CN108342445A - A kind of method and beta carotene of Blakeslea trispora fermentation production beta carotene - Google Patents
A kind of method and beta carotene of Blakeslea trispora fermentation production beta carotene Download PDFInfo
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Abstract
It ferments the present invention relates to a kind of Blakeslea trispora and produces the method and bata-carotene of bata-carotene, belong to field of fermentation engineering.The above method includes：The positive bacterium of culture Blakeslea trispora and Blakeslea trispora bear bacterium respectively, are inoculated with simultaneously, ferment, and fermentation process controls the pH value of fermentation system as 5.8 6.2 with conditioning agent, Semi-continuous cultivation, blowing when fermentation starts 60h, feed supplement.Conditioning agent is to contain PO4 3+And not acidic materials of metal ion and salt.The fermentation medium used in fermentation process contains starch phosphate Ester.The method is simple, of low cost, by with containing PO4 3+And the acidic materials of metal ion and salt do not control fermentation pH value and provide PO4 3+, avoid bringing excessive K into+, it is used cooperatively starch phosphate Ester in addition, to improve the yield of bata-carotene.Thus bata-carotene yield obtained by is higher, is suitable for industrialized production.
Description
Technical field
The present invention relates to field of fermentation engineering, and more particularly to a kind of method of Blakeslea trispora fermentation production beta carotene
And beta carotene.
Background technology
Existing fermentation process is usually using KH2PO4Or K2HPO4Deng containing PO4 3+Salt promote phosphorylation level, carry simultaneously
K is supplied+, enhance permeability of cell membrane, enter cell beneficial to nutriment.Use KH2PO4Or K2HPO4Deng containing PO4 3+Salt amount mistake
At least phosphorylation level is inadequate, significantly reduces Carotenoid Metabolism flux, and a large amount of K can at most be brought by crossing+So that thalline
Permeability of cell membrane becomes excessively strong, and beta carotene is easily by intracellular transport to extracellular so that and product accumulation rate is accordingly slack-off,
Production of units efficiency declines.
Meanwhile existing Blakeslea trispora ferments beta carotene based on single batch fermentation, in view of its natural limits throughput, originally
Body fermentation period is longer, and the technique that needs to develop Semi-continuous cultivation in technique or continuously cultivate is produced into further decreasing
This.
Therefore, the method for producing beta carotene that need to ferment to existing Blakeslea trispora is improved.
Invention content
The purpose of the present invention is to provide a kind of methods of Blakeslea trispora fermentation production beta carotene, and the method is simple, at
This is cheap, by with containing PO4 3+And the acidic materials of metal ion and salt do not control fermentation pH value and provide PO4 3+, avoid
Bring excessive K into+, it is used cooperatively starch phosphate Ester in addition, to improve the yield of beta carotene.
The second object of the present invention is to provide a kind of beta carotene, is fermented by above-mentioned Blakeslea trispora and produce β-carrot
The method of element is produced and is obtained, and the beta carotene yield is higher, is suitable for industrialized production.
The present invention solves its technical problem using following technical scheme to realize：
The present invention proposes a kind of method of Blakeslea trispora fermentation production beta carotene, includes the following steps：
The positive bacterium of culture Blakeslea trispora and Blakeslea trispora bear bacterium respectively, are inoculated with, ferment, fermentation process is with conditioning agent simultaneously
The pH value for controlling fermentation system is 5.8-6.2, blowing when fermentation starts 60h, feed supplement and Semi-continuous cultivation.
Conditioning agent is to contain PO4 3+And not acidic materials of metal ion and salt.
The fermentation medium used in fermentation process contains starch phosphate Ester.
The present invention also proposes a kind of beta carotene, the method production for the production beta carotene that fermented by above-mentioned Blakeslea trispora
And it obtains.
Preferably, the yield of the beta carotene of gained is 4-5g/L.
