A method of improving acarbose fermentation unit
Technical field
The present invention relates to a kind of method for improving fermentation unit more particularly to a kind of sides for improving acarbose fermentation unit
Method belongs to pharmaceutical technology field.
Background technique
Acarbose is by false four sugar substance of actinoplanes (Actinoplanessp) fermenting and producing, and Chinese is not
Name: O-4,6- double deoxidation -4 [[(1S, 4R, 5S, 6S) 4,5,6- trihydroxy -3- (hydroxymethyl) -2- cyclohexene] amino]-(- D-
Glycopyranosyl (1 → 4)-O-)-D- glycopyranosyl (1 → 4)-D- glucopyranose.Acarbose energy and alpha-glucosidase
Competitive inhibition (Wehmeier UF, Piepersberg W.Biotechology and molecular occurs
Biology of the α-glucisidase inhibitor acarbose [J] .Appl Microbiol Biotechnol,
2004,63:613-625) inhibit the polysaccharide of food to decompose, slow down the absorption of sugar accordingly, to reduce postprandial hyperglycemia, cooperate
Dietary therapy diabetes.
The metabolic pathway of microorganism be completed by many associated metabolic pathways collaborations, and its metabolic activity energy according to
The variation of external environment carries out height adjustment (referring to storage torch, the Beijing Li Yourong modern industry fermentation control [M]: chemical work
Industry publishing house, 2002).The physiological property of strain is key factor for the generation of purpose product, but fermentation condition, such as matrix
The control of ingredient and metabolite and its concentration, temperature, pH value, dissolved oxygen is not only related to the growth of microorganism, but also shadow
Ring the yield of metabolite.In fermentation process, the composition and condition of culture of culture medium are very big to yield effect, it is necessary to carry out optimal
Change control.(referring to the Beijing Luo Li new microbial fermentation physiology [M]: Chemical Industry Press, 2009).
Existing zymotechnique is accessed in slant medium using strain and carries out inclined-plane culture.During traditional inclined-plane culture
Generally be protected from light or natural lighting using whole process, the duration and intensity to illumination without control, it is low there are fermentation unit the problems such as.
Summary of the invention
The object of the invention is to overcome the defect of the prior art, provide a kind of side for improving acarbose fermentation unit
Method, this method improve slant strains by light irradiation time, intensity of illumination and the lighting color during control inclined-plane culture
Quality, to improve fermentation unit.
Technical problem of the present invention is solved by following technical scheme.
A method of acarbose fermentation unit being improved, by the inclined-plane actinoplanes Actinoplanes sp. bacterium
Kind culture illumination condition is controlled the method to improve fermentation unit;
The illumination condition control refers to that using incandescent lamp be light source, when intensity of illumination is adjusted to 30-300lux, illumination
Between control in 0-24 hours/day.
The method of above-mentioned raising acarbose fermentation unit, the intensity of illumination are 120-180lux, light control time
At 12-20h/ days.
The method of above-mentioned raising acarbose fermentation unit, the light control time is 12-16h/ days, intensity of illumination
150-180lux。
The method of above-mentioned raising acarbose fermentation unit, the slant strains culture, the group of slant medium become
Sucrose 30g/L, peptone 5g/L, KC1 0.5g/L, KH2P041.0g/L, l-tyrosine 1g/L, MgS040.5g/L, agar
20g/L, solvent are water, and initial pH is 7.0.
The present invention has been surprisingly found that illumination to actinoplanes Actinoplanes sp. slant strains in R&D process
It is affected, the illumination in adjustable inclined surface apparatus incubation can have an impact fermentation unit.According to experimental result it can be seen that
It the use of incandescent lamp is light source during Actinoplanes sp. inclined-plane culture, light application time is controlled in 12-20h/ days, illumination
Fermentation unit, especially light control time be can be improved when 120-180lux in 12-16h/ days, intensity of illumination 150-180lux
When fermentation unit significantly improve.
Detailed description of the invention
Fig. 1 is the influence curve figure of light irradiation time and illumination to lab scale fermentation shake flask unit.
Specific embodiment
Invention is further described in detail With reference to embodiment.
The test of 1 illumination condition of embodiment
(1) seed bottle seed culture
Every cultured fresh inclined plane inoculating is in the 500m1 triangular flask that seed culture medium loading amount is 20%, 27 DEG C of temperature
Under the conditions of degree, with the speed oscillation culture 48h of 250r/min on shaking table, shake-flask seed liquid is made;
(2) shake flask fermentation
Cultured female bottle seed liquor is connected to the 500m1 triangular flask that fermentation medium loading amount is 10% by 15% inoculum concentration
In, with the speed oscillation culture 168h of 250r/min under the conditions of 27 DEG C of temperature;
Experimental result is shown in Fig. 1 to use incandescent lamp as seen from Figure 1 during Actinoplanes sp. inclined-plane culture
For light source, light application time is controlled in 12-20h/ days, illumination 120-180lux, more preferably 12-16h/ days, illumination 150-180lux.
2 comparative experiments of embodiment
Prepare slant medium, culture medium composition it is as follows: sucrose 30g/L, peptone 5g/L, KC1 0.5g/L,
KH2P04 1.0g/L, l-tyrosine 1g/L, MgS04 0.5g/L, agar 20g/L, solvent are water, and initial pH is 7.0.121℃
Sterilizing 30 minutes.
Seed bottle culture medium is prepared, culture medium composition is as follows: starch 10g/L, soybean cake powder 10g/L, CaCO3 2g/L、
Glycerol 20g/L, solvent are water, and initial pH is 7.0.121 DEG C sterilize 30 minutes.
Fermentation shake flask culture medium is prepared, culture medium composition is as follows: maltose 60g/L, glucose 20g/L, soybean cake powder
15g/L、FeCl3 0.2g/L、CaCL2 2g/L、CaCO34g/L, sodium glutamate 2g/L, KH2P04 1g/L, solvent are that water is initial
PH is 7.0.121 DEG C sterilize 30 minutes.
Slant strains preparation: sterile transfer low temperature glycerol tube bacterial strain is in fresh, sterile slant medium.One group of (control
Group is 1) shading culture 6 days at 28 DEG C.Indoor Natural illumination (illumination 80-150lux) culture 6 at second group (control group 2) 28 DEG C
It.It is irradiated 16h/ days, is cultivated 6 days using the incandescent light of 150lux at 28 DEG C of third group (experimental group 1).4th group of (experimental group
2) it is irradiated 12h/ days using the incandescent light of 180lux at 28 DEG C, culture 6 days.
It picks them separately two groups of slant strains to be inoculated in seed culture medium, under the conditions of 27 DEG C of temperature, with 250r/ on shaking table
Shake-flask seed liquid is made in the speed oscillation culture 48h of min;Cultured female bottle seed liquor is connected to fermentation by 15% inoculum concentration
In the 500m1 triangular flask that culture medium loading amount is 10%, with the speed oscillation culture 168h of 250r/min under the conditions of 27 DEG C of temperature.It surveys
Determining fermentation unit the results are shown in Table 1.
1 fermentation unit measurement result of table
As seen from the results in Table 1, by adjusting the illumination condition of actinoplanes Actinoplanes sp. inclined-plane culture, energy
Fermentation unit is had a huge impact.It the use of incandescent lamp is light source, light application time is controlled in 12-20h/ days, illumination 120-
When 180lux, compared with using Indoor Natural illumination condition, fermentation unit, especially light control time can be improved in 12-
16h/ days, intensity of illumination be 150-180lux when, fermentation unit improve it is more significant.