CN102942638A - Preparation method for dextran with weight average molecular weight of 3000-6000Da - Google Patents
Preparation method for dextran with weight average molecular weight of 3000-6000Da Download PDFInfo
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Abstract
The present invention utilizes dextran enzyme to direct degrade dextran with a weight average molecular weight of 7000-80000Da including micro-molecular dextran (i.e., dextran 10), small molecular dextran (i.e., dextran 20), low molecular dextran (i.e., dextran 40), middle molecular dextran (i.e. dextran70) to special micro-molecular dextran with a weight average molecular weight of 3000-6000Da. The special micro-molecular dextran can be used as a raw material for preparation of special micro-molecular iron dextran. The preparation method is mild and safe in reaction conditions, less in three wastes, easy to control the weight average molecular weight of the final product dextran, narrow in molecular weight distribution, simple in process, rare in dextran loss, and low in production costs. The product after degradation does not need to be processed, and can be directly used for oxidation and complexation to produce high-quality special micro-molecular iron dextran.
Description
Technical field
The present invention relates to a kind of dextran, also claim the preparation method of dextran, specifically weight-average molecular weight is the preparation technology of the special scintilla dextran of 3000-6000Da, and it mainly as the main raw material of mending the chalybeate Iron Dextran, belongs to field of fine chemical.
Background technology
The present invention relates to raw materials for production---the preparation method of special scintilla dextran (weight-average molecular weight is 3000-6000Da) of the special scintilla Iron Dextran of medicine of a kind of prevention, treatment piglet and livestock hypoferric anemia.
Studies show that in a large number, the 50 milligrams of iron of only having an appointment in piglet when birth body need 7 milligrams of iron could satisfy its growth needs every day, and piglet can only obtain 1 milligram of iron every day from the sow breast milk, therefore, after the piglet birth 7 days, if iron fails to be replenished, then can't satisfy its growth needs and produce hypoferric anemia, thereby cause its growth retardation, lose weight, disease resistance is low, and mortality ratio increases etc.Therefore, rear 1-4 days of piglet birth, must replenish enough iron to satisfy its needs in whole lactation to piglet.At present, generally acknowledge that both at home and abroad piglet benefit chalybeate is iron dextran inj preferably, be the iron dextran inj of 150mg/ml by disposable intramuscular injection 1ml iron level in 1-4 days after piglet birth, can satisfy piglet whole lactation to the needs of iron, prevention piglet hypoferric anemia, promote piglet growth, reduce the piglet mortality ratio.But the weight-average molecular weight of the iron dextran inj of at present domestic production is all larger, mostly is 7000-24000Da, this to intramuscular injection after body totally unfavorable to the absorption of medicine, cause the injection site that the necrosis region of black is arranged more and affect the carcase quality of pig.And be 3000-5000Da from the weight-average molecular weight of the iron dextran inj of certain brand of European import, be preferably iron dextran inj of the quality of generally acknowledging both at home and abroad at present.Although it mainly is that (molecular weight is about 5 * 108Da) because domestic dextran manufacturer goes out high molecular dextran with Production by Enzymes that domestic manufacturer can't produce the reason of so special scintilla Iron Dextran, but with acid hydrolyzation high molecular dextran is degraded, obtain again the dextran of different molecular weight with ethanol precipitation, its production cost is high, and arrive scarcely ever special scintilla dextran, thereby special scintilla dextran is provided for downstream manufacturer (being the manufacturer of Iron Dextran).
For solving the preparation problem of special scintilla dextran (weight-average molecular weight is 3000-6000Da), China's CN101205256A patent has proposed to use the membrane sepn classification in the production technique of Dextran 5 (weight-average molecular weight is 4500-5500Da), replaced with alcohol fraction and separated, although in production technique without alcohol, but its dextran molecule amount degraded is still processed with the method for acid heating, the shortcoming of this method is to need after the acid treatment to neutralize with alkali, this process will product waste liquid---sodium chloride solution, increase the processing of " three wastes ", and, the large usage quantity of soda acid, increase production cost, acid hydrolyzation needs to carry out under higher temperature simultaneously, and degradation time is also long, and energy consumption is high, the molecular weight distribution of the dextran product that this method obtains is also wider, obtain the narrower dextran of molecular weight distribution, then the loss of dextran is larger, and output is relatively low.China's CN1041762A patent has proposed with the based on alkaline hydrolysis larger dextran of molecular weight of degrading, and the shortcoming of the method is substantially the same with the acid hydrolyzation of Chinese CN101205256A patent proposition." control of Molecular Weight of Dextran Produced by Enzymatic Synthesis " that the 10th phase of the 24th volume " fine chemistry industry " October in 2007 publishes has proposed to add the high molecular dextran that the dextran enzyme liberating generates in the synthetic dextran process of the dextransucrase that produces with leuconostoc mesentroides, but the degradation time of this method is longer, it is 24 hours, and the weight-average molecular weight of the dextran product that obtains is larger, reaches 1.0 * 105-2.0 * 104.
