CN110195023A - A kind of Wine brewing yeast strain and its application - Google Patents
A kind of Wine brewing yeast strain and its application Download PDFInfo
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- CN110195023A CN110195023A CN201910567660.0A CN201910567660A CN110195023A CN 110195023 A CN110195023 A CN 110195023A CN 201910567660 A CN201910567660 A CN 201910567660A CN 110195023 A CN110195023 A CN 110195023A
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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
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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
- C12P23/00—Preparation of compounds containing a cyclohexene ring having an unsaturated side chain containing at least ten carbon atoms bound by conjugated double bonds, e.g. carotenes
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- C12R2001/865—Saccharomyces cerevisiae
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Abstract
The present invention relates to microorganisms technical field, a kind of Wine brewing yeast strain and its application are disclosed.The present invention provides the mutagenesis Wine brewing yeast strain of one plant of high-yield astaxanthin, the astaxanthin yield of the mutagenic strain is significantly improved relative to starting strain, wherein shake flask fermentation astaxanthin yield improves 3.98 times, 65.93mg/L is reached, astaxanthin ratio shared in carotenoid improves 1.48 times, has reached 68.56%.And by the optimization to technological condition for fermentation, the astaxanthin yield in 5L fermentor is made to be up to 404.78mg/L.
Description
Technical field
The present invention relates to microorganisms technical field, more particularly to a kind of Wine brewing yeast strain and its application.
Background technique
Astaxanthin (Astaxanthin, C40H52O4, molecular weight 596.84) is that one kind is widely present in microorganism and sea
The highest level product of natural carotenoid and carotenogenesis in foreign biology, according to two chiral carbons in β-
Existence form on ionone ring can be divided into 3 kinds of configurations (3S, 3 ' S), (3R, 3 ' S), (3R, 3 ' R).Astaxanthin has very strong
Oxidation resistance, can prevent chain reaction, anti-lipid peroxidation come protective film knot by the active oxygen in scavenger-cell
Structure, to play the effect for preventing oxidative damage.Unlike other carotenoid, astaxanthin connects the interior of cell simultaneously
Film and outer membrane, can clean the active oxygen of intracellular outer membrane simultaneously, thus astaxanthin have than other carotenoid it is stronger
Inoxidizability, oxidation resistance are ascorbic 65 times, 54 times of beta carotene, canthaxanthin, zeaxanthin and lutein
10 times, 100 times of vitamin E.Exactly because astaxanthin has unique chemical structure and property, decline so that it has to slow down
Always, the functions such as antitumor, strengthen immunity, prevention cardiovascular disease, in the row such as food, medicine, health care product and aquaculture
Industry has huge market value.The third-largest carotenoid of beta carotene and lutein is only second to as market scale,
The market share of astaxanthin has accounted for the 29% of carotenoid, it is contemplated that market scale reaches 25.7 hundred million dollars within 2025.It is wide
The market demand make the research of astaxanthin and production that there is bright prospect.
Currently, mainly there are chemical synthesis, algae and red phaffia rhodozyma biosynthesis in the source of astaxanthin.Chemically synthesized shrimp
Green element is different with natural astaxanthin configuration, and oxidation resistance is only the 1/3 of natural astaxanthin, and in addition people also worry to synthesize shrimp
The safety of the different stereoisomers and some potential synthetic intermediates of green element.Haematococcus pluvialis (Haematococcus
Pluvialis) and phaffiafhodozyma (Xanthophyllomyces dendrorhous) is industrially to produce astaxanthin at present
Main bacteria seed.However the haematococcus pluvialis growing period is slow, cell density is low, causes economic cost high.Red hair husband's ferment
Mother as the microorganism for naturally producing astaxanthin present in nature, but due to phaffiafhodozyma production astaxanthin for (3R,
3 ' R) configuration, oxidation resistance is lower, limits its application in the industrial production.Therefore, sight is turned to it by researcher
His Microbial cell factories are used to Biosynthesis of Astaxanthin.2017, Kildegaard et al. constructed shrimp in solution rouge yeast
Green element synthesis path, and the copy number by increasing CrtZ, balance metabolic flux improve the production of astaxanthin, finally make to solve rouge ferment
Astaxanthin yield has reached 54.6mg/L in mother.2018, Park et al. navigated to truncated BKT gene (trCrBKT) carefully
After birth is overexpressed ispD the and ispF gene identified by FVSEOF software, and optimization of fermentation conditions, makes large intestine in 5L fermentor
The astaxanthin yield of bacillus has reached 385.04mg/L.
