CN108342445B - Method for producing beta-carotene by Blakeslea trispora fermentation and beta-carotene - Google Patents
Method for producing beta-carotene by Blakeslea trispora fermentation and beta-carotene Download PDFInfo
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Abstract
The invention relates to a method for producing beta-carotene by Blakeslea trispora fermentation and beta-carotene, belonging to the field of fermentation engineering. The method comprises the following steps: respectively culturing positive Blakeslea trispora and negative Blakeslea trispora, simultaneously inoculating, fermenting, controlling the pH value of the fermentation system to be 5.8-6.2 by using a regulator in the fermentation process, semi-continuously culturing, discharging materials and supplementing materials when the fermentation is started for 60 hours. The regulator is PO-containing4 3+And does not contain metal ions and acidic substances of salts. The fermentation medium adopted in the fermentation process contains starch phosphate substances. The method is simple and low in cost, and the PO is used4 3+And an acidic substance free of metal ions and salts for controlling fermentation pH and providing PO4 3+Avoid bringing excessive K+In addition, starch phosphate is used to increase the yield of beta-carotene. The yield of the beta-carotene obtained by the method is high, and the method is suitable for industrial production.
Description
Technical Field
The invention relates to the field of fermentation engineering, and particularly relates to a method for producing beta-carotene by fermenting Blakeslea trispora and beta-carotene.
Background
KH is commonly used in the existing fermentation method2PO4Or K2HPO4Iso-containing PO4 3+Salts to increase phosphorylation while providing K+It can enhance the permeability of cell membrane and promote the entry of nutrients into cells. Use KH2PO4Or K2HPO4Iso-containing PO4 3+The phosphorylation level is not enough when the salt content is too low, thereby greatly reducing the metabolic flux of the carotenoid, and the salt content is too lowMany will carry a large number of K+The cell membrane permeability of the thalli is over strong, the beta-carotene is easy to be transferred from the inside of the cell to the outside of the cell, the accumulation rate of the product is correspondingly slowed, and the unit production efficiency is reduced.
Meanwhile, the existing beta-carotene fermented by Blakeslea trispora is mainly fermented in a single tank, and in view of limited natural yield and longer fermentation period, a semi-continuous culture or continuous culture process needs to be developed in the process so as to further reduce the production cost.
Therefore, improvement of the existing method for producing beta-carotene by Blakeslea trispora fermentation is needed.
Disclosure of Invention
The invention aims to provide a method for producing beta-carotene by Blakeslea trispora fermentation, which is simple and low in cost and adopts PO-containing fermentation4 3+And an acidic substance free of metal ions and salts for controlling fermentation pH and providing PO4 3+Avoid bringing excessive K+In addition, starch phosphate is used to increase the yield of beta-carotene.
The second purpose of the invention is to provide beta-carotene, which is produced by the method for producing the beta-carotene by fermenting the Blakeslea trispora, has high yield and is suitable for industrial production.
The technical problem to be solved by the invention is realized by adopting the following technical scheme:
the invention provides a method for producing beta-carotene by Blakeslea trispora fermentation, which comprises the following steps:
respectively culturing positive Blakeslea trispora and negative Blakeslea trispora, simultaneously inoculating, fermenting, controlling the pH value of the fermentation system to be 5.8-6.2 by using a regulator in the fermentation process, discharging when the fermentation is started for 60h, supplementing materials and carrying out semi-continuous culture.
The regulator is PO-containing4 3+And does not contain metal ions and acidic substances of salts.
The fermentation medium adopted in the fermentation process contains starch phosphate substances.
The invention also provides beta-carotene produced by the method for producing the beta-carotene by fermenting the Blakeslea trispora.
Preferably, the yield of beta-carotene obtained is 4-5 g/L.
The method for producing the beta-carotene by the fermentation of the Blakeslea trispora and the beta-carotene provided by the preferred embodiment of the invention have the beneficial effects that:
(1) the product has multiple functions, and the production cost is reduced.
