A kind of method for preparing acarbose

A technology for acarbose and starch saccharification liquid, applied in the field of medicine, can solve problems such as high cost and poor effect, and achieve the effects of low production cost, high fermentation level, and reduced extraction and purification pressure.

Active Publication Date: 2015-11-25
LIVZON NEW NORTH RIVER PHARMA
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  • Description
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Problems solved by technology

[0004] For industrial production, cost is the priority. Maltose is the most nutrient component in the composition of acarbose fermentation medium. Direct purchase of maltose and high maltose syrup is relatively expensive.
At the same time, we have also conducted investigations. The maltose and high maltose syrup we purchased were directly used for the fermentation of actinomycetes. The concentration of acarbose in the product is not high, especially the effect of pure maltose is even worse.
The fermentation of acarbose requires compound maltose, mainly maltobiose and maltotriose, supplemented by a small amount of maltotetraose and glucose. The higher the content of compound maltose, the higher the fermentation level. The impurities in the fermentation broth, especially The content of the key impurity A is also easy to control at a low level (there is currently no good extraction method for impurity A, which is mainly controlled by fermentation)

Method used

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  • A kind of method for preparing acarbose

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Weigh 7.5kg of cornstarch (sugar content 82%), add tap water to 37.5L (concentration: 20%), adjust the pH to 5.6 after dissolving, add 3.75g of high temperature resistant a-amylase (0.05%, calculated as cornstarch) ), 3.75g of anhydrous calcium chloride (0.05%), heated to 105°C to start timing, maintained for 10 minutes, took a sample to measure DE=17, cooled to 60°C, added 0.06% of starch mass of pullulanase, 0.06% Fungal enzyme, 0.25% β-amylase, maintained at 59°C for 8 hours, sampling and testing, the total amount of compound maltose was 5.3kg, and the conversion rate was 86.2% relative to the total sugar content of corn starch. Preparation of fermentation medium: take 29L starch saccharification liquid (equivalent to 4.1kg maltose, ratio 6.8%), other nutrients include 2.6% glucose, 1.0% soybean cake powder, 0.5% monosodium glutamate, 0.7% glycerin, 0.3% C a CO 3 and 0.03%F e Cl 3 , put it into a 100L fermenter, prepare 60L of fermentation medium with pH 6.5, inse...

Embodiment 2

[0020] Weigh 12kg of potato starch (sugar content 70%), add tap water to 40L (concentration is 30%), adjust the pH to 6.2 after dissolving, add 0.2% (calculated as corn starch) high temperature resistant a-amylase, 0.02% anhydrous Calcium chloride, start timing when heated to 110°C, maintain for 5 minutes, take a sample to measure DE=11, cool down to 60°C, add 0.2% pullulanase, 0.2% fungal enzyme, 0.45% β-amylase by starch quality, 55°C After maintaining for 4 hours, sampling and testing showed that the total amount of maltose in the starch saccharification solution was 7.98kg, the conversion rate was 95.0%, and the amount of glucose in the saccharification solution was 97g. Preparation of fermentation medium: Take 20L starch saccharification liquid (equivalent to 6.4kg maltose, ratio 10.6%), other nutrients include 2.4% glucose, 1.0% soybean cake powder, 0.5% monosodium glutamate, 0.6% glycerin, 0.5% C a CO 3 and 0.05%F e Cl 3 , put it into a 100L fermenter, prepare 60L of...

Embodiment 3

[0022] Weigh 5500kg cornstarch (85% sugar content) and add tap water to 13.75m 3 (Concentration is 40%), adjust the pH to 6.0 after dissolving, add 0.1% (calculated by corn starch) high temperature resistant a-amylase 5.5kg, 0.08% anhydrous calcium chloride 4.4kg, heat to 105°C to start timing, maintain For 5 minutes, take a sample to measure DE=13, cool down to 60°C, add 0.1% pullulanase, 0.1% fungal enzyme, and 0.3% β-amylase by starch quality, maintain at 58°C for 4 hours, take a sample for detection, maltose in the starch saccharification solution The total amount is 4146.8kg, relative to the total sugar content of cornstarch, the conversion rate is 88.7%, and the glucose content in the saccharification solution is 62kg. Preparation of fermentation medium: take 8m 3 Starch saccharification liquid (equivalent to 3402.5kg maltose, ratio 8.5%), other nutrients include 1.8% glucose, 1.0% soybean cake powder, 0.5% monosodium glutamate, 0.6% glycerin, 0.5% C a CO 3 and 0.05%F...

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Abstract

The invention discloses a method for preparing acarbose, which adopts four enzymes to cooperatively carry out starch liquefaction and saccharification, and comprises the following steps that: (1) starch emulsion is liquefied with alpha-amylase at appropriate pH and temperature, and the DE (Dextrose Equivalent) value is controlled between 11 and 18; (2) the starch liquefied liquid is cooled to an appropriate temperature, the appropriate pH is adjusted, pullulanase, fungal enzyme and beta-amylase are added, certain time is maintained, and the composite maltose conversion rate can reach 85-95%; and (3) the liquefied starch is used for acarbose fermentation after being sterilized. According to the starch liquefaction method provided by the invention, the compound maltose content is high, and the high yield and the low impurities of acarbose fermentation are ensured. The method is simple, is easy to operate and is low in cost, and is suitable for the industrial production of acarbose fermentation.

Description

technical field [0001] The invention belongs to the technical field of medicine, and in particular relates to a method for preparing acarbose. technical background [0002] Acarbose is a potent glucosidase inhibitor used as an oral antidiabetic drug for the treatment of diabetes by motile actinomycetes ( Actinoplane sutahensis ) obtained by fermentation. The molecular structure of acarbose is as follows, which is composed of 4 monosaccharides. Its biosynthesis is mainly sugar metabolism, and the composition of carbon source (sugar source) in the fermentation medium is very critical. The biggest feature of acarbose fermentation is that the productivity of the producing bacteria significantly depends on the osmotic pressure concentration of the culture solution (the osmotic control fermentation method for preparing acarbose, patent publication number: CN1178837, Bayer Company), the osmotic pressure Control is mainly achieved by maintaining a high concentration of substrates ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P19/26C12R1/045
Inventor 李小兵唐阳刚李敬辉魏青时伟陆敏姚清国
Owner LIVZON NEW NORTH RIVER PHARMA
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