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Preparation methods and application of streptomyces parvulus OUCMDZ-2554 bacterial strain and product actinomycin D thereof

A technology of actinomycin and pH 7.5, applied in the field of preparation and application of Streptomyces parvulus OUCMDZ-2554 strain and its product actinomycin D, to achieve the effects of mass production, high yield and simple separation process

Inactive Publication Date: 2014-03-26
OCEAN UNIV OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Streptomyces parvulus OUCMDZ-2554 is a high-yield single compound actinomycin D. This compound has long synthesis steps, low yield, expensive raw materials, high toxicity, and poor safety. The solvent corrosion used does not meet the requirements of green chemistry (Tanaka T , Nakajima K, Okawa K. Studies on 2-aziridinecarboxylic acid. 1V. Total synthesis of actinomycin D via ring-opening reaction of aziridine. Bull Chem Soc Jpn, 1980, 53(5): 1352-1355)

Method used

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  • Preparation methods and application of streptomyces parvulus OUCMDZ-2554 bacterial strain and product actinomycin D thereof
  • Preparation methods and application of streptomyces parvulus OUCMDZ-2554 bacterial strain and product actinomycin D thereof
  • Preparation methods and application of streptomyces parvulus OUCMDZ-2554 bacterial strain and product actinomycin D thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] [Example 1] Isolation and Identification of OUCMDZ-2554 Strain

[0022] 1. Strain isolation

[0023] Actinomycetes were isolated by the plate dilution method. Take the sea mud (from the Yellow River Delta) out of the refrigerator and place it for a period of time, thaw it, take 1g of the sea mud sample, and dilute it step by step with sterile seawater to 10 -2 , take 0.3mL each and spread evenly on the separation plate (yeast extract 0.2%, 2% agar, pH7.5), apply 3 plates for each dilution, culture upside down at 28°C, pick the colony of the mountain to streak and purify , the purified bacterial strain access slant medium [slant medium composition (%): soluble starch 2%, KNO 3 0.1%, K 2 HPO 4 0.05%, MgSO 4 ·7H 2 O0.05%, NaCl0.05%, FeSO 4 0.001%, pH7.5, agar 2%], stored in a refrigerator at 4°C.

[0024] 2. Identification of OUCMDZ-2554 strain

[0025] The actinomycete strain OUCMDZ-2554 was isolated from sea mud collected from the Yellow River Delta in Dongying,...

Embodiment 2

[0029] [Example 2] Preparation and Application of Actinomycin D

[0030] 1. Preparation of pure actinomycin D and establishment of standard curve

[0031] Take an appropriate amount of S.parvulus OUCMDZ-2554 cultured on a slant for 3 days, inoculate it into a 500mL Erlenmeyer flask containing 150mL of culture solution (yeast extract 0.2%, pH7.5), and place it on a shaker at 28°C and 180 rpm , shaker for 7 days. The fermentation broth was extracted three times with an equal volume of ethyl acetate, and the combined ethyl acetate extracts were concentrated under reduced pressure to obtain 48 mg of crude extract. The obtained extract was separated by LH-20 gel column chromatography (eluted with methanol) to obtain 35 mg of high-purity actinomycin D.

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Abstract

The invention relates to preparation methods and application of streptomyces parvulus OUCMDZ-2554 bacterial strain and a product actinomycin D thereof. The preservation number of the bacterial strain is CGMCC7907. The streptomyces parvulus OUCMDZ-2554 has high yield of single compound actinomycin D, and the yield is 0.25mg / mL. The bacterial strain obtained at home for the firstly time has high yield, the culture medium for the bacterial strain only uses one nutritional ingredient, so that the cost is low, the preparation method is simple and easy to operate, the fermentation period is short, a large quantity of active substances can be produced by fermentation in large scale, the problem of medicine source is fundamentally solved, and the bacterial strain can offer a precursor for structural modification of the compounds and study on structure-activity relationship.

Description

Technical field: [0001] The present invention relates to Streptomyces parvulus OUCMDZ-2554 (derived from the Yellow River Delta) and its method and application for preparing actinomycin D (strain preservation number: CGMCC 7907, preservation date: July 10, 2013, preservation place: No. 3, No. 1 Courtyard, Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences). Background technique: [0002] Cancer has always been a serious threat to human health and life, and the demand for developing highly effective and low-toxic antitumor drugs has become increasingly urgent. The secondary metabolites of microorganisms, especially Streptomyces, have always been an important source of new drug discovery. With the development of a large number of terrestrial microorganisms, in recent years, people have gradually begun to turn to the secondary metabolites of microorganisms in special habitats. [0003] The sources of microorganisms in the Yel...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/06C12N1/20C12P21/02C12R1/465
Inventor 朱伟明王聪王乂刘培培
Owner OCEAN UNIV OF CHINA
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