The Blakeslea trispora fermentation production method of beta carotene and having for beta carotene that present pre-ferred embodiments provide
Beneficial effect is：
(1) multi-purpose content reduces production cost.
Contain PO4 3+And the acidic materials of metal ion and salt not can be used for regulating and controlling pH, while providing a large amount of PO4 3+,
To improve phosphorylation level.Starch phosphate Ester can also improve phosphorylation level while providing the sources C.It uses simultaneously
The above substance, can be by KH2PO4Or K2HPO4Dosage be down to optimal level, avoid K+It is excessive.
(2) simplify operation, exempt the sugared technique of control.
Starch phosphate Ester is as the sources effect C late, it is not necessary to control feed supplement stream and add glucose.
(3) simple for process, process easy-regulating.
(4) yield/cost ratio is improved.
Beta carotene yield obtained by present pre-ferred embodiments and uses half simultaneously compared with the yield higher of the prior art
Continuous culture process further decreases energy consumption cost while improving total yield.
Specific implementation mode
It in order to make the object, technical scheme and advantages of the embodiment of the invention clearer, below will be in the embodiment of the present invention
Technical solution be clearly and completely described.The person that is not specified actual conditions in embodiment, builds according to normal condition or manufacturer
The condition of view carries out.Reagents or instruments used without specified manufacturer is the conventional production that can be obtained by commercially available purchase
Product.
Have below to the method and beta carotene of the Blakeslea trispora of embodiment of the present invention fermentation production beta carotene
Body explanation.
The method of Blakeslea trispora fermentation production beta carotene provided in an embodiment of the present invention includes the following steps：
The positive bacterium of culture Blakeslea trispora and Blakeslea trispora bear bacterium respectively, and culture can refer to such as under type：By original three spore
The mould positive bacterium of Bradley and original Blakeslea trispora bear bacterium and are inoculated in the inclined-plane containing PDA culture medium respectively, in 26-30 DEG C (preferably 28
DEG C) under conditions of cultivate 4-8 days (preferably 6 days), then transfer in seed culture medium again, in 26-30 DEG C, 200-240rpm
Under conditions of cultivate 44-52h, be passed through air by the flow of 1-2vvm in incubation.Preferably, it is inoculated in seed culture medium
Afterwards, it cultivates 48h under conditions of 28 DEG C, 220rpm, air is passed through by the flow of 1.5vvm in incubation.
Optionally, above-mentioned seed culture medium for example may include the corn steep liquor of the glucose of 1-3wt%, 2-4wt%
Dry powder, the yeast extract of 0.5-1.5wt%, the potassium dihydrogen phosphate of 0.04-0.1wt%, 0.005-0.015wt% magnesium sulfate,
The sodium glutamate of 0.02-0.04wt% and the sunflower oil solution of 2-4wt%.
Preferably, above-mentioned seed culture medium may include the glucose of 2wt%, the Dried Corn Steep Liquor Powder of 3wt%, 1wt% ferment
The certain herbaceous plants with big flowers of female medicinal extract, the potassium dihydrogen phosphate of 0.07wt%, the magnesium sulfate of 0.01wt%, the sodium glutamate of 0.03wt% and 3wt%
Flower seed oil solution.
By after culture the positive bacterium of Blakeslea trispora and Blakeslea trispora bear bacterium simultaneously inoculation fermentation.Inoculation simultaneously can for example incite somebody to action
It is 1 that the positive bacterium of Blakeslea trispora of culture and the Blakeslea trispora of culture, which bear bacterium by weight,：3-7 is inoculated in simultaneously containing fermentation
In the fermentation tank of culture medium.Preferably, the weight ratio that the positive bacterium of above-mentioned Blakeslea trispora bears bacterium with Blakeslea trispora is 1：5, pass through
The positive bacterium of Blakeslea trispora and Blakeslea trispora bear bacterium in the sexual combination of this ratio, can generate more sex hormone under more other ratios
Trisporic acid improves the metabolic flux of beta carotene.
