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A method for improving the yield of streptavidin

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: 2017-08-11
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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  • A method for improving the yield of streptavidin
  • A method for improving the yield of streptavidin
  • A method for improving the yield of streptavidin

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 250mL shaker flask contain...

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. Add 200 mL of proline mother liquor to a 5 L fermenter w...

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 method for increasing the yield of streptolycin, which comprises the following steps: (1) preparation of culture medium and amino acid mother liquor; (2) seed cultivation; (3) addition of proline for fermentation: adding secondary seeds Centrifuge to obtain mycelium, resuspend the mycelium with fermentation medium, then inoculate it into a fermenter with fermentation medium at an inoculation density of 5% to 30% by volume, then add proline mother liquor, The stirring speed is 200-600rpm, the ventilation rate is 1-3vvm, and the pH is 6-8, and it is fermented for 80-150h to obtain a fermentation liquid; (4) Determination of the content of streptavidin. By adding proline in the fermentation process, the invention greatly increases the yield of the streptolycin in the fermentation process, thereby laying a foundation for further research and application of the streptolycin.

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 Patents(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