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Preparation method for malformin C

A technology of seed and microbial fermentation, applied in the field of medicine

Inactive Publication Date: 2014-08-06
SHENYANG PHARMA UNIVERSITY +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In addition, Yasuhiro Kojimal etc. synthesized malformin C by solid-phase synthesis, and the yield was only 15% (Yasuhiro Kojimal, Toshiaki Sunazukal, Kenichiro Nagail, et al.Solid-phase synthesis and biological activity of malformin C and its derivatives.The Journal of Antibiotics 2009, 62: 681-686)

Method used

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  • Preparation method for malformin C
  • Preparation method for malformin C
  • Preparation method for malformin C

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1: The characteristics of the fungus SYP2612 of the present invention on different media.

[0027]

[0028] See attached figure 1 : Colony morphology of strain SYP2612.

[0029] Characteristics of strain SYP2612 under light microscope:

[0030] The conidia head is spherical to radial; the conidiophore wall is smooth, the top capsule is spherical, the spore-forming structure is double-layered, and the conidia are spherical, with rough or verrucous projections. See attached figure 2 :Sporangium and spore morphology of strain SYP2612.

[0031] Use molecular biology methods to extract the above-mentioned fungal genomic DNA, using ITS1 and ITS4 (ITS1: 5'-TCC GTC GGT GAA CCT GCG G-3'; ITS4: 5'-TCC TCC GCT TAT TGA TAT GC-3') as The primers were amplified by PCR, sequenced, and the measured sequence was compared with the rRNA homologous sequence in Genbank. 11 strains with higher homology were selected, and Sphingobacterium siyangensis was used as the reference, and the bacte...

Embodiment 2

[0032] Example 2: Method for preparing malformin C by aerobic microbial shake flask fermentation

[0033] 1. Medium

[0034] 1. Slant medium (g / L)

[0035] PDA medium: Potato 200, glucose 10, agar 20, distilled water to prepare pH 7.2-7.4 121 ℃ 30min

[0036] Preparation of potato juice: Take 200g of peeled potatoes, cut into small pieces, boil them in 1000ml of water for one hour, and filter, and make up the filtrate to 1000ml with distilled water.

[0037] 2. Shake flask seed medium (g / L)

[0038]

[0039] 3. Shake flask and fermenter fermentation medium (g / L) Is the medium of shake flask and fermentor the same?

[0040]

[0041] 2. Aerobic microbial fermentation process

[0042] 1. Slope activation of strains: open the freeze-dried tube of strain skimmed milk, pick a small amount of powder and evenly spread it on the above-mentioned sterilized slant, cultivate it at 28℃ for 5-7 days, the lawn grows dense and cloth. Full of sloping surface, no bacteria, store in refrigerator at 4℃.

[00...

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Abstract

The invention discloses a biological fermentation method for preparing a strain of malformin C and a method for separating and purifying malformin C from metabolite of the strain. The strain is separated from soil acquired in Phuket Island of Thailand on June, 2007; the bacterial colony of the strain is black on a PDA medium; and the stain is identified as Aspergillus tubingensis with a laboratory accession number of SYP2612 and a CGMCC accession number of XXX. The methods provided by the invention are advanced, employ low-cost raw materials, are suitable for large-scale industrial production, do not discharge harmful substances during production, have no pollution and no public hazard to a surrounding environment and accord with environmental protection requirements.

Description

Technical field: [0001] The invention relates to the technical field of medicines, that is, a method for producing malformin C by using biological fermentation, which provides a strain for preparing malformin C by microbial fermentation and a method for producing malformin C using its fermentation broth. technical background: [0002] The cyclic peptide compound malformin C is produced by a strain of Aspergillus niger found on moldy rice by Steenbergen S et al. in 1968. Its fermentation yield is extremely low, only 200ug / ml (Steenbergen, S., and ED Weinberg. Trace metal). requirements for malformin biosynthesis. Growth. 1968, 32: 125-134). In addition, Yasuhiro Kojimal et al. used solid-phase synthesis to synthesize malformin C with a yield of only 15% (Yasuhiro Kojimal, Toshiaki Sunazukal, Kenichiro Nagail, et al. Solid-phase synthesis and biological activity of malformin C and its derivatives. The Journal of Antibiotics 2009, 62: 681-686). Keiichi HAGIMORI et al. described th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P21/04C12R1/66
Inventor 张怡轩吴春福周骅
Owner SHENYANG PHARMA UNIVERSITY
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