Containing PO4 3+And the acidic substance without metal ions and salts can be used for regulating pH and providing a large amount of PO4 3+Thereby increasing the level of phosphorylation. The starch phosphate substance can provide a C source and can also improve the phosphorylation level. The KH can be prepared by using the above materials simultaneously2PO4Or K2HPO4The dosage of (A) is reduced to a proper level to avoid K+In excess.
(2) The operation is simplified, and the sugar control process is avoided.
The starch phosphate is used as a delayed C source, and the glucose is not required to be added in a controlled feeding flow.
(3) The process is simple and the process is easy to regulate.
(4) The yield/cost ratio is improved.
The yield of the beta-carotene obtained by the preferred embodiment of the invention is higher than that of the prior art, and the semi-continuous culture process is adopted, so that the total yield is improved, and the energy consumption cost is further reduced.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. The examples, in which specific conditions are not specified, were conducted under conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products available commercially.
The method for producing beta-carotene by fermentation of Blakeslea trispora and beta-carotene according to the embodiment of the present invention will be described in detail below.
The method for producing the beta-carotene by the fermentation of the Blakeslea trispora provided by the embodiment of the invention comprises the following steps:
respectively culturing positive Blakeslea trispora and negative Blakeslea trispora according to the following modes: respectively inoculating original Blakeslea trispora positive bacteria and original Blakeslea trispora negative bacteria to a slant containing PDA culture medium, culturing for 4-8 days (preferably 6 days) under the condition of 26-30 ℃ (preferably 28 ℃), then transferring to a seed culture medium, culturing for 44-52 hours under the conditions of 26-30 ℃ and 200-. Preferably, after inoculation into the seed culture medium, the seed culture medium is cultured for 48 hours at 28 ℃ and 220rpm, and air is introduced at a flow rate of 1.5vvm during the culture.
Alternatively, the seed medium may include, for example, 1 to 3wt% of glucose, 2 to 4wt% of corn steep liquor dry powder, 0.5 to 1.5wt% of yeast extract, 0.04 to 0.1wt% of monopotassium phosphate, 0.005 to 0.015wt% of magnesium sulfate, 0.02 to 0.04wt% of sodium glutamate, and 2 to 4wt% of sunflower seed oil solution.
Preferably, the seed culture medium comprises 2wt% of glucose, 3wt% of corn steep liquor dry powder, 1wt% of yeast extract, 0.07wt% of monopotassium phosphate, 0.01wt% of magnesium sulfate, 0.03wt% of sodium glutamate and 3wt% of sunflower seed oil solution.
Inoculating and fermenting the cultured positive Blakeslea trispora and negative Blakeslea trispora simultaneously. Simultaneously inoculating, for example, the cultured positive blakeslea trispora and the cultured negative blakeslea trispora in a weight ratio of 1:3-7 are simultaneously inoculated in a fermentation tank containing a fermentation medium. Preferably, the weight ratio of the positive blakeslea trispora to the negative blakeslea trispora is 1: and 5, by the sexual combination of the positive Blakeslea trispora and the negative Blakeslea trispora according to the proportion, more sex hormone trisporac acid can be generated in comparison with other proportions, and the metabolic flux of the beta-carotene is improved.
Preferably, the biomass dry weight of the positive Blakeslea trispora and the negative Blakeslea trispora used for inoculation is not lower than 10g/L, so as to ensure that the fermentation process can produce a larger amount of sex hormone trisporac acid.
Further, after the simultaneous inoculation, dry compressed air may be introduced into the fermentation tank at a flow rate of 1-2vvm (preferably 1.5 vvm), and then the fermentation may be carried out at a stirring speed of 180-220rpm (200 rpm). The biomass dry weight of the fermentation system in the fermentation process is preferably not more than 50g/L to meet the aerobic requirement.
Because the pH value of the fermentation system is increased in the fermentation process, if the pH value is too high, the production of beta-carotene is not facilitated. Therefore, in the embodiment of the invention, the regulator is added in the fermentation process to control the pH value of the fermentation system to be 5.8-6.2.