Preferably, the biomass dry weight of the inoculation negative bacterium of the positive bacterium of Blakeslea trispora and Blakeslea trispora used is not less than
10g/L, to ensure that fermentation process can generate larger amount of sex hormone trisporic acid.
Further, while after inoculation, drying can be passed through into fermentation tank by the flow of 1-2vvm (preferably 1.5vvm)
Then compressed air is fermented under the speed of agitator of 180-220rpm (200rpm).The life of fermentation system in fermentation process
Object amount dry weight is preferably more than 50g/L, to meet aerobic demand.
Since in fermentation process, the pH value of fermentation system can increase, if pH value is excessively high, can be unfavorable for generating β-Hu Luo instead
Bu Su.Therefore, conditioning agent is added in the embodiment of the present invention, in fermentation process to control the pH value of fermentation system as 5.8-6.2.
Preferably, conditioning agent is to contain PO4 3+And the not acidic materials of metal ion and salt, such as phosphoric acid or phytic acid, it is excellent
It is selected as phytic acid.By using containing PO4 3+And the not acidic materials of metal ion and salt, especially phytic acid, it can effectively control hair
Ferment pH value simultaneously provides PO4 3+, avoid bringing excessive K into+, improve phosphorylation level.
It is worth noting that the pH value in fermentation process needs whole detection that adjusting is then replenished in time when pH value is more than 6.2
Agent.
Optionally, above-mentioned fermentation medium contains starch phosphate Ester, it is preferable that starch phosphate Ester
For phosphorylation PASELLI EASYGEL.On the one hand, starch phosphate Ester can also improve while providing the sources C (sources effect C late)
Phosphorylation level, because the addition of conditioning agent (phytic acid) is little, starch phosphate Ester can be used as the supplement of phytic acid.It is another
Aspect, starch phosphate Ester contain starch, alternative common starch, and its hydrolyzable is grape during the fermentation
Sugar, to reduce the dosage of common glucose sugar.In addition, conditioning agent and starch phosphate Ester are used in conjunction with, it can be by KH2PO4
Or K2HPO4Dosage be down to optimal level, avoid K+It is excessive.
Preferably, the content of P can maintain 150ppm or more in the zymotic fluid of the embodiment of the present invention, to keep substrate phosphorylation
High-caliber material base.
Optionally, above-mentioned fermentation medium can for example contain 0.8-1.2wt% glucose, 1.0-3.0wt% phosphorus
It is acidified PASELLI EASYGEL, 0.8-1.2wt% yeast extracts, 1.8-2.2wt% Dried Corn Steep Liquor Powders, 0.5-0.9wt% di(2-ethylhexyl)phosphates
Hydrogen potassium, 0.8-1.2wt% magnesium sulfate and 1.8-2.2wt% sunflower oils.
Preferably, fermentation medium can contain 1wt% glucose, 2wt% phosphorylations PASELLI EASYGEL, 1wt% yeast
Medicinal extract, 2wt% Dried Corn Steep Liquor Powders, 0.7wt% potassium dihydrogen phosphates, 1wt% magnesium sulfate and 2wt% sunflower oils.
Fermentation starts to start blowing when 60h, and the blowing amount during blowing for example can be 5-60vt%, or
30-50vt%.Preferably, biomass is not less than 30g/L in fermentation system when blowing and content beta-carotene is not less than
6wt%.
Accordingly, feed supplement is carried out to remaining fermentation system after blowing, by feed supplement, to meet the battalion needed for growth
Substance etc. is supported, it is made constantly to grow.Blowing is combined with feed supplement, on the one hand can extend cultivation cycle, on the other hand can be dropped
Low production cost.
During the present invention is implemented, feed supplement is to be complemented at the kind liquid of the negative bacterium of the kind liquid and Blakeslea trispora of the positive bacterium of Blakeslea trispora
In fermentation system, supplement ratio can be 1:3-7, preferably 1:5.
