CN107400644A - A kind of culture medium and its application in hydrogenlike silicon ion fermentation and a kind of preparation method of sorbic acid - Google Patents
A kind of culture medium and its application in hydrogenlike silicon ion fermentation and a kind of preparation method of sorbic acid Download PDFInfo
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- CN107400644A CN107400644A CN201710617502.2A CN201710617502A CN107400644A CN 107400644 A CN107400644 A CN 107400644A CN 201710617502 A CN201710617502 A CN 201710617502A CN 107400644 A CN107400644 A CN 107400644A
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Abstract
The present invention relates to technical field of microbial fermentation, more particularly to a kind of culture medium and its application in hydrogenlike silicon ion fermentation and a kind of preparation method of sorbic acid.Using culture medium provided by the invention, by way of stream plus sorbaldehyde, ferment hydrogenlike silicon ion, sorbaldehyde can be converted into sorbic acid, conversion ratio is up to 74.98%~79.94%, and the concentration in nutrient solution is up to 7.55g/L~8.05g/L.Preparation method provided by the invention, safely and environment will not be polluted, the yield of gained sorbic acid is larger, and fermentation time is shorter.
Description
Technical field
The present invention relates to technical field of microbial fermentation, more particularly to a kind of culture medium and its in hydrogenlike silicon ion fermentation
Application and a kind of preparation method of sorbic acid.
Background technology
Sorbic acid is a kind of unrighted acid, the entitled Sorbic acid of English, also known as 2,4- hexadiene acid, 2- third
Alkenyl acrylic acid.As the aliphatic acid natural with other, sorbic acid participates in metabolic processes in human body, and by human consumption
And absorption, produce carbon dioxide and water.Sorbic acid has inhibitory action to yeast, mould, aerobic bacteria, bacterium, der Pilz, moreover it is possible to
Prevent clostridium botulinum, staphylococcus aureus, the growth and breeding of salmonella.From the aspects of security, sorbic acid is a kind of
The preservative of internationally recognized safety (GRAS), security are very high.FAO (Food and Agriculture Organization of the United Nation), the World Health Organization, U.S. FDA are all right
Its security gives affirmative.Therefore, sorbic acid and its salt are because being widely used in food, tobacco, cosmetic field, as anti-
Rotten agent or antistaling agent.
The preparation of sorbic acid and its salt is mainly completed by chemical method at present, but the technique of chemical synthesis sorbic acid is very
Complicated, control difficulty height simultaneously can produce substantial amounts of waste material, and ecological environment is caused seriously to pollute.And manufacturing enterprise of China generally deposits
The problem of be：Synthetic technology backwardness, technique imperfection, production cost are high.Therefore, backward sorb acid production process is not
The sorbic acid dosage expanded day by day can be met.Therefore, the production method of sorbic acid is further studied, improves the yield of sorbic acid simultaneously
The pollution to environment is reduced, is urgent problem to be solved.
The content of the invention
In view of this, the technical problem to be solved in the present invention is to provide a kind of culture medium and its fermented in hydrogenlike silicon ion
In application and a kind of preparation method of sorbic acid；The growth of the suitable hydrogenlike silicon ion of culture medium provided by the invention, and it is suitable
Fermentation condition substantially increase the yield of sorbic acid.
Culture medium provided by the invention include water and：
In some embodiments of the invention, culture medium include water and：
In some embodiments of the invention, culture medium include water and：
In some embodiments of the invention, culture medium include water and：
In the present invention, FeCl3、CuSO4、MnSO4、ZnCl2Using hydrate or non-hydrate all can, using hydrate
When, as long as the content of inorganic salts is consistent with above-mentioned culture medium prescription in nutrient solution, it is all in the protection model of the present invention
Within enclosing.
Culture medium provided by the invention is more suitable for the culture of hydrogenlike silicon ion, and experiment shows, using other culture mediums pair
Hydrogenlike silicon ion is cultivated, and thalli growth is bad, is influenceed sorbaldehyde and is converted to sorbic acid.
Culture medium provided by the invention prepares the application in sorbic acid for hydrogenlike silicon ion fermentation.
