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Actinomycete Streptomycesbottropensis and application thereof

A technology of Streptomyces cholerae and actinomycetes is applied in the field of actinomycete strains with high production of phospholipase D, which can solve the problems of complex production process of phosphatidylserine, chemical synthesis is limited to laboratory scale, etc., and achieves mild implementation conditions, The effect of simple operation process

Inactive Publication Date: 2013-12-04
SHENYANG PHARMA UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the isolation and extraction of phosphatidylserine from nature and the production process of chemical synthesis are very complicated, and the chemical synthesis is limited to the laboratory scale.

Method used

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  • Actinomycete Streptomycesbottropensis and application thereof
  • Actinomycete Streptomycesbottropensis and application thereof
  • Actinomycete Streptomycesbottropensis and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] The characteristics of the microbial strain Streptomyces bottropensis used for fermenting and preparing phospholipase D according to the present invention:

[0063]

[0064] The strain was cultured on the culture medium for 7 days, and the strain was identified as Streptomyces bottropensis according to its culture characteristics and mycelial morphology.

[0065] Concrete steps for screening high-yield phospholipase D bacterial strains of the present invention:

[0066] (1) Activation of strains for screening.

[0067] The strains used for screening were inoculated from the original slant to the fresh Gaoshi No. 1 solid slant with bamboo sticks, and cultured in a 28°C incubator for 7 days.

[0068] (2) Preparation of screening plate

[0069] Pour 200mL of the sterilized Gaussian culture medium into a sterile assay plate to make a lower plate; add 2mL of 98% substrate lecithin solution to the 100mL separation medium cooled to about 55°C after sterilization to make i...

Embodiment 2

[0086] Detection of transesterification activity of phospholipase D prepared by microbial fermentation of the present invention

[0087] Dissolve an appropriate amount of soybean-derived phosphatidylcholine (PC) in 10 mL of chromatographically pure chloroform (final concentration 5 mg / mL) as the organic phase; take another 10 mL of fermented crude enzyme solution, and add L-serine (final concentration 50 mg / mL) to it , TritonX-100 (v / v, 2.4%), CaCl 2 (final concentration 80mM) is the aqueous phase; mix the organic phase and the aqueous phase 1:1 in a Erlenmeyer flask, place on a shaking shaker at 20-45°C for 1-10h, and perform biotransformation. After the reaction, the two liquid phases were statically separated, and phosphatidylserine was present in the organic phase of the lower chloroform layer. The organic phase was taken for HPLC detection. Chromatographic conditions: Kromasil5μC18 chromatographic column, mobile phase ratio is methanol: acetonitrile: water = 16:2:3, ELS...

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Abstract

The invention relates to actinomycete for production of phospholipase D and application thereof and belongs to the field of biotechnology. The phospholipase D produced by the actinomycetes after fermentation has catalytic action on soybean phosphatidylcholine and L-serine to efficiently produce phosphatidylserine through transesterification. The actinomycete is collected from a wellhead NO.1066 in Dongfeng town, Dawa County, Panjin city, Liaoning province, and is identified as Streptomycesbottropensis, preserved in the key laboratory for screening of new drugs in Liaoning province under the number of SYP-A-6632, and is preserved in CGMCC under the number of CGMCC No.6993. A screening method of the Streptomycesbottropensis includes: 1, activating strains to be activated; 2, preparing a screening plate; 3, screening and decomposing lecithin strains by a plate method; 4, detecting transesterification activity. The Streptomycesbottropensis has the advantages that the operation process is simple, implementation conditions are mild, phosphatidylserine can be bio-converted efficiently and quickly, and basis of industrial production of phosphatidylserine in future is laid.

Description

technical field [0001] The invention belongs to the technical field of biopharmaceuticals, in particular to an actinomycete Streptomyces bottropensis strain and its application, in particular to an actinomyces strain with high phospholipase D production and its application. Background technique [0002] Phosphatidylserine (phosphatidylserine, PS), also known as serine phospholipid or diacylglyceryl phosphatidylserine, is a natural phospholipid, which was first extracted and named by Jordi Folch in 1942. Phosphatidylserine is not a single component, but a group of compounds, which is caused by the large variation of lipoacetyl residues in products extracted from raw materials from different sources. PS uses glycerol as the main skeleton, the 1st and 2nd carbon atoms are connected to fatty acids to form its non-polar tail, and the 3rd carbon atom is connected to a phospholipid with serine to form its polar head. [0003] [0004] There are many preparation methods for phos...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N9/16C12P13/06C12R1/465
Inventor 张怡轩
Owner SHENYANG PHARMA UNIVERSITY
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