Preferably, to make in the fermentation process of different phase, the nutriment that strain is obtained is more sufficient and uniform, this
Feed supplement is using feedback material formula feed supplement method in inventive embodiments, namely fills into fermentating liquid volume at that time into fermentation tank at interval of 10h and be
The kind liquid and Blakeslea trispora of the positive bacterium of Blakeslea trispora of 4-6vt% bear the kind liquid of bacterium.
Then, Semi-continuous cultivation puts tank to 180-240h.
By the above method compared with the prior art for, the unit content of beta carotene can be made to be increased to 12% from 8% or so
Left and right, yield are promoted to 4.5g/L or so (such as 3.7-4.8g/L) from 3.2g/L or so.
The feature and performance of the present invention are described in further detail with reference to embodiments.
Embodiment 1
The positive bacterium of original Blakeslea trispora and original Blakeslea trispora are born bacterium to be inoculated in respectively containing the oblique of PDA culture medium
Face cultivates 6 days under conditions of 28 DEG C, then transfers in seed culture medium, 48h is cultivated under conditions of 28 DEG C, 220rpm, training
During supporting air is passed through by the flow of 1.5vvm.
It is 1 that the positive bacterium of the Blakeslea trispora of culture and Blakeslea trispora, which are born bacterium by weight,：5 ratio is inoculated in containing hair
In the fermentation tank of ferment culture medium.The biomass dry weight that the positive bacterium of Blakeslea trispora bears bacterium with Blakeslea trispora when inoculation is 15g/L.
After being inoculated with simultaneously, drying compressed air is passed through into fermentation tank by the flow of 1.5vvm, then stirring in 200rpm
It ferments under mix rotating speed.The biomass dry weight of fermentation system can reach 45g/L or so in fermentation process.It is added in fermentation process
Phytic acid is to control the pH value of fermentation system as 5.8.
It is that 30vt% carries out blowing by blowing amount, at this point, biomass is 30g/L, β-Hu in fermentation system when fermentation 60h
Radish cellulose content is 6.02wt%.Additionally fill at interval of 10h the 5vt%'s of fermentating liquid volume at that time after blowing into fermentation tank
Blakeslea trispora kind liquid, it is 1 that this Blakeslea trispora kind liquid, which contains weight ratio,：The 5 positive strain liquid of Blakeslea trispora and three spore Bradleys
Mould negative strain liquid.Semi-continuous cultivation puts tank to 180h.
Wherein, seed culture medium contains 2wt% glucose, 3wt% Dried Corn Steep Liquor Powders, 1wt% yeast extracts, 0.07wt%
Potassium dihydrogen phosphate, 0.01wt% magnesium sulfate, 0.03wt% sodium glutamates and 3wt% sunflower oil solution.
Fermentation medium contains 1wt% glucose, 2wt% phosphorylations PASELLI EASYGEL, 1wt% yeast extracts, 2wt%
Dried Corn Steep Liquor Powder, 0.7wt% potassium dihydrogen phosphates, 1wt% magnesium sulfate and 2wt% sunflower oils.
Embodiment 2
The positive bacterium of original Blakeslea trispora and original Blakeslea trispora are born bacterium to be inoculated in respectively containing the oblique of PDA culture medium
Face cultivates 8 days under conditions of 26 DEG C, then transfers in seed culture medium, 52h is cultivated under conditions of 26 DEG C, 200rpm, training
During supporting air is passed through by the flow of 1vvm.
It is 1 that the positive bacterium of the Blakeslea trispora of culture and Blakeslea trispora, which are born bacterium by weight,：3 ratio is inoculated in containing hair
In the fermentation tank of ferment culture medium.The biomass dry weight that the positive bacterium of Blakeslea trispora bears bacterium with Blakeslea trispora when inoculation is 12g/L.
