Mucor circinelloides bacterial strain, cultivating method and application of the same in C21, C19 sterides and aza sterides biotransformation

A kind of technology of Mucor curcumae and steroid organisms, which is applied in the field of hydroxylated derivatives, can solve the problems such as no reports on the transformation of steroid substances, and achieves easy cultivation and preservation, reduced synthesis steps, and simple and convenient preparation method. Effect

Inactive Publication Date: 2008-07-16
ZHENGZHOU UNIV
View PDF0 Cites 10 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The present invention screens out a fungus that has a hydroxylation effect on steroidal compounds. At present, there are no reports of its use in the transformation of steroidal substances at home and abroad.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Mucor circinelloides bacterial strain, cultivating method and application of the same in C21, C19 sterides and aza sterides biotransformation
  • Mucor circinelloides bacterial strain, cultivating method and application of the same in C21, C19 sterides and aza sterides biotransformation
  • Mucor circinelloides bacterial strain, cultivating method and application of the same in C21, C19 sterides and aza sterides biotransformation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Collect soil samples from the northern suburbs of Zhengzhou City, Luoyang Luanchuan Mountain, Songshan and other places, weigh 10g of various soil samples, respectively suspend them in 0.9% NaCl solution, filter with 8 layers of gauze to remove larger impurity particles, inoculate and enrich Medium (yeast extract 0.15%, peptone 0.12%, glucose 3%, potassium dihydrogen phosphate 1%, pH 4.5, steroid mixture 0.5%), after 48 hours of culture, take 10 ml of culture solution and use the same method for secondary enrichment. Spread the selective plate (the composition is the same as the enrichment medium, containing 3% agar). After preliminary screening, 36 strains that can grow on the plate containing the substrate were obtained, and they were picked out and stored on the slant made by transferring the PDA medium and used for the transformation test.

[0028]Respectively inoculate 36 strains obtained from the above preliminary screening into transformation medium (yeast extract 0....

Embodiment 2

[0030] Optimize the fermentation medium for the selected Mucor circinelloides lusitanicus AY234877 CGMCC.NO.2256, and optimize the conditions such as carbon source, nitrogen source, metal ions, initial pH value, inoculation amount, liquid volume, etc. The composition of the fermentation medium was determined to be: maltose 50‰; peptone 18‰; yeast extract 1.5‰; magnesium sulfate 0.9‰; potassium dihydrogen phosphate 0.9‰; pH 6.0; inoculation volume 5%; liquid volume 80ml / 500ml bottle.

Embodiment 3

[0032] Configure 1L liquid medium according to the medium formula described in Example 2. After inserting the bacteria, culture at 28°C for 48h (shaker speed 220rpm), add 1.0g / L of substrate pregnenolone I, and then at 28°C Transformation for 96h. After TLC monitors the conversion termination (the raw material point disappears), the mycelium is removed by filtration, the fermentation broth is extracted 4 times with ethyl acetate, the organic phases are combined, concentrated under reduced pressure, and then washed 3 times with saturated sodium bicarbonate solution and saturated chlorinated Wash with sodium solution once, wash with water 3 times, and then dry with anhydrous sodium sulfate, and evaporate the solvent under reduced pressure to obtain 1.05 g of white solid, which is separated by column chromatography (ethyl acetate:chloroform 1:1) to obtain 1,256 mg of hydroxylated product , The yield is 24.38%, the compound 2 is 487mg, the yield is 46.38%, the structure is shown in th...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention discloses a Mucor circinelloides lusitanicus AY234877 strain which has the stereoselective hydroxylase activity, which is collected in the China general microbiological culture collection center on November 20, 2007, the collection number is CGMCC.NO.2256, and the invention is characterized in that the positions of 6 Beta, 7 Alpha, 11 Alpha and 14 Alpha of C21 and C19 steroid substrates can be carried out a hydroxylation. The invention further discloses a method for preparing the 6 Beta, 7 Alpha, 11 Alpha and 14 Alpha hydroxylated derivatives of the C21 and C19 steroids and aza steroids. The method has simple operation, low cost and better yield and stereoselectivity; furthermore, the invention does not cause environmental pollution. The strain can carry out the hydroxylation reaction of the C21 and C19 steroids and aza steroid compounds which have the structural characteristics of 4-en-3-one or 5-en-3-ol, and the obtained hydroxylated derivatives are the important intermediates for the synthesis of a plurality of steroid compounds. Compared with the steroid hydroxylation chemical synthesis, the method can simplify drug synthetic steps, reduce pollution, reduce cost and have important application value for promoting the development process of the steroid drug intermediates of China.

Description

Technical field [0001] The present invention relates to a microbial strain, its culture method and its application in steroid biotransformation, in particular to a strain with stereoselective hydroxylated steroid substrate and an optimized transformation medium and the use of the strain to have 4 -Method for converting C21 and C19 steroidal substrates with structural characteristics of en-3-one and 5-en-3-ol into hydroxylated derivatives. Background technique [0002] Steroids, also known as steroids, are organic compounds with cyclopentane polyhydrophenanthrene nucleus, which are widely present in biological tissues, such as sterols, vitamin D, bile acids, many sex hormones, adrenal cortex Hormones, certain carcinogenic hydrocarbons, steroidal saponin and steroidal alkaloids fall into this category. Due to the differences in the substituents, double bond positions or stereo configurations on the nucleus, a series of compounds with unique physiological functions have been formed....

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/14C12P33/10C12P33/06C12R1/785
Inventor 刘宏民单丽红李永红曹晨张洪杰
Owner ZHENGZHOU UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap