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Method for extracting biological products by reverse adsorption

A biological product, adsorption and extraction technology, applied in the field of biochemistry, can solve the problems of low adsorption capacity and inconspicuous chromatographic distribution, and achieve the effect of simplifying the production process

Active Publication Date: 2020-02-07
HEZE RUIZHI TECH DEV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The upper column liquid enters from the bottom of the resin column and discharges from the upper part, which is called reverse upper column. It is generally believed that the reverse upper column has the defects of unobvious chromatographic distribution and low adsorption capacity. Therefore, whether the reverse upper column method is used in production or No reports of current practical applications have been found in scientific research

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] ⑴Preparation of biotransformation liquid: Cultivate C. spinosa in 30L fermentation medium for 18h, add 80g of aseptically treated solid dehydroepiandrosterone (prasterone), ventilation flow rate 1.8Nm³ / h, temperature 29°C, biological After about 30 hours of transformation, TLC detection was carried out. After the transformation was completed, the cultivation was terminated to obtain a biotransformation solution. HPLC detection of biotransformation fluid, dehydroepiandrosterone was not detected, 7-hydroxydhydroepiandrosterone was not detected, and dihydroxyprasterone (7α, 15α-dihydroxyandrostenolone) was 2.21mg / ml; Add 60L of deionized water to the above-mentioned biotransformation liquid for dilution, then carry out the first crushing with a colloid mill, and then carry out the second crushing with a high-pressure homogenizer, wherein the homogenizing pressure is 60MP, so that the dihydroxyprasterone product is from The mycelium is fully released to obtain the biologica...

Embodiment 2

[0036] (1) Preparation of the biotransformation liquid: the same as the preparation method of the biotransformation liquid in Example 1.

[0037] (2) The method for extracting biological products by reverse adsorption, comprising the following steps:

[0038] Step 1): Take 4000ml of the biological product liquid to be tested prepared in Example 2, filter through a 100-mesh filter, and adjust the pH to 7.15 with liquid caustic soda;

[0039] Step 2), use a resin column with a specification of θ22×400, fix a 40-mesh stainless steel filter at the bottom of the resin column, and then load 70ml of non-polar macroporous adsorption resin on the top of the stainless steel filter, and adsorb on the non-polar macroporous Reserve a space of 60mm height on the upper part of the resin, and then fix another 40-mesh stainless steel filter; in this step, use rubber plugs and rubber tubes to connect the above two same resin columns in series, and connect the top of the upper resin column with ...

Embodiment 3

[0044] ⑴Preparation of biotransformation liquid: recombine the 7-β dehydrogenase compound glucose dehydrogenase gene into engineering Escherichia coli, culture, and collect the bacteria by centrifugation; take 100g of the bacteria in parts by mass and suspend them in 400g of deionized water, and ultrasonically Crushed to obtain the whole cell enzyme liquid, added 25g of glucose, 10g of 7-ketolithocholic acid, and catalyzed the reaction for 8 hours to obtain 627ml of ursodeoxycholic acid reaction solution, the content of ursodeoxycholic acid was 15.15mg / ml;

[0045] (2) The method for extracting biological products by reverse adsorption, comprising the following steps:

[0046] Step 1): Take 300ml of ursodeoxycholic acid reaction solution, add 300ml of deionized water, adjust the pH to 9.0 with liquid caustic soda, and filter through an 80-mesh filter;

[0047] Step 2), use a resin column with a specification of θ22×400, fix a 40-mesh stainless steel filter at the bottom of the...

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Abstract

The present invention discloses a method for extracting biological products by reverse adsorption. A macroporous adsorption resin is used, a reverse column adsorption manner is used to directly extract weakly polar or non-polar products from biological product liquid containing cells and mycelial fragments, and besides, the extracted products has high purity. The method overcomes a series of disadvantages caused by use of organic solvent extraction in extraction of traditional biological products and also resolves contradiction of strict requirements for column liquid in traditional resin separation. The method is simple in technical processes, has strong selective adsorbability for the products, enables the products to be good in quality, and produces less production waste; and at the same time, online extraction and continuous production of some biological products are realized.

Description

technical field [0001] The invention relates to the field of biochemical technology, in particular to a method for extracting biological products by reverse adsorption. Background technique [0002] Biotransformation and enzymatic catalysis are two important methods for the production of drugs and drug intermediates. Because the above methods have the characteristics of less pollution and high stereoselectivity, the chemical synthesis methods of many products are being gradually replaced. Although, the technology of immobilized cells and immobilized enzymes in the prior art has been applied in the production of some biological products, and some liquid enzymes are also used in the form of clarified liquids, but based on the instability of biocatalysts, therefore, most Part of the biocatalyst participates in the biological reaction in the form of whole cells or whole bacteria broken liquid, and it becomes very difficult to extract the product from the reaction system after th...

Claims

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Application Information

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IPC IPC(8): B01D15/18B01D15/16B01D15/42
CPCB01D15/1807B01D15/1871B01D15/168B01D15/424
Inventor 彭继先于海勤刘艳平黄明华
Owner HEZE RUIZHI TECH DEV
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