Preferably, the regulator is PO-containing4 3+And an acidic substance free of metal ions and salts, such as phosphoric acid or phytic acid, preferably phytic acid. By using a composition containing PO4 3+And the acidic substance without metal ions and salts, especially phytic acid, can effectively control the fermentation pH value and provide PO4 3+Avoid bringing excessive K+Increasing the level of phosphorylation.
It is worth to say that the pH value in the fermentation process needs to be detected in the whole process, and when the pH value is more than 6.2, the regulator is supplemented in time.
Optionally, the fermentation medium contains a starch phosphate, preferably a phosphorylated distarch phosphate. On one hand, the starch phosphate substance can provide a C source (a slow-acting C source) and simultaneously improve the phosphorylation level, and the starch phosphate substance can be used as the supplement of the phytic acid due to the small addition amount of the regulator (the phytic acid). On the other hand, the starch phosphate ester substance contains starch, can replace common starch, and can be hydrolyzed into glucose in the fermentation process, thereby reducing the using amount of the common glucose. In addition, KH can be obtained by using regulator together with starch phosphate2PO4Or K2HPO4The dosage of (A) is reduced to a proper level to avoid K+In excess.
Preferably, the level of P in the fermentation broth of the present example is maintained above 150ppm to maintain a material basis for high levels of substrate phosphorylation.
Alternatively, the fermentation medium may contain, for example, 0.8 to 1.2wt% of glucose, 1.0 to 3.0wt% of phosphorylated distarch phosphate, 0.8 to 1.2wt% of yeast extract, 1.8 to 2.2wt% of corn steep liquor dry powder, 0.5 to 0.9wt% of monopotassium phosphate, 0.8 to 1.2wt% of magnesium sulfate, and 1.8 to 2.2wt% of sunflower seed oil.
Preferably, the fermentation medium may contain 1wt% glucose, 2wt% phosphorylated distarch phosphate, 1wt% yeast extract, 2wt% corn steep liquor dry powder, 0.7wt% monopotassium phosphate, 1wt% magnesium sulfate and 2wt% sunflower seed oil.
The material is discharged when 60 hours is started from the fermentation, and the discharge amount in the discharging process can be, for example, 5-60vol% or 30-50 vol%. Preferably, the biomass in the fermentation system is not less than 30g/L and the beta-carotene content is not less than 6wt% when discharging.
Correspondingly, feeding the rest fermentation system after discharging, and continuously growing the strains by feeding the nutrients and the like required by the growth of the strains. The combination of discharging and feeding can prolong the culture period and reduce the production cost.
In the implementation of the invention, the supplement is to supplement the liquid of the positive blakeslea trispora and the liquid of the negative blakeslea trispora in the fermentation system, and the supplement proportion can be 1:3-7, and preferably 1: 5.
Preferably, in order to ensure that nutrient substances obtained by the strains are sufficient and uniform in the fermentation processes of different stages, the feeding method is adopted in the embodiment of the invention, namely, the fermentation tank is fed with the liquid of the positive blakeslea trispora and the liquid of the negative blakeslea trispora at the time of fermentation liquor volume of 4-6vol% every 10 h.
Then, semi-continuously culturing for 240h at 180 ℃ and placing the culture tank.
Compared with the prior art, the method can increase the unit content of the beta-carotene from about 8 percent to about 12 percent, and the yield is increased from about 3.2g/L to about 4.5g/L (such as 3.7-4.8 g/L).
The features and properties of the present invention are described in further detail below with reference to examples.
Example 1
Respectively inoculating original Blakeslea trispora positive bacteria and original Blakeslea trispora negative bacteria on a slant containing a PDA culture medium, culturing for 6 days at 28 ℃, then transferring to a seed culture medium, culturing for 48 hours at 28 ℃ and 220rpm, and introducing air at a flow rate of 1.5vvm in the culture process.
The weight ratio of the cultured positive blakeslea trispora to the cultured negative blakeslea trispora is 1:5 in a fermenter containing the fermentation medium. The biomass dry weight of the positive Blakeslea trispora and the negative Blakeslea trispora during inoculation is 15 g/L.
