A kind of method utilizing microbial transformation to prepare Boldenone

A technology of Boldenone and transformation process, which is applied in the field of preparation of Boldenone by microbial transformation, which can solve the problems of only 0.15% feed concentration, low commercial value, unfavorable separation and purification, etc., to improve feed concentration and conversion rate , short reaction route, rapid growth effect

Active Publication Date: 2021-08-27
ZHEJIANG XIANJU PHARMA
View PDF4 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since this type of enzyme is a relatively complex oxidoreductase, and the enzyme activity in most microorganisms is not high, it is difficult to form a competitive commercial process; in addition, the enzyme is usually regulated by the nutritional status of the microorganism, and will Strengthen the reverse reaction and re-oxidize the reduced hydroxyl group to ketone group, making the final conversion rate low, which is not conducive to separation and purification
In the Chinese patent CN102168099, the inventor even transferred the steroid 1,2-position dehydrogenase gene into Pichia pastoris, and constructed the dehydrogenation-keto reductase catalytic route for 4-androstenedione (AD) in vivo , but the feeding concentration is only 0.15%, and the commercial value is not great

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 kind of method utilizing microbial transformation to prepare Boldenone
  • A kind of method utilizing microbial transformation to prepare Boldenone
  • A kind of method utilizing microbial transformation to prepare Boldenone

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0067] Prepare seed liquid shake flask, glucose 4.5%, corn steep liquor 2.0%, adjust pH value to 4-4.5. Use a 500ml triangular flask with a liquid volume of 100ml, sterilize at 115°C for 15 minutes, after cooling down to room temperature, insert an inoculation loop of Saccharomyces cerevisiae, and culture it on a shaker at 30°C and 160 rpm for 24 hours; prepare a shaker flask for fermentation broth , glucose 5.5%, corn steep liquor 2.0%, yeast extract 0.9%, potassium dihydrogen phosphate 0.68%, sodium hydroxide 0.13%, pH 6.2-6.4, use a 500ml triangular flask with a liquid volume of 100ml, 115°C for 15 minutes Sterilize. Cool to room temperature.

[0068] Move 5 milliliters of strain liquid into each fermentation broth shaker bottle, and cultivate it on a shaker at 30° C. at 160 rpm for 16 hours. Put 0.25 grams of 1,4-androstenedione through a 100-mesh sieve after crushing into each bottle, and continue to ferment for 24 Hours, inactivated at 80°C for 30 minutes. The ferment...

Embodiment 2

[0070] Prepare seed liquid shake flask, glucose 4.5%, corn steep liquor 2.0%, adjust pH value to 4-4.5. Use a 500ml triangular flask with a liquid volume of 100ml, sterilize at 115°C for 15 minutes, after cooling down to room temperature, insert an inoculation loop of Saccharomyces cerevisiae, and culture it on a shaker at 30°C and 180 rpm for 24 hours; prepare a shaker flask for fermentation broth , glucose 5.5%, corn steep liquor 2.0%, yeast extract 0.9%, potassium dihydrogen phosphate 0.68%, sodium hydroxide 0.13%, pH 6.2-6.4, use a 500ml triangular flask with a liquid volume of 100ml, 115°C for 15 minutes Sterilize. Cool to room temperature.

[0071] Transfer 10 ml of strain solution into each shaker bottle of fermentation broth, culture in a shaker at 30°C and 180 rpm for 20 hours, add 0.25 g of 1,4-androstenedione dissolved in 2 times of ethanol, and dissolve again in 2 times of ethanol after 12 hours 0.25 grams of 1,4-androstenedione, supplemented with 1% nutrient glu...

Embodiment 3

[0073] Prepare seed liquid shake flask, glucose 4.5%, corn steep liquor 2.0%, adjust pH value to 4-4.5. Use a 500ml triangular flask with a liquid volume of 100ml, sterilize at 115°C for 15 minutes, and after cooling down to room temperature, insert an inoculation loop of grape juice yeast, and culture it on a shaker at 30°C at 200 rpm for 16 hours; Bottle, glucose 5.5%, corn steep liquor 2.0%, yeast extract 0.9%, potassium dihydrogen phosphate 0.68%, sodium hydroxide 0.13%, pH6.2-6.4, use a 500ml triangular flask with a liquid volume of 100ml, 115°C for 15 minutes to sterilize. Cool to room temperature.

[0074] Transfer 20 milliliters of strain liquid into each fermentation broth shaker bottle, cultivate it in a shaker at 30°C at 200 rpm for 14 hours, put in 0.25 grams of 1,4-androstenedione that has been crushed and passed through a 100-mesh sieve, and crushed again after 12 hours. 0.25 g of 1,4-androstenedione through a 100-mesh sieve, supplemented with 1% nutritional gl...

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 method for preparing boldenone, which uses yeast to convert 1,4-androstenedione (ADD) into boldenone, and the yeast is Saccharomyces cerevisiae Saccharomyces cerevisiae CGMCC 2.346 or grape juice yeast Saccharomyces uvarum One of CGMCC 3.4636. The beneficial effect that the present invention reaches is: 1, utilize biotransformation to effectively avoid the strong reductant selectivity difference in the chemical method, the defect that the process can't meet the environmental safety requirement of production process; 3, the process of the present invention prepares boldenone with a short reaction route, excellent product quality, and is suitable for industrial production.

Description

technical field [0001] The invention belongs to the field of steroid biotechnology, in particular to a method for preparing boldenone through microbial transformation. Background technique [0002] Boldenone (17β-hydroxy-1,4-androstadien-3-one, CAS:846-48-0), also known as Boldenone, is a kind of protein anabolic agent, which has the ability to enhance blood vessel oxygen supply , strengthen muscles, highlight blood vessels, increase appetite and so on. The structural formula is: [0003] [0004] As for the preparation method of Boldenone, at present, most domestic enterprises use chemical synthesis methods to prepare it from 4-androstenedione (AD) and 1,4-androstenedione (ADD). [0005] Route 1: Starting from 4-androstenedione (AD) to prepare boldenone, it is necessary to reduce the 17th position with a potassium borohydride reducing agent to prepare testosterone (TS), and then use microbial or chemical methods to 1, The 2-position dehydrogenation produces Boldenone....

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): C12P33/00C12R1/865C12R1/85
Inventor 张小强曹桂阳刘建
Owner ZHEJIANG XIANJU PHARMA
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