Saccharomyces cerevisiae is as a kind of generally acknowledged safe mode microorganism, and genetic background understands, molecule manipulation is simple, is making
It is widely used in the production such as wine, food baking, drug, health care product.Compared to Escherichia coli, its thallus vitamin, protein
Content is high, can eat medicinal and fodder yeast；Compared to algae, its growth cycle is shorter and is easier to cultivate.Therefore, shrimp is realized
High yield of the green element in saccharomyces cerevisiae will show great competitiveness in carotenoid industrialization.However, up to now
Astaxanthin yield in the saccharomyces cerevisiae of open report is still not up to commercial production level.2017, Yu Hongwei seminar
Increase the copy number between precursor supply and adjustment rate-limiting enzyme by mutation GGPP synthase and balance the methods of metabolic flux, will make
The astaxanthin yield of brewer yeast is improved to the higher level of 47.18mg/L.
Summary of the invention
In view of this, the purpose of the present invention is to provide a kind of by mutagenic obtained Wine brewing yeast strain, keep its shrimp green
Plain yield is significantly improved；
Another object of the present invention is that providing the Wine brewing yeast strain after above-mentioned mutagenesis in production related chemicals
In application；
The Wine brewing yeast strain production astaxanthin after above-mentioned mutagenesis is utilized another object of the present invention is that providing
Method.
For achieving the above object, the invention provides the following technical scheme:
A kind of Wine brewing yeast strain, deposit number are CGMCC No.17913, are deposited in China on June 10th, 2019
Microbiological Culture Collection administration committee common micro-organisms center, classification naming are Saccharomyces Cerevisiae in S accharomyces
cerevisiae。
The Saccharomyces Cerevisiae in S yBE_Sc14CY10 that the present invention produces astaxanthin using one plant is starting strain, with atmospheric pressure at room
Plasma induced-mutation technique (Atmospheric and room temperature plasma, ARTP) is to SyBE_Sc14CY10
Carry out physical mutagenesis, obtain saccharomyces cerevisiae mutated library, astaxanthin yield is then measured by shake flask fermentation and HPLC, yield compared with
Three high plant mutant bacterium are the object advanced optimized；Then based on the higher three plant mutants bacterium of astaxanthin yield, pass through
The oxidative pressure that hydrogen peroxide applies carries out chemical mutagenesis (ChemicalMutagenesis, CM), and continuous mutagenesis 21 is on behalf of a week
Phase obtains saccharomyces cerevisiae mutated library again, the yield of astaxanthin is equally measured by shake flask fermentation and HPLC, and yield is higher
Three plant mutant bacterium are as the object advanced optimized；It recycles according to this, with each period, physically or chemically mutagenic obtained astaxanthin is produced
Based on measuring higher three plants of yeast, alternately physics or chemical mutagenesis are finally obtained luring for one plant of high-yield astaxanthin
Become Wine brewing yeast strain, is named as SyBE_Sc04030020.
It is verified by fermentation test, SyBE_Sc04030020 astaxanthin yield and ratio are significantly improved, wherein shaking flask
Ferment 3.98 times of astaxanthin output increased, has reached 65.93mg/L, and astaxanthin ratio shared in carotenoid improves
1.48 times, reach 68.56%.And by the optimization to technological condition for fermentation, make the astaxanthin yield in 5L fermentor
It can reach 404.78mg/L.Above-mentioned each yield data belongs to most high yield in the saccharomyces cerevisiae astaxanthin yield being currently known
Amount.
Based on above-mentioned technical effect, the present invention provides the Wine brewing yeast strains to be in production astaxanthin or with astaxanthin
Application in the other chemicals of intermediate chemical；And in the microorganism formulation of preparation production astaxanthin or preparation production with shrimp
Green element is the application in the microorganism formulation of the other chemicals of intermediate chemical.