The preparation method of sorbic acid provided by the invention is, using sorbaldehyde as substrate, is sent out with culture medium of the present invention
Ferment hydrogenlike silicon ion, obtain the zymotic fluid containing sorbic acid.
The hydrogenlike silicon ion that the hydrogenlike silicon ion that the present invention uses is CGMCC1.3368 for deposit number.
In the present invention, the fermentation includes thalli growth step and sorbaldehyde step of converting；
The thalli growth step is：Thalline is seeded into culture medium of the present invention to rise, temperature is 28~35 DEG C, is stirred
Mix rotating speed is 300~500rpm, and throughput 3mL/min, pH are 6.8~7.4；Cultivate to the OD of bacterium solution600It is worth for 9~12.5；
The sorbaldehyde step of converting is：To OD600Value, which reaches to flow in 9~12.5 bacterium solution, adds sorbaldehyde, and speed of agitator is
300~500rpm, 30 DEG C, throughput is 3~7mL/min, and fermentation time is 60~84h.
The thalline refers to the seed liquor of activation, and the volume fraction of inoculation is 10%~15%；Preferably 10%, 13% or
15%.
Cultivate to bacterium solution OD600It is worth for 9~12.5 96~120h of needs.
In thalli growth step, the mode for maintaining pH value is stream acid liquid.Specially stream plus hydrochloric acid solution, some embodiments
In, stream plus the hydrochloric acid solution that concentration is 2M.
In the embodiment of the present invention, the final concentration of 10mL/L of stream plus sorbaldehyde sorbaldehyde into nutrient solution.
In some embodiments, the thalli growth step is：Thalline is seeded into culture medium of the present invention to rise, temperature
For 35 DEG C, speed of agitator 300rpm, throughput 3mL/min, pH 7.0；Cultivate to the OD of bacterium solution600It is worth for 9~12.5；
The sorbaldehyde step of converting is：To OD600Value, which reaches to flow in 9~12.5 bacterium solution, adds sorbaldehyde, and speed of agitator is
300rpm, 30 DEG C, throughput 7mL/min, fermentation time 72h.
In some embodiments, the thalli growth step is：Thalline is seeded into culture medium of the present invention to rise, temperature
For 30 DEG C, speed of agitator 500rpm, throughput 3mL/min, pH 6.8；Cultivate to the OD of bacterium solution600It is worth for 9~12.5；
The sorbaldehyde step of converting is：To OD600Value, which reaches to flow in 9~12.5 bacterium solution, adds sorbaldehyde, and speed of agitator is
400rpm, 30 DEG C, throughput 3mL/min, fermentation time 72h.
In some embodiments, the thalli growth step is：Thalline is seeded into culture medium of the present invention to rise, temperature
For 35 DEG C, speed of agitator 300rpm, throughput 3mL/min, pH 7；Cultivate to the OD of bacterium solution600It is worth for 9~12.5；
The sorbaldehyde step of converting is：To OD600Value, which reaches to flow in 9~12.5 bacterium solution, adds sorbaldehyde, and speed of agitator is
500rpm, 30 DEG C, throughput 5mL/min, fermentation time 72h.
In some embodiments, the thalli growth step is：Thalline is seeded into culture medium of the present invention to rise, temperature
For 28 DEG C, speed of agitator 400rpm, throughput 3mL/min, pH 7.4；Cultivate to the OD of bacterium solution600It is worth for 9~12.5；
The sorbaldehyde step of converting is：To OD600Value, which reaches to flow in 9~12.5 bacterium solution, adds sorbaldehyde, and speed of agitator is
300rpm, 30 DEG C, throughput 7mL/min, fermentation time 72h.
In some embodiments, the thalli growth step is：Thalline is seeded into culture medium of the present invention to rise, temperature
For 30 DEG C, speed of agitator 500rpm, throughput 3mL/min, pH 6.8；Cultivate to the OD of bacterium solution600It is worth for 9~12.5；
The sorbaldehyde step of converting is：To OD600Value, which reaches to flow in 9~12.5 bacterium solution, adds sorbaldehyde, and speed of agitator is
400rpm, 30 DEG C, throughput 3mL/min, fermentation time 72h.
