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Actinomycetes culture medium and application thereof

A culture medium, actinomycetes technology, applied in bacteria, microorganism-based methods, microorganisms, etc., can solve the problem of low xylosidic bond activity, achieve high enzyme production efficiency, reduce environmental pollution, and shorten the effect of synthetic pathways

Inactive Publication Date: 2018-09-07
DALIAN UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In order to overcome the above-mentioned problems existing in the prior art field, the object of the present invention is to provide a kind of actinomycetes medium formula, solve the activity of hydrolyzing DAXP C-7 xylosidic bond of original Micromonosporaceae and Cellulomonasaceae bacterial strains relatively low problem

Method used

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  • Actinomycetes culture medium and application thereof
  • Actinomycetes culture medium and application thereof
  • Actinomycetes culture medium and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0019] (1) Prepare medium: ① 10g yeast extract, 4g peptone, 4g dipotassium hydrogen phosphate, 14g microcrystalline cellulose, add distilled water to 1000ml; ② Mix the ingredients in the formula evenly, adjust the pH to 7.0, and pack the medium Add 2mL 2.5g / L CaCl to 25mL to 100mL Erlenmeyer flask 2 , 1g / L ZnSO 4 ·7H 2 O, 0.25g / L CuSO 4 ·5H 2 O, 5g / L FeSO 4 ·7H 2 O, 1g / L MnSO 4 4H 2 O, 50g / L Na 2 SO 4 , 12.5g / LMgSO 4 ·7H 2 O. Mixture; ③ Sterilize the aliquoted culture medium under high temperature and high pressure for 20 minutes, and store it at 4°C for later use.

[0020] (2) Experimental operation: ① Add 10 μL strain F16 to each bottle of culture medium, and incubate at 30°C for 48 hours at 150 rpm / min; HCl), 2 μL CaCl2 (1M), 30 μL Sample, 10 μL DAXP; reaction conditions: metal bath at 30°C 200 rpm / min, react for 2 hours; after the reaction, add 400 μL methanol to terminate the reaction); ③Take the supernatant of the sample after the reaction , using the HPLC-U...

Embodiment 2

[0025] (1) Prepare medium: ①10g yeast extract, 4g peptone, 4g dipotassium hydrogen phosphate, 14g microcrystalline cellulose, add distilled water to 1000ml; ②Mix the ingredients in the formula evenly, adjust the pH to 7.0, and dispense 25mL to 100mL In the Erlenmeyer flask, add 2mL 0g / L, 0.001g / L, 0.005g / L, 0.01g / L, 0.05g / L, 0.1g / L, 0.5g / L, 1g / L, 5g / L, 10g / L CuSO 4 ·5H 2 O solution; ③ Sterilize the aliquoted culture medium under high temperature and high pressure for 20 minutes, and store it at 4°C for later use.

[0026] (2) Experimental operation: ① Add 10 μL strain F16 to each bottle of medium, and incubate at 150 rpm / min at 30°C for 48 hours; HCl), 2 μL CaCl 2 (1M), 30 μL Sample, 10 μL DAXP; reaction conditions: metal bath at 30°C 200 rpm / min, react for 2 hours; after the reaction, add 400 μL methanol to terminate the reaction); ③ take the supernatant of the sample after the reaction, and use HPLC-UV Enzyme activity was determined by method (HPLC-UV method as shown in ...

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Abstract

The invention belongs to the industrial biotechnology field and provides an actinomycetes culture medium and application thereof. The actinomycetes culture medium consists of the following basic compositions: a yeast extract, peptone, Ca<2+>, Zn<2+>, Cu<2+>, Fe<2+> and a carbon source; the mass ratio of the above compositions is (2-10):(1-8):(0.01-0.2):(0.002-0.03):(0.0002-0.003):(2-14); distilledwater is dissolved and diluted, and the concentration of the yeast extract is controlled to be 2-12 g / L. The actinomycetes culture medium and the application thereof provided by the invention have the benefits that micromonospora and fiberomonas bacterial strains cultured by the actinomycetes culture medium can produce fibrous body analogues; one of enzymes has the activity of hydrolyzing a DAXPC-7 glycosidic bond of a taxol precursor; the enzyme production efficiency is relatively high, so that the synthetic pathway of taxol can be greatly shortened, the cost is reduced, the environmental pollution is reduced, and the like; therefore, the actinomycetes culture medium is an excellent enzyme preparation for preparation of a pharmaceutical intermediate for industrial microbial transformation.

Description

technical field [0001] The invention relates to an actinomycetes culture medium and its application, belonging to the technical field of industrial biology. Background technique [0002] Actinomycetes are a class of microorganisms with important economic value and multiple uses. Nearly 70% of the more than 8,000 microbial active substances found in microorganisms are produced by actinomycetes, and many of them have been widely used in clinical practice. According to the World Health Organization (WHO) report on cancer, cancer has become the second leading cause of death in the world, so the development and production of anticancer drugs is very urgent. [0003] Today, about 60% of anti-cancer agents are derived from natural products, and more than 30 compounds derived from natural products have been used in clinical treatment research of various stages of cancer. Paclitaxel (Taxol) compound is one of the existing, most excellent, high-efficiency and low-toxic broad-spectru...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12R1/04
CPCC12N1/20
Inventor 窦同意陈静刘成林
Owner DALIAN UNIV OF TECH
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