CN114606282B - Method for reducing 4' -oxygen-isovaleryl tylosin component by changing dissolved oxygen in fermentation process of tylosin - Google Patents
Method for reducing 4' -oxygen-isovaleryl tylosin component by changing dissolved oxygen in fermentation process of tylosin Download PDFInfo
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Abstract
The invention relates to a method for changing dissolved oxygen in a fermentation process of tylosin to reduce 4' -oxygen-isovaleryl tylosin components, which comprises the steps of carrying out fermentation production of the tylosin in a fermentation tank by taking streptomyces thermotolerans as a production strain, regulating and controlling pH at the initial stage of fermentation, then supplementing tylosin dry powder and L-leucine, and then supplementing glucose solution until the fermentation is completed; the dissolved oxygen amount of different fermentation time is controlled in the whole fermentation process, the dissolved oxygen amount is more than 50% in 0-35 h, the dissolved oxygen amount is 20-30% in 35-70 h, and the dissolved oxygen amount is more than 70% in the period from 70h to tank discharge. The invention changes the air distribution form of the tylosin fermentation process, controls the dissolved oxygen in different fermentation time periods, solves the problems of mycelium aging autolysis and large foam caused by the pressure increase of the independent stirrer, effectively reduces the content of 4' -oxygen-isovaleryl tylosin components in the fermentation liquor, further improves the content of the tylosin in the fermentation liquor, and realizes that the proportion of the tylosin components in the fermentation liquor is higher than 84%.
Description
Technical Field
The invention relates to a method for reducing 4' -oxygen-isovaleryl tylosin components by changing dissolved oxygen in a fermentation process of tylosin, belonging to the technical field of fermentation.
Background
The telavancin, also called acetylisovaleryl tylosin (AIV) and 3-O-acetyl-4' -O-isovaleryl tartrate, has a structure shown in a formula I and is a macrolide antibiotic special for livestock. As a brand-new macrolide antibiotic, the telavancin overcomes the defects of other macrolide medicines, has the advantages of high efficiency, low toxicity, low residue, no drug resistance among macrolide medicines and the like, and is popular in the market.
In the prior art, various tylosin derivatives have been produced mainly by adding tylosin and different acyl donors, wherein the tylosin derivatives comprise the acetyl isovaleryl tylosin, namely the tylosin, which is a third-generation upgrading product of tylosin. By selectively acylating the 3' -OH and 4' -OH positions of tylosin, the synthesis of acetylisovaleryl tylosin genes has been elucidated to include two genes, acyA and acyB, responsible for the acylation function based on tylosin, wherein acyB comprises Bl and B2, wherein gene acyA is mainly responsible for the acetylation function and gene acyB is mainly responsible for the isovalerylation function, B1 gene is responsible for isovalerylation gene on tylosin 4' -OH, and B2 is a positive regulatory gene activating B1. The specific biosynthetic pathway is shown below:
fermentation biosynthesis pathway of AIV
The composition of the fermentation broth of the tylosin is complex, 3-O-acetyl-4 ' -O-isovaleryl tylosin (T2 i 5) is taken as a main component, but the fermentation broth further contains intermediate products such as 3-O-acetyltylosin (T20), 4' -O-isovaleryl tylosin (T0 i 5), 3-O-acetyl-4 ' -O-isovaleryl Lei Luojun (R2 i 5) and the like, and related substances of the acetylisovaleryl tylosin are shown in the following table.