In some embodiments, the thalli growth step is：Thalline is seeded into culture medium of the present invention to rise, temperature
For 28 DEG C, speed of agitator 400rpm, throughput 3mL/min, pH 7.4；Cultivate to the OD of bacterium solution600It is worth for 9~12.5；
The sorbaldehyde step of converting is：To OD600Value, which reaches to flow in 9~12.5 bacterium solution, adds sorbaldehyde, and speed of agitator is
500rpm, 30 DEG C, throughput 5mL/min, fermentation time 72h.
The step of also including seed activation in the present invention, before the fermentation, the culture medium that the seed activation uses include：
Water and peptone 15g/L；Yeast extract 5g/L；Glucose 5g/L.
In the present invention, the temperature of activation is 28~35 DEG C, and speed of agitator is 150~200rpm, and the time is 24~48h.
In specific embodiment, activation is divided into two steps, and the strain that glycerine freezes is seeded to activation medium, 30 DEG C of shaking tables
After 150rpm cultures 48h, gained bacterium solution is seeded to 35 DEG C, rotating speed 150rpm of activation medium, incubation time 36 hours again.
The inoculum concentration being inoculated with again is 1.5%.
The invention provides a kind of culture medium, and using the culture medium, by way of stream plus sorbaldehyde, fermentation class ball is red thin
Bacterium, sorbaldehyde can be converted into sorbierite, for conversion ratio up to 74.98%~79.94%, the concentration in nutrient solution is reachable
7.55g/L~8.05g/L.Preparation method provided by the invention, safely and environment will not be polluted, the production of gained sorbierite
Measure larger, fermentation time is shorter.
Brief description of the drawings
Fig. 1 shows that embodiment 1 measures the chromatogram of sorbic acid acid content, and wherein abscissa is the time, and ordinate is height；
Fig. 2 shows that embodiment 2 measures the chromatogram of sorbic acid acid content, and wherein abscissa is the time, and ordinate is height；
Fig. 3 shows that embodiment 3 measures the chromatogram of sorbic acid acid content, and wherein abscissa is the time, and ordinate is height；
Fig. 4 shows that embodiment 4 measures the chromatogram of sorbic acid acid content, and wherein abscissa is the time, and ordinate is height；
Fig. 5 shows that embodiment 5 measures the chromatogram of sorbic acid acid content, and wherein abscissa is the time, and ordinate is height；
Fig. 6 shows that embodiment 6 measures the chromatogram of sorbic acid acid content, and wherein abscissa is the time, and ordinate is height；
Fig. 7 shows that comparative example 1 measures the chromatogram of sorbic acid acid content, and wherein abscissa is the time, and ordinate is height；
Fig. 8 shows that comparative example 2 measures the chromatogram of sorbic acid acid content, and wherein abscissa is the time, and ordinate is height.
Embodiment
Application and a kind of preparation of sorbic acid the invention provides a kind of culture medium and its in hydrogenlike silicon ion fermentation
Method, those skilled in the art can use for reference present disclosure, be suitably modified technological parameter realization.In particular, institute
Have similar replacement and change it is apparent to those skilled in the art, they are considered as being included in the present invention.
The method of the present invention and application are described by preferred embodiment, and related personnel can be not substantially being departed from the present invention
Hold, methods herein and application be modified or suitably changed with combining in spirit and scope, to realize and using the present invention
Technology.
With reference to embodiment, the present invention is expanded on further：
Embodiment 1
First, pre-process
The hydrogenlike silicon ion of conventional glycerol stocks is inoculated in the triangular flask equipped with seed culture medium by 1% inoculum concentration
In, it is placed in 30 DEG C of shaking table 150rpm cultures 48h.Percent by volume (g/L) is counted by weight, and the composition of seed culture medium is albumen
Peptone 15g/L, yeast extract 5g/L, glucose 5g/L, surplus are water, pH 7.0.
