CN106167814B - A method of improving acarbose fermentation unit - Google Patents

A method of improving acarbose fermentation unit Download PDF

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Publication number
CN106167814B
CN106167814B CN201610793405.4A CN201610793405A CN106167814B CN 106167814 B CN106167814 B CN 106167814B CN 201610793405 A CN201610793405 A CN 201610793405A CN 106167814 B CN106167814 B CN 106167814B
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fermentation unit
illumination
actinoplanes
days
culture
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CN106167814A (en
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刘军旗
王俊刚
李英然
何茹
曹艳霞
赵宇
李珍
贾伟娜
刘汉忠
李雪然
袁玉伟
张敬坤
马蕙
宋娅娅
张蕾
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CSPC GROUP SECRET SNOW GLUCOSE Co.,Ltd.
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HEBEI HUARONG PHARMACEUTICAL CO Ltd
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    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12PFERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P19/00Preparation of compounds containing saccharide radicals
    • C12P19/26Preparation of nitrogen-containing carbohydrates

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  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)

Abstract

The present invention relates to a kind of methods for improving acarbose fermentation unit, improve fermentation unit by being controlled actinoplanes Actinoplanes sp. slant strains culture illumination condition.According to experimental result it can be seen that during Actinoplanes sp. inclined-plane culture, it the use of incandescent lamp is light source, light application time control can be improved fermentation unit at 12-20h/ days, illumination 120-180lux, and especially light control time fermentation unit when 12-16h/ days, intensity of illumination is 150-180lux significantly improves.

Description

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.

Claims (2)

1. a kind of method for improving acarbose fermentation unit, which is characterized in that by actinoplanes Actinoplanes Sp. the method that slant strains culture illumination condition is controlled to improve fermentation unit;
The illumination uses incandescent lamp, and the light control time at 12-16h/ days, intensity of illumination 150-180lux, cultivates 6 days.
2. the method according to claim 1 for improving acarbose fermentation unit, which is characterized in that the slant strains training It supports, the group of slant medium becomes 30 g/L of sucrose, peptone 5 g/L, KCl 0. 5 g/L, KH2P04 1. 0 g/L 、 L-tyrosine 1 g/L, MgS040. 5 g/L, 20 g/L of agar, solvent are water, and initial pH is 7.0.
CN201610793405.4A 2016-08-31 2016-08-31 A method of improving acarbose fermentation unit Active CN106167814B (en)

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CN115161365B (en) * 2022-09-08 2023-01-06 上海现代制药股份有限公司 Fermentation process for increasing acarbose yield

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CA2282735A1 (en) * 1997-02-28 1998-09-03 Bayer Aktiengesellschaft Acarbose (acb) cluster from actinoplanes sp. se 50/110
CN101603066A (en) * 2008-06-13 2009-12-16 上海医药工业研究院 A kind of preparation method of acarbose
CN102140485A (en) * 2010-12-25 2011-08-03 浙江工业大学 Method for preparing acarbose through microbial fermentation
CN102978261A (en) * 2012-08-24 2013-03-20 河北华荣制药有限公司 High malt syrup and method for improving acarbose fermentation unit by using high malt syrup

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CA2282735A1 (en) * 1997-02-28 1998-09-03 Bayer Aktiengesellschaft Acarbose (acb) cluster from actinoplanes sp. se 50/110
CN101603066A (en) * 2008-06-13 2009-12-16 上海医药工业研究院 A kind of preparation method of acarbose
CN102140485A (en) * 2010-12-25 2011-08-03 浙江工业大学 Method for preparing acarbose through microbial fermentation
CN102978261A (en) * 2012-08-24 2013-03-20 河北华荣制药有限公司 High malt syrup and method for improving acarbose fermentation unit by using high malt syrup

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