Method for preparing 11alpha, 17alpha-hydroxyprogesterone by conversion of immobilized hydroxylase

A technology for immobilizing hydroxylase and hydroxyprogesterone, which is applied to immobilized enzymes, microorganism-based methods, biochemical equipment and methods, etc. question

Active Publication Date: 2021-04-30
SHANGHAI INST OF TECH
View PDF4 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The technical problems to be solved by the present invention are: the existing fermentation method for preparing 11α, 17α-hydroxyprogesterone has low feed concentration, large amount of fermentation waste water, complex extraction and purification process of the product, and low final yield of the product, etc.

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

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Aspergillus ochraceus was cultured on a slant to obtain the strains required for fermentation.

[0033] 1. Preparation of Aspergillus ochraceus slant strain and seed solution

[0034] (1) Incline cultivation

[0035] Incline medium: peeled potato 200g / L, glucose 20g / L, agar 20g / L, natural pH, sterilized at 121℃ for 30min.

[0036] The bacterial species preserved in the glycerol tube is aseptically drawn a certain amount of bacterial liquid and spread evenly on the slant medium, and cultured at 28°C for 4-6 days.

[0037] (2) Seed cultivation

[0038] The weight portion composition of primary medium comprises (g): glucose 5, yeast extract 2, K 2 HPO4 1.0, NaCl 0.1, water 1000, and adjust the pH to 5.0, sterilize by high-pressure steam at 121°C for 30 minutes, and cool down for use.

[0039] With the strain cultivated in step (1), under sterile conditions, use an inoculation loop to pick 6 ring spores into 100mL seed medium, inoculate 4 bottles in the same way, and c...

Embodiment 2

[0054] Aspergillus ochraceus was cultured on a slant to obtain the strains required for fermentation.

[0055] 1. Preparation of Aspergillus ochraceus slant strain and seed solution

[0056] (1) Incline cultivation

[0057] Incline medium: peeled potato 200g / L, glucose 20g / L, agar 20g / L, natural pH, sterilized at 121℃ for 30min.

[0058] The bacterial species preserved in the glycerol tube is aseptically drawn a certain amount of bacterial liquid and spread evenly on the slant medium, and cultured at 28°C for 4-6 days.

[0059] (2) Seed cultivation

[0060] The weight portion composition of primary medium comprises (g): glucose 7, yeast extract 5, K 2 HPO 4 1.5, NaCl 0.4, water 1000, and adjust the pH to 5.5, sterilize by high-pressure steam at 121°C for 30 minutes, and cool for later use.

[0061] With the strain cultivated in step (1), under aseptic conditions, use an inoculation loop to pick 6 ring spores into 100mL seed medium, inoculate 4 bottles in the same way, an...

Embodiment 3

[0076] Aspergillus ochraceus was cultured on a slant to obtain the strains required for fermentation.

[0077] 1. Preparation of Aspergillus ochraceus slant strain and seed solution

[0078] (1) Incline cultivation

[0079] Incline medium: peeled potato 200g / L, glucose 20g / L, agar 20g / L, natural pH, sterilized at 121℃ for 30min.

[0080] Draw a certain amount of bacterial solution evenly on the slant medium under aseptic operation to absorb the bacterial species preserved in the glycerol tube, and culture at 28°C for 4-6 days at a constant temperature.

[0081] (2) Seed cultivation

[0082] The composition by weight of primary culture medium comprises (g): glucose 10, yeast extract 8, K 2 HPO 4 2.0, NaCl 1.0, water 1000, and adjust the pH to 6.8, sterilize by high-pressure steam at 121°C for 30 minutes, and cool down for use.

[0083] With the strain cultivated in step (1), under aseptic conditions, use an inoculation loop to pick 6 ring spores into 100mL seed medium, in...

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

PropertyMeasurementUnit
wavelengthaaaaaaaaaa
Login to view more

Abstract

The invention discloses a method for preparing 11alpha, 17alpha-hydroxyprogesterone by utilizing immobilized hydroxylase conversion, which is characterized in that 17alpha-hydroxyprogesterone is used as a substrate, the 11alpha, 17alpha -hydroxyprogesterone is prepared by utilizing the immobilized hydroxylase conversion, and the method comprises the following steps of: preparing a culture medium, culturing aspergillus ochraceus in the culture medium, inducing hydroxylase, preparing the immobilized hydroxylase, preparing a conversion system, converting a substrate, extracting and refining a product and the like. The method has the advantages that the feeding concentration is high, the substrate conversion rate is high, the conversion period is short, the product is easy to separate, and the immobilized hydroxylase can be repeatedly used. The feeding concentration of 17 alpha-hydroxyprogesterone in a single enzyme reaction is 40 g/L, the molar conversion rate can reach 93%, and meanwhile, the immobilized hydroxylase can be repeatedly recycled for more than three times. The preparation method is simple in operation process and relatively low in production cost, and has a relatively high industrial application prospect.

Description

technical field [0001] The invention relates to a method for preparing 11α, 17α-hydroxyprogesterone by conversion of immobilized hydroxylase, and belongs to the technical field of preparation of steroid intermediates. Background technique [0002] Steroids, also known as steroids, are a class of compounds containing cyclopentane polyhydrophenanthrene nuclei. Its basic structure consists of three six-membered rings and one five-membered ring, which are called A, B, C, and D rings respectively. Generally, there are angular methyl groups (-CH 3 ), there may be a hydroxyl group (-OH) or a keto group (-C=O) at the 3rd and 11th positions, some double bonds exist in the A ring or the B ring, and there are side chains of different lengths at the 17th position. A series of compounds with unique physiological functions are formed due to the differences in substituents, double bond positions or stereo configurations on the steroidal core. Steroid drugs are the second largest class of...

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): C12N11/10C12N11/084C12N9/02C12N1/14C12P33/10C12R1/66
CPCC12N1/14C12N9/0083C12N11/10C12P33/10C12N11/084
Inventor 管世敏荣绍丰蔡宝国李茜茜
Owner SHANGHAI INST OF TECH
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