A Penicillium simplex capable of biosynthesizing 1,2-dihydrotestolactone (testololactone) and its synthetic method

A simple penicillium and microorganism technology is applied in the field of bioengineering to achieve the effects of high conversion rate, few by-products, and simple and convenient preparation method

Active Publication Date: 2017-05-24
TIANJIN INST OF IND BIOTECH CHINESE ACADEMY OF SCI +1
View PDF1 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are no reports on the direct synthesis of 1,2-dihydrotestolactone by one-step transformation of Penicillium simpleum at home and abroad. By optimizing the culture conditions of the strain and the transformation conditions, a higher substrate feeding concentration can be achieved, and a high yield can be obtained. target product

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
  • A Penicillium simplex capable of biosynthesizing 1,2-dihydrotestolactone (testololactone) and its synthetic method
  • A Penicillium simplex capable of biosynthesizing 1,2-dihydrotestolactone (testololactone) and its synthetic method
  • A Penicillium simplex capable of biosynthesizing 1,2-dihydrotestolactone (testololactone) and its synthetic method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Embodiment 1: Screening and identification of strains

[0027] Soil samples were collected from Qingcheng Mountain in Dujiangyan, Sichuan Province, Jixian Mountain in Tianjin City, Weifang, Shandong and other places, and 5g of each soil sample was weighed and suspended in sterile water, oscillated and mixed, and then left to stand. The supernatant was diluted and applied on the PDA solid medium, the mold on the plate was separated, and the obtained single bacteria were transferred to the slant prepared by the PDA medium for preservation, and used for the transformation experiment.

[0028] Inoculate the individual molds obtained above into liquid transformation medium (glucose 30g / L, corn steep liquor dry powder 10g / L, NaNO 3 2g / L, KCl0.5g / L, K 2 HPO 4 ·3H 2 O2g / L, KH 2 PO 4 1g / L, MgSO 4 ·7H 2 O0.5g / L, FeSO 4 ·7H 2 (00.02g / L, pH6.2), 30°C, 200r / min shaking on a shaker for 24-48h, adding 0.5g / L17α-hydroxyprogesterone to start the reaction, and taking samples for...

Embodiment 2

[0030] Embodiment 2: The method for synthesizing 1,2-dihydrotestolactone (testololactone) with pregnane-4-ene-3,20-dione as substrate

[0031] 1 Preparation of seed solution

[0032] The strain of Penicillium simplicissimum WY134-2 was inoculated in the primary seed culture medium, cultured at 30°C for 16 hours, and the shaker speed was 300r / min to obtain the seed culture solution.

[0033] The composition of the seed medium is: glucose 30g / L, corn steep liquor dry powder 10g / L, NaNO 3 2g / L, KCl0.5g / L, K 2 HPO 4 ·3H 2 O2g / L, KH 2 PO 4 1g / L, MgSO 4 ·7H 2 O0.5g / L, FeSO 4 ·7H 2 O0.02g / L, pH6.2.

[0034] 2 secondary fermentation culture

[0035] The seed culture solution obtained in step 1 was transferred to a fresh fermentation medium according to the inoculum amount of 3%. The culture conditions were: 30°C, the rotating speed of the shaker was 250r / min, and the fermentation time was 20h, and the cell fermentation solution was obtained.

[0036] The composition of the...

Embodiment 3

[0042] Embodiment 3: The method for the synthesis of 1,2-dihydrotestolactone (testololactone) with 17β-hydroxyl-androst-4-en-3-one as substrate

[0043] 1 Preparation of seed solution

[0044] The strain of Penicillium simplicissimum WY134-2 was inoculated in the primary seed culture medium, cultured at 30°C for 20 hours, and the shaker speed was 200r / min to obtain the seed culture solution.

[0045] The composition of the seed medium is as shown in Example 2.

[0046] 2 secondary fermentation culture

[0047] The seed culture solution obtained in step 1 was transferred to fresh fermentation medium according to the inoculum amount of 4%. The culture conditions were: 30°C, the rotating speed of the shaker was 220r / min, and the fermentation time was 18h, and the cell fermentation solution was obtained.

[0048] The composition of the fermentation medium is as shown in Example 2.

[0049] 3 Transformation reactions

[0050] 17β-Hydroxy-androst-4-en-3-one II, which was solubil...

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 strain of Penicillium simplicissimum WY134-2 and a method for producing 1,2-dihydrotestolactone (testololactone) by fermenting and transforming C21 or C19 steroid compounds with the strain. The strain was deposited on August 12, 2013 in the General Microbiology Center of the China Committee for the Collection of Microorganisms, with a preservation number of CGMCC NO.8017. The production method is as follows: using Penicillium simplicissimum WY134‑2 as the bacterial species, through primary seed cultivation and secondary fermentation expansion cultivation, putting pulverized steroid compounds into the bacterial fermentation broth, acidifying, and extracting with organic solvents , and silica gel column chromatography to obtain the product 1,2-dihydrotestolactone with a yield of 56-96%. The method is simple and convenient, has mild conditions, few by-products, and does not pollute the environment.

Description

technical field [0001] The invention relates to the field of bioengineering, and the invention relates to a bacterial strain and a synthesis method for biosynthesizing 1,2-dihydrotestolactone (testololactone). Background technique [0002] [0003] 1,2-Dihydrotestolactone, the English name is testololactone, the chemical formula is C 19 h 24 o 3 , molecular weight 302.41, white or off-white powder, CAS4416-57-3. 1,2-dihydrotestolactone, as a class of hormone drugs, is the first generation aromatase inhibitor. It can inhibit the synthesis of estrogen such as estradiol and estrone in the body, reduce the production of estrogen, and reduce the level of estrogen in the blood, and is used for the treatment of pseudoprecocious puberty. In addition, its derivatives are clinically used as cellular anticancer drugs and in the treatment of diseases such as male infertility (Dao T L. Estrogen synthesis in human breast tumor and its inhibition by testololactone and bromoandrostene...

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 Patents(China)
IPC IPC(8): C12N1/14C12P33/20C12R1/80
CPCC12N1/14C12P33/12C12P33/20C12N1/145C12R2001/80
Inventor 吴洽庆王玉杨贝贝陈曦朱敦明马延和
Owner TIANJIN INST OF IND BIOTECH CHINESE ACADEMY OF SCI
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