After being inoculated with simultaneously, drying compressed air is passed through into fermentation tank by the flow of 1vvm, then in the stirring of 180rpm
It ferments under rotating speed.The biomass dry weight of fermentation system is 40g/L in fermentation process.Phytic acid is added in fermentation process to control
The pH value of fermentation system is 6.2.
It is that 50vt% carries out blowing by blowing amount, at this point, biomass is 35.1g/L, β-in fermentation system when fermentation 60h
Carotene carotene content is 6.5wt%.Additionally fill at interval of 10h the 4vt%'s of fermentating liquid volume at that time after blowing into fermentation tank
Blakeslea trispora kind liquid, it is 1 that this Blakeslea trispora kind liquid, which contains weight ratio,：The 3 positive strain liquid of Blakeslea trispora and three spore Bradleys
Mould negative strain liquid.Semi-continuous cultivation puts tank to 180h.
Wherein, seed culture medium contain 1wt% glucose, 2wt% Dried Corn Steep Liquor Powders, 0.5wt% yeast extracts,
0.04wt% potassium dihydrogen phosphates, 0.005wt% magnesium sulfate, 0.02wt% sodium glutamates and 2wt% sunflower oil solution.
Fermentation medium contain 0.8wt% glucose, 1wt% phosphorylations PASELLI EASYGEL, 0.8wt% yeast extracts,
1.8wt% Dried Corn Steep Liquor Powders, 0.5wt% potassium dihydrogen phosphates, 0.8wt% magnesium sulfate and 1.8wt% sunflower oils.
Embodiment 3
The positive bacterium of original Blakeslea trispora and original Blakeslea trispora are born bacterium to be inoculated in respectively containing the oblique of PDA culture medium
Face cultivates 4 days under conditions of 30 DEG C, then transfers in seed culture medium, 44h is cultivated under conditions of 30 DEG C, 240rpm, training
During supporting air is passed through by the flow of 2vvm.
It is 1 that the positive bacterium of the Blakeslea trispora of culture and Blakeslea trispora, which are born bacterium by weight,：7 ratio is inoculated in containing hair
The biomass dry weight that the positive bacterium of Blakeslea trispora bears bacterium with Blakeslea trispora when being inoculated in the fermentation tank of ferment culture medium is 10g/L.
After being inoculated with simultaneously, drying compressed air is passed through into fermentation tank by the flow of 2vvm, then in the stirring of 220rpm
It ferments under rotating speed.The biomass dry weight of fermentation system is 50g/L in fermentation process.Phytic acid is added in fermentation process to control
The pH value of fermentation system is 6.0.
It is that 40vt% carries out blowing by blowing amount, at this point, biomass is 32.4g/L, β-in fermentation system when fermentation 60h
Carotene carotene content is 6.3wt%.Additionally fill at interval of 10h the 6vt%'s of fermentating liquid volume at that time after blowing into fermentation tank
Blakeslea trispora kind liquid, it is 1 that this Blakeslea trispora kind liquid, which contains weight ratio,：The 7 positive strain liquid of Blakeslea trispora and three spore Bradleys
Mould negative strain liquid.Semi-continuous cultivation puts tank to 180h.
Wherein, seed culture medium contain 3wt% glucose, 4wt% Dried Corn Steep Liquor Powders, 1.5wt% yeast extracts,
0.1wt% potassium dihydrogen phosphates, 0.015wt% magnesium sulfate, 0.04wt% sodium glutamates and 4wt% sunflower oil solution.
Fermentation medium contain 1.2wt% glucose, 3wt% phosphorylations PASELLI EASYGEL, 1.2wt% yeast extracts,
2.2wt% Dried Corn Steep Liquor Powders, 0.9wt% potassium dihydrogen phosphates, 1.2wt% magnesium sulfate and 2.2wt% sunflower oils.
Embodiment 4
The embodiment of the present invention and embodiment 1 difference lies in：Conditioning agent is phosphoric acid.
