CN102978261B - A kind of high maltose syrup and the method using its raising acarbose fermentation unit - Google Patents
A kind of high maltose syrup and the method using its raising acarbose fermentation unit Download PDFInfo
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- CN102978261B CN102978261B CN201210503105.XA CN201210503105A CN102978261B CN 102978261 B CN102978261 B CN 102978261B CN 201210503105 A CN201210503105 A CN 201210503105A CN 102978261 B CN102978261 B CN 102978261B
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Abstract
A kind of high maltose syrup and the method using its raising acarbose fermentation unit, the interior high maltose syrup includes maltose, trisaccharide, tetrose and the above ingredient of tetrose, and glucose content is 0 in the high maltose syrup.The present invention also provides the methods for improving acarbose fermentation unit, it is cultivated in the medium using high maltose syrup of the invention；The culture medium is slant medium, female bottle seed culture medium and fermentation flask culture medium.The present invention passes through the adjusting to wherein trisaccharide, tetrose and the above components, effectively raise fermentation level, the progress for being more advantageous to because of the improper disadvantage for being suppressed thallus biosynthesis of carbon source concentration of component and improving acarbose biosynthetic metabolism is avoided, fermentation index is improved.
Description
Technical field
The present invention relates to a kind of high maltose syrup and the methods for improving acarbose fermentation unit using it, belong to medical science
Field.
Background technique
Acarbose is the secondary metabolite that actinoplanes (Actinoplanes sp.) is generated.Inhibit because it has
The activity of Q- glucuroide can reduce the decomposition of polysaccharide and oligosaccharides in human body intestinal canal, and then reduce blood sugar concentration, therefore
The clinically drug as treatment diabetes.
In fermentation process, carbon source is the important element for constituting drive member, while being also to generate various metabolism
The primary raw material of product and intracellular reserve substance.Acarbose molecule has false tetrose structure, this shows its biosynthesis
There is close relationship with the catabolism of carbon source material.To acarbose biosynthesis pathway studies have shown that forming 1 molecule
Acarbose, theoretically need the glucose of two molecules and the maltose of a molecule.Fermentation produces the carbon source control of acarbose
Technique, it is necessary to which the content of strict control glucose simultaneously keeps high concentration of the maltose in fermentation liquid to improve the conjunction of acarbose
At.Culture medium carbon source ingredient has great importance for acarbose fermenting and producing in fermentation process.
Due in industrial mass production, generally using high maltose syrup and glucose as carbon in acarbose fermenting and producing
Source, high maltose syrup are added different types of enzyme by starch milk and are process, since the type of enzyme and the difference of dosage will lead to
Each component changes in high maltose syrup, and high maltose syrup typically contains maltose, glucose, trisaccharide and tetrose and tetrose
Above sugar composition, thus its compound composition and each component ratio synthesize generation to the growth of acarbose producing strains and acarbose
It has thanked to direct effect, has not had been reported that research acarbose fermenting carbon source, ratio pair more than trisaccharide, tetrose and tetrose temporarily at present
The detailed test or creation data that acarbose fermentation yield influences, this is a blank.
Summary of the invention
The technical problem to be solved by the present invention is to overcome the defect of the prior art to provide a kind of high maltose syrup, the high wheat
Bud syrup is conducive to the progress of acarbose biosynthetic metabolism, to improve fermentation index.In addition, the present invention further provides
The method for improving acarbose fermentation unit using the high maltose syrup.
Technical problem of the present invention is realized by the following technical solutions.
A kind of high maltose syrup includes maltose, trisaccharide, tetrose and the above ingredient of tetrose, glucose content zero.
Above-mentioned high maltose syrup, the total reducing sugar quality % after the trisaccharide Zhan Gaomai syrup is given money as a gift is 3.0-9.0%, preferably
6.0-8.0%；The tetrose and the above ratio of tetrose are 6-12%, preferably 8-10%；The ratio is the chromatographically pure after giving money as a gift
Degree.
A method of acarbose fermentation unit is improved, is carried out in the medium using above-mentioned high maltose syrup.
The method of above-mentioned raising acarbose fermentation unit, the culture medium are slant medium, female bottle seed culture medium
With fermentation flask culture medium.
The slant medium forms (g/L): sucrose 30g/L, peptone 5g/L, KC10.5g/L, K2HPO41.0g/L、
L-tyrosine 1g/L, MgSO40 .5g/L, agar 20g/L, solvent are water, and initial pH is 7.0；Mother's bottle culture medium composition:
Starch 10g/L, soybean cake powder 10g/L, CaCO32g/L, glycerol 20g/L, solvent are water, and initial pH is 7.0；The fermentation flask
Culture medium composition: maltose 60g/L, glucose 20g/L, soybean cake powder 15g/L, FeCl30.5 g/L、CaCl2 2g/L、CaCO3
4g/L, sodium glutamate 2g/L, K2HPO41g/L, solvent are water, and initial pH is 7.0；Maltose used in the fermentation flask is
Amount after malt saccharic composition in high wheat syrup is pure, pure calculation method are as follows: high wheat syrup quality × sugar concentration × maltose
Content=sterling maltose quality.