The method of two kinds of fermenting and producing astaxanthins is provided in the present invention, and the method for the first production astaxanthin is that will protect
The Wine brewing yeast strain that hiding number is CGMCC No.17913, which is inoculated into SC solid medium, to be activated, and is then transferred to SC liquid again
Seed culture fluid is prepared in body culture medium, finally seed culture fluid is transferred in YPDG fermentation medium and is fermented, until cell
OD600Basicly stable, chemical activators are not further added by.
In the specific implementation process, the Wine brewing yeast strain that deposit number is CGMCC No.17913 is lined into SC solid
Culture medium carries out activation culture, is then seeded into the culture of SC fluid nutrient medium to OD600=5-8, then press OD600=0.2 is transferred to
The culture of SC fluid nutrient medium is to OD600=6.5-7.5 is prepared into seed culture fluid, and seed culture fluid is finally pressed end OD600=0.1
30 DEG C, 250rpm shaking table oscillation and fermentation are transferred in YPDG fermentation medium, until cell OD600Basicly stable, chemical activators are not
It is further added by.It is detected by HPLC, in shake flask fermentation, the yield of astaxanthin is 65.93mg/L；
The Wine brewing yeast strain inoculation that it is CGMCC No.17913 that the method for second of production astaxanthin, which is by deposit number,
It is activated into SC solid medium, is then transferred in SC fluid nutrient medium again and prepares seed culture fluid, finally by seed culture
Liquid, which is transferred in YPD fermentation medium, to ferment, until cell OD600Basicly stable, chemical activators are not further added by；
In the specific implementation process, the Wine brewing yeast strain that deposit number is CGMCC No.17913 is lined into SC solid
Culture medium carries out activation culture, is then seeded into the culture of SC fluid nutrient medium to OD600=6-7, then be transferred to by 4% inoculum concentration
SC fluid nutrient medium culture to logarithmic growth phase mid-term is prepared into seed culture fluid, and seed culture fluid is finally pressed 10% inoculum concentration
It is transferred in YPD fermentation medium and carries out ferment tank, until cell OD600Basicly stable, chemical activators are not further added by.
During fermentation, glucose is added as carbon source with fed-batch mode, and is controlled within the scope of 0-1g/L and (be free of 0), bacterium
When body grows into deadtime, D- (+)-galactolipin inducer is added, stream plus glucose are simultaneously stopped, into chemical activators rank
Section；Carbon source is switched to ethyl alcohol in fermentor at this time, same to be added by fed-batch mode, and controls the concentration of alcohol in fermentor and exist
5g/L or less；
Wherein, an important factor for influencing fermentation biomass, there are also the nitrogen sources in fermentation medium.The present invention is preferably with yeast
Leaching powder is nitrogen source, in order to avoid nitrogen source feed supplement is excessive, the fed-batch mode successively decreased using concentration gradient to adding yeast extract,
The yeast extract mother liquor that certain volume is added into fermentor at regular intervals (is soaked with yeast in initial YPD fermentation medium
The concentration of powder is 1.0 ×).Specific feed supplement process is as follows: be added 1.2 × yeast extract be added 1.0 in the 10h of fermentation, 65h ×
0.8 × yeast extract is added in yeast extract, 75h, and 0.3 × yeast extract is added in 120h, and the later period maintains always 0.3 × concentration,
180h (cell grows into deadtime) stops feed supplement.Start every liter when fermenting 10h and add 1.2 × yeast extract, is in total
30g yeast extract, mother liquid concentration are 400g/L, and 30g yeast extract volume is 75ml.Deadtime is grown into 75h cell,
OD600Growth slows down, reduce nitrogen source be added to 0.8 ×.Ferment 120h, strain growth growth slow down once again, reduce nitrogen source be added to
0.3×。
Fed batch fermentation test result shows that astaxanthin yield reaches 226.19mg/L, compares shake flask fermentation yield
(65.93mg/L) improves 2.43 times.
In order to further increase astaxanthin yield, continue to optimize ferment tank condition.In addition to fermentation medium
Proportion composition and addition manner, temperature be also influence strain growth and metabolism key factor.In above-mentioned fed batch fermentation
On the basis of, D- (+)-galactolipin inducer is added as separation, the fermentation temperature before being added is 30 DEG C, is most had under this condition
Conducive to cell growth.Equal cells grow into deadtime, start at this time plus D- (+)-galactolipin induces chemical activators path base
Because starting to express, into fermentation second stage, 20 DEG C are cooled the temperature to, to be conducive to the accumulation of astaxanthin, final astaxanthin is produced
Amount has reached 404.78mg/L, relative to 5.14 times of shake flask fermentation output increased.
From the above technical scheme, the present invention provides the mutagenesis Wine brewing yeast strain of one plant of high-yield astaxanthin, the mutagenesis
The astaxanthin yield of bacterial strain is significantly improved relative to starting strain, and wherein shake flask fermentation astaxanthin yield improves 3.98 times,
65.93mg/L is reached, astaxanthin ratio shared in carotenoid improves 1.48 times, reached 68.56%.And
By the optimization to technological condition for fermentation, the astaxanthin yield in 5L fermentor is made to be up to 404.78mg/L.
Biological deposits explanation
SyBE_Sc04030020, classification naming: Saccharomyces Cerevisiae in S accharomyces cerevisiae, in June, 2019
It is deposited within 10th China Committee for Culture Collection of Microorganisms's common micro-organisms center, address is Chaoyang District, Beijing City North Star west
The institute 3 of road 1, Institute of Microorganism, Academia Sinica, deposit number are CGMCC No.17913.
Detailed description of the invention
Fig. 1 show the shake flask fermentation astaxanthin yield of mutagenic strain and starting strain of the present invention；
Fig. 2 show the accounting of each chemicals in the shake flask fermentation carotenoid of mutagenic strain and starting strain of the present invention
Figure；
Fig. 3 show the testing result of mutagenic strain ferment tank of the present invention；
Fig. 4 show the testing result of mutagenic strain ferment tank of the present invention after optimization temperature.
Specific embodiment
The invention discloses a kind of Wine brewing yeast strain and its application, those skilled in the art can use for reference present disclosure,
It is suitably modified realization of process parameters.In particular, it should be pointed out that all similar substitutions and modifications carry out those skilled in the art
Say it is it will be apparent that they are considered as being included in the present invention.Wine brewing yeast strain of the present invention and application have passed through
Preferred embodiment is described, related personnel obviously can not depart from the content of present invention, in spirit and scope to described herein
Wine brewing yeast strain and application be modified or appropriate changes and combinations, carry out implementation and application the technology of the present invention.
Below with reference to embodiment, the present invention is further explained.
Embodiment 1: shake flask fermentation test
Fermentation: the mutagenesis Wine brewing yeast strain SyBE_Sc04030020 that -80 DEG C of glycerol saves is lined into SC solid culture
Base carries out activation culture, is then seeded into 5mlSC fluid nutrient medium, cultivates 18h to OD600=5-8, then press OD600=0.2 turn
It is connected in secondary seed medium test tube.OD after 14-16h600After reaching 7 or so, take seed culture fluid by whole OD600=0.1 turn
It is connected in 50mL YPDG culture medium and ferments, 30 DEG C, 250rpm shaking table shaken cultivation 96h；
HPLC detection: 1.5mLEP pipe is added in the bacterium solution after taking 500uL to ferment, and 12000r is centrifuged 2min, outwells supernatant, add
Enter 1ml sterile water and cell is resuspended, cell is resuspended in the centrifugation addition of falling supernatant 1ml 3M HCl, and boiling water bath 3min, ice bath 3min are repeated
3 times, supernatant is abandoned in twice of centrifugation washing, and after 1ml acetone resuspension cell is added, a small amount of quartz sand vortex oscillation 10min is added, will
After acetone extract 12000r is centrifuged 5min, after 0.2 μm of organic membrane filtration, takes 200 μ L to be added in sample bottle and carry out
HPLC high performance liquid chromatography detection.Use acetone gradient dilution to 50mg/l, 25mg/L, 12.5mg/ respectively the mark product of astaxanthin
L,6.25mg/L,3.125mg/L.The setting of location parameter sets wavelength are as follows: 450nm (lycopene), 470nm (astaxanthin).