2nd, seed liquor
By strain obtained by above-mentioned culture according to 1% inoculum concentration it is sterile be transferred in the triangular flask equipped with seed culture medium, put
24h is cultivated in 30 DEG C of shaking table 200rpm.Percent by volume (g/L) is counted by weight, and the composition of seed culture medium is peptone 15g/
L, yeast extract 5g/L, glucose 5g/L, surplus are water, pH 7.0.
3rd, production fermentation
1st, 6L fermentation mediums, fermentation medium components are put into 10L fermentation tanks：By weight percentage, peptone：
30g/L, yeast extract：5g/L, molasses 7g/L, FeCl3 45mg/L、CuSO4 16mg/L、MnSO4 94mg/L、ZnCl2
50mg/L, sodium molybdate 40mg/L, surplus are water.
2nd, seed liquor is inoculated in fermentation tank according to 10% inoculum concentration；
Thalli growth：Cultivation temperature is 35 DEG C, stir speed (S.S.) 300rpm, and keeps persistently being filled with for air 3mL/min.Training
PH value is maintained 7 when supporting, cultivates 120h, OD600Reach 11.25.
3rd, fermenting and producing：After thalline culture 120h, stream plus sorbaldehyde make its final concentration of 10ml/L, 30 DEG C, agitation revolution
For 300rpm, throughput 7ml/min, ferment 72h, and supernatant, high performance liquid chromatography measurement Determination of sorbic (sample is collected by centrifugation
Product dilute 50 times), Determination of sorbic is 7.9g/L (Fig. 1), conversion ratio 78.45%.
Embodiment 2
First, pre-process
The hydrogenlike silicon ion of conventional glycerol stocks is inoculated in the triangular flask equipped with seed culture medium by 1% inoculum concentration
In, it is placed in 30 DEG C of shaking table 150rpm cultures 48h.Percent by volume (g/L) is counted by weight, and the composition of seed culture medium is albumen
Peptone 15g/L, yeast extract 5g/L, glucose 5g/L, surplus are water, pH 7.0.
2nd, seed liquor
By strain obtained by above-mentioned culture according to 1% inoculum concentration it is sterile be transferred in the triangular flask equipped with seed culture medium, put
24h is cultivated in 30 DEG C of shaking table 200rpm.Percent by volume (g/L) is counted by weight, and the composition of seed culture medium is peptone 15g/
L, yeast extract 5g/L, glucose 5g/L, surplus are water, pH 7.0.
3rd, production fermentation
1st, 6L fermentation mediums, fermentation medium components are put into 10L fermentation tanks：By weight percentage, peptone：
20g/L, yeast extract：25g/L, molasses 1g/L, FeCl3 27mg/L、CuSO410mg/L、MnSO4 56mg/L、ZnCl2
30mg/L, sodium molybdate 24mg/L, surplus are water.
2nd, seed liquor is inoculated in fermentation tank according to 15% inoculum concentration；
Thalli growth：Cultivation temperature is 30 DEG C, stir speed (S.S.) 500rpm, and keeps persistently being filled with for air 3mL/min.Training
PH value is maintained 6.8 when supporting, cultivates 108h, OD600Reach 10.35.
3rd, fermenting and producing：After thalline culture 108h, stream plus sorbaldehyde make its final concentration of 10ml/L, 30 DEG C, agitation revolution
For 400rpm, throughput 3ml/min, ferment 72h, and supernatant, high performance liquid chromatography measurement Determination of sorbic (sample is collected by centrifugation
Product dilute 50 times), Determination of sorbic is 7.8g/L (Fig. 2), conversion ratio 77.46%.
Embodiment 3
First, pre-process
The hydrogenlike silicon ion of conventional glycerol stocks is inoculated in the triangular flask equipped with seed culture medium by 1% inoculum concentration
In, it is placed in 30 DEG C of shaking table 150rpm cultures 48h.Percent by volume (g/L) is counted by weight, and the composition of seed culture medium is albumen
Peptone 15g/L, yeast extract 5g/L, glucose 5g/L, surplus are water, pH 7.0.
