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Streptolydigin output improving method

A technology for the production of streptolycin, which is applied in the field of increasing the production of streptolycin in the fermentation process, can solve the problems of low production of streptolycin and the impact on the further research, development and utilization of streptolycin, and achieves The effect of increasing yield

Active Publication Date: 2014-07-23
TIANJIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Documents at home and abroad show that the yield of streptolycin is low, which affects people's in-depth research and further development and utilization of streptolycin

Method used

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  • Streptolydigin output improving method
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  • Streptolydigin output improving method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] (1) Preparation of culture medium and amino acid mother solution:

[0024] ①Seed medium: starch 30g, glucose 5g, yeast extract powder 2g, peptone 4g, MgSO 4 ·7H 2 O0.5g, NaCl0.5g, K 2 HPO 4 1.5g, add water to make up to 1L, steam sterilize at 121℃ for 20min;

[0025] ② Fermentation medium: starch 40g, glucose 5g, peptone 2g, MgSO 4 ·7H 2 O0.5g, NaCl0.5g, K 2 HPO 4 1g, add water to make up to 1L, pH6.5, steam sterilization at 121℃ for 20min;

[0026] ③Proline mother liquor: Weigh 0.3g of proline, dilute to 100mL with ultrapure water, and pass through a 0.2μm sterile filter to obtain sterile proline mother liquor;

[0027] (2) Seed culture: Streptomyces lydicus (Streptomyces lydicus) E9CGMCC No.3075 slant was inoculated in a 250mL shaker flask containing 50mL seed medium, and cultured for 48h at a shaker speed of 200rpm and a temperature of 28°C to obtain First-class seeds: Inoculate the first-class seeds with a volume ratio of 10% in a 250-mL shaker flask contai...

Embodiment 2

[0036] (1) Preparation of culture medium and amino acid mother solution:

[0037] ①Seed medium: starch 10g, glucose 6g, yeast extract powder 5g, peptone 5g, MgSO 4 ·7H 2 O1g, NaCl1g, K 2 HPO 4 1g, add water to make up to 1L, steam sterilize at 121℃ for 20min;

[0038] ② Fermentation medium: starch 30g, glucose 8g, peptone 3g, MgSO 4 ·7H 2 O1g, NaCl1g, K 2 HPO 4 1g, add water to make up to 1L, steam sterilize at 121℃ for 20min;

[0039] ③Proline mother liquor: Weigh 1g of proline, dilute to 200mL with ultrapure water, and pass through a 0.3μm sterile filter to obtain sterile proline mother liquor;

[0040] (2) Seed cultivation: same as step (2) of Example 1;

[0041] (3) Fermentation by adding proline: Centrifuge 200ml of the secondary seeds at 4000rpm for 10min to obtain mycelium, resuspend the mycelium with 200ml of fermentation medium, and inoculate to the container at an inoculation density of 10% by volume. In a 5L fermenter with a fermentation medium, add 200mL ...

Embodiment 3

[0047] (1) Preparation of culture medium and amino acid mother solution:

[0048] ①Seed medium: 50g starch, 1g glucose, 1g yeast extract powder, 1g peptone, MgSO 4 ·7H 2 O0.1g, NaCl0.1g, K 2 HPO 4 0.1g, add water to make up to 1L, steam sterilize at 121℃ for 20min;

[0049] ② Fermentation medium: starch 10g, glucose 1g, peptone 1g, MgSO 4 ·7H 2 O0.1g, NaCl0.1g, K 2 HPO 4 0.1g, add water to make up to 1L, steam sterilize at 121℃ for 20min;

[0050] ③Proline mother liquor: Weigh 0.1g of proline, dilute to 50mL with ultrapure water, and pass through a 0.1μm sterile filter to obtain sterile proline mother liquor;

[0051] (2) Seed cultivation: same as step (2) of Example 1;

[0052] (3) Fermentation by adding proline: Centrifuge 100ml of the secondary seeds at 3000rpm for 20min to obtain mycelium, resuspend the mycelium with 100ml of fermentation medium, and inoculate to the container at an inoculation density of 5% by volume. In a 5L fermenter with a fermentation medium...

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Abstract

The invention discloses a streptolydigin output improving method, which comprises the following steps: (1) preparing of a culture medium and an amino acid mother liquid; (2) seed culture; (3) adding of proline for fermentation, to be more specific, centrifuging two stage seeds to obtain mycelium, using a fermentation medium for resuspending the mycelium, inoculating the mycelium into a fermentation tank provided with the fermentation medium according to the inoculum density of 5% to 30% in volume ratio, then adding a proline mother liquid for 80-150h of fermentation under the conditions of the speed of agitator of 200-600rpm, the ventilation volume of 1-3vvm and the pH of 6-8 to obtain a fermentation broth; and (4) determination of streptolydigin content. According to the streptolydigin output improving method, by adding of the proline in the fermentation process, the streptolydigin output in the fermentation process can be greatly improved, and a foundation is laid for the development and utilization of streptolydigin.

Description

technical field [0001] The invention belongs to the field of industrial microorganisms, and relates to a method for increasing the yield of streptavidin in the fermentation process. Background technique [0002] Liddistreptacin is a typical Tetramic acid antibiotic with good antibacterial, antitumor and anti-HIV protease activities. The latest research shows that streptolycin is an important inhibitor of RNA polymerase, which has been discussed in three related papers in Cell and MolCell, which has attracted widespread attention. Improving the level of secondary metabolites is an important research task in the development of new microbial drugs and industrial microbial fermentation, including strain improvement and process optimization control. Documents at home and abroad show that the yield of streptolycin is low, which affects people's in-depth research and further development and utilization of streptolycin. At present, the methods used to increase the yield of antibio...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P17/18C12R1/465
Inventor 程景胜陈恳元英进梁英权崔少飞
Owner TIANJIN UNIV
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