After the simultaneous inoculation, dry compressed air was introduced into the fermenter at a flow rate of 1.5vvm and the fermentation was then carried out at a stirring speed of 200 rpm. The biomass dry weight of the fermentation system in the fermentation process can reach about 45 g/L. Adding phytic acid during the fermentation process to control the pH value of the fermentation system to be 5.8.
Discharging the materials according to the discharge amount of 30vol% when the fermentation is carried out for 60h, wherein the biomass in the fermentation system is 30g/L, and the content of beta-carotene is 6.02 wt%. And additionally supplementing Blakeslea trispora seed solution which is 5vol% of the volume of the fermentation liquor at intervals of 10h after discharging, wherein the Blakeslea trispora seed solution contains 1: and 5. positive Blakeslea trispora strain liquid and negative Blakeslea trispora strain liquid. Semi-continuously culturing for 180h, and placing in a tank.
Wherein the seed culture medium contains 2wt% of glucose, 3wt% of corn steep liquor dry powder, 1wt% of yeast extract, 0.07wt% of monopotassium phosphate, 0.01wt% of magnesium sulfate, 0.03wt% of sodium glutamate and 3wt% of sunflower seed oil solution.
The fermentation medium contains 1wt% of glucose, 2wt% of phosphorylated distarch phosphate, 1wt% of yeast extract, 2wt% of corn steep liquor dry powder, 0.7wt% of monopotassium phosphate, 1wt% of magnesium sulfate and 2wt% of sunflower seed oil.
Example 2
Respectively inoculating original Blakeslea trispora positive bacteria and original Blakeslea trispora negative bacteria on a slant containing a PDA culture medium, culturing for 8 days at 26 ℃, then transferring to a seed culture medium, culturing for 52 hours at 26 ℃ and 200rpm, and introducing air at a flow rate of 1vvm in the culture process.
The weight ratio of the cultured positive blakeslea trispora to the cultured negative blakeslea trispora is 1:3 in a fermenter containing the fermentation medium. The biomass dry weight of the positive Blakeslea trispora and the negative Blakeslea trispora during inoculation is 12 g/L.
After the simultaneous inoculation, dry compressed air was introduced into the fermenter at a flow rate of 1vvm, and the fermentation was then carried out at a stirring speed of 180 rpm. The biomass dry weight of the fermentation system during fermentation was 40 g/L. Adding phytic acid during the fermentation process to control the pH value of the fermentation system to be 6.2.
Discharging the materials according to the discharge amount of 50vol% when the fermentation is carried out for 60h, wherein the biomass in the fermentation system is 35.1g/L, and the content of beta-carotene is 6.5 wt%. And additionally supplementing 4vol% of Blakeslea trispora liquid of the volume of the fermentation broth at intervals of 10h after discharging, wherein the Blakeslea trispora liquid contains 1:3 and the positive strain liquid and the negative strain liquid of the blakeslea trispora. Semi-continuously culturing for 180h, and placing in a tank.
Wherein the seed culture medium contains 1wt% of glucose, 2wt% of corn steep liquor dry powder, 0.5wt% of yeast extract, 0.04wt% of monopotassium phosphate, 0.005wt% of magnesium sulfate, 0.02wt% of sodium glutamate and 2wt% of sunflower seed oil solution.
The fermentation medium contains 0.8wt% of glucose, 1wt% of phosphorylated distarch phosphate, 0.8wt% of yeast extract, 1.8wt% of corn steep liquor dry powder, 0.5wt% of monopotassium phosphate, 0.8wt% of magnesium sulfate and 1.8wt% of sunflower seed oil.
Example 3
Respectively inoculating original Blakeslea trispora positive bacteria and original Blakeslea trispora negative bacteria on a slant containing a PDA culture medium, culturing for 4 days at 30 ℃, then transferring to a seed culture medium, culturing for 44h at 30 ℃ and 240rpm, and introducing air at a flow rate of 2vvm in the culture process.
The weight ratio of the cultured positive blakeslea trispora to the cultured negative blakeslea trispora is 1: 7 when inoculated in a fermentation tank containing a fermentation medium, the dry weight of the biomass of the positive Blakeslea trispora and the dry weight of the biomass of the negative Blakeslea trispora are both 10 g/L.
After the simultaneous inoculation, dry compressed air was introduced into the fermenter at a flow rate of 2vvm and the fermentation was carried out at a stirring speed of 220 rpm. The biomass dry weight of the fermentation system during fermentation was 50 g/L. Adding phytic acid during the fermentation process to control the pH value of the fermentation system to be 6.0.
Discharging the materials according to the discharge amount of 40vol% when the fermentation is carried out for 60h, wherein the biomass in the fermentation system is 32.4g/L, and the content of beta-carotene is 6.3 wt%. And additionally supplementing Blakeslea trispora seed solution which is 6vol% of the volume of the fermentation liquor at intervals of 10h after discharging, wherein the Blakeslea trispora seed solution contains 1: 7 positive strain liquid and negative strain liquid of Blakeslea trispora. Semi-continuously culturing for 180h, and placing in a tank.
Wherein the seed culture medium contains 3wt% of glucose, 4wt% of corn steep liquor dry powder, 1.5wt% of yeast extract, 0.1wt% of monopotassium phosphate, 0.015wt% of magnesium sulfate, 0.04wt% of sodium glutamate and 4wt% of sunflower seed oil solution.
The fermentation medium contains 1.2wt% of glucose, 3wt% of phosphorylated distarch phosphate, 1.2wt% of yeast extract, 2.2wt% of corn steep liquor dry powder, 0.9wt% of monopotassium phosphate, 1.2wt% of magnesium sulfate and 2.2wt% of sunflower seed oil.
Example 4
The embodiment of the present invention is different from embodiment 1 in that: the regulator is phosphoric acid.
Example 5
The embodiment of the present invention is different from embodiment 1 in that: the amount of the discharged material was 5 vol%.
Example 6
The embodiment of the present invention is different from embodiment 1 in that: the discharge amount was 15 vol%.
Example 7
The embodiment of the present invention is different from embodiment 1 in that: after feeding, semi-continuous culture was continued for 200 h.
Example 8
The embodiment of the present invention is different from embodiment 1 in that: after feeding, the semi-continuous culture was continued for 240 h.
Test example 1
The above examples 1-8 were repeated to obtain sufficient beta-carotene. The results of comparing the unit content and yield of beta-carotene obtained using beta-carotene produced by a conventional general fermentation method as a control are shown in table 1.
TABLE 1 beta-carotene Unit content (%) and yield (g/L)
| Example 1 | Example 2 | Example 3 | Example 4 | Example 5 | Example 6 | Example 7 | Example 8 | Control group | |
| Unit content | 10.3 | 10.3 | 9.4 | 11.8 | 9.2 | 8.9 | 11.2 | 10.7 | 8.7 |
| Yield of the product | 4.8 | 4.3 | 4.6 | 4.2 | 3.8 | 3.7 | 4.4 | 4.4 | 3.1 |
As can be seen from Table 1, the beta-carotene obtained in examples 1-8 is higher than that obtained in the control group in both unit content and yield, which indicates that the method for producing beta-carotene by fermentation of Blakeslea trispora provided in the examples of the present invention has better effect.
Comparing example 1 with examples 4-6, it can be seen that the unit content and yield of beta-carotene obtained in example 1 are higher than those obtained in examples 4-6, indicating that the production conditions of example 1 are optimal. Comparing example 1 with examples 7-8, it can be seen that the unit content and yield of beta-carotene obtained by the three are not very different, which indicates that the semi-continuous culture can reach 240h after feeding.
Test example 2
Taking example 1 as an example, comparison groups 1-4 were provided, comparison group 1 and example 1 were discharged at 20 hours from the start of fermentation, comparison group 2 and example 1 were discharged at 40 hours from the start of fermentation, comparison group 3 and example 1 were discharged at 80 hours from the start of fermentation, comparison group 4 and example 1 were discharged at 100 hours from the start of fermentation, and the results are shown in table 2.
TABLE 2 beta-carotene Unit content (%) and yield (g/L)
| Example 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 of the product | 4.8 | 2.8 | 3.0 | 3.6 | 3.2 |
As can be seen from Table 2, the beta-carotene obtained in example 1 was higher in both unit content and yield than those obtained in the control groups 1 to 4, indicating that the discharge start time was set to be optimal at 60 hours after the start of fermentation within the same total fermentation period.
Test example 3
Taking example 1 as an example, controls 5 and 6 were set, and the control 5 was different from example 1 in that phytic acid was not added during the fermentation, and the control 6 was different from example 1 in that phosphorylated distarch phosphate was not contained in the fermentation medium, and the unit content and yield of the obtained β -carotene were compared, and the results are shown in table 3.
TABLE 3 beta-carotene Unit content (%) and yield (g/L)
| Example 1 | Control group 5 | Control group 6 | |
| Unit content | 10.3 | 8.7 | 8.6 |
| Yield of the product | 4.8 | 3.1 | 2.8 |
As can be seen from Table 3, the beta-carotene obtained in example 1 was higher in both unit content and yield than those obtained in controls 5 and 6, which indicates that the addition of phytic acid during the fermentation process and the presence of phosphorylated distarch phosphate in the fermentation medium in the examples of the present invention are both beneficial to increasing the unit content and yield of beta-carotene.
Test example 4
Taking example 1 as an example, control groups 7 and 8 were set, the difference between the control group 7 and example 1 was that the dry weight of the biomass of the positive and negative bacteria, i.e., B.trispora used for inoculation was 8g/L, the difference between the control group 8 and example 1 was that the dry weight of the biomass of the fermentation system during fermentation was 55g/L, and the results of comparing the unit content and yield of the obtained beta-carotene are shown in Table 4.
TABLE 4 beta-carotene Unit content (%) and yield (g/L)
| Example 1 | Control group 7 | Control group 8 | |
| Unit content | 10.3 | 6.0 | 5.8 |
| Yield of the product | 4.8 | 2.4 | 3.2 |
As can be seen from Table 4, the beta-carotene obtained in example 1 is higher in both unit content and yield than those obtained in the control groups 7 and 8, which shows that the biomass dry weight of the positive and negative Blakeslea trispora used for inoculation is controlled to be not less than 10g/L, and the biomass dry weight of the fermentation system in the fermentation process is controlled to be not more than 50g/L, which is beneficial to improving the unit content and yield of the beta-carotene.
Test example 5
Taking example 1 as an example, control groups 9 to 14 were set, control group 9 differing from example 1 in the amount of blowdown of 3vol%, control group 10 differing from example 1 in the amount of blowdown of 5vol%, control group 11 differing from example 1 in the amount of blowdown of 15vol%, control group 12 differing from example 1 in the amount of blowdown of 45vol%, control group 13 differing from example 1 in the amount of blowdown of 60vol%, and control group 14 differing from example 1 in the amount of blowdown of 70 vol%. The results of comparing the unit contents and yields of the obtained beta-carotene are shown in Table 5.
TABLE 5 beta-carotene Unit content (%) and yield (g/L)
| Example 1 | Control group 9 | Control group 10 | Control group 11 | Control group 12 | Control group 13 | Control group 14 | |
| Unit content | 10.3 | 8.9 | 9.9 | 9.8 | 10.1 | 10.0 | 8.6 |
| Yield of the product | 4.8 | 3.7 | 4.0 | 3.9 | 4.3 | 4.3 | 3.4 |
As can be seen from Table 5, the beta-carotene obtained in the examples 1, the control group 12 and the control group 13 are higher in both unit content and yield than the control groups 10 and 11, and the control groups 10 to 11 are higher than the control group 9 and the control group 14, which shows that the control of the discharge amount in the examples of the present invention to 30-60vol% is more favorable for improving the unit content and yield of the beta-carotene than to 5-30vol%, and the influence of the discharge amount less than 5vol% on the improvement of the unit content and yield of the beta-carotene is not obvious.