The main fermentation method of the tylosin at present comprises the following steps: after inoculation of the fermenter, the pH is naturally raised to 7.50, then continuously fed with acid, the off-line is controlled between 7.45 and 7.55, the pH of the fermentation liquor naturally metabolizes and falls below 6.9 when the fermentation time is 35-45 h, then the substrate tylosin dry powder and L-leucine are fed in, and then the glucose solution is fed in to enable the pH to be between 6.95 and 7.25 until the fermentation is completed. In the prior art, the fermentation tank stirring paddle combination is generally three layers of stirring, wherein the bottom layer adopts a parabolic stirrer, the middle layer adopts a straight blade disc stirrer, and the upper layer adopts an axial flow type stirring paddle. Although the combination has higher energy consumption and gas-liquid dispersing effect, the combination is unfavorable for controlling the liquid level of fermentation, and larger foam is easy to be generated after the surface layer of the liquid level is in stirring contact with the upper layer, thereby further affecting the feeding and tank discharging volumes. Meanwhile, the existing fermentation technology only analyzes the liquid phase results of T20 and T2i5 in the process of the metabolism of the tylosin, so as to control the feeding rate of the next step, but for the tylosin, hyphae are easy to break due to mechanical stress such as stirring and the like to generate foam, so that secondary metabolism is influenced, the volume is controlled below 85% (near the upper stirring), the rolling of the fermentation liquid is aggravated when the fermentation liquid surface is contacted with the upper stirring in the subsequent stage, the foam is rapidly increased, the fermentation state is uncontrollable, and the content of 4' -oxygen-isovaleryl tylosin is too high.
Disclosure of Invention
Aiming at the defects of the prior art, the invention provides a method for reducing 4' -oxygen-isovaleryl tylosin component by changing dissolved oxygen in the fermentation process of the tylosin.
The technical scheme of the invention is as follows:
a method for changing dissolved oxygen in the fermentation process of tylosin to reduce 4' -oxygen-isovaleryl tylosin components comprises the steps of carrying out fermentation production of tylosin in a fermentation tank by taking streptomyces thermotolerans as a production strain, regulating and controlling the pH of fermentation liquor to 7.00-7.50 at the initial stage of fermentation, then feeding tylosin dry powder and L-leucine as substrates in a fed-batch manner, and then feeding glucose solution to enable the pH of the fermentation liquor to be between 6.95 and 7.25 until the fermentation is completed;
the dissolved oxygen of the fermentation liquid is controlled to be more than 50% in 0-35 h, 20-30% in 35-70 h and more than 70% in 70-tank discharging period.
According to the invention, preferably, the fermentation tank comprises a fermentation tank body, wherein a stirring device is arranged in the fermentation tank body, and an annular air distributor is arranged below the stirring device; the annular air distributor comprises an annular pipe and an air inlet pipe, wherein the annular pipe is communicated with the air inlet pipe, and 4 large air outlet holes and 30 small air outlet holes are uniformly formed in the annular pipe; the air inlet pipe extends out of the fermentation tank body, and a discharge hole is formed in the bottom of the fermentation tank body.
Further preferably, the stirring device is fixed on a motor seat at the top end of the fermentation tank body, a motor is arranged on the motor seat, the side surface of the motor is connected with a speed reducer, and a sealing ring, a stirring shaft penetrating through the top cover of the fermentation tank, a shaft coupling, a straight blade disc stirrer, a middle bearing and a parabolic stirrer are sequentially connected below the speed reducer.
Further preferably, the distance between the parabolic stirrer and the annular air distributor is 5-20 cm, and the width of the parabolic stirrer is the same as the diameter of the annular air distributor.
Further preferably, the annular tube wall thickness phi 125 x 4mm is fixed on the fermenter by a flange.
Further preferably, the size of the large air outlet holes is phi 22mm by 4, and the size of the small air outlet holes is phi 18mm by 30.
Further preferably, the sum of the areas of the large air outlet holes and the small air outlet holes is less than or equal to 80% of the cross section area of the annular tube.
According to the invention, the ratio of the material culture medium adopted in the fermentation production is as follows: 10 to 20g/L of starch, 1 to 10g/L of yeast extract, 0.1 to 0.8g/L of fish extract and K 2 HPO 4 0.01～0.5g/L。
According to the invention, the fermentation is carried out at the initial stage by adopting a mixture of glycerol and citric acid to adjust the pH;
further preferably, the mass ratio of glycerin to citric acid is 1:1.
According to the invention, the mass ratio of the tylosin dry powder to the L-leucine is (5-6) 1, wherein the tylosin dry powder is added according to 800-1000 mug/mL.
The beneficial effects are that:
1. the invention changes the air distribution form of the tylosin fermentation process, effectively controls the dissolved oxygen in different fermentation time periods, solves the problems of mycelium aging autolysis and large foam caused by the pressure increase of the tylosin due to the single stirrer, effectively reduces the component proportion of 4' -oxygen-isovaleryl tylosin in the fermentation broth, further improves the component proportion of the tylosin in the fermentation broth, and realizes that the component proportion of the tylosin in the fermentation broth is more than 84%.