Therefore, solving the problem that special scintilla dextran raw material difficulty that Iron Dextran manufacturer produces special scintilla Iron Dextran is asked, is to need not to process the production technique that is directly used in the special scintilla Iron Dextran of preparation behind the special scintilla dextran in the urgent need to the dextran of exploring the larger molecular weight of can directly degrading.
Summary of the invention
The object of the invention is to: will be that the dextran (comprise Dextran 10 or Dextran 20 or Dextran 40 or macrodex one or more) of 7000-80000Da is degraded to the special scintilla dextran that weight-average molecular weight is 3000-6000Da from the weight-average molecular weight of buying back on the market with dextranase, and avoid using acid hydrolyzation or based on alkaline hydrolysis to cause and be difficult to control the molecular weight of the finished product and the emission problem of " three wastes ".Foundation of the present invention is based on the narrow spectrum principle of enzyme, and namely dextran is the single-minded substrate of dextranase, and single-minded α-1,6 glycosidic link that interrupts dextran of dextranase is realized.The reaction conditions of this method is both gentle, safety, be easy to again control the molecular weight of product, can be directly that the dextran of larger molecular weight is degraded to molecular weight distribution is narrower, weight-average molecular weight is the special scintilla dextran of 3000-6000Da, and this method or a kind of production technique of cleaning, do not need acid adding or alkali, need not to process sodium-chlor, technique is simple, the dextran loss is few, production cost is low, need not after the degraded to process and to be directly used in oxidation, high-quality special scintilla Iron Dextran is produced in complexing, can solve the raw material problem that the special scintilla dextran difficulty of Iron Dextran manufacturer is asked.
The present invention is achieved in that
Prepare the method for special scintilla dextran (weight-average molecular weight is 3000-6000Da), principle is: be that the dextran (comprise Dextran 10, Dextran 20 or Dextran 40 or macrodex one or more) of 7000-80000Da is degraded to special scintilla dextran (weight-average molecular weight is 3000-6000Da) with dextranase with weight-average molecular weight.
Specific practice is as follows: be the dextran of 7000-80000Da by adding 5-100 grammes per square metre average molecular weight in per 100 gram water, fully stir, regulate pH to 3.5-9.0, then add 200-10000 unit (U) dextranase, keep solution temperature between 35-65 ℃, preferably 45-55 ℃, reacted 5-40 minute, be preferably 15-25 minute, then directly be warmed up to 70-100 ℃, stirred 5-20 minute, and with the dextran enzyme-deactivating, namely obtained special scintilla dextran solution.
Obtain need not to process after the degraded of special scintilla dextran solution can be directly used in oxidation, high-quality special scintilla Iron Dextran is produced in complexing.
The invention has the advantages that:
Be that the dextran of 7000-80000Da is (such as Dextran 10 with dextran enzyme liberating weight-average molecular weight, 20,40,70) be special scintilla dextran (weight-average molecular weight is 3000-6000Da), the reaction conditions of this method is both gentle, safety, be easy to again control the molecular weight of product, can be directly that the dextran of larger molecular weight is degraded to molecular weight distribution is narrower, weight-average molecular weight is the special scintilla dextran of 3000-6000Da, and this method or a kind of production technique of cleaning, do not need acid adding or alkali, need not to process sodium-chlor, technique is simple, the dextran loss is few, production cost is low, need not after the degraded to process and to be directly used in oxidation, high-quality special scintilla Iron Dextran is produced in complexing, can solve the raw material problem that the special scintilla dextran difficulty of Iron Dextran manufacturer is asked, but suitability for industrialized production.
Embodiment
Further specify the present invention below by embodiment, but content not thereby limiting the invention.