2nd, seed liquor
By strain obtained by above-mentioned culture according to 1.5% inoculum concentration it is sterile be transferred in the triangular flask equipped with seed culture medium,
It is placed in 35 DEG C of shaking table 150rpm cultures 36h.Percent by volume (g/L) is counted by weight, and the composition of seed culture medium is peptone
15g/L, yeast extract 5g/L, glucose 5g/L, surplus are water, pH 7.0.
3rd, production fermentation
1st, 6L fermentation mediums, fermentation medium components are put into 10L fermentation tanks：By weight percentage, peptone：
10g/L, yeast extract：12g/L, molasses 15g/L, FeCl3 9mg/L、CuSO43mg/L、MnSO4 19mg/L、ZnCl2 10mg/
L, sodium molybdate 8mg/L, surplus are water.
2nd, seed liquor is inoculated in fermentation tank according to 13% inoculum concentration；
Thalli growth：Cultivation temperature is 35 DEG C, stir speed (S.S.) 300rpm, and keeps persistently being filled with for air 3mL/min.Training
PH value is maintained 7 when supporting, cultivates 120h, OD600Reach 12.13.
3rd, fermenting and producing：After thalline culture 120h, stream plus sorbaldehyde make its final concentration of 10ml/L, 30 DEG C, agitation revolution
For 500rpm, throughput 5ml/min, ferment 72h, and supernatant, high performance liquid chromatography measurement Determination of sorbic (sample is collected by centrifugation
Product dilute 50 times), Determination of sorbic is 8.05g/L (Fig. 3), conversion ratio 79.94%.Empirical tests, the conversion ratio of the present embodiment
Other each embodiments are significantly better than, the effect has statistical significance, p<0.05.
Embodiment 4
First, pre-process
The hydrogenlike silicon ion of conventional glycerol stocks is inoculated in the triangular flask equipped with seed culture medium by 1% inoculum concentration
In, it is placed in 30 DEG C of shaking table 150rpm cultures 48h.Percent by volume (g/L) is counted by weight, and the composition of seed culture medium is albumen
Peptone 15g/L, yeast extract 5g/L, glucose 5g/L, surplus are water, pH 7.0.
2nd, seed liquor
By strain obtained by above-mentioned culture according to 1.5% inoculum concentration it is sterile be transferred in the triangular flask equipped with seed culture medium,
It is placed in 35 DEG C of shaking table 150rpm cultures 36h.Percent by volume (g/L) is counted by weight, and the composition of seed culture medium is peptone
15g/L, yeast extract 5g/L, glucose 5g/L, surplus are water, pH 7.0.
3rd, production fermentation
1st, 6L fermentation mediums, fermentation medium components are put into 10L fermentation tanks：By weight percentage, peptone：
20g/L, yeast extract：25g/L, molasses 1g/L, FeCl3 27mg/L、CuSO410mg/L、MnSO4 56mg/L、ZnCl2
30mg/L, sodium molybdate 24mg/L, surplus are water.
2nd, seed liquor is inoculated in fermentation tank according to 15% inoculum concentration；
Thalli growth：Cultivation temperature is 28 DEG C, stir speed (S.S.) 400rpm, and keeps persistently being filled with for air 3mL/min.Training
PH value is maintained 7.4 when supporting, cultivates 96h, OD600Reach 9.96.
3rd, fermenting and producing：After thalline culture 96h, stream plus sorbaldehyde make its final concentration of 10ml/L, 30 DEG C, and agitation revolution is
300rpm, throughput 7ml/min, ferment 72h, and supernatant, high performance liquid chromatography measurement Determination of sorbic (sample is collected by centrifugation
50 times of dilution), Determination of sorbic is 7.55g/L (Fig. 4), conversion ratio 74.98%.
Embodiment 5
First, pre-process
The hydrogenlike silicon ion of conventional glycerol stocks is inoculated in the triangular flask equipped with seed culture medium by 1% inoculum concentration
In, it is placed in 30 DEG C of shaking table 150rpm cultures 48h.Percent by volume (g/L) is counted by weight, and the composition of seed culture medium is albumen
Peptone 15g/L, yeast extract 5g/L, glucose 5g/L, surplus are water, pH 7.0.