2. According to the invention, the mode that air is introduced from the bottom of the fermentation tank through the straight pipe in the traditional fermentation process is changed, the axial-flow stirring paddles at the upper layer in the fermentation tank are removed, the straight pipe air distributor is changed into the annular air distributor, the injection pressure is improved after air is split, and the air is discharged through the air distributor provided with the large air outlet holes and the small air outlet holes, so that the diameter of air bubbles is far smaller than that of the straight pipe injection type air distributor. And meanwhile, the air outlet is sprayed downwards, after being stirred and contacted with the lower layer, the air outlet is smashed by the stirring paddle, an air pocket is formed on the back surface of the paddle, and when the air pocket is smashed, an enriched small bubble area is formed, so that the contact area of gas and liquid is further increased, and the dissolved oxygen content in the later fermentation liquid is improved.
3. The invention removes the upper layer stirring, realizes 5-10 percent reduction of the electricity consumption of a single fermentation tank, and effectively reduces the production cost.
Drawings
FIG. 1 is a schematic diagram of the structure of the fermenter according to the present invention.
Fig. 2 is a schematic view of the structure of the annular air distributor of the present invention.
FIG. 3 is the effect of different stirring frequencies on the composition of tylosin 4 "-O-isovaleryl tylosin (T0 i 5).
FIG. 4 shows the change in cell concentration under different dissolved oxygen conditions.
FIG. 5 is a graph showing the change in 4 "-O-isovaleryltylosin (T0 i 5) under different dissolved oxygen conditions.
FIG. 6 is a change in 3-O-acetyl-4' -O-isovaleryl tylosin (T2 i 5) under the same dissolved oxygen conditions.
FIG. 7 is a comparison of titers under different dissolved oxygen conditions.
In the figure: 1. the fermentation tank body 2, the motor seat 3, the motor 4, the speed reducer 5, the sealing ring 6, the fermentation tank top cover 7, the stirring shaft 8, the coupling 9 and the straight leaf disc stirrer, 10, an intermediate bearing, 11, a parabolic stirrer, 12, an air inlet pipe, 13, an annular air distributor, 14, a discharge port, 15, an annular pipe, 16, a large air outlet hole, 17 and a small air outlet hole.
Detailed Description
The technical scheme of the present invention will be further described with reference to examples and drawings, but the scope of the present invention is not limited thereto. The reagents and medicines related to the examples are common commercial products unless specified; the experimental procedures referred to in the examples, unless otherwise specified, are conventional in the art.
The equipment used in the examples is conventional existing equipment.
The raw materials used in the examples were starch from the company Jin He, nerngtou county, glucose from the company Nerngtou Gu Fu, biotechnology, inc., and fish meat extract from the company Jinan Asia Kang Linuo, bio-engineering, inc., and tylosin dry powder was Qilu pharmaceutical (Nernou) Co.
As shown in fig. 1-2, a fermentation tank used in the fermentation process of the invention comprises a fermentation tank body 1, wherein a stirring device is arranged in the fermentation tank body 1, the stirring device comprises a motor seat 2 fixed at the top end of the fermentation tank body 1, a motor 3 is arranged on the motor seat 2, and the side surface of the motor 3 is connected with a speed reducer 4.
The lower part of the speed reducer 4 is sequentially connected with a sealing ring 5, a stirring shaft 7 penetrating through a top cover 6 of the fermentation tank, a coupler 8, a straight blade disc stirrer 9, an intermediate bearing 10 and a parabolic stirrer 11. An annular air distributor 13 is arranged 10cm below the parabolic stirrer 11; the annular air distributor 13 comprises an annular pipe 15 and an air inlet pipe 12, wherein the annular pipe 15 is communicated with the air inlet pipe 12, and 4 large air outlet holes 16 and 30 small air outlet holes 17 are uniformly formed in the annular pipe 15; the air inlet pipe 12 extends out of the fermentation tank body 1, and a discharge port 14 is arranged at the bottom of the fermentation tank body 1.