The method of above-mentioned raising acarbose fermentation unit, concrete operations carry out as follows:
(1) female bottle seed culture
Female bottle using cultured fresh inclined-plane carry out dig block method be inoculated with, under the conditions of 27 DEG C of temperature, on shaking table with
Mother's bottle seed liquor is made in the speed oscillation culture of 280r/min 2-3 days；
(2) shake flask fermentation
Cultured female bottle seed liquor is connected in the fermentation medium after sterilizing, inoculum concentration 15%, fermentation shake flask is 27
With the speed oscillation culture 168h of 280r/min under the conditions of DEG C temperature.
The present invention has studied in detail in acarbose fermentation production process, trisaccharide and tetrose and four in the high maltose syrup
The ratio of the above component of sugar, is conducive to the control in practical mass production to carbon source in acarbose fermentation medium.The present invention is logical
The adjusting to wherein trisaccharide, tetrose and the above components is crossed, fermentation level is effectively increased, is avoided because of carbon source concentration of component not
The disadvantage for being properly suppressed thallus biosynthesis is more advantageous to the progress for improving acarbose biosynthetic metabolism, improves
Fermentation index.
Acarbose is made of aminocyclitol, 42 amino, 24,62 dideoxy glucose and a molecule maltose three parts.Through
Inventor tests discovery maltose and glucose is best mixed carbon source.In research with the mutation Actinoplanes of actinoplanes
Bacterium CKD2485216 is the process that acarbose generates strain, just has the formation of acarbose in fermentation early stage, concentration with
The growth of thallus and increase, in fermentation latter stage, side components C starts to accumulate.Glucose and maltose be both energy substance again
It is the precursor substance of acarbose.Glucose is just depleted in fermentation early stage, and maltose is slow in entire fermentation process
Ground is utilized, and after glucose exhausts, serves as energy substance.Because maltose can be directly incorporated into the structure of acarbose
In, so consumption of the glucose in early stage will lead to the reduction of later period maltose content, and then affect the yield of acarbose.
Therefore it must be strictly controlled the content of glucose and the high concentration that keeps maltose in fermentation liquid improve the conjunction of acarbose
At.
Specific embodiment
High maltose syrup is to be made from starch, by the syrup made of enzymatic hydrolysis based on maltose.High malt
Syrup is that a kind of maltose content is higher (>=70%), the starch syrup of glucose content lower (≤10%), is a kind of colourless
Transparent sticky liquid, low sugariness have malt flavor.The production process of high malt sugar are as follows: starch (or starch milk) --- it sizes mixing
--- primary injection --- liquefaction ----secondary injection ----saccharification ----activity carbon decoloring ----slagging-off ----high malt sugar.
The invention will be further described with reference to embodiments.
Embodiment 1 gropes the high maltose syrup component ratio of suitable acarbose actinomycete fermentation using orthogonal experiment, obtains
To suitable high maltose syrup trisaccharide and the above ratio of tetrose and tetrose.
The orthogonal experiment optimizes high maltose syrup component ratio in the fermentation medium of acarbose actinomyces
The step of are as follows:
(1) high malt sugar that trisaccharide, tetrose and the above ingredient of tetrose containing different proportion and glucose content are 0 is prepared
Slurry.Trisaccharide ratio is made in 3-21%, tetrose and tetrose ratio in 3- with the syrup of different component ratio and sterling maltose
21% high maltose syrup does the orthogonal reality that trisaccharide in high maltose syrup, tetrose and the above component of tetrose influence fermentation unit
Test (sugared ratio is the chromatographic purity after giving money as a gift, and unit is quality %):
Fermentation unit is detected after carrying out fermented and cultured 168h with above-mentioned syrup:
；It can be seen that the ratio of trisaccharide in high maltose syrup more than 3-9%, tetrose and tetrose in 6-12% according to upper table
It is preferred.
(2) high malt sugar that trisaccharide, tetrose and the above ingredient of tetrose containing different proportion and glucose content are 0 is prepared
Slurry.Trisaccharide ratio is made in 3-9%, tetrose and tetrose ratio 6-12% with the syrup of different component ratio and sterling maltose
High maltose syrup, do the orthogonal experiment that trisaccharide in high maltose syrup, tetrose and the above component of tetrose influence fermentation unit:
；It can be seen that each component optimal proportion is trisaccharide 6-8%, tetrose and tetrose or more in high maltose syrup according to upper table
In 8-10%.