2nd, seed liquor
By strain obtained by above-mentioned culture according to 2% inoculum concentration it is sterile be transferred in the triangular flask equipped with seed culture medium, put
48h is cultivated in 28 DEG C of shaking table 180rpm.Percent by volume (g/L) is counted by weight, and the composition of seed culture medium is peptone 15g/
L, yeast extract 5g/L, glucose 5g/L, surplus are water, pH 7.0.
3rd, production fermentation
1st, 6L fermentation mediums, fermentation medium components are put into 10L fermentation tanks：By weight percentage, peptone：
10g/L, yeast extract：12g/L, molasses 15g/L, FeCl3 9mg/L、CuSO43mg/L、MnSO4 19mg/L、ZnCl2 10mg/
L, sodium molybdate 8mg/L, surplus are water.
2nd, seed liquor is inoculated in fermentation tank according to 13% inoculum concentration；
Thalli growth：Cultivation temperature is 30 DEG C, stir speed (S.S.) 500rpm, and keeps persistently being filled with for air 3mL/min.Training
PH value is maintained 6.8 when supporting, cultivates 108h, OD600Reach 10.48.
3rd, fermenting and producing：After thalline culture 108h, stream plus sorbaldehyde make its final concentration of 10ml/L, 30 DEG C, agitation revolution
For 400rpm, throughput 3ml/min, ferment 72h, and supernatant, high performance liquid chromatography measurement Determination of sorbic (sample is collected by centrifugation
Product dilute 50 times), Determination of sorbic is 7.70g/L (Fig. 5), conversion ratio 76.47%.
Embodiment 6
First, pre-process
The hydrogenlike silicon ion of conventional glycerol stocks is inoculated in the triangular flask equipped with seed culture medium by 1% inoculum concentration
In, it is placed in 30 DEG C of shaking table 150rpm cultures 48h.Percent by volume (g/L) is counted by weight, and the composition of seed culture medium is albumen
Peptone 15g/L, yeast extract 5g/L, glucose 5g/L, surplus are water, pH 7.0.
2nd, seed liquor
By strain obtained by above-mentioned culture according to 2% inoculum concentration it is sterile be transferred in the triangular flask equipped with seed culture medium, put
48h is cultivated in 28 DEG C of shaking table 180rpm.Percent by volume (g/L) is counted by weight, and the composition of seed culture medium is peptone 15g/
L, yeast extract 5g/L, glucose 5g/L, surplus are water, pH 7.0.
3rd, production fermentation
1st, 6L fermentation mediums, fermentation medium components are put into 10L fermentation tanks：By weight percentage, peptone：
30g/L, yeast extract：5g/L, molasses 7g/L, FeCl3 45mg/L、CuSO4 16mg/L、MnSO4 94mg/L、ZnCl2
50mg/L, sodium molybdate 40mg/L, surplus are water.
2nd, seed liquor is inoculated in fermentation tank according to 10% inoculum concentration；
Thalli growth：Cultivation temperature is 28 DEG C, stir speed (S.S.) 400rpm, and keeps persistently being filled with for air 3mL/min.Training
PH value is maintained 7.4 when supporting, cultivates 96h, OD600Reach 10.01.
3rd, fermenting and producing：After thalline culture 96h, stream plus sorbaldehyde make its final concentration of 10ml/L, 30 DEG C, and agitation revolution is
500rpm, throughput 5ml/min, ferment 72h, and supernatant, high performance liquid chromatography measurement Determination of sorbic (sample is collected by centrifugation
50 times of dilution), Determination of sorbic is 7.65g/L (Fig. 6), conversion ratio 75.97%.
Comparative example 1
First, pre-process
The hydrogenlike silicon ion of conventional glycerol stocks is inoculated in the triangular flask equipped with seed culture medium by 1% inoculum concentration
In, it is placed in 30 DEG C of shaking table 150rpm cultures 48h.Percent by volume (g/L) is counted by weight, and the composition of seed culture medium is albumen
Peptone 15g/L, yeast extract 5g/L, glucose 5g/L, surplus are water, pH 7.0.
