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Natamycin fermentation technology based on addition of exogenous saturated fatty acid

A fermentation process and natamycin technology, applied in the field of industrial biology, can solve problems such as cytotoxicity, and achieve the effects of promoting product synthesis, strong application value, and reducing the load of detection and feeding operations.

Active Publication Date: 2019-06-28
山东元泰生物工程有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The present invention aims at the problems of cytotoxicity and feedback inhibition that are easily produced by directly adding precursor substances such as acetic acid, and develops a kind of Streptomyces gilvosporeus Z8 (Streptomyces gilvosporeus Z8) as a production bacterium, which utilizes glucose and saturated fatty acid dual carbon sources to carry out sodium Synthetic method of tamycin

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] Example 1: Preliminary screening of lipid substrates that can be used in natamycin biosynthesis

[0052] In a 250mL Erlenmeyer flask, a liquid medium was prepared according to the natamycin fermentation medium formula. The carbon source was 80g / L glucose (control), 50g / L animal oil (lard, sheep oil, tallow, fish oil), 50g / L vegetable oil (corn oil, peanut oil, rapeseed oil, soybean oil), 50g / L oleic acid, 50g / L palmitic acid, 50g / L stearic acid, add 20-30 to each bottle The glass beads were sealed with 8 layers of gauze, sterilized in a high-temperature sterilization pot at 115°C for 15 minutes, and then taken out immediately, and shaken at 220 rpm in a 30°C shaker until the culture medium was cooled to obtain a fermentation broth with dispersed particles. ; The mature fermented seeds were connected to the above fermentation medium with 8% inoculum and cultured in a constant temperature shaking incubator at 29°C for 4 days. After the end, the concentration of natamycin in ...

Embodiment 2

[0053] Example 2: Glucose-oil substrate compound addition test

[0054] In a 250mL Erlenmeyer flask, a liquid medium was prepared according to the natamycin fermentation medium formula. The carbon source was 80g / L glucose (control), 80g / L glucose + 10g / L animal oil (lard, sheep oil, beef Oil, fish oil), 80g / L glucose+10g / L vegetable oil (corn oil, peanut oil, rapeseed oil, soybean oil), 80g / L glucose+10g / L glycerol, 80g / L glucose+10g / L oleic acid, 80g / L glucose+10g / L palmitic acid, 80g / L glucose+10g / L stearic acid, 20-30 glass beads are added to each bottle, sealed with 8 layers of gauze, in a high-temperature sterilization pot Sterilize at 115°C for 15 minutes and take it out immediately, shake at 220 rpm in a 30°C shaker until the culture medium is cooled to obtain an oily substrate fermentation broth with dispersed particles; the mature fermented seeds are connected to the above fermentation at 8% inoculum The culture medium was cultured in a constant temperature shaking inc...

Embodiment 3

[0055] Example 3: Particle size optimization of saturated fatty acids

[0056] Sterilize palmitic acid and stearic acid in a high-temperature sterilization pot at 115°C for 15 minutes, take them out and grind them in a pre-sterilized mortar, and divide them into particle sizes of about 0.7mm, 0.15mm, 0.45mm, 1.3mm, and 1.8mm. Of saturated fatty acid particles. In a 250mL Erlenmeyer flask, prepare a liquid medium according to the natamycin fermentation medium formula, add 20-30 glass beads to each bottle, seal with 8 layers of gauze, and sterilize in a high-temperature sterilization pot at 115°C for 15 minutes. After cooling, add 50g / L palmitic acid (about 0.15mm in diameter), 50g / L palmitic acid (about 0.45mm in diameter), and 50g / L palmitic acid (about 0.45mm in diameter) prepared above into different shake flasks. 1.3mm), 50g / L palmitic acid (diameter about 1.8mm), 50g / L stearic acid (diameter about 0.15mm), 50g / L stearic acid (diameter about 0.45mm), 50g / L stearic acid ( (Ab...

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Abstract

The invention discloses a natamycin fermentation technology based on addition of exogenous saturated fatty acid. By adding a slow-release precursor source, namely the saturated fatty acid, which has no toxic or side effects on cells, when beta-oxidation utilization is conducted in the cells, the exogenous saturated fatty acid generates a large quantity of acetyl CoA precursors and NADH, thus supply of natamycin macrolide precursors and ATP is guaranteed, and product synthesis is promoted. Since the saturated fatty acid is in a solid state under normal temperature and is not dissolved in water,the concentration of the saturated fatty acid cannot influence cell growth and metabolism in a fermentation liquor, and therefore, in the technology, rigid control over the concentration of the saturated fatty acid is not needed. By means of the technology, the biosynthesis level of the natamycin can be effectively improved, the detection and material supply operation load of precisely regulatingthe precursor concentration in the fermentation liquor is greatly lowered, and the technology has important application value in the industry.

Description

Technical field [0001] The invention relates to a method for promoting the synthesis of natamycin, in particular to a natamycin fermentation process based on the addition of exogenous saturated fatty acids, belonging to the field of industrial biotechnology. Background technique [0002] Natamycin (Natamycin) is a polyene macrolide antifungal agent. At present, its production is mainly produced by Streptomyces producing bacteria (Streptomyces Chattanooga, Streptomyces chrysosporium, Streptomyces lydia and Streptomyces natalensis, etc.) is achieved through liquid submerged fermentation. Natamycin can bind to the sterol groups on the fungal cell membrane, thereby causing the fungal cell membrane to deform and rupture, so it has a strong inhibitory effect on fungi, yeast and other fungi. In the past 30 years, Natamycin has played an important role in the treatment of fungal diseases, food preservation and preservation with its extremely high safety (FDA, §172.155, 1994), and has a ...

Claims

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Application Information

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IPC IPC(8): C12P19/62C12N1/20C12R1/465
Inventor 曾昕李峰曾化伟徐大勇张标信丙越李珊珊
Owner 山东元泰生物工程有限公司
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