Example 1:
Adding the 18g weight-average molecular weight in the 500ml there-necked flask of agitator and heating in water bath is dextran and the 360ml distilled water of 7000Da, start agitator, be warmed up to 35 ℃, pH value of solution is transferred to 3.5, add 720 unit dextranases, isothermal reaction 5 minutes, then directly be warmed up to 80 ℃, stirred 10 minutes, with the dextran enzyme-deactivating, namely obtain special scintilla dextran solution, high pressure liquid chromatography is shown as single narrow peak, weight-average molecular weight is 3553Da, and Breadth parameter of molecular weight distribution is 1.23.
The special scintilla dextran solution of aforesaid method preparation need not to process to be directly used in produces special scintilla iron dextran inj.
Example 2:
In with the 500ml there-necked flask of agitator and heating in water bath, add 18g Dextran 40 (weight-average molecular weight is 40000Da) and 55ml distilled water, start agitator, be warmed up to 50 ℃, pH value of solution is 5.0, add 3250 unit dextranases, isothermal reaction 15 minutes, then directly be warmed up to 90 ℃, stirred 10 minutes, with the dextran enzyme-deactivating, namely obtain special scintilla dextran solution, high pressure liquid chromatography is shown as single narrow peak, weight-average molecular weight is 4560Da, and Breadth parameter of molecular weight distribution is 1.21.
The special scintilla dextran solution of aforesaid method preparation need not to process to be directly used in produces special scintilla iron dextran inj.
Example 3:
In with the 500ml there-necked flask of agitator and heating in water bath, add 50g macrodex (weight-average molecular weight is 76000Da) and 50ml distilled water, start agitator, be warmed up to 65 ℃, pH value of solution is transferred to 9.0, add 5000 unit dextranases, isothermal reaction 40 minutes, then directly be warmed up to 100 ℃, stirred 10 minutes, with the dextran enzyme-deactivating, namely obtain special scintilla dextran solution, high pressure liquid chromatography is shown as single narrow peak, weight-average molecular weight is 5020Da, and Breadth parameter of molecular weight distribution is 1.25.
The special scintilla dextran solution of aforesaid method preparation need not to process to be directly used in produces special scintilla iron dextran inj.
Example 4:
In with the 500ml there-necked flask of agitator and heating in water bath, add 18g Dextran 20 (weight-average molecular weight is 18000Da) and 360ml distilled water, start agitator, be warmed up to 45 ℃, pH value of solution is transferred to 3.5, add 720 unit dextranases, isothermal reaction 25 minutes, then directly be warmed up to 70 ℃, stirred 10 minutes, with the dextran enzyme-deactivating, namely obtain special scintilla dextran solution, high pressure liquid chromatography is shown as single narrow peak, weight-average molecular weight is 3860Da, and Breadth parameter of molecular weight distribution is 1.20.
The special scintilla dextran solution of aforesaid method preparation need not to process to be directly used in produces special scintilla iron dextran inj.
Example 5:
In with the 500ml there-necked flask of agitator and heating in water bath, add 50g Dextran 10 (weight-average molecular weight is 10000Da) and 50ml distilled water, start agitator, be warmed up to 55 ℃, pH value of solution is transferred to 6.0, add 5000 unit dextranases, isothermal reaction 40 minutes, then directly be warmed up to 80 ℃, stirred 10 minutes, with the dextran enzyme-deactivating, namely obtain special scintilla dextran solution, high pressure liquid chromatography is shown as single narrow peak, weight-average molecular weight is 4201Da, and Breadth parameter of molecular weight distribution is 1.18.
The special scintilla dextran solution of aforesaid method preparation need not to process to be directly used in produces special scintilla iron dextran inj.
Example 6:
Adding the 18g weight-average molecular weight in the 500ml there-necked flask of agitator and heating in water bath is dextran and the 55ml distilled water of 80000Da, start agitator, be warmed up to 45 ℃, pH value of solution is transferred to 5.5, add 720 unit dextranases, isothermal reaction 20 minutes, then directly be warmed up to 80 ℃, stirred 10 minutes, with the dextran enzyme-deactivating, namely obtain special scintilla dextran solution, high pressure liquid chromatography is shown as single narrow peak, weight-average molecular weight is 4565Da, and Breadth parameter of molecular weight distribution is 1.24.
The special scintilla dextran solution of aforesaid method preparation need not to process to be directly used in produces special scintilla iron dextran inj.
