Method for preparing recombined human atrial natriuretic peptide rhANP by using ferment in high density

A high-density fermentation, human-friendly technology, applied in fermentation and other directions, can solve the problems of limiting the industrial application of ANP, difficulty in ANP expression and purification, etc.

Active Publication Date: 2005-10-26
SHENZHEN STATE BIOCHEM ENG TECHDEV CENT
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  • Abstract
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Problems solved by technology

However, due to the difficulty in the expression and purifi

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  • Method for preparing recombined human atrial natriuretic peptide rhANP by using ferment in high density

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Embodiment Construction

[0033] The present invention is further described below in conjunction with embodiment.

[0034] The preparation method of the present embodiment comprises the following process steps:

[0035] A. The slant strains are expanded and cultivated to make seed liquid

[0036] The glycerol-producing strains preserved at -70°C were inoculated in a test tube at a ratio of 1% (3ml LB / Amp+medium), and slant strains were made under the culture conditions of 30°C, 300rpm, and a culture cycle of 15 hours; Inoculate in a small conical flask (20ml LB / Amp+medium) at a ratio of 1%, culture at 30°C and 300rpm for 8 hours; then inoculate a large conical flask at a ratio of 1% (1500ml 2-YT / Amp+medium) , 30° C., 300 rpm for 15 hours.

[0037] B. Fermentation

[0038] The above seed solution was inoculated into 30L fermentation medium at a ratio of 3%. The composition of the fermentation medium is:

[0039] Glycerin 5g / L, tryptone 7g / L, yeast powder 3g / L, KH 2 PO 4 7g / L,

[0040] K 2 HPO...

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Abstract

This invention relates to microorganism fermentation domain, is a method of preparing rhANP by the way of high density fermentation. This invention uses PCW112-ANP/DH5 alpha , through expand seed culture medium solution talus bacterium, high density fermentation, separation and purity e.g. craft procedure, we prepares the rhANP. This invention dissolves the problem of ANP express and difficulty of purity which has imprisoned the industrial application of it. Its production condition is easy to control and the material of bacterium is easy to obtain, with good repeat activity, low cost, and product can be applied to biology pharmacy.

Description

technical field [0001] The invention belongs to the technical field of biological fermentation, in particular to a method for preparing recombinant human natriuretic natriuretic peptide (rhANP) by high-density fermentation. Background technique [0002] Human natriuretic peptide (ANP) is a polypeptide hormone produced and secreted by atrial myocytes, and has three forms: α, β, and γ. α-ANP has the strongest biological activity and is composed of 28 amino acids. Its structure is as follows: [0003] [0004] The biological effects of ANP mainly include four aspects: excretion of sodium and diuresis; expansion of blood vessels; lowering of blood pressure; anti-arrhythmia. Clinically, ANP is currently mainly used for the treatment of acute heart failure. [0005] In 1986, my country officially listed the recombinant human natriuretic peptide project as a national key research project. However, due to the difficulties in the expression and purification of ANP, the industri...

Claims

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Application Information

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IPC IPC(8): C12P21/02
Inventor 林枫于志江王玉树周兆平章刚陈旭梁四五姚小建骆晓栋欧阳藩
Owner SHENZHEN STATE BIOCHEM ENG TECHDEV CENT
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