NO.2
It is shown.
It is another object of the present invention to: the preparation method of above-mentioned tea seeds saccharomycete is provided, this method is to take fresh tea leaf
Seed benevolence is put into container in right amount, and 25 DEG C of warm water of doubling dose are added, and constant temperature impregnates 10~16h or so, dips soak with collarium is connect
And in the flat lining out culture of MRS, cultivation temperature is 25 DEG C, after growing bacterium colony, selects independent bacterium colony, and with transfer needle picking
Sample is placed on glass slide, with methylene blue, sees whether to be saccharomycete, if it is saccharomycete, is inoculated with the expanding propagation bacterium colony,
Again row identification breeding, 4 times repeatedly, make be same form bacterium colony on final plate until, then the bacterium colony is tea seeds saccharomycete.
It is another object of the present invention to: application of the above-mentioned tea seeds saccharomycete in vegetable oil production is provided.
The production stage of above-mentioned vegetable oil are as follows: weigh tea seeds benevolence and water is added to impregnate 16~20h under the conditions of 25 DEG C, will impregnate
Add water after good tea seeds cleaning, wherein tea seeds: then water=1:3 carries out 2 wheel mashing, filters to get filtrate, filters according to every liter
Tea seeds saccharomycete is added into filtrate for the ratio that 1~5g tea seeds yeast is added in liquid, stirs evenly, is put into container and is placed in 35 DEG C
After ferment at constant temperature 8h, filtrate will appear white top, pulls white top heating out, tea seeds crude oil can be obtained.
It is another object of the present invention to: application of the above-mentioned tea seeds saccharomycete in production black tea is provided.
The production stage of above-mentioned black tea are as follows: take two leaves and a bud dark brownish green, rub through kneading machine to there is juice exudation, according to every public affairs
Tea seeds yeast, 28 DEG C of ferment at constant temperature, when smelling tealeaves are added to juice in the ratio that 4~8g tea seeds yeast is added in jin fresh tea passes
It is the black tea with tea seeds yeast fragrance by drying tea leaves when with strong yeast fragrance.
Compared with the prior art, the advantages of the present invention are as follows: in now all of tea seed oil production technology, tea seed oil
Be directly to be extracted from the mixture of tea seeds constituent, these tea seeds constituents to the recovery rate of tea seed oil,
The constituent of tea seed oil and the flavor of tea seed oil impact.And the present invention utilizes tea seeds yeast fermentation technique, it can
To extract tea seeds grease body first from the mixture of tea seeds constituent, after purification by tea seeds grease body, then from oil
Tea seed oil is extracted in rouge body to come out；Other constituent residual quantities of tea seeds are minimum in the tea seed oil extracted in this way,
It ensure that the quality of tea seed oil, and enormously simplify the work of tea seed oil following process.It has been proved by practice that the present invention reaches
The ideal production targets such as production cost is low, yield is high, oil quality is good, tea seed oil flavor is dense.
Detailed description of the invention:
Fig. 1 is the microphoto of tea seeds yeast colony form；
Fig. 2 is the microphoto of tea seeds yeast personal feature and situation of sprouting；
Fig. 3 is position of the tea seeds yeast in Pichia system development tree.
Specific embodiment
In order to keep the object of the invention, technical solution and advantage clearer, below with reference to examples and drawings to the present invention
It is described in further detail.
Embodiment 1: the screening and preservation of bacterial strain
Tea seeds saccharomycete of the invention the preparation method comprises the following steps: weigh fresh tea passes seed benevolence 100g, add 25 DEG C of warm water 200g, it is permanent
After temperature impregnates 10h, soak is dipped and in the flat lining out culture of MRS with collarium is connect, cultivation temperature is 25 DEG C, grows bacterium colony
Afterwards, independent bacterium colony is selected, is placed on glass slide with transfer needle picking sample, with methylene blue, sees whether to be saccharomycete, such as
Fruit is saccharomycete, is inoculated with the expanding propagation bacterium colony, then row identification breeding, 4 times repeatedly, makes to be same form bacterium colony on final plate
Until, then the bacterium colony is tea seeds saccharomycete.Picking single colonie connects bacterium on MRS slant medium, 25 DEG C of culture 48h, in case
Preservation and identification use.
Attached drawing 1 is shown in the morphologic observation of bacterial strain: the bacterium colony of tea seeds saccharomycete is in butyrous, and milky, surface is calm, smooth
It is reflective.The individual morphology of 28 DEG C of tea seeds saccharomycete culture 72h is as shown in Figure 2: cell is subsphaeroidal, oval or rodlike, size
2.0-5.5 μm of 1.3-2.0 μ m has pseudohypha, budding breeding.
The physiological and biochemical property of tea seeds saccharomycete is referring to table 1: referring to " primary Jie Shi Bacteria Identification handbook ", with the tea of purification
Leaf seed bacterial strain is test material, is tested respectively to carbon-source cpd listed in table, the results showed that, in addition to inositol and D- lactose
(Niu Yuan) outside, remaining carbon-source cpd is positive, and above-mentioned test is by " in China National Academy of Food & Fermentation Industries's microorganism detection
The heart " is completed.