Embodiment 5
The embodiment of the present invention and embodiment 1 difference lies in：Blowing amount is 5vt%.
Embodiment 6
The embodiment of the present invention and embodiment 1 difference lies in：Blowing amount is 15vt%.
Embodiment 7
The embodiment of the present invention and embodiment 1 difference lies in：After feed supplement, continue Semi-continuous cultivation to 200h.
Embodiment 8
The embodiment of the present invention and embodiment 1 difference lies in：After feed supplement, continue Semi-continuous cultivation to 240h.
Test example 1
It repeats to implement above-described embodiment 1-8, obtains enough beta carotenes.To be given birth to by existing universal fermentation process
Beta carotene obtained by production is control, compares the unit content and yield of the beta carotene of gained, the results are shown in Table 1.
The unit content (%) and yield (g/L) of 1 beta carotene of table
As can be seen from Table 1, the beta carotene obtained by embodiment 1-8 either exists compared with the beta carotene obtained by control group
Equal higher in unit content or yield illustrates the side of Blakeslea trispora fermentation production beta carotene provided in an embodiment of the present invention
Method effect is preferable.
Comparative example 1 and embodiment 4-6 can be seen that the unit content and yield of the beta carotene of 1 gained of embodiment
It is above embodiment 4-6, illustrates that the working condition of embodiment 1 is optimal.Comparative example 1 and embodiment 7-8 can be seen that three
The unit content and yield of the beta carotene of gained are not much different, and illustrate that the Semi-continuous cultivation after feed supplement can be to 240h.
Test example 2
By taking embodiment 1 as an example, control group 1-4 is set, and the difference blowing of control group 1 and embodiment 1 is when fermentation starts 20h
It carries out, difference lies in blowings to carry out when the fermentation beginning 40h for control group 2 and embodiment 1, the difference of control group 3 and embodiment 1
Be that blowing carries out when fermentation starts 80h, control group 4 and embodiment 1 difference lies in blowings when fermentation starts 100h into
Row compares the unit content and yield of the beta carotene of gained, and the results are shown in Table 2.
The unit content (%) and yield (g/L) of 2 beta carotene of table
| Embodiment 1 | Control group 1 | Control group 2 | Control group 3 | Control group 4 | |
| Unit content | 10.3 | 7.7 | 8.4 | 9.8 | 8.9 |
| Yield | 4.8 | 2.8 | 3.0 | 3.6 | 3.2 |
As can be seen from Table 2, embodiment 1 is either still produced in unit content compared with the beta carotene obtained by control group 1-4
Equal higher in amount illustrates that within identical fermentation total period, it is best that the blowing time started is set to the 60h after fermentation starts.
Test example 3
By taking embodiment 1 as an example, control group 5 and 6 is set, and difference lies in do not add in fermentation process with embodiment 1 for control group 5
Enter phytic acid, difference lies in phosphorylation PASELLI EASYGEL, comparison gained are free of in fermentation medium with embodiment 1 for control group 6
Beta carotene unit content and yield, the results are shown in Table 3.
The unit content (%) and yield (g/L) of 3 beta carotene of table
| Embodiment 1 | Control group 5 | Control group 6 | |
| Unit content | 10.3 | 8.7 | 8.6 |
| Yield | 4.8 | 3.1 | 2.8 |
As can be seen from Table 3, embodiment 1 compared with control group 5 and 6 gained beta carotene either unit content still
Equal higher in yield illustrates to be added in phytic acid and fermentation medium in fermentation process of the embodiment of the present invention and form sediment containing phosphorylation two
Powder phosphate is conducive to improve the unit content and yield of beta carotene.
