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Fermentation preparation process of Streptomyces marinensis and secondary metabolite Streptochlorin thereof

A secondary metabolite, marine Streptomyces technology, applied in fermentation, microbial-based methods, biochemical equipment and methods, etc., can solve problems such as low yield, low Streptochlorin output, unfavorable industrial development, etc., to achieve fast flow rate, The effect of large injection volume and high purity

Inactive Publication Date: 2016-06-08
NINGBO UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] There are few reports on the fermentation of Streptochlorin in literature at home and abroad. In the experiment, we used the general fermentation conditions and preparation conditions to find that the output of Streptochlorin was low and the yield was low. This became a limiting factor for further research on this compound and was not conducive to later industrial development.

Method used

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  • Fermentation preparation process of Streptomyces marinensis and secondary metabolite Streptochlorin thereof
  • Fermentation preparation process of Streptomyces marinensis and secondary metabolite Streptochlorin thereof
  • Fermentation preparation process of Streptomyces marinensis and secondary metabolite Streptochlorin thereof

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Experimental program
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Effect test

specific Embodiment 1

[0033] A marine streptomyces, the bacteria is SYYLWHS-1-4 strain, named as Streptomyces sp., and the preservation number is CGMCC No.11468, which was preserved in China Microorganism Culture Collection Management Committee on September 30, 2015. Microbiology Center. The colony was pale yellow and translucent at the initial stage of plate culture, and the colony was round with a smooth and moist surface. As the culture time prolongs, the aerial hyphae developed and covered the entire surface of the colony, showing white. The concrete steps of this marine Streptomyces fermentation production secondary metabolite Streptochlorin are as follows:

[0034] (1) Solid culture

[0035] The marine streptomyces with the preservation number CGMCCNo.11468 stored in a -80°C refrigerator was coated on a solid plate medium, and cultured in a 27.5°C incubator for 48 hours to obtain plate strains; the formula of the solid plate medium was as follows: Pure water 1L, yeast extract 1.889g, solubl...

specific Embodiment 2

[0055] 1. Determination of significant influencing factors of culture conditions

[0056] Yeast powder for medium (X 1 ), soluble starch (X 2 ), MgSO 4 (X 3 ), K 2 HPO 4 (X 4 ), CaCl 2 (X 5 ), initial pH (X 6 ), mediumvolume(X 7 ), temperature (X 8 ), marinum salt (X 9 ) nine factors for variable analysis. Each variable is set to a high (+1) and a low (-1) level (see Table 1 for specific values). Use Minitab software to generate 12 factors and 20 Plackett-Burman experiments (where X 10 、X 11 and x 12 is a dummy variable), and Streptochlorin output (mg / L) was used as the response value (Y), the effect value of each factor was analyzed by Mintab software, and the factor with a relatively high impact effect was selected for further investigation. The experimental design is shown in Table 1 and the results are shown in Table 2, and the analysis results are shown in Table 3.

[0057] The level of each variable in the medium formula of table 1

[0058]

[0059] ...

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Abstract

The invention discloses a fermentation preparation process of Streptomyces marinensis and a secondary metabolite thereof. The strain is characterized by having a preservation number of CGMCC No. 11468. The fermentation process specifically includes the steps of: inoculating Streptomyces marinensis to PDA, culturing in an incubator at 27.5 DEG C for 2 d; picking a single colony, inoculating to a seed culture medium, and culturing at 27.5 DEG C for 2 d; inoculating 5% by volume of a seed liquid to a fermentation medium, culturing at 27.5 DEG C for 11 d under the condition of 150 r / min; extracting a metabolite; and finally, separating and purifying a sampling liquid by a high-speed countercurrent chromatography to obtain a product. The process has the advantages of high preparation speed, and high product purity and yield.

Description

technical field [0001] The invention relates to the technical field of microbial fermentation and extraction and separation of natural products, in particular to a fermentation preparation process of marine streptomyces and its secondary metabolite Streptochlorin. Background technique [0002] Streptochlorin is an indole antibiotic. Jae et al. isolated it from marine-derived Streptomyces 04DH110 in 2007, and found that the compound had antiproliferative activity on human cells (Jae, SH.; Jeong, HS.; Lee , HS.; Park, SK.; Kim, HM.; Kwon, HJ. J. Microbiol. Biotechnol. 2007, 17, 1403). In studies in recent years, the compound has been confirmed to have good antitumor and antiallergic activities (Lee, SH.; Shin, HJ.; Kim, DY.; Shim, DW.; Kim, TJ.; Ye, SK .;Won,HS.;Koppula,S.;Kang,TB.;Lee,KH.PLoS One.2013,8(9),e74194), anti-inflammatory activity (Shim,DW.;Shin,HJ.;Han,JW .;Shin,WY.;Sun,X.;Shim,EJ.;Kim,TJ.;Kang,TB.;Lee,KH.Int.J.Mol.Sci.2015.16,6902), antibacterial activity (Zhan...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12P17/18C12R1/465
CPCC12P17/182C12N1/205C12R2001/465
Inventor 何山严小军励林
Owner NINGBO UNIV
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