The annular air distributor 13 has a wall thickness phi 125 mm by 4mm and is fixed on the fermentation tank through a flange. The size of the large air outlet hole 16 is phi 22mm, and the size of the small air outlet hole 17 is phi 18mm. The sum of the areas of the large air outlet hole 16 and the small air outlet hole 17 is less than or equal to 80% of the sectional area of the annular tube 15.
Example 1
A method for changing dissolved oxygen in the fermentation process of tylosin to reduce 4' -oxygen-isovaleryl tylosin components comprises the steps of carrying out fermentation production of tylosin in a fermentation tank by taking streptomyces thermotolerans as a production strain, adopting a mixture of glycerol and citric acid to adjust and regulate pH to 7.0 at the initial stage of fermentation, feeding tylosin dry powder and L-leucine as substrates, leading to metabolic change due to the feeding of the substrates, starting to rise the fermentation pH, and feeding glucose solution to lead the pH of fermentation liquor to be 7.2 until the fermentation is completed;
the growth condition of hypha is combined in the whole fermentation process, stirring frequency is regulated to step by step control the dissolved oxygen metabolism of different fermentation time, wherein the stirring frequency is controlled to be 0-30 Hz within 0-35 h, the dissolved oxygen amount of fermentation liquor is controlled to be 55%, the stirring frequency is controlled to be 50 Hz within 35-70 h so as to maintain the dissolved oxygen amount of fermentation liquor to be 30%, and the stirring frequency is controlled to be 40-50 Hz during the period from 70h to tank discharge to be 75%.
The material culture medium adopted in the fermentation production comprises the following components in proportion: 15g/L starch, 5g/L yeast extract, 0.5g/L fish extract, K 2 HPO 4 0.3g/L. The mass ratio of the glycerol to the citric acid is 1:1.
The mass ratio of the tylosin dry powder to the L-leucine is 5:1, wherein the tylosin dry powder is added according to 900 mug/mL.
The fermented broth after completion of fermentation was sampled and assayed at a titer of 24265. Mu.g/mL, a composition ratio of tylosin (T2 i 5) of 84.17% and a composition ratio of 4 "-O-isovaleryltylosin (T0 i 5) of 4%.
Example 2
A method for changing dissolved oxygen in the fermentation process of tylosin to reduce 4' -oxygen-isovaleryl tylosin components comprises the steps of carrying out fermentation production of tylosin in a fermentation tank by taking streptomyces thermotolerans as a production strain, adopting a mixture of glycerol and citric acid to adjust and regulate pH to 7.0 at the initial stage of fermentation, feeding substrate tylosin dry powder and L-leucine, then beginning to raise the pH, and then adding glucose solution to enable the pH of fermentation liquor to be 7.1 until the fermentation is completed;
the growth condition of hypha is combined in the whole fermentation process, stirring frequency is regulated to step by step control the dissolved oxygen metabolism of different fermentation time, wherein the stirring frequency is controlled to be 0-30 Hz within 0-35 h, the dissolved oxygen amount of fermentation liquor is controlled to be 60%, the stirring frequency is controlled to be 50 Hz within 35-70 h so as to maintain the dissolved oxygen amount of fermentation liquor to be 20%, and the stirring frequency is controlled to be 40-50 Hz during the period from 70h to tank discharge to be 80%.
The material culture medium ratio adopted in the fermentation production comprises: 15g/L starch, 5g/L yeast extract, 0.5g/L fish extract, K 2 HPO 4 0.3g/L. The mass ratio of the glycerol to the citric acid is 1:1.
The mass ratio of the tylosin dry powder to the L-leucine is 6:1, wherein the tylosin dry powder is added according to 800 mug/mL.
Example 3
A method for changing dissolved oxygen in the fermentation process of tylosin to reduce 4' -oxygen-isovaleryl tylosin components comprises the steps of carrying out fermentation production of tylosin in a fermentation tank by taking streptomyces thermotolerans as a production strain, adopting a mixture of glycerol and citric acid to adjust and regulate pH to 7.2 at the initial stage of fermentation, feeding substrate tylosin dry powder and L-leucine, and feeding glucose solution to enable the pH of fermentation liquor to be 7.0 until the fermentation is completed;
the growth condition of hypha is combined in the whole fermentation process, stirring frequency is regulated to step by step control the dissolved oxygen metabolism of different fermentation time, wherein the stirring frequency is controlled to be 0-30 Hz within 0-35 h, the dissolved oxygen amount of fermentation liquor is controlled to be 65%, the stirring frequency is controlled to be 50 Hz within 35-70 h so as to maintain the dissolved oxygen amount of fermentation liquor to be 25%, and the stirring frequency is controlled to be 40-50 Hz during the period from 70h to tank discharge to be 85%.
The material culture medium ratio adopted in the fermentation production comprises: 15g/L starch, 5g/L yeast extract, 0.5g/L fish extract, K 2 HPO 4 0.3g/L. The mass ratio of the glycerol to the citric acid is 1:1.
The mass ratio of the tylosin dry powder to the L-leucine is 6:1, wherein the tylosin dry powder is added according to 1000 mug/mL.
Comparative example 1
A method for changing dissolved oxygen in a fermentation process of tylosin to reduce 4' -O-isovaleryl tylosin components is different from example 1 in that an upper axial flow stirring paddle in the prior art is reserved.
Sampling and measuring the fermentation broth after fermentation, wherein the titer is 20116 mug/mL, the component ratio of the telavancin (T2 i 5) is 81.98%, and the component ratio of the 4' -O-isovaleryltylosin (T0 i 5) is 6.59%.
Comparative example 2
A method for changing dissolved oxygen in a fermentation process of tylosin to reduce 4' -oxygen-isovaleryl tylosin components is the same as in example 1, and is different in that stirring frequency is adjusted so that dissolved oxygen amounts from 0 to 35 hours and 70 hours of the fermentation process to a tank discharging period are 40 to 60 percent.
Sampling and measuring the fermented liquid with the titer of 22018 mug/mL, the component proportion of the telavancin (T2 i 5) of 83.52% and the component proportion of the 4' -O-isovaleryl tylosin (T0 i 5) of 4.42%.
Comparative example 3
A method for changing dissolved oxygen in a fermentation process of tylosin to reduce 4' -oxygen-isovaleryl tylosin components is the same as in example 1, and is different in that stirring frequency is adjusted so that the dissolved oxygen amount in the whole fermentation process is 20-40%.
Sampling to determine the fermentation broth after fermentation, the titer is 10708 mug/mL, the component ratio of the telavancin (T2 i 5) is 72.25%, and the component ratio of the 4' -O-isovaleryltylosin (T0 i 5) is 8.24%
Comparative example 4
A method for changing dissolved oxygen in a fermentation process of tylosin to reduce 4' -oxygen-isovaleryl tylosin components is the same as in example 1, an upper-layer axial-flow stirring paddle in the prior art is reserved, and stirring frequency is adjusted so that the dissolved oxygen in the whole fermentation process is 0-20%.
Sampling and measuring fermentation broth with titer of 9164 μg/mL, composition ratio of telavancin (T2 i 5) of 32.36%, composition ratio of 4 "-O-isovaleryltylosin (T0 i 5) of 15.11%
Comparative example 5
The existing three-layer stirring fermentation tank is adopted for fermentation production of the teicoplanin.
The specific method comprises the following steps: the method is characterized in that heat-resistant streptomycete is used as a production strain, ventilation stirring culture is carried out in a liquid fermentation culture medium containing starch, yeast powder, fish extract, yeast extract and inorganic salt, when the pH of fermentation liquor is reduced by 7.00-7.20, substrate tylosin dry powder and L-leucine are fed in, in the middle and later period (100-160 h) of the fermentation process, the change of dissolved oxygen in the fermentation process is regulated by regulating stirring batches for 40-50 Hz, three batches are carried out, the unstable problem of T0i5 is found, the rotating speed is reduced through process control, the insufficient dissolved oxygen has a direct relation, and the T0i5 and the dissolved oxygen form a negative correlation.
Sampling and measuring 01 batch fermentation broth with titer of 21098 μg/mL, composition ratio of telavancin (T2 i 5) of 82.68%, composition ratio of 4 "-O-isovaleryltylosin (T0 i 5) of 5.62%
The test results for example 1 and comparative examples 1 to 5 are shown in table 1 below and fig. 3 to 7:
TABLE 1
As can be seen from table 1 and fig. 3, when using the existing three-layer stirring fermenter to ferment and produce teicoplanin, in the same fermentation period, the control effect of air flow on dissolved oxygen is low, and the control of fermentation is difficult possibly due to foam, the control effect of stirring rotation speed on dissolved oxygen is highest, the problem of instability of T0i5 is found to be in negative correlation with the dissolved oxygen due to the fact that the dissolved oxygen is insufficient by reducing frequency, and three batches of experiments prove that the problem of instability of T0i5 is in direct relation with the reduction of rotation speed of process control and the insufficient dissolved oxygen is in negative correlation with the dissolved oxygen. Therefore, the dissolved oxygen change in the fermentation process can be controlled to effectively control the isovalerylation pathway to be reduced to below 4.5 percent by controlling the rotating speed. Compared with comparative example 5, the invention adopts the annular air distributor to adjust the air flow and the stirring speed at different fermentation time so as to achieve the mode of controlling the dissolved oxygen level, and removes the upper stirring blade, thereby effectively reducing the component proportion of 4' -oxygen-isovaleryl tylosin.
As can be seen from FIGS. 4 to 7, in example 1 of the present invention, compared with comparative examples 2 to 4, the 4 "-O-isovaleryl tylosin (T0 i 5) fraction, the 3-O-acetyl-4" -O-isovaleryl tylosin (T2 i 5) fraction and the fermentation titer can be significantly reduced without changing the feeding means by the dissolved oxygen amount in different time periods during the fermentation. Comparative examples 2 to 4 caused poor growth of teicoplanin mycelia in the early stage due to low control of dissolved oxygen in the process, and showed a remarkable negative correlation of 4 "-O-isovaleryl tylosin with dissolved oxygen control.
Compared with comparative example 1, the upper stirring is only changed, but the components of 4 '-oxygen-isovaleryl tylosin in a tank are higher than those of example 1, and the potency and the components of 3-O-acetyl-4' -O-isovaleryl tylosin are reduced, because the shearing force of the upper stirring causes the mycelium to be broken seriously in the middle and later stages of fermentation, the mycelium generates cavitation bubbles, and the foam is increased uncontrollably, so that the substrate addition is affected. In comparative example 2, the control of the decrease in dissolved oxygen had less effect on the 3-O-acetyl-4 "-O-isovaleryl tylosin component, but the 4" -O-isovaleryl tylosin component increased, increasing the difficulty of the extraction process.
Therefore, the invention effectively slows down the later mycelium fracture autolysis by modifying and adjusting the fermentation tank and controlling the dissolved oxygen amount in the fermentation process by stages through stirring frequency, reduces the content of 4' -oxygen-isovaleryl tylosin component in fermentation, improves the conversion rate, improves the total titer of the tank by 5% -10%, improves the AIV component of fermentation liquor by more than or equal to 84%, and simultaneously removes stirring leaves at the upper layer, thereby reducing the electricity consumption of single-batch fermentation tanks by 5% -10%, and realizing new economic benefit.
Claims (10)
1. A method for changing dissolved oxygen in a fermentation process of tylosin to reduce 4' -oxygen-isovaleryl tylosin components is characterized in that fermentation production of the tylosin is carried out in a fermentation tank by taking streptomyces thermotolerans as a production strain, the pH of fermentation liquor is regulated and controlled to 7.00-7.50 at the initial stage of fermentation, then substrate tylosin dry powder and L-leucine are fed in, and then glucose solution is fed in to enable the pH of the fermentation liquor to be 6.95-7.25 until the fermentation is completed;
the dissolved oxygen amount of the fermentation liquid is controlled to be more than 50% in 0-35 h, 20-30% in 35-70 h and more than 70% in 70-tank discharging period.