Test example 6
Taking example 1 as an example, a control group 15-16 was set, the difference between the control group 15 and example 1 was that the biomass in the fermentation system was 25g/L when discharging, and the difference between the control group 16 and example 1 was that the beta-carotene content was 5wt% when discharging. The results of comparing the unit contents and yields of the obtained beta-carotene are shown in Table 6.
TABLE 6 beta-carotene Unit content (%) and yield (g/L)
| Example 1 | Control group 15 | Control group 16 | |
| Unit content | 10.3 | 9.1 | 8.2 |
| Yield of the product | 4.8 | 3.6 | 4.0 |
As can be seen from Table 6, the beta-carotene obtained in example 1 was higher in both unit content and yield than those obtained in the control groups 15 and 16, which indicates that the biomass in the fermentation system was not less than 30g/L and the beta-carotene content was not less than 6wt% during discharging in the examples of the present invention, which is favorable for increasing the unit content and yield of beta-carotene.
In summary, the method for producing beta-carotene by Blakeslea trispora fermentation provided by the embodiment of the invention is simple and low in cost, and adopts PO-containing fermentation4 3+And an acidic substance free of metal ions and salts for controlling fermentation pH and providing PO4 3+Avoid bringing excessive K+In addition, starch phosphate is used to increase the yield of beta-carotene. The yield of the beta-carotene produced by the method for producing the beta-carotene by fermenting the Blakeslea trispora is higher than that of the beta-carotene produced by fermenting the Blakeslea trisporaHigh efficiency and suitability for industrial production.
The embodiments described above are some, but not all embodiments of the invention. The detailed description of the embodiments of the present invention is not intended to limit the scope of the invention as claimed, but is merely representative of selected embodiments of the invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Claims (7)
1. A method for producing beta-carotene by Blakeslea trispora fermentation is characterized by comprising the following steps:
respectively culturing positive Blakeslea trispora and negative Blakeslea trispora, simultaneously inoculating, fermenting, controlling the pH value of a fermentation system to be 5.8-6.2 by using a regulator in the fermentation process, semi-continuously culturing, discharging materials and supplementing materials when the fermentation is started for 60 hours;
the regulator is phytic acid or phosphoric acid;
the fermentation medium adopted in the fermentation process contains phosphorylated distarch phosphate, and the phosphorylated distarch phosphate accounts for 1.0-3.0wt% of the fermentation medium.
2. The method for producing beta-carotene by Blakeslea trispora fermentation according to claim 1, wherein the biomass dry weight of said positive Blakeslea trispora and said negative Blakeslea trispora used for inoculation is not less than 10 g/L.
3. The method for producing beta-carotene by Blakeslea trispora fermentation according to claim 1, wherein the biomass dry weight of said fermentation system during fermentation does not exceed 50 g/L.
4. The method for producing beta-carotene by Blakeslea trispora fermentation according to claim 1, wherein the amount of the discharged material during discharging is 30-60 vol%.
5. The method for producing beta-carotene by Blakeslea trispora fermentation according to claim 1, wherein the biomass in said fermentation system is not less than 30g/L and the beta-carotene content is not less than 6wt% when discharging.
6. The method for producing beta-carotene by Blakeslea trispora fermentation according to claim 1, wherein the feed is supplemented with the strain of Blakeslea trispora positive strain and/or the strain of Blakeslea trispora negative strain in the fermentation system.
7. The method for producing beta-carotene by Blakeslea trispora fermentation according to claim 6, wherein the feeding is carried out by a feed feeding method, and the strain of Blakeslea trispora positive strain and the strain of Blakeslea trispora negative strain are fed at intervals of 10 hours, wherein the volume of the fermentation broth is 4-6 vol%.
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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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| Publication number | Priority date | Publication date | Assignee | Title |
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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 |
| 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 |
| 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 |
| 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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