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Synthetic medium for erythromycin fermentation

A technology for synthesizing culture medium and fermentation culture medium, which is applied in the field of synthetic culture medium for erythromycin fermentation, and can solve the problems of low erythromycin yield and accuracy discount.

Active Publication Date: 2018-08-31
EAST CHINA UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] However, in this field, the erythromycin yield of the synthetic medium used in the current research process is very low, and the accuracy of the research on the synthesis mechanism of erythromycin A under such conditions may be greatly compromised

Method used

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  • Synthetic medium for erythromycin fermentation
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  • Synthetic medium for erythromycin fermentation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0061] Example 1. Erythromycin A production detection-high-throughput biological method to determine erythromycin production

[0062] Strain used for detection: Bacillus pumilus CMCC(B)63202 was purchased from Guangdong Huankai Microbial Technology Co., Ltd.

[0063] Slope culture medium for detection of bacteria: peptone 0.5%, beef extract 0.3%, dipotassium hydrogen phosphate 0.3%, glucose 1%, sodium chloride 0.5%, agar 2%, pH=7. This medium is used to cultivate the test bacteria (Bacillus pumilus).

[0064] Detection medium: peptone 0.5%, beef extract 0.3%, dipotassium hydrogen phosphate 0.3%, glucose 1%, sodium chloride 0.5%, pH=7.

[0065] Operation steps to determine the yield of erythromycin:

[0066] (1) Eggplant bottle culture: Take out the strains stored in the glycerol tube from the refrigerator at -80°C, draw a line with a bamboo stick, and cultivate it in an incubator at 37°C for 5-6 days.

[0067] (2) Preparation of bacterial suspension: Wash the Bacillus pumilus from the e...

Embodiment 2

[0069] Example 2: Optimization of erythromycin A synthetic medium

[0070] In order to find substances that promote the synthesis of erythromycin, the inventors conducted a large number of screenings in order to find out from a large number of candidates that promote the synthesis of erythromycin. The inventors selected 38 substances (including carbon sources, nitrogen sources, phosphorus sources, inorganic salts, amino acids, precursors, chelating agents, and trace elements) as the basic medium. The used substances and their amounts are listed in Table 1. .

[0071] Regarding the aforementioned preliminary selection of candidate substances, the inventors removed one substance in the medium one by one, and the obtained medium (referred to as medium x) was subjected to the fermentation experiment of red Saccharopolyspora saccharomyces (for the fermentation method, see the previous "Cultivation Method- —48-well plate fermentation culture" part); the final erythromycin yield (C x ) A...

Embodiment 3

[0082] Example 3 Plackett-Burman experiment

[0083] In Example 2, the obtained medium contained 19 components. In order to further obtain the components that have the greatest influence on erythromycin, the present inventors analyzed which of the 19 components have a greater influence on the synthesis of erythromycin through the Plackett-Burman experiment. The Plackett-Burman experimental design is shown in Table 5, and the concentration of each factor is shown in Table 4, where 1, -1 represents the highest and lowest concentration of each component. Saccharopolyspora erythraea was fermented for 120h to determine the concentration of erythromycin in each culture medium. According to Design Expert software, the product concentration is analyzed, and each substance is increased and decreased. Increasing concentrations include: glucose, arginine, aspartic acid, cysteine, leucine, trisodium citrate, cobalt chloride, sodium tetraborate, ferric chloride, calcium carbonate; decreasin...

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Abstract

The invention belongs to the field of biological fermentation engineering, and relates to a synthetic medium for erythromycin fermentation. The synthetic medium comprises glucose, alanine, arginine, aspartic acid, cysteine, leucine, isoleucine, serine, threonine, dipotassium hydrogen phosphate, dihydrogen phosphate potassium, magnesium sulfate heptahydrate, trisodium citrate, cobalt chloride hexahydrate, sodium tetraborate, ferric chloride, copper chloride, ammonium molybdate and calcium carbonate. The synthetic medium medium for the erythromycin fermentation is suitable for the growth of erythromycin producing bacteria such as saccharopolyspora erythraea, and can very obviously improve the yield of erythromycin.

Description

Technical field [0001] The present invention belongs to the field of biological fermentation engineering. More specifically, the present invention relates to a synthetic medium for erythromycin fermentation. Background technique [0002] Erythromycin A is a widely used broad-spectrum macrolide antibiotic. It is mainly used to treat diseases caused by Gram-positive bacteria [1]. Erythromycin (Erythromycin, EM for short) is a broad-spectrum antibiotic of the macrolide class, which has small side effects and other factors, and has become one of the main antibiotic products. [0003] In the past few decades, researchers have conducted a lot of research on the synthesis of erythromycin A, and continuously improved the production of industrial erythromycin A through continuous fermentation optimization [2,3]. However, fermentation optimization has reached the limit for the improvement of erythromycin A synthesis. In order to further understand the relationship between erythromycin A sy...

Claims

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Application Information

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IPC IPC(8): C12P19/62C12N1/20C12R1/01
CPCC12N1/20C12P19/62
Inventor 储炬廖建国洪铭黄明志庄英萍牟翰杨含
Owner EAST CHINA UNIV OF SCI & TECH
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