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Streptomyces strain and application thereof in production of staurosporine

A technology of staurosporine and streptomyces, applied in the field of microbial engineering, can solve the problems of low fermentation unit level and unsuitability for industrialization, and achieve the effect of industrialized production and high production capacity

Active Publication Date: 2018-10-19
ZHEJIANG HISUN PHARMA CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In summary, although there are many staurosporine-producing bacteria in published patents and literatures, the strains are relatively primitive and the level of fermentation units is low, which is not suitable for industrialization

Method used

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  • Streptomyces strain and application thereof in production of staurosporine

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Experimental program
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Effect test

Embodiment 1

[0034] Embodiment 1: strain source

[0035]The Streptomyces HS-HY-153 (CGMCC NO. 14806) of the present invention is an original strain isolated from a soil sample in Tiantai Mountain, Zhejiang, and then obtained through mutagenesis.

[0036] After the original strain was cultured in ISP2 slant medium at 28°C for 7-10 days, the mycelium was scraped off with an inoculation spatula under aseptic conditions to obtain a bacterial suspension for NTG (nitrosoguanidine) mutagenesis treatment.

[0037] Weigh 2 mg of NTG crystals and dissolve in 2 mL of sterile Tris buffer (pH 6.0). Use a pipette to draw 1mL NTG solution and add it to 1mL bacterial suspension, and shake it on a rotary or reciprocating shaker at 28°C for 1 hour. The remaining specific steps are as follows:

[0038] (1) Preparation and cultivation of mycelia

[0039] Solid medium formula: ISP2, sterilized at 121°C for 20 minutes, cooled to 50-60°C and poured onto the plate, properly diluted the mutagenized bacterial su...

Embodiment 2

[0045] Example 2: Morphology, cultural characteristics, physiological and biochemical characteristics of Streptomyces HS-HY-153 (CGMCC NO.14806).

[0046] The experiments were carried out with reference to the relevant content in the "Streptomyces Identification Manual", "Classification and Identification of Actinomycetes", "Common Bacterial System Identification Manual" and other books: the color judgment was compared with the color in the RAL K7 color card.

[0047] 1. Morphological characteristics of the strain: Inoculate the strain HS-HY-153 (CGMCC NO.14806) in the ISP2 medium for insert culture, and after culturing at 28°C for 3-5 days, take the cover glass on the slide and place it on the optical Observed under a microscope at 400× magnification, the results are shown in figure 1 .

[0048] 2. The cultural characteristics of the strain: After the strain HS-HY-153 (CGMCC NO.14806) was cultured on the ISP2 medium at 28°C for 7-10 days, the colony was round in shape, with ...

Embodiment 3

[0073] Embodiment 3: strain identification

[0074] 1. 16S rDNA sequence analysis of Streptomyces HS-HY-153 (CGMCC NO.14806)

[0075] The experiments were carried out referring to the relevant content in the book "Molecular Cloning Experiment Guide". The mycelium was collected, and then the total DNA was extracted with an Actinomycetes DNA extraction kit. The 16S rDNA sequence was amplified using the universal primer 27F (SEQ ID NO: 1) / 1495R (SEQ ID NO: 2). The amplification system and PCR reaction program are shown in Table 8. PCR products were detected by 0.8% agarose gel electrophoresis , The PCR product was purified and recovered using the SanPrep column PCR purification product kit, and the purified PCR product was directly sent to Nanjing GenScript Biotechnology Co., Ltd. for sequence determination.

[0076] Table 8 PCR amplification system and reaction program

[0077]

[0078] After the 16S rDNA sequence of strain HS-HY-153 (CGMCC NO.14806) was checked, it was co...

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Abstract

The invention discloses a novel Streptomyces strain and application thereof. The strain is named as streptomyce sp. HS-HY-153, and the preservation number thereof is CGMCC NO. 14806; the invention also discloses a method for preparing Staurosporine by culturing the strain HS-HY-153.

Description

technical field [0001] The invention belongs to the technical field of microbial engineering, and in particular relates to a Streptomyces strain and its application in preparing staurosporine. Background technique [0002] Staurosporine (staurosporine, structural formula shown in formula 1), originally found in the fermentation product of Streptomycessaurosporeus (AM-2282) strain, has antibacterial, antifungal, hypotensive, platelet aggregation, antitumor and neuroprotective properties. This biological activity, especially as an effective inhibitor of protein kinase C, is of great research and development value in the treatment of cancer. On April 28, 2017, the FDA approved its semi-synthetic drug Midostaurin (Midostaurin, trade name Rydapt, structural formula shown in Formula 2) in combination with chemotherapy for newly diagnosed acute myeloid leukemia (acutemyeloid leukemia, AML ) for the treatment of adult patients, for patients with specific gene mutation FLT3. At the...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12P17/18C12R1/46
CPCC12N1/20C12P17/188C12N1/205C12R2001/46
Inventor 方佳双张辉项仁鑫邓爱文王微芬姜南应灵萍王继栋滕云
Owner ZHEJIANG HISUN PHARMA CO LTD
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