2. The method for changing dissolved oxygen in a fermentation process of tylosin to reduce 4 "-O-isovaleryl tylosin component according to claim 1, wherein the fermentation tank comprises a fermentation tank body, a stirring device is arranged in the fermentation tank body, and an annular air distributor is arranged below the stirring device; the annular air distributor comprises an annular pipe and an air inlet pipe, wherein the annular pipe is communicated with the air inlet pipe, and 4 large air outlet holes and 30 small air outlet holes are uniformly formed in the annular pipe; the air inlet pipe extends out of the fermentation tank body, and a discharge hole is formed in the bottom of the fermentation tank body.
3. The method for changing dissolved oxygen in the fermentation process of the tylosin to reduce 4' -oxygen-isovaleryl tylosin components according to claim 2, wherein the stirring device is fixed on a motor seat at the top end of a fermentation tank body, a motor is arranged on the motor seat, a speed reducer is connected to the side surface of the motor, and a sealing ring, a stirring shaft penetrating through a top cover of the fermentation tank, a shaft coupling, a straight leaf disc stirrer, a middle bearing and a parabolic stirrer are sequentially connected below the speed reducer.
4. The method for reducing the dissolved oxygen of a fermentation process of tylosin to reduce 4 "-O-isovaleryl tylosin component according to claim 3, wherein a distance between the parabolic stirrer and the annular air distributor is 5-20 cm, and a width of the parabolic stirrer is the same as a diameter of the annular air distributor.
5. The method of altering the dissolved oxygen of a fermentation process of tylosin to reduce the 4 "-O-isovaleryl tylosin composition according to claim 2, wherein the annular tube wall thickness phi 125 x 4mm is secured to the fermenter by a flange.
6. The method of altering the dissolved oxygen of a fermentation process of tylosin to reduce 4 "-O-isovaleryl tylosin composition according to claim 2, wherein the large exit holes are of size phi 22mm x 4 and the small exit holes are of size phi 18mm x 30.
7. The method of altering the dissolved oxygen of a fermentation process of tylosin to reduce 4 "-O-isovaleryl tylosin composition of claim 2, wherein the sum of the areas of the large gas outlet holes and the small gas outlet holes is less than or equal to 80% of the cross-sectional area of the annular tube.
8. The method for changing dissolved oxygen in a fermentation process of tylosin to reduce 4' -oxygen-isovaleryl tylosin components according to claim 1, wherein the material culture medium adopted in the fermentation production is prepared by the following steps: 10-20 g/L of starch, 1-10 g/L of yeast extract, 0.1-0.8 g/L of fish extract and K 2 HPO 4 0.01~0.5 g/L。
9. The method of altering the dissolved oxygen of a fermentation process of tylosin to reduce 4 "-O-isovaleryl tylosin component according to claim 1, wherein the initial stage of fermentation is the adjustment of pH with a mixture of glycerol and citric acid; the mass ratio of the glycerol to the citric acid is 1:1.
10. The method for reducing the dissolved oxygen in the fermentation process of tylosin to reduce 4' -O-isovaleryl tylosin components according to claim 1, wherein the mass ratio of tylosin dry powder to L-leucine is (5-6): 1, and the tylosin dry powder is added according to 800-1000 mug/mL.
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| CN103224872A (en) * | 2013-04-27 | 2013-07-31 | 齐鲁制药（内蒙古）有限公司 | Fermentation tank for improving tylosin production level |
| CN103276031A (en) * | 2013-06-28 | 2013-09-04 | 宁夏泰瑞制药股份有限公司 | Culture medium for producing acetylisovaleryltylosin by fermenting streptomyces thermotolerans and fermentation method |
| CN112126667A (en) * | 2020-09-30 | 2020-12-25 | 宁夏泰益欣生物科技有限公司 | Fermentation method for improving titer of tulathromycin |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN103224872A (en) * | 2013-04-27 | 2013-07-31 | 齐鲁制药（内蒙古）有限公司 | Fermentation tank for improving tylosin production level |
| CN103276031A (en) * | 2013-06-28 | 2013-09-04 | 宁夏泰瑞制药股份有限公司 | Culture medium for producing acetylisovaleryltylosin by fermenting streptomyces thermotolerans and fermentation method |
| CN112126667A (en) * | 2020-09-30 | 2020-12-25 | 宁夏泰益欣生物科技有限公司 | Fermentation method for improving titer of tulathromycin |
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