2nd, seed liquor
By strain obtained by above-mentioned culture according to 1.5% inoculum concentration it is sterile be transferred in the triangular flask equipped with seed culture medium,
It is placed in 35 DEG C of shaking table 150rpm cultures 36h.Percent by volume (g/L) is counted by weight, and the composition of seed culture medium is peptone
15g/L, yeast extract 5g/L, glucose 5g/L, surplus are water, pH 7.0.
3rd, production fermentation
1st, 6L fermentation mediums, fermentation medium components are put into 10L fermentation tanks：By weight percentage, peptone：
10g/L, yeast extract：12g/L, sucrose 15g/L, surplus are water.
2nd, seed liquor is inoculated in fermentation tank according to 13% inoculum concentration；
Thalli growth：Cultivation temperature is 35 DEG C, stir speed (S.S.) 300rpm, and keeps persistently being filled with for air 3mL/min.Training
PH value is maintained 7 when supporting, cultivates 120h, OD600Reach 8.9.
3rd, fermenting and producing：After thalline culture 120h, stream plus sorbaldehyde make its final concentration of 10ml/L, 30 DEG C, agitation revolution
For 500rpm, throughput 5ml/min, ferment 72h, and supernatant, high performance liquid chromatography measurement Determination of sorbic (sample is collected by centrifugation
Product dilute 50 times), Determination of sorbic is 4.8g/L (Fig. 7), conversion ratio 47.67%.
Comparative example 2
First, pre-process
The hydrogenlike silicon ion of conventional glycerol stocks is inoculated in the triangular flask equipped with seed culture medium by 1% inoculum concentration
In, it is placed in 30 DEG C of shaking table 150rpm cultures 48h.Percent by volume (g/L) is counted by weight, and the composition of seed culture medium is albumen
Peptone 15g/L, yeast extract 5g/L, glucose 5g/L, surplus are water, pH 7.0.
2nd, seed liquor
By strain obtained by above-mentioned culture according to 1.5% inoculum concentration it is sterile be transferred in the triangular flask equipped with seed culture medium,
It is placed in 35 DEG C of shaking table 150rpm cultures 36h.Percent by volume (g/L) is counted by weight, and the composition of seed culture medium is peptone
15g/L, yeast extract 5g/L, glucose 5g/L, surplus are water, pH 7.0.
3rd, production fermentation
1st, 6L fermentation mediums, fermentation medium components are put into 10L fermentation tanks：By weight percentage, peptone：
10g/L, yeast extract：12g/L, glucose 15g/L, FeCl3 9mg/L、CuSO43mg/L、MnSO4 19mg/L、ZnCl2
10mg/L, sodium molybdate 8mg/L, surplus are water.
2nd, seed liquor is inoculated in fermentation tank according to 13% inoculum concentration；
Thalli growth：Cultivation temperature is 35 DEG C, stir speed (S.S.) 300rpm, and keeps persistently being filled with for air 3mL/min.Training
PH value is maintained 7 when supporting, cultivates 120h, OD600Reach 9.2.
3rd, fermenting and producing：After thalline culture 120h, stream plus sorbaldehyde make its final concentration of 10ml/L, 30 DEG C, agitation revolution
For 500rpm, throughput 5ml/min, ferment 72h, and supernatant, high performance liquid chromatography measurement Determination of sorbic (sample is collected by centrifugation
Product dilute 50 times), Determination of sorbic is 6.05g/L (Fig. 8), conversion ratio 60.08%.
The result of embodiment 1~6 and comparative example 1~2 shows that the method that embodiment 1~6 provides prepares sorbic acid, its turn
Rate is higher than comparative example 1~2, and the effect has statistical significance, p<0.05.And in embodiments, the conversion ratio of embodiment 3
Other each embodiments are significantly better than, the effect has statistical significance, p<0.05.
It the above is only the preferred embodiment of the present invention, it is noted that come for those skilled in the art
Say, under the premise without departing from the principles of the invention, some improvements and modifications can also be made, these improvements and modifications also should be regarded as
Protection scope of the present invention.
Claims (8)
- A kind of 1. culture medium, it is characterised in that including water and：
- 2. application of the culture medium in hydrogenlike silicon ion fermentation prepares sorbic acid described in claim 1.
