Method of increasing yield of spinosad by improving fermentation condition of saccharopolyspora spinosa based on metabonomics

A technology of Saccharopolyspora spinosyn and spinosad, which is applied in the field of improving the fermentation conditions of Saccharopolyspora spinosa to increase the yield of spinosad based on metabolomics, and can solve the problems such as unclear metabolic mechanism

Inactive Publication Date: 2015-03-25
TIANJIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Studies have shown that increasing dissolved oxygen in the fermentation process can help increase the yield of spinosad, but the related metabolic mechanism is still unclear, which has also become a stumbling block to the industrial production of spinosad to a certain extent.

Method used

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  • Method of increasing yield of spinosad by improving fermentation condition of saccharopolyspora spinosa based on metabonomics
  • Method of increasing yield of spinosad by improving fermentation condition of saccharopolyspora spinosa based on metabonomics
  • Method of increasing yield of spinosad by improving fermentation condition of saccharopolyspora spinosa based on metabonomics

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Experimental program
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Embodiment 1

[0075] The method for improving the fermentation conditions of Saccharopolyspora spinosa based on metabolomics to increase the yield of spinosad comprises the following steps:

[0076] (1) Determination of metabolites in Saccharopolyspora spinosa:

[0077] ① Cell collection and quenching:

[0078] Saccharopolyspora spinosa was fermented and cultivated in medium 1, and six time points were taken in the logarithmic phase 62h, 86h, 110h and the stationary phase 134h, 158h and 182h of Saccharopolyspora spinosa fermentation, and 3 samples were taken at each time point. For each sample, 5ml of fermentation broth, mix the sample with 18mL of quenching solution and place it for 5min, centrifuge at 3000rpm, 4°C for 3min, discard the supernatant; wash with distilled water at 4°C to remove the culture medium components on the cells ;3000rpm, 4°C, centrifuge for 3min, discard the supernatant, keep the bacteria; wash repeatedly 3 times;

[0079] The quenching solution is a methanol aqueo...

Embodiment 2

[0145] The method for improving the fermentation conditions of Saccharopolyspora spinosa based on metabolomics to increase the yield of spinosad comprises the following steps:

[0146] (1) Determination of metabolites in Saccharopolyspora spinosa:

[0147] ① Cell collection and quenching:

[0148] Saccharopolyspora spinosa was fermented and cultivated in medium 1, at six time points of log phase 62h, 86h, 110h and stationary phase 134h, 158h and 182h of Saccharopolyspora spinosa fermentation, and 4 samples were taken at each time point For each sample, 8ml of fermentation broth, mix the sample with 19mL of quenching solution, place it for 8min, centrifuge at 4000rpm, 4°C for 3min, discard the supernatant; wash with distilled water at 4°C to remove the culture medium components on the cells ;4000rpm, 4°C, centrifuge for 4min, discard the supernatant, keep the bacteria; wash twice;

[0149] Quenching solution is the same as Example 1, and medium 1 is the same as Example 1;

...

Embodiment 3

[0177] The method for improving the fermentation conditions of Saccharopolyspora spinosa based on metabolomics to increase the yield of spinosad comprises the following steps:

[0178] (1) Determination of metabolites in Saccharopolyspora spinosa:

[0179] ① Cell collection and quenching:

[0180] Saccharopolyspora spinosa was fermented and cultured in medium 1, and at six time points of logarithmic phase 62h, 86h, 110h and stationary phase 134h, 158h and 182h of Saccharopolyspora spinosa fermentation, each time point was sampled 5 For each sample, 10ml of fermentation broth, mix the sample with 20mL of quenching solution and place it for 10min, centrifuge at 5000rpm, 4°C for 5min, discard the supernatant; wash with 4°C distilled water to remove the medium components on the cells ;5000rpm, 4°C, centrifuge for 5min, discard the supernatant, keep the bacteria; wash 4 times repeatedly;

[0181] The quenching solution is a methanol aqueous solution with a volume concentration of...

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Abstract

The invention discloses a method of increasing the yield of spinosad by improving the fermentation condition of saccharopolyspora spinosa based on metabonomics. The method comprises the following steps: (1) determining intracellular metabolite of the saccharopolyspora spinosa; (2) analyzing the capability of the saccharopolyspora spinosa of producing the spinosad; (3) carrying out PLS analysis; and (4) carrying out process analysis. According to the method disclosed by the invention, the metabolite change law in the process of producing the spinosad by virtue of fermentation of the saccharopolyspora spinosa is revealed by utilizing the metabonomics means and combining multivariate statistics, and a metabolic change mechanism related to production of the spinosad is further explained; by analyzing the spinosad production capability and the metabolic level of the saccharopolyspora spinosa in different growth environments, the metabolin related to the production of the spinosad is found, thus providing a direction for optimizing the culture process of the saccharopolyspora spinosa and increasing the fermentation yield of the spinosad. The invention also provides new thinking and method for study in other culture technological processes of producing macrolides microorganisms.

Description

technical field [0001] The invention belongs to the field of biological pesticides, and in particular relates to a method for improving the fermentation conditions of Saccharopolyspora spinosa based on metabolomics to increase the yield of spinosad. Background technique [0002] Spinosyn, also known as spinosyn or spinosyn, is a macrolide antibiotic produced by secondary metabolism of Saccharopolyspora spinosa under aerobic fermentation conditions, mainly including spinosyn A (approx. 85%-90%) and spinosyn D (about 10%-15%) two compounds. Spinosyn, as a high-efficiency broad-spectrum macrolide insecticidal active substance, has a unique mode of action, wide insecticidal spectrum, short half-life, high insecticidal activity, no drug resistance, low residue, less environmental pollution and Some unique physical and chemical properties, biological characteristics and novel mechanism of action, such as being harmless to humans and animals, are called a new generation of broad-s...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P19/62G01N30/02C12R1/645
Inventor 卢文玉尹静张传波
Owner TIANJIN UNIV
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