Test example 4
By taking embodiment 1 as an example, control group 7 and 8 is set, and difference lies in inoculation three spores used with embodiment 1 for control group 7
It is 8g/L that the mould positive bacterium of Bradley and Blakeslea trispora, which bear the biomass dry weight of bacterium, and difference lies in fermentations with embodiment 1 for control group 8
The biomass dry weight of fermentation system is 55g/L in the process, compares the unit content and yield of the beta carotene of gained, result
As shown in table 4.
The unit content (%) and yield (g/L) of 4 beta carotene of table
| Embodiment 1 | Control group 7 | Control group 8 | |
| Unit content | 10.3 | 6.0 | 5.8 |
| Yield | 4.8 | 2.4 | 3.2 |
As can be seen from Table 4, embodiment 1 compared with control group 7 and 8 gained beta carotene either unit content still
Equal higher in yield illustrates that the embodiment of the present invention will be inoculated with the positive bacterium of Blakeslea trispora used and Blakeslea trispora bears the biology of bacterium
Dry weight control is measured to be not less than 10g/L and controlling the biomass dry weight of fermentation system in fermentation process no more than 50g/
L is conducive to improve the unit content and yield of beta carotene.
Test example 5
By taking embodiment 1 as an example, control group 9-14 is set, control group 9 is 3vt% difference lies in blowing amount with embodiment 1,
Control group 10 and embodiment 1 are 5vt% difference lies in blowing amount, and difference lies in blowing amounts to be for control group 11 and embodiment 1
15vt%, control group 12 and embodiment 1 are 45vt% difference lies in blowing amount, control group 13 and embodiment 1 difference lies in
Blowing amount is 60vt%, and control group 14 is 70vt% difference lies in blowing amount with embodiment 1.Compare the beta carotene of gained
Unit content and yield, the results are shown in Table 5.
The unit content (%) and yield (g/L) of 5 beta carotene of table
As can be seen from Table 5, the beta carotene of embodiment 1, control group 12 and control group 13 compared with control group 10 and 11 gained
The equal higher either in unit content or yield, and control group 10-11 illustrates this compared with 14 higher of control group 9 and control group
Inventive embodiments by blowing amount control 30-60vt% compared with 5-30vt% be more advantageous to improve beta carotene unit content and
Yield, blowing amount is less than 5vt% to the unit content and yield effect unobvious of raising beta carotene.
Test example 6
By taking embodiment 1 as an example, control group 15-16 is set, and difference lies in fermentation bodies when blowing with embodiment 1 for control group 15
Biomass is 25g/L in system, and control group 16 is 5wt% difference lies in content beta-carotene when blowing with embodiment 1.Comparison
The unit content and yield of the beta carotene of gained, the results are shown in Table 6.
The unit content (%) and yield (g/L) of 6 beta carotene of table
| Embodiment 1 | Control group 15 | Control group 16 | |
| Unit content | 10.3 | 9.1 | 8.2 |
| Yield | 4.8 | 3.6 | 4.0 |
As can be seen from Table 6, embodiment 1 compared with control group 15 and 16 gained beta carotene either unit content also
It is equal higher in yield, illustrates that biomass in fermentation system when blowing is not less than 30g/L and β-carrot by the embodiment of the present invention
Cellulose content is conducive to improve the unit content and yield of beta carotene not less than 6wt%.
In conclusion the method for Blakeslea trispora fermentation production beta carotene provided in an embodiment of the present invention, the method letter
It is single, it is of low cost, by with containing PO4 3+And the acidic materials of metal ion and salt do not control fermentation pH value and provide PO4 3+,
It avoids bringing excessive K into+, it is used cooperatively starch phosphate Ester in addition, to improve the yield of beta carotene.By above-mentioned three
Beta carotene yield is higher obtained by the method production of the mould fermentation production beta carotene of spore Bradley, is suitable for industrialized production.
Embodiments described above is a part of the embodiment of the present invention, instead of all the embodiments.The reality of the present invention
The detailed description for applying example is not intended to limit the range of claimed invention, but is merely representative of the selected implementation of the present invention
Example.Based on the embodiments of the present invention, those of ordinary skill in the art are obtained without creative efforts
Every other embodiment, shall fall within the protection scope of the present invention.