Chromatographic column C18 column.Mobile phase A: second eyeball: water=9:1；Mobile phase B: methanol: isopropanol=3:2.25 DEG C of column temperature, sample temperature 20
℃。
As a result as depicted in figs. 1 and 2, astaxanthin yield 65.93mg/L improves 3.98 times relative to bacterium germination out, shrimp
Green element ratio shared in carotenoid improves 1.48 times, has reached 68.56%.
The test of embodiment 2:5L ferment tank
Fermentation basal medium be initial glucose concentration be 20g/L YPD culture medium (1% yeast extract, 2% peptone,
2% glucose), fed batch fermentation test is carried out to SyBE_Sc04030020 in the 5L fermentor of laboratory.First order seed training
It supports: taking SyBE_Sc04030020 glycerol stock 200ul to be seeded in the test tube equipped with 5mL SC seed culture medium, 30 DEG C, 250rpm
It cultivates to OD600=6-7；Secondary seed culture: first order seed is forwarded to by 4% inoculum concentration equipped with 50mL SC culture medium
In 250mL shaking flask, 30 DEG C, under the conditions of 250rpm culture to mid log phase (OD600=5-6).
Fermentor inoculation: with 10% inoculum concentration (secondary seed of 250ml, initial OD600About 0.5) secondary seed is transferred
Start to ferment into the 5L fermentor equipped with 2.25L fermentation medium；The initial liquid amount of fermentor is 2.5L, and initial medium is
YPD containing 2% glucose.
Fermentor parameter setting: fermentation temperature is 30 DEG C, and pH is controlled 5.8, and ventilatory capacity is set as 2.5vvm, DO 40%,
Stirring is simultaneously associated with DO by range of speeds 400-600rpm.Entire fermentation process is divided into strain growth stage and production of astaxanthin rank
Section, D- (+)-galactolipin inducer addition moment is two stage separation.
The strain growth stage is the lasting supply for guaranteeing carbon source, and glucose is added with fed-batch mode, and is controlled in 0-1g/L
In range, to be reduced as far as the generation of by-product, because glucose extra in culture medium can be in yeast flow metabolism
The by-products such as glycerol, ethyl alcohol and acetic acid are converted into, so as to cause cell yield reduction.When thalli growth enters deadtime, add
Enter final concentration of 20g/L D- (+)-galactolipin inducer, stream plus glucose is simultaneously stopped, into the chemical activators stage.At this time
Carbon source is switched to ethyl alcohol in fermentor, same to pump fed-batch mode addition by feed supplement, and controls the concentration of alcohol in fermentor and exist
5g/L or less.
In addition, an important factor for influencing fermentation biomass, there are also the nitrogen sources in fermentation medium.The present invention is with yeast extract
For nitrogen source, in order to avoid nitrogen source feed supplement is excessive, the fed-batch mode successively decreased using concentration gradient adds yeast into fermentor
Powder is soaked, the yeast extract mother liquor of certain volume is added into fermentor at regular intervals (with yeast extract in initial medium
Concentration be 1.0 ×).Specific feed supplement process is as follows: 1.2 × yeast extract is added in the 10h of fermentation, 65h, 1.0 × ferment is added
0.8 × yeast extract is added in mother's leaching powder, 75h, and 0.3 × yeast extract is added in 120h, and the later period maintains always 0.3 × concentration,
180h stops feed supplement.Start every liter when fermenting 10h and add 1.2 × yeast extract, in total i.e. 30g yeast extract, mother liquid concentration
It is 400g/L, 30g yeast extract volume is 75ml.Deadtime, OD are grown into 75h cell600Growth slows down, and reduces nitrogen source
Be added to 0.8 ×.Ferment 120h, and strain growth growth slows down once again, reduce nitrogen source be added to 0.3 ×.
