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Method for producing natamycin by using streptomyces gilvosporeus through fermentation

A production method and a natamycin technology are applied in the field of fermentation production of natamycin by Streptomyces lividans, and can solve the problems of affecting growth activity, imbalance of carbon-nitrogen ratio, increase in production cost, etc., and achieve the improvement of growth activity, The effect of shortening the fermentation cycle and shortening the lag period

Active Publication Date: 2017-05-17
SHANDONG LUKANG PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In the existing solutions, although increasing the amount of nutrients in the fermentation broth will improve the fermentation level to a certain extent, excessive nutrients will make the carbon-nitrogen ratio unbalanced, and the mycelium will affect other nutrients in the excessive nutrients. Growth activity, thereby affecting the concentration of fermentation bacteria and product content; and increasing the dynamic conditions of fermentation initiation, such as increasing ventilation, increasing stirring, etc., will greatly increase production costs

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Activation of the spore liquid of Streptomyces lutea lutea: The spore liquid of Streptomyces lutea lutea with the preservation number CGMCCNO. 2 O for dilution, the dilution factor is 10 -5 , spread on a plate containing 20ml of slant culture medium sterilized at 120°C for 35min, and cultivate at 26°C for 8 days for activation culture. The slant medium includes: glucose 11.0g / L, malt extract powder 2.5g / L, Yeast extract powder 3.5g / L, peptone 5.0g / L, agar 16.0g / L and the rest of water, the initial pH value is 6.9.

[0047] Seed culture: Add 40ml of seed culture medium sterilized at 130°C for 35min in a 500ml shake flask, pick and inoculate the activated Streptomyces chrysanthemum spores in the culture medium, the inoculation amount is 1.9%, at 210rpm, 25 Cultivate at ℃ for 26 hours to obtain 10.5% Streptomyces chrysanthemum chrysanthemum. Sodium 9.0g / L, potassium dihydrogen phosphate 0.4g / L, calcium carbonate 5.0g / L and the rest of water, the initial pH value is 6.9. ...

Embodiment 2

[0050] Activation of the spore liquid of Streptomyces lutea lutea: The spore liquid of Streptomyces lutea lutea with the preservation number CGMCCNO. 2 O for dilution, the dilution factor is 10 -5 , spread it on a plate containing 30ml of slant medium sterilized at 130°C for 25min, and cultivate it at 30°C for 6 days for activation culture. The slant medium includes: glucose 9.0g / L, malt extract powder 3.5g / L, Yeast extract powder 3.0g / L, peptone 5.5g / L, agar 14.0g / L and the rest of water, the initial pH value is 7.1.

[0051] Seed culture: Add 60ml of seed culture medium sterilized at 130°C for 25min in a 500ml shake flask, pick the activated Streptomyces chrysanthemum spores and inoculate them in the culture medium, the inoculation amount is 2.1%, at 230rpm, 28 Cultivate at ℃ for 20 hours to obtain Streptomyces chrysanthemum with a concentration of 10.3%. The seed medium includes: glucose 19.0g / L, yeast powder 3.0g / L, peptone 4.5g / L, corn steep liquor 4.5g / L, Sodium 10.5g / ...

Embodiment 3

[0054] Activation of the spore liquid of Streptomyces lutea lutea: The spore liquid of Streptomyces lutea lutea with the preservation number CGMCCNO. 2 O for dilution, the dilution factor is 10 -5 , spread it on a plate containing 25ml of slant culture medium sterilized at 121°C for 30 minutes, and cultivate it at 28°C for 6 days for activation culture. The slant medium includes: glucose 10.0g / L, malt extract powder 3.0g / L, Yeast extract powder 3.0g / L, peptone 5.0g / L, agar 15.0g / L and the rest of water, the initial pH value is 7.0.

[0055] Seed culture: Add 50ml of seed medium sterilized at 121°C for 30min in a 500ml shake flask, pick and inoculate the activated Streptomyces chrysanthemum spores in the medium, the inoculation amount is 2.0%, at 220rpm, 28 Cultivate at ℃ for 24 hours to obtain a concentration of 10.4% Streptomyces chrysanthemum, the seed medium includes: glucose 20.0g / L, yeast powder 3.0g / L, peptone 5.0g / L, corn steep liquor 5.0g / L, Sodium 10.0g / L, potassium...

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Abstract

The invention provides a method for producing natamycin by using streptomyces gilvosporeus through fermentation. The method comprises the following steps of: 1) activating a streptomyces gilvosporeus spore liquid; 2) inoculating the streptomyces gilvosporeus spore liquid activated in the step 1) into a seed culture medium, and performing seed culture so as to obtain streptomyces gilvosporeus; and 3) inoculating the streptomyces gilvosporeus obtained in the step 2) into a fermentation culture medium, and performing aerobic fermentation culture, thereby obtaining natamycin. According to the method, an appropriate culture medium ratio is adopted, a synergist is added into basic materials, the growth activity of mycelia inn environment with a balanced carbon-to-nitrogen ratio and inorganic salts can be greatly improved, power conditions do not need to be increased merely, the mycelia can be relatively rapidly adaptive to the environment of a culture liquid, the lag phase can be shortened, and thus the fermentation cycle can be shortened. The whole fermentation cycle is 79-82 hours, the final concentration of the mycelia of streptomyces gilvosporeus is 44-45%, and the content of natamycin is 6.9-7g / L.

Description

technical field [0001] The invention relates to the technical field of microbial fermentation, in particular to a fermentation production method of Streptomyces chrysanthemum chrysanthemum natamycin. Background technique [0002] Natamycin is a polyene macrolide antibiotic, a secondary metabolite produced by Streptomyces genus Streptomyces Natal, Streptomyces chrysanthemum and other bacteria under aerobic conditions, which can inhibit yeast The growth of bacteria and filamentous fungi and the production of mycotoxins have been widely used in food preservation (food grade) and treatment of diseases caused by fungi (medical grade). At present, only nisin, natamycin and polylysine are allowed to be used in the world as biological preservatives. Among them, natamycin is an efficient, safe, non-toxic and low-dose biological preservative. It has been used by more than 50 countries around the world, including the United States, and has become a food additive widely used in the wor...

Claims

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Application Information

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IPC IPC(8): C12P19/62C12R1/465
CPCC12N1/20C12P19/626
Inventor 金伟郑长春王博慕东卢春玲马艳茹张兴灿张玲陈君
Owner SHANDONG LUKANG PHARMA
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