Table 1: tea seeds yeast utilization of carbon source
The determination and analysis of sequence of tea seeds saccharomycete ITSrDNA and 26SrDNA
The sequencing results of tea seeds saccharomycete ITSrDNA and 26SrDNA such as SEQ ID NO.1 and SEQ ID
Shown in NO.2: the ITSrDNA sequence of tea seeds saccharomycete is 516bp, and 26SrDNA sequence is 533bp.Referring to subordinate list
2, sequence homology retrieval is carried out using Blest, the saccharomycete that similarity reaches 90% or more there are 7 kinds, wherein with Meyerozyma
The similarity of caribbica has reached 100%.
Table 2: tea seeds saccharomycete ITSrDNA and 26SrDNA sequence and nearly edge yeast kind tetraploid rice result
Based on the homology of ITSrDNA and 26SrDNA, phylogenetic analysis is carried out with MEGA5.0 software, as a result
Phylogenetic tree as shown in Fig. 3 is arrived.Phylogenetic analysis the result shows that, Meyerozyma in the genealogical tree
caribbica CBS9966TIt (FUNCBS/AY187283) is individual outer group's kind, tea seeds saccharomycete is independent with this kind
Constitute a branch；The similitude of the two sequence is maximum, and genetic distance is minimum；Therefore, the relationship of tea seeds saccharomycete and this kind
Relationship is nearest；Above-mentioned test is by " China National Academy of Food & Fermentation Industries's microorganism detection " center " is completed.
In summary morphologic observation, mode of reproduction, pseudohypha, carbon source reaction, ITSrDNA and 26SrDNA sequence alignment,
And the building and analysis of phylogenetic tree, " China National Academy of Food & Fermentation Industries's microorganism detection " center " is by tea seeds yeast
The dientification of bacteria are as follows: Meyerozymacaribbica, the number that we give tea seeds saccharomycete is JJZ11；Therefore, tea seeds ferment
Female bacterium is defined as: Meyerozyma caribbica JJZ11.By tea seeds saccharomycete (or Meyerozyma caribbica
JJZ11 it) was deposited in China typical culture collection center (CCTCC) on September 9th, 2016, deposit number is CCTCC M
2016470。
Embodiment 2: the production applied to vegetable oil
By taking camellia seed oil as an example: weighing oil tea (Camellia oleifera) seed benevolence 1000g, add water under the conditions of 20 DEG C
Impregnate 12h, will add water after the cleaning of soaked tea seed, wherein tea seed: then water=1:3 carries out 2 wheel mashing, filters
Tea seeds saccharomycete is added into filtrate according to the ratio that 5g tea seeds yeast is added in every liter of filtrate, stirs evenly, is put into for filtrate
After container is placed in 35 DEG C of ferment at constant temperature 10h, filtrate will appear white top, pulls white top heating out, can produce oil tea
Seed crude oil.
Embodiment 3: tea seeds saccharomycete is for producing black tea
It takes two leaves and a bud dark brownish green, rubs through kneading machine to there is juice exudation, 4g tea seeds are added according to per kilogram fresh tea passes
Tea seeds yeast dry powder is added to juice in the ratio of yeast, stirs evenly, 28 DEG C of ferment at constant temperature, has strong ferment when smelling tealeaves
It is the black tea with tea seeds yeast characteristic by drying tea leaves when female fragrance；The black tea is the decomposition that undergoes microbial fermentation
Teas, it is not only right rich in a variety of amino acid needed by human, natural anti-oxidation polypeptide, tea polyphenols, vitamin, acetic acid and lactic acid
Human body has health-care effect, and there are also special yeast fragrance.
Embodiment and application field described above are merely a preferred embodiment of the present invention, and the simultaneously feasible implementation of non-present invention
Exhaustion.For persons skilled in the art, it is made under the premise of without departing substantially from the principle of the invention and spirit
Any obvious change and application field support it is wide, should all be contemplated as falling with claim protection model of the invention
Within enclosing.