Fed batch fermentation test result is as shown in figure 3, final 5L fermentor astaxanthin yield reaches 226.19mg/L, phase
2.43 times are improved than shake flask fermentation yield (65.93mg/L).
Embodiment 3: the 5L ferment tank test after fermentation temperature optimization
In order to further increase astaxanthin yield, continue to optimize ferment tank condition.In addition to fermentation medium
Proportion composition and addition manner, temperature is also to influence the key factor of strain growth and metabolism, therefore on the basis of embodiment 2
The temperature of fermentation process is optimized.Specific implementation step: fermentation process is divided into two stages, the temperature control of first stage
It is 30 DEG C, is grown under this condition most beneficial for cell.Equal cells grow into deadtime, start at this time plus D- (+)-galactolipin lures
It leads chemical activators path gene to start to express, into fermentation second stage, 20 DEG C is cooled the temperature to, to be conducive to astaxanthin
Accumulation, fermentation process is as shown in figure 4, final astaxanthin yield has reached 404.78mg/L, relative to shake flask fermentation output increased
5.14 times.
Therefore, high-yield astaxanthin mutant strain SyBE_Sc04030020 is in 5L fermentor through process optimization, optimum temperature
Condition control are as follows: using D- (+)-galactolipin inducer addition moment as the strain growth stage of fermentation process and production of astaxanthin rank
The separation of section, strain growth phase temperature are 30 DEG C, and production of astaxanthin phase temperature is 20 DEG C.
The above is only a preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art
For member, various improvements and modifications may be made without departing from the principle of the present invention, these improvements and modifications are also answered
It is considered as protection scope of the present invention.
Claims (10)
1. a kind of Wine brewing yeast strain, which is characterized in that deposit number is CGMCC No.17913.
2. the Wine brewing yeast strain that deposit number is CGMCC No.17913 is intermediate chemistry in production astaxanthin or with astaxanthin
Application in the other chemicals of product.
3. the Wine brewing yeast strain that deposit number is CGMCC No.17913 is in the microorganism formulation or system of preparation production astaxanthin
Standby production is using astaxanthin as the application in the microorganism formulation of the other chemicals of intermediate chemical.
4. a kind of method for producing astaxanthin, which is characterized in that the S. cervisiae for being CGMCC No.17913 by deposit number
Strain, which is inoculated into SC solid medium, to be activated, and is then transferred in SC fluid nutrient medium again and is prepared seed culture fluid, finally will kind
Sub- culture solution, which is transferred in YPDG fermentation medium, to ferment, until cell OD600Basicly stable, chemical activators are not further added by.
5. method according to claim 4, which is characterized in that the S. cervisiae for being CGMCC No.17913 by deposit number
Strain lines SC solid medium and carries out activation culture, is then seeded into the culture of SC fluid nutrient medium to OD600=5-8, then press
OD600=0.2 is transferred to the culture of SC fluid nutrient medium to OD600=6.5-7.5 is prepared into seed culture fluid, finally by seed culture
Liquid presses end OD600=0.1 is transferred in YPDG fermentation medium 30 DEG C, 250rpm shaking table oscillation and fermentation, until cell OD600Substantially steady
Fixed, chemical activators are not further added by.
6. a kind of method for producing astaxanthin, which is characterized in that the S. cervisiae for being CGMCC No.17913 by deposit number
Strain, which is inoculated into SC solid medium, to be activated, and is then transferred in SC fluid nutrient medium again and is prepared seed culture fluid, finally will kind
Sub- culture solution, which is transferred in YPD fermentation medium, to ferment, until cell OD600Basicly stable, chemical activators are not further added by；
During fermentation, glucose is added as carbon source with fed-batch mode, and is controlled within the scope of 0-1g/L, and thalli growth, which enters, to stop
When the demurrage, D- (+)-galactolipin inducer is added, stream plus glucose are simultaneously stopped, into the chemical activators stage；It ferments at this time
Carbon source is switched to ethyl alcohol in tank, same to be added by fed-batch mode, and controls the concentration of alcohol in fermentor in 5g/L or less.
