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Microorganism strain and method for converting cephalosporin C into deacetylate cephalosporin C by using same

A technology for deacetylating cephalosporins and cephalosporins, which is applied in the directions of fungi and fermentation, can solve the problems of low yield, high cost, unsuitable for industrial-scale production and the like, and achieves the effect of simple and easy preparation method.

Inactive Publication Date: 2006-12-27
NORTHEAST PHARMA GRP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0016] 1, purpose of the invention: the present invention provides a kind of microbial new bacterial strain that can effectively transform CPC into DCPC, i.e. Rhodotorula. , high cost, unsuitable for industrial-scale production, etc.

Method used

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  • Microorganism strain and method for converting cephalosporin C into deacetylate cephalosporin C by using same
  • Microorganism strain and method for converting cephalosporin C into deacetylate cephalosporin C by using same
  • Microorganism strain and method for converting cephalosporin C into deacetylate cephalosporin C by using same

Examples

Experimental program
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Effect test

example 1

[0061] Example 1: R-92 bacterium culture medium: glucose 0.5%, peptone 1.2%, beef extract 0.1%, magnesium sulfate 0.05%, all the other are distilled water, adjust pH5.0, every 250ml Erlenmeyer flask is filled with 10ml prepared medium, Steam sterilize at 115°C for 30 minutes, inoculate R-92 strains, culture at 25-27°C with shaking at 100r / min for 72 hours, and centrifuge the culture solution at 4000r / min to obtain R-92 wet bacteria.

[0062] 0.5 g of CPCNa was dissolved in 50 ml of buffer solution of pH 5.8, 0.5 g of the wet thalline prepared above was added, transformed at 27° C. and 200 r / min for 38 hours, and the conversion rate was determined by HPLC to be 80%, and the transformed liquid was centrifuged at 4000 r / min. The centrifugate was filtered with a 0.8 μm membrane, concentrated under reduced pressure at 40°C to 1 / 5 of the original volume, added 4 times the amount of ethanol, stirred for crystallization, suction filtered, and dried to obtain 0.22 g of a light yellow po...

example 2

[0063] Example 2: Bacteria culture medium: 1% glucose, 1% peptone, 0.3% beef extract, 0.02% magnesium sulfate, and the rest are distilled water, adjust the pH of the medium to 6.0, fill 50ml of medium in every 250ml Erlenmeyer flask, and steam out at 115°C Bacteria were incubated for 30 minutes, inserted into R-92, shaken at 240r / min, 30°C for 48 hours, and centrifuged to obtain the bacteria.

[0064] CPCNa 1.5g was dissolved in 50ml pH5.8 phosphate buffer, 2g of wet bacteria was added, 28-30°C, 200r / min shaking transformation, HPLC determined that the transformation was completed in 28 hours, and the transformation rate was 100%. The transformation solution was centrifuged at 4000r / min to remove bacteria, the centrifugate was filtered through a 0.8μm membrane, concentrated under reduced pressure at 40°C to 1 / 3 of the original volume, 4 times the volume of ethanol was added, stirred to crystallize, suction filtered, and dried to obtain 1.1g of a yellowish powder.

example 3

[0065] Example 3: Same as Example 2, the difference is that the pH of the transformation solution is 4.5, and the transformation time is 50 hours. The conversion rate measured by HPLC is 49%. After concentration and crystallization, 0.36 g of DCPC solid was obtained.

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Abstract

This invention relates to a new Rhodotorula.sp used to prepare DCPC by biotransformation method and the method of using this strain to prepare DCPC. One kind of microorganism strain, its features is that the strain is rhodotorula category, its name is Rhodotorula R-92, its number in preservation center is CGMCC NO.1277, strain is used as biotransformation agent in this transform method. Organism transformation substrate is cephalosporin C salt water solution. Transformation agent is added to substrate to transform or blending ferment, then white or tiny yellow DCPC powder is got after resin proposal, concentration and solvent crystallization. The goal of this invention is to solve the problem of low yield and high cost and not fit to industrial scale generating in the generation of DCPC.

Description

[0001] One, technical field: the present invention relates to a kind of new red yeast fungus (Rhodotorula.sp) that is used for biotransformation method production preparation deacetyl cephalosporin C (DCPC) and utilizes this bacterial strain to prepare deacetyl cephalosporin C method. 2. Background technology: [0002] Cephalosporin C (Cephalosporin C, referred to as CPC) is a fermentation product, its structural formula is: [0003] [0004] When the acetyl group is removed from the 3-position of CPC, deacetyl cephalosporin C (Deactyl Cephalosporin C, referred to as DCPC) is obtained, and its structural formula is: [0005] [0006] DCPC is an impurity in the fermentation production of CPC. Due to the small amount and the trouble of recovery, it is not worth it as a by-product. However, as an intermediate of semi-synthetic cephalosporins, it has considerable value and can be used in the chemical synthesis of various cephalosporins, such as cefotaxime sodium, ceftriaxon...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/16C12P35/02
Inventor 卢法章冯丽艳张凤云冯德成郭凤尧刘辉杨国宏安静娴
Owner NORTHEAST PHARMA GRP
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