SEQUENCE LISTING
Sequence table
<110>Guizhou college of education
<120>a kind of tea seeds saccharomycete and its application
<130> nm:
<160> 2
<170> PatentIn version
<210>SEQ ID NO:1
<211> 516
<212> DNA
<213>tea seeds saccharomycete
<220>
<221> ITSrDNA
<400> 1
TACAGTATTC TTTTGCCAGC GCTTAACTGC GCGGCGAAAA ACCTTACACA 50
CAGTGTCTTT TTGATACAGA ACTCTTGCTT TGGTTTGGCC TAGAGATAGG 100
TTGGGCCAGA GGTTTAACAA AACACAATTT AATTATTTTT ATTGATAGTC 150
AAATTTTGAA TTAATCTTCA AAACTTTCAA CAACGGATCT CTTGGTTCTC 200
GCATCGATGA AGAACGCAGC GAAATGCGAT AAGTAATATG AATTGCAGAT 250
TTTCGTGAAT CATCGAATCT TTGAACGCAC ATTGCGCCCT CTGGTATTCC 300
AGAGGGCATG CCTGTTTGAG CGTCATTTCT CTCTCAAACC CCCGGGTTTG 350
GTATTGAGTG ATACTCTTAG TCGAACTAGG CGTTTGCTTG AAAAGTATTG 400
GCATGGGTAG TACTGGATAG TGCTGTCGAC CTCTCAATGT ATTAGGTTTA 450
TCCAACTCGT TGAATGGTGT GGCGGGATAT TTCTGGTATT GTTGGCCCGG 500
CCTTACAACA ACCAAA 516
<210>SEQ ID NO:2
<211> 533
<212> DNA
<213>tea seeds saccharomycete
<220>
<221> 26SrDNA
<400> 2
GCTCAAATTT GAAATCTGGC ACCTTCGGTG TCCGAGTTGT AATTTGAAGA 50
TTGTAACCTT GGGGTTGGCT CTTGTCTATG TTTCTTGGAA CAGGACGTCA 100
CAGAGGGTGA GAATCCCGTG CGATGAGATG CCCAATCCTA TGTAAGGTGC 150
TTTCGAAGAG TCGAGTTGTT TGGGAATGCA GCTCTAAGTG GGTGGTAAAT 200
TCCATCTAAA GCTAAATATT GGCGAGAGAC CGATAGCGAA CAAGTACAGT 250
GATGGAAAGA TGAAAAGAAC TTTGAAAAGA GAGTGAAAAA GTACGTGAAA 300
TTGTTGAAAG GGAAGGGTTT GAGATCAGAC TCGATATTTT GTGAGCCTTG 350
CCTTCGTGGC GGGGTGACCC GCAGCTTATC GGGCCAGCAT CGGTTTGGGC 400
GGTAGGATAA TGGCGTAGGA ATGTGACTTT GCTTCGGTGA AGTGTTATAG 450
CCTGCGTTGA TGCTGCCTGC CTAGACCGAG GACTGCGATT TTATCAAGGA 500
TGCTGGCATA ATGATCCCAA ACCGCCCGTC TTG 533
Claims (7)
1. a kind of tea seeds saccharomycete, it is characterised in that: its deposit number are as follows: CCTCC M 2016470.
2. tea seeds saccharomycete according to claim 1, it is characterised in that: the ITSrDNA sequence of the tea seeds yeast is such as
Shown in SEQ ID NO.1,26SrDNA sequence is as shown in SEQ ID NO.2.
3. a kind of preparation method of tea seeds saccharomycete as described in claim 1, it is characterised in that: take fresh tea leaf seed benevolence suitable
Amount is put into container, and 25 DEG C of warm water of doubling dose are added, and constant temperature impregnates 10~16h, dips soak and in MRS using collarium is connect
Flat lining out culture, cultivation temperature are 25 DEG C, after growing bacterium colony, select independent bacterium colony, and put with transfer needle picking sample
It on glass slide, with methylene blue, sees whether to be saccharomycete, if it is saccharomycete, is inoculated with the expanding propagation bacterium colony, then row mirror
Fixed breeding, 4 times repeatedly, make be same form bacterium colony on final plate until, then the bacterium colony is tea seeds saccharomycete.
4. a kind of application of tea seeds saccharomycete as described in claim 1 in production vegetable oil.
5. application of the tea seeds saccharomycete according to claim 4 in production vegetable oil, which is characterized in that vegetable oil
Production stage are as follows: weigh tea seeds benevolence and water is added to impregnate 16~20 h under the conditions of 25 DEG C, will add after the cleaning of soaked tea seeds
Water, wherein tea seeds: then water=1:3 carries out 2 wheel mashing, filters to get filtrate, 1~5g tea seeds ferment is added according to every liter of filtrate
Tea seeds saccharomycete is added into filtrate for female ratio, stirs evenly, and is put into after container is placed in 35 DEG C of ferment at constant temperature 8h, filtrate meeting
There is white top, pulls white top heating out, tea seeds crude oil can be obtained.
6. a kind of application of tea seeds saccharomycete as described in claim 1 in production black tea.
7. application of the tea seeds saccharomycete according to claim 6 in production black tea, which is characterized in that the production of black tea
Step are as follows: take two leaves and a bud dark brownish green, rub through kneading machine to there is juice exudation, 4~8g tealeaves is added according to per kilogram fresh tea passes
Tea seeds yeast is added to juice in the ratio of seed yeast, and 28 DEG C of ferment at constant temperature will when smelling tealeaves with strong yeast fragrance
Drying tea leaves are the black tea with tea seeds yeast fragrance.
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| CN114480151B (en) * | 2022-02-24 | 2023-08-01 | 福建春伦集团有限公司 | Yeast and application thereof in tea tree quality improvement and efficacy enhancement |
| CN114680213B (en) * | 2022-04-27 | 2024-06-21 | 贵州泰和现代生态农业科技有限公司 | Latifolia fermented tea and preparation method thereof |
| CN115627239B (en) | 2022-12-21 | 2023-03-21 | 广东省农业科学院蚕业与农产品加工研究所 | Sulfur-reducing yeast and application thereof in fruit and vegetable desulfurization |
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| CN101755940A (en) * | 2009-12-14 | 2010-06-30 | 冯磊 | Processing method and application for yeast black tea |
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