Embodiment 2 improves acarbose fermentation unit using high maltose syrup
Fermentor verifying (seeding tank material liquid volume 1000L, fermentor material liquid volume are carried out with different high wheat syrup
6200L, inventory are that formula equal proportion expands)
3, high maltose syrup forms:
；3 groups of parallel laboratory test results:
Two, high maltose syrup forms:
；3 groups of parallel laboratory test results:
Three, high maltose syrup forms:
；3 groups of parallel laboratory test results:
Four, high maltose syrup forms:
；3 groups of parallel laboratory test results:
Five, high maltose syrup forms:
；3 groups of parallel laboratory test results:
Six, high maltose syrup forms:
；3 groups of parallel laboratory test results:
Claims (4)
1. a kind of method for improving acarbose fermentation unit, which is characterized in that the method is using culture medium to Actinoplanes
Bacterium is cultivated；The culture medium is slant medium, female bottle seed culture medium and fermentation flask culture medium；The fermentation flask culture
Base is to be prepared and formed using high maltose syrup；The high maltose syrup is by maltose, trisaccharide, tetrose and the above ingredient of tetrose
Composition, the trisaccharide Zhan Gaomai syrup give money as a gift after total reducing sugar quality % 3.0-9.0%；The tetrose and tetrose are with the % that improves quality
Ratio is 6-12%；The ratio is the chromatographic purity after giving money as a gift.
2. the method according to claim 1, wherein the slant medium forms: sucrose 30g/L, peptone
5g/L、KC1 0.5g/L、K2HPO41.0g/L, l-tyrosine 1g/L, MgSO40.5g/L, agar 20g/L, solvent are water, just
Beginning pH is 7.0；Mother's bottle seed culture medium composition: starch 10g/L, soybean cake powder 10g/L, CaCO32g/L, glycerol 20g/L,
Solvent is water, and initial pH is 7.0；The fermentation flask culture medium composition: maltose 60g/L, glucose 20g/L, soybean cake powder
15g/L、FeCl3 0.5g/L、CaCl2 2g/L、CaCO34g/L, sodium glutamate 2g/L, K2HPO41g/L, solvent are water, initially
PH is 7.0；
Maltose used in the fermentation flask culture medium is the amount after the malt saccharic composition in high wheat syrup is pure, pure calculating
Method are as follows: high wheat syrup quality × sugar concentration × maltose content=sterling maltose quality.
3. according to the method described in claim 2, it is characterized in that, operation carries out as follows:
(1) female bottle seed culture
By 1 × 1.5cm2Cultured fresh inclined-plane is seeded to female bottle seed culture medium dress with block method is dug by the inoculum concentration of/female bottle
In the 500mL triangular flask that amount is 20%, under the conditions of 27 DEG C of temperature, with the speed oscillation culture 48h of 280r/min, system on shaking table
Obtain female bottle of seed liquor；
(2) shake flask fermentation
Cultured female bottle seed liquor is connected to the 500m1 triangular flask that fermentation flask culture medium loading amount is 10% by 15% inoculum concentration
In, with the speed oscillation culture 168h of 280r/min under the conditions of 27 DEG C of temperature.
4. according to the method described in claim 3, it is characterized in that, the trisaccharide quality % ratio is 6.0-8.0%；Described four
Sugared and tetrose is with % ratio of improving quality for 8-10%；The ratio is the chromatographic purity after giving money as a gift.
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| CN201210503105.XA CN102978261B (en) | 2012-08-24 | 2012-11-30 | A kind of high maltose syrup and the method using its raising acarbose fermentation unit |
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| CN201210503105.XA CN102978261B (en) | 2012-08-24 | 2012-11-30 | A kind of high maltose syrup and the method using its raising acarbose fermentation unit |
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| CN106167814B (en) * | 2016-08-31 | 2019-08-09 | 河北华荣制药有限公司 | A method of improving acarbose fermentation unit |
| CN108192944A (en) * | 2018-01-22 | 2018-06-22 | 协赛(上海)生物科技有限公司 | A kind of production method of microbial biomass |
| CN109355333B (en) * | 2018-12-05 | 2020-10-27 | 山东鲁抗医药股份有限公司 | Preparation method of acarbose |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DE19821038A1 (en) * | 1998-05-11 | 1999-11-18 | Bayer Ag | New acarviosin glycoside, prepared by biotransformation of acarbose, used as saccharase inhibitor for treating diabetes |
| 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 |
| CN102220396A (en) * | 2011-04-27 | 2011-10-19 | 江西农业大学 | Simple fermentation method for acarbose |
-
2012
- 2012-11-30 CN CN201210503105.XA patent/CN102978261B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DE19821038A1 (en) * | 1998-05-11 | 1999-11-18 | Bayer Ag | New acarviosin glycoside, prepared by biotransformation of acarbose, used as saccharase inhibitor for treating diabetes |
| 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 |
| CN102220396A (en) * | 2011-04-27 | 2011-10-19 | 江西农业大学 | Simple fermentation method for acarbose |
Non-Patent Citations (1)
| Title |
|---|
| 阿卡波糖生物合成和发酵工艺研究进展;顾觉奋;《国外医药（抗生素分册）》;20060531;第27卷(第3期);122-125 |
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