The special scintilla dextran solution of aforesaid method preparation is directly used in the special scintilla iron dextran inj of production, concrete steps are seen CN102429865A: special scintilla dextran is after peroxidation, complexing, desalination, concentrated preparation, filtration and embedding sterilization, can obtain the iron dextran inj product, test-results is as follows:
Claims (4)
1. the preparation method of a special scintilla dextran is characterized in that: adopting dextranase is that the dextran of 7000-80000Da directly is degraded to special scintilla dextran with weight-average molecular weight, and its preparation method may further comprise the steps:
Be that the dextran of 7000-80000Da is added to the water with weight-average molecular weight, stir, the pH to 3.5-9.0 of regulator solution, under 35-65 ℃ of temperature, add dextranase, isothermal reaction 5-40 minute, be warmed up to again 70-100 ℃, stirring namely obtains special scintilla dextran solution with the dextran enzyme-deactivating;
Described special scintilla dextran refers to that weight-average molecular weight is the dextran of 3000-6000Da;
Described weight-average molecular weight is the dextran of 7000-80000Da: water: the ratio of dextranase is 5-100g: 100g: 200-10000U.
2. the preparation method of a kind of special scintilla dextran as claimed in claim 1, it is characterized in that: described weight-average molecular weight is the dextran of 7000-80000Da: comprise in the macrodex that weight-average molecular weight is the Dextran 10 of 8000-12000Da, Dextran 20 that weight-average molecular weight is 16000-24000Da, weight-average molecular weight is 32000-42000Da Dextran 40 or weight-average molecular weight be 64000-76000Da one or more.
3. the preparation method of a kind of special scintilla dextran as claimed in claim 1, it is characterized in that: the pH regulator of solution is to 3.5-6.0, and under 45-55 ℃ of temperature, the isothermal reaction time is 15-25 minute.
4. the preparation method of a kind of special scintilla dextran as claimed in claim 1 is characterized in that: need not to process after the special scintilla dextran degraded that makes, its solution can be directly used in oxidation, special scintilla Iron Dextran is produced in complexing.
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103993052A (en) * | 2014-04-02 | 2014-08-20 | 广州甘蔗糖业研究所 | Method for fermentation production of micro molecular weight dextran |
| CN106543294A (en) * | 2015-09-18 | 2017-03-29 | 瑞普（天津）生物药业有限公司 | A kind of preparation method of iron-dextrin |
| CN107988284A (en) * | 2017-12-07 | 2018-05-04 | 中国科学院过程工程研究所 | A kind of method for producing Dextran 10 and products thereof and purposes |
| CN110183548A (en) * | 2019-06-28 | 2019-08-30 | 瑞普（天津）生物药业有限公司 | A kind of preparation method and applications of low molecular weight dextran iron |
| CN110218267A (en) * | 2019-07-04 | 2019-09-10 | 汉邦环宇多糖生物科技（河源）有限公司 | A kind of Preparation method and use of iron-dextrin |
| CN114990173A (en) * | 2021-03-01 | 2022-09-02 | 广州华汇生物实业有限公司 | Preparation method of alpha-glucan |
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| CN102690857A (en) * | 2012-05-28 | 2012-09-26 | 广西民族大学 | Method for preparing medium-low molecular weight dextran by using biological method |
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| 梁达奉: "《中国博士学位论文全文数据库》", 15 June 2012 * |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103993052A (en) * | 2014-04-02 | 2014-08-20 | 广州甘蔗糖业研究所 | Method for fermentation production of micro molecular weight dextran |
| CN103993052B (en) * | 2014-04-02 | 2016-07-13 | 广州甘蔗糖业研究所 | A kind of method of fermenting and producing scintilla amount dextran |
| CN106543294A (en) * | 2015-09-18 | 2017-03-29 | 瑞普（天津）生物药业有限公司 | A kind of preparation method of iron-dextrin |
| CN107988284A (en) * | 2017-12-07 | 2018-05-04 | 中国科学院过程工程研究所 | A kind of method for producing Dextran 10 and products thereof and purposes |
| CN110183548A (en) * | 2019-06-28 | 2019-08-30 | 瑞普（天津）生物药业有限公司 | A kind of preparation method and applications of low molecular weight dextran iron |
| CN110218267A (en) * | 2019-07-04 | 2019-09-10 | 汉邦环宇多糖生物科技（河源）有限公司 | A kind of Preparation method and use of iron-dextrin |
| CN114990173A (en) * | 2021-03-01 | 2022-09-02 | 广州华汇生物实业有限公司 | Preparation method of alpha-glucan |
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