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A method for efficiently fermenting and producing macrolide compound fw05328-1

A technology for macrocyclic lactams and compounds, which is applied in the field of high-efficiency fermentation to produce macrocyclic lactam compound FW05328-1, which can solve the problems of low fermentation titer and product ratio

Active Publication Date: 2021-08-03
FUJIAN INST OF MICROBIOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0007] For this reason, the technical problem to be solved by the present invention is to provide a method for efficiently fermenting and producing the macrocyclic lactam compound FW05328-1, so as to solve the problem of low fermentation titer and product ratio of the FW05328-1 compound in the prior art

Method used

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  • A method for efficiently fermenting and producing macrolide compound fw05328-1
  • A method for efficiently fermenting and producing macrolide compound fw05328-1
  • A method for efficiently fermenting and producing macrolide compound fw05328-1

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] The preserved Micromonospora marine strain Micromonospora sp.FIM-MA181224 was cultured on a high asparagine solid medium at 35°C for 15 days to obtain a fresh slant of Micromonospora marine micromonospora sp.FIM-MA181224.

[0037] Inoculate the fresh slanted pieces (0.5cm×0.5cm) of Micromonospora marine Micromonospora sp.FIM-MA181224 in the shake flask seed medium, and cultivate them at 28°C and 220r / min for 72h to obtain the shake flask seeds. Inoculate the shake flask fermentation medium with 4% inoculum amount, culture it at 28°C and 220r / min for 6 days, put it into the bottle, and measure the content and component ratio of FW05328-1 in the fermentation broth by HPLC method.

[0038] The seed medium preparation method: cornstarch 6g, peptone 5g, soybean meal 5g, yeast powder 3g, K 2 HPO 4 ·3H 2 O 0.5g, CaCO 3 1g, 16g sea salt, 1L tap water, adjust the pH value to pH7.2.

[0039] Fermentation medium preparation method: soluble starch 10g, peptone 5g, soybean cake...

Embodiment 2

[0055] In this embodiment, the fermentation medium in the fermentation method is further optimized, and a more suitable seed liquid medium and fermentation medium are screened out.

[0056] The fermentation method of the compound FW05328-1 described in this example is the same as in Example 1, the only difference being:

[0057] Described seed medium composition and content are as follows: soluble starch 18g, glucose 6g, peptone 4g, yeast powder 5g, (NH 4 ) 2 SO 4 0.5g, K 2 HPO 4 ·3H 2 O 0.5g, CaCO 3 2.0g, sea salt 16g, tap water 1L, adjust the pH value to pH7.2;

[0058] The composition and content of the fermentation medium are as follows: soluble starch 16g, glucose 4g, peptone 6g, soybean cake powder 2.5g, yeast powder 2g, K 2 HPO 4 ·3H 2 O 0.5g, (NH 4 ) 2 SO 4 0.5g, CaCO 3 1.0g, sea salt 10g, 3-aminobutyric acid 0.1g, tap water 1L, adjust the pH value to 6.6.

Embodiment 3

[0067] Embodiment 3 10L fermentation tank fermentation culture

[0068]The preserved Micromonospora marine strain Micromonospora sp.FIM-MA181224 was cultured on a high asparagine solid medium at 35°C for 15 days to obtain a fresh slant of Micromonospora marine micromonospora sp.FIM-MA181224.

[0069] Inoculate the shake flask seed medium with fresh slanted excavated pieces (0.5 cm×0.5 cm) of Micromonospora sp. FIM-MA181224, and culture them at 28° C. and 220 r / min for 72 hours to obtain shake flask seeds.

[0070] Inoculate the above-mentioned cultivated shake flask seeds into a 10L fermenter with an inoculum amount of 1.5%, the medium capacity of the fermenter is 7L, the fermentation temperature is 28°C, the tank pressure is 0.03Mpa, and the rotation speed is controlled at 180r / min for 0-48 hours of fermentation. The volume is 1:1.0vvm. With the growth of mycelia, when the bacterial concentration reaches 7% after about 48 hours of fermentation, increase the speed to 200r / min,...

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Abstract

The invention belongs to the technical field of microbial fermentation, and in particular relates to a method for efficiently fermenting and producing a macrocyclic lactam compound FW05328-1. The method for efficiently fermenting and producing the macrocyclic lactam compound FW05328-1 described in the present invention is based on the existing reported biosynthetic pathway, by optimizing the fermentation medium and adding 3-aminobutyric acid for product induction, and establishing Adapted to the fermentation medium and culture process of the strain producing the target product, in the 10L-500L fermenter fermentation experiment, the fermentation production capacity of FW05328-1 was increased from 249.8mg / L to 654.2mg / L, which greatly improved the production capacity of the target product ( Increased by 261.9%); and, the proportion of FW05328-1 compound components in the fermentation broth increased from 22.3% to 74.8% (HPLC method), and the increase reached 335.4%, thus providing a basis for the large-scale preparation of FW05328-1.

Description

technical field [0001] The invention belongs to the technical field of microbial fermentation, and specifically relates to a method for efficiently fermenting and producing a macrocyclic lactam compound FW05328-1, and more specifically relates to a method for increasing the fermentation yield of a macrocyclic lactam compound FW05328-1 based on amino acid induction. Background technique [0002] With the continuous advancement of medical technology, more and more medical problems have been overcome, and human life expectancy has been greatly extended. However, cancer is still one of the major medical problems that humans need to overcome. Whether in developed or developing countries, cancer is the disease with the highest mortality rate, and its incidence and mortality are still increasing. At present, the main treatments for cancer include surgery, radiotherapy and antineoplastic drug therapy, especially antineoplastic drug therapy. The continuous growth of the global cancer...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P17/10C12N1/20C12R1/29
CPCC12N1/20C12P17/10
Inventor 孙菲周剑赵薇江红
Owner FUJIAN INST OF MICROBIOLOGY
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