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CRM197 strain culture medium, preparing method and fermentation culture method

A technology of CRM197 and strain culture medium, which is applied in the fields of biopharmaceuticals and microbial cultivation, to achieve the effects of reducing production costs, ensuring uniformity, and simple and convenient operation

Active Publication Date: 2019-11-15
ZHEJIANG VACIN BIO PHARMA LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] CRM197 is an avirulent form of diphtheria toxin, but it is immunologically indistinguishable from diphtheria toxin

Method used

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  • CRM197 strain culture medium, preparing method and fermentation culture method
  • CRM197 strain culture medium, preparing method and fermentation culture method
  • CRM197 strain culture medium, preparing method and fermentation culture method

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Embodiment Construction

[0058] The present invention will be described in further detail below in conjunction with the accompanying drawings.

[0059] 1. Raw material preparation

[0060] 1.1. CRM197 strain: The CRM197 strain in this application comes from the American Type Culture Collection (ATCC), and the strain number is ATCC39255. In addition, the CRM197 strain of the present application can also be selected from strain numbers ATCC39526, ATCC11049, ATCC51926, ATCC51280 and other strains.

[0061] Before using the above strains, they were resuscitated and activated with 1 mL of CRM197 liquid medium.

[0062] 1.2. Formula and preparation method of basic nutrient solution:

[0063] Table 1 Formula table of basic nutrient solution

[0064] Reagent Material name factory Formula g / L 1 arginine Shanghai Concord Amino Acid Co., Ltd. 2 2 Asparagine Shanghai Lingfeng Chemical Reagent Co., Ltd. 2 3 aspartic acid Hubei Bafeng Pharmaceutical Co., Ltd. 0.5 4 ...

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Abstract

The invention relates to the technical field of bio-pharmaceuticals and microculture, in particular to a CRM197 strain culture medium, a preparing method and a fermentation culture method. The CRM197strain culture medium is a liquid culture medium. Each L of culture medium comprises 250 mL of a basic nutrient solution, 7.2 mL of acetic acid, 30 g of maltulose monohydrate, 2 mL of sodium lactate,0.5 g of tyrosine, 1.96 g of monopotassium phosphate, 5.1 g of dipotassium phosphate, 6.13 mL of a growth factor a, 8 mL of a growth factor b, 8 mL of a growth factor c and 1.3 g of calcium chloride.All the components can be rapidly dissolved when the culture medium is prepared, and the uniformity of the culture medium is ensured. By means of the step-by-step amplification fermentation culture amplification mode, the CRM197 strain fermentation culture method has the advantages of being rapid, convenient to implement, low in cost and high in efficiency, the large-scale amplification of a CRM197 strain is facilitated, and therefore a high-expression-amount fermentation bacteria liquid is obtained.

Description

technical field [0001] The invention relates to the technical fields of biopharmaceuticals and microorganism culture, in particular to a CRM197 strain culture medium, a preparation method and a fermentation culture method. Background technique [0002] Diphtheria toxin (DT) is a protein toxin synthesized and secreted by toxin-producing strains of Corynebacterium diphtheriae, which is produced in the form of a single polypeptide, and the polypeptide is easily spliced ​​and cleaved at residues 190, 192 or 193 , resulting in the formation of two subunits A fragment and B fragment linked by a disulfide bond (Moskaug et al. Biol. Chem. 264: 15709-15713, 1989). Fragment A is the catalytically active part, which is an NAD-dependent ADP-ribosyltransferase, which specifically targets the protein synthesis factor EF-2, thereby inactivating EF-2, and the protein synthesis in the cell stops. [0003] CRM197 is an avirulent form of diphtheria toxin, but it is immunologically indistingui...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12R1/16
CPCC12N1/20
Inventor 马伟郭阳漾王丹雅
Owner ZHEJIANG VACIN BIO PHARMA LTD
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