7. method according to claim 6, which is characterized in that the S. cervisiae for being CGMCC No.17913 by deposit number
Strain lines SC solid medium and carries out activation culture, is then seeded into the culture of SC fluid nutrient medium to OD600=6-7, then press
4% inoculum concentration is transferred to SC fluid nutrient medium culture to logarithmic growth phase mid-term and is prepared into seed culture fluid, finally trains seed
Nutrient solution is transferred in YPD fermentation medium by 10% inoculum concentration and carries out ferment tank, until cell OD600It is basicly stable, astaxanthin
Synthesis is not further added by.
8. according to claim 6 or 7 the method, which is characterized in that the initial glucose concentration of the YPD fermentation medium is
20g/L。
9. method according to claim 6, which is characterized in that during fermentation, the fed-batch side successively decreased using concentration gradient
Formula adds yeast extract into YPD fermentation medium.
10. method according to claim 9, which is characterized in that during fermentation, 1.2 times of yeast extracts are added when 10h, when 65h
It is added 1.0 times of yeast extracts, when 75h is added 0.8 times of yeast extract, and 0.3 times of yeast extract is added in when 120h, and the later period maintains always
In 0.3 times of concentration, stop feed supplement when cell grows into deadtime；
1.2 times of yeast extracts, concentration are 400g/L, volume 75ml.
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| CN111235044A (en) * | 2019-12-31 | 2020-06-05 | 天津大学 | Recombinant saccharomyces cerevisiae strain for synthesizing delta-tocotrienol, construction method and application |
| CN111979132A (en) * | 2020-08-20 | 2020-11-24 | 宜昌东阳光生化制药有限公司 | Fermentation medium for high-yield astaxanthin and application thereof |
| CN112481144A (en) * | 2020-12-11 | 2021-03-12 | 广东省微生物研究所（广东省微生物分析检测中心） | Yeast mutant strain for producing carotenoid and application of mutant site thereof |
| CN113699052A (en) * | 2020-05-20 | 2021-11-26 | 万华化学(四川)有限公司 | Recombinant saccharomyces cerevisiae for producing astaxanthin and application thereof |
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Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN111235044A (en) * | 2019-12-31 | 2020-06-05 | 天津大学 | Recombinant saccharomyces cerevisiae strain for synthesizing delta-tocotrienol, construction method and application |
| CN111235044B (en) * | 2019-12-31 | 2022-01-04 | 天津大学 | Recombinant saccharomyces cerevisiae strain for synthesizing delta-tocotrienol, construction method and application |
| CN113699052A (en) * | 2020-05-20 | 2021-11-26 | 万华化学(四川)有限公司 | Recombinant saccharomyces cerevisiae for producing astaxanthin and application thereof |
| CN113699053A (en) * | 2020-05-20 | 2021-11-26 | 万华化学(四川)有限公司 | Recombinant saccharomyces cerevisiae for producing astaxanthin and application thereof |
| CN113699052B (en) * | 2020-05-20 | 2023-08-11 | 万华化学(四川)有限公司 | Recombinant saccharomyces cerevisiae for producing astaxanthin and application thereof |
| CN113699053B (en) * | 2020-05-20 | 2023-08-11 | 万华化学(四川)有限公司 | Recombinant saccharomyces cerevisiae for producing astaxanthin and application thereof |
| CN111979132A (en) * | 2020-08-20 | 2020-11-24 | 宜昌东阳光生化制药有限公司 | Fermentation medium for high-yield astaxanthin and application thereof |
| CN112481144A (en) * | 2020-12-11 | 2021-03-12 | 广东省微生物研究所（广东省微生物分析检测中心） | Yeast mutant strain for producing carotenoid and application of mutant site thereof |
| CN112481144B (en) * | 2020-12-11 | 2022-04-12 | 广东省科学院微生物研究所(广东省微生物分析检测中心) | Yeast mutant strain for producing carotenoid and application of mutant site thereof |
| CN114574376A (en) * | 2022-05-09 | 2022-06-03 | 暨南大学 | Saccharomyces cerevisiae Delta-M3 for high-yield carotenoid production and application thereof |
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