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High density fermentation culture medium of restructured human interferon alpha2b

A recombinant human interferon, high-density fermentation technology, applied in the field of biomedicine, to achieve high-density growth and reduce the effect of inhibition

Active Publication Date: 2011-08-31
BEIJING KAWIN TECH SHARE HLDG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, most of the high-density fermentation media for the production of recombinant human interferon α2b use glucose as a carbon source, which can easily produce acetic acid during the cultivation process, while the high-density fermentation media for the production of recombinant human interferon α2b use composite carbon sources (glycerol+ Glucose) by adding some amino acids (glycine, methionine) and some appropriate concentrations of trace elements (VB1, MgSO4.7H2 sub>O, FeSO4.7H2O, MnSO4 CaCl2) have not yet been used as acetic acid inhibitors see the report

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Example 1 High-density fermentation of recombinant human interferon α2b

[0021] Formula 1: Peptone 6g / L, yeast extract 2g / L, glucose 0.02%, glycerol 0.6%, Na 2 HPO 4 11g / L, NaH 2 PO 4 1g / L, KH 2 PO 4 4g / L, NaCl 2g / L, FeSO 4 .7H 2 O 0.01g / L, MgSO 4 .7H 2 O 0.02g / L, CaCl 2 0.001g / L, VB 1 0.01g / L, MnSO 4 0.01g / L, glycine 0.2g / L, and methionine 0.01g / L.

[0022] The method for preparing a high-density fermentation medium of recombinant human interferon α2b of the present invention includes the following steps:

[0023] (1) Weigh 100L of components according to the above formula;

[0024] (2) Dissolve in a specified amount of purified aqueous solution;

[0025] (3) Adjust the PH value to 7.0 with sodium hydroxide solution;

[0026] (4) Sterilize in a fermentation tank of corresponding volume (121°C, 30 minutes).

[0027] (5) After accessing suitable recombinant E. coli for cultivation, 1762 g of bacteria containing recombinant human interferon α2b was obtained, and the expression l...

Embodiment 2

[0028] Example 2: High-density fermentation of recombinant human interferon α2b

[0029] Formula 2: Peptone 10g / L, yeast extract 5g / L, glucose 0.5%, glycerol 1%, Na 2 HPO 4 15g / L, NaH 2 PO 4 4g / L, KH 2 PO 4 6g / L, NaCl 3g / L, FeSO 4 .7H 2 O 0.5g / L, MgSO 4 .7H 2 O 0.5g / L, CaCl 2 0.02g / L, VB 1 0.03g / L, MnSO 4 0.05g / L, glycine 0.5g / L, methionine 0.06g / L.

[0030] The method for preparing a high-density fermentation medium of recombinant human interferon α2b of the present invention includes the following steps:

[0031] (1) Weigh 100L of components according to the above formula;

[0032] (2) Dissolve in a specified amount of purified aqueous solution;

[0033] (3) Adjust the PH value to 7.0 with sodium hydroxide solution;

[0034] (4) Sterilize in a fermentation tank of corresponding volume (121°C, 30 minutes).

[0035] (5) After accessing a suitable recombinant E. coli for cultivation, 1863g of bacteria containing recombinant human interferon α2b is obtained, and the expression level o...

Embodiment 3

[0036] Example 3: High-density fermentation of recombinant human interferon α2b

[0037] Formula 3: Peptone 18g / L, yeast extract 8g / L, glucose 2%, glycerol 4%, Na 2 HPO 4 27g / L, NaH 2 PO 4 7.5g / L, KH 2 PO 4 6.5g / L, NaC16.5g / L, FeSO 4 .7H 2 O 0.08g / L, MgSO 4 .7H 2 O 1.2g / L, CaCl 2 0.01g / L, VB 1 0.1g / L, MnSO 4 0.1g g / L, glycine 1.2g / L, and methionine 0.1g g / L.

[0038] The method for preparing a high-density fermentation medium of recombinant human interferon α2b of the present invention includes the following steps:

[0039] (1) Weigh 100L of components according to the above formula;

[0040] (2) Dissolve in a specified amount of purified aqueous solution;

[0041] (3) Adjust the PH value to 7.0 with sodium hydroxide solution;

[0042] (4) Sterilize in a fermentation tank of corresponding volume (121°C, 30 minutes).

[0043] (5) After accessing suitable recombinant E. coli for cultivation, 1547 g of bacteria containing recombinant human interferon α2b was obtained, and the expressi...

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Abstract

The invention provides a high density fermentation culture medium of restructured human interferon alpha2b, the culture medium can effectively reduce the generation amount of acetic acid, reduce the inhibiting effect of acetic acid, and improve growing speed rate of restructured fungus and the generation rate of restructured protein.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a high-density fermentation medium of recombinant human interferon α2b. Background technique [0002] Escherichia coli is currently the most widely used engineering bacteria in industrial production. The basic process of using Escherichia coli to produce protein is to first insert the target gene into a suitable plasmid, then transform the plasmid into Escherichia coli, cultivate the Escherichia coli, and extract the target protein from the Escherichia coli. Among them, the cultivation of Escherichia coli and the high-density fermentation of engineering bacteria are very important, which directly determines the amount of target protein obtained. High cell-density culture technology (High cell-density culture, HCDC), that is, high-density fermentation technology, increases the fermentation density of the bacteria, and finally increases the specific productivity of the product (the out...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/21C12P21/02C07K14/435C12R1/19
Inventor 张春丽周德胜吉春
Owner BEIJING KAWIN TECH SHARE HLDG
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