- 3. a kind of preparation method of sorbic acid, it is characterised in that using sorbaldehyde as substrate, with the culture medium described in claim 1 Ferment hydrogenlike silicon ion, obtains the zymotic fluid containing sorbic acid.
- 4. preparation method according to claim 3, it is characterised in that the hydrogenlike silicon ion is that deposit number is CGMCC1.3368 hydrogenlike silicon ion.
- 5. preparation method according to claim 3, it is characterised in that the fermentation includes thalli growth step and sorbaldehyde Step of converting；The thalli growth step is：Thalline is seeded to the culture medium described in claim 1, temperature is 28~35 DEG C, stirring Rotating speed is 300~500rpm, and throughput 3mL/min, pH are 6.8~7.4；Cultivate to the OD of bacterium solution600It is worth for 9~12.5；The sorbaldehyde step of converting is：To OD600Value, which reaches to flow in 9~12.5 bacterium solution, adds sorbaldehyde, speed of agitator 300 ~500rpm, 30 DEG C, throughput is 3~7mL/min, and fermentation time is 60~84h.
- 6. preparation method according to claim 5, it is characterised in that the stream plus sorbaldehyde sorbaldehyde into nutrient solution Final concentration of 10mL/L.
- 7. preparation method according to claim 3, it is characterised in that the step of also including seed activation before the fermentation, The culture medium that the seed activation uses includes：Water and peptone 15g/L；Yeast extract 5g/L；Glucose 5g/L.
- 8. preparation method according to claim 3, it is characterised in that the temperature of the activation is 28~35 DEG C, and stirring turns Speed is 150~200rpm, and the time is 24~48h.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN108220203A (en) * | 2018-03-06 | 2018-06-29 | 上海海洋大学 | A kind of fermentation medium of hydrogenlike silicon ion |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1986774A (en) * | 2005-12-23 | 2007-06-27 | 王栋 | Industrial preparing process, fermented liquid and use of spheroidal erythrocyte |
| CN101348770A (en) * | 2007-07-18 | 2009-01-21 | 烟台天泰生物工程有限公司 | Rhodobacter sphaeroides, microbial preparation containing viable bacteria or zymocyte liquid of Rhodobacter sphaeroides and use of Rhodobacter sphaeroides |
| CN101928689A (en) * | 2010-07-13 | 2010-12-29 | 王栋 | Method for industrially preparing Rhodobacter sphaeroides, fermentation liquor of Rhodobacter sphaeroides and application |
-
2017
- 2017-07-26 CN CN201710617502.2A patent/CN107400644B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1986774A (en) * | 2005-12-23 | 2007-06-27 | 王栋 | Industrial preparing process, fermented liquid and use of spheroidal erythrocyte |
| CN101348770A (en) * | 2007-07-18 | 2009-01-21 | 烟台天泰生物工程有限公司 | Rhodobacter sphaeroides, microbial preparation containing viable bacteria or zymocyte liquid of Rhodobacter sphaeroides and use of Rhodobacter sphaeroides |
| CN101928689A (en) * | 2010-07-13 | 2010-12-29 | 王栋 | Method for industrially preparing Rhodobacter sphaeroides, fermentation liquor of Rhodobacter sphaeroides and application |
Non-Patent Citations (3)
| Title |
|---|
| GLAESER ET AL.: "Photo-oxidative stress in Rhodobacter sphaeroides: protective role of carotenoids and expression of selected genes", 《MICROBIOLOGY》 * |
| 徐丽丽等: "光合细菌最佳生长条件筛选", 《生物技术》 * |
| 汪多仁等: "《精细化工中间体》", 31 January 2008, 海洋出版社 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108220203A (en) * | 2018-03-06 | 2018-06-29 | 上海海洋大学 | A kind of fermentation medium of hydrogenlike silicon ion |
| CN108220203B (en) * | 2018-03-06 | 2021-03-02 | 上海海洋大学 | Fermentation medium of rhodobacter sphaeroides and application of fermentation medium in fermentation production of rhodobacter sphaeroides |
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