Claims (10)
1. a kind of method of Blakeslea trispora fermentation production beta carotene, which is characterized in that include the following steps：
The positive bacterium of culture Blakeslea trispora and Blakeslea trispora bear bacterium respectively, are inoculated with, ferment simultaneously, fermentation process is controlled with conditioning agent
The pH value of fermentation system is 5.8-6.2, and Semi-continuous cultivation blowing and coordinates feed supplement when fermentation starts 60h；
The conditioning agent is to contain PO4 3+And not acidic materials of metal ion and salt；
The fermentation medium used in fermentation process contains starch phosphate Ester.
2. the method for Blakeslea trispora fermentation production beta carotene according to claim 1, which is characterized in that the adjusting
Agent is phytic acid or phosphoric acid；
Preferably, the conditioning agent is phytic acid.
3. the method for Blakeslea trispora fermentation production beta carotene according to claim 1, which is characterized in that the starch
Phosphoric acid ester substance is phosphorylation PASELLI EASYGEL；
Preferably, the phosphorylation PASELLI EASYGEL accounts for the 1.0-3.0wt% of the fermentation medium.
4. the method for Blakeslea trispora fermentation production beta carotene according to claim 1, which is characterized in that used in inoculation
The positive bacterium of the Blakeslea trispora and the Blakeslea trispora bear bacterium biomass dry weight be not less than 10g/L.
5. the method for Blakeslea trispora fermentation production beta carotene according to claim 1, which is characterized in that fermentation process
Described in fermentation system biomass dry weight be no more than 50g/L.
6. the method for Blakeslea trispora fermentation production beta carotene according to claim 1, which is characterized in that blowing process
In blowing amount be 5-60vt%；
Preferably, the blowing amount is 30-50vt%.
7. the method for Blakeslea trispora fermentation production beta carotene according to claim 1, which is characterized in that institute when blowing
It states in fermentation system biomass and is not less than 6wt% not less than 30g/L and content beta-carotene.
8. the method for Blakeslea trispora according to claim 1 fermentation production beta carotene, which is characterized in that feed supplement be by
The kind liquid that the kind liquid of the positive bacterium of Blakeslea trispora and/Blakeslea trispora bear bacterium is complemented in the fermentation system.
9. the method for Blakeslea trispora fermentation production beta carotene according to claim 8, which is characterized in that feed supplement uses
Present material formula feed supplement method, at interval of 10h fill into fermentating liquid volume be 4-6vt% the positive bacterium of the Blakeslea trispora described kind of liquid and
The Blakeslea trispora bears the described kind of liquid of bacterium.
10. a kind of beta carotene, which is characterized in that the beta carotene is by such as three spore of claim 1-9 any one of them
The method of the mould fermentation production beta carotene of Bradley is produced and is obtained；
Preferably, the yield of the beta carotene is 4-5g/L.
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| CN102787158A (en) * | 2011-05-20 | 2012-11-21 | 浙江医药股份有限公司新昌制药厂 | Method for producing natural beta-carotene by fermentation and application |
| CN105441370A (en) * | 2016-01-29 | 2016-03-30 | 山东和田旺生物科技有限公司 | Microbial agent and preparing method thereof |
| CN105506048A (en) * | 2015-12-31 | 2016-04-20 | 内蒙古金达威药业有限公司 | Fermentation method for preparing beta-carotin by using Blakeslea trispora |
| CN107604036A (en) * | 2017-09-22 | 2018-01-19 | 嘉必优生物技术（武汉）股份有限公司 | A kind of method for preparing bata-carotene and bata-carotene product |
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| CN102787158A (en) * | 2011-05-20 | 2012-11-21 | 浙江医药股份有限公司新昌制药厂 | Method for producing natural beta-carotene by fermentation and application |
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