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A Two-Stage Dissolved Oxygen Control Method for Improving the Production of Spinosad

A spinosad and stage technology, applied in the fields of fermentation engineering and biopharmaceuticals, can solve the problems of stages and control of dissolved oxygen of Saccharopolyspora spinosa that have not yet been seen

Inactive Publication Date: 2015-10-28
HUAZHONG UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no research report on the stage-by-stage control of dissolved oxygen in the fermentation process of Saccharopolyspora spinosa

Method used

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  • A Two-Stage Dissolved Oxygen Control Method for Improving the Production of Spinosad

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] A kind of two-stage dissolved oxygen control method provided by the invention improves spinosad output, and its specific steps are as follows:

[0032] 1. Spore Preparation of S. spinosa

[0033] The freeze-dried powder in the freeze-drying tube of S. spinosa was transferred into the slant medium, and activated at 28° C. for 10 days to obtain Saccharopolyspora spinosa spores.

[0034] The slant medium per liter (unit: gram / liter) contains glucose: 10.0; casein: 2.0; yeast powder: 1.0; agar: 15.0.

[0035] 2. Preparation of Primary Seed Solution of Saccharopolyspora spinosa

[0036] Inoculate the S. spinosa spores obtained in step 1 into the S. spinosa primary seed medium, and culture at 30° C. for 72 hours to obtain the S. spinosa primary seed liquid.

[0037] Described per liter primary seed culture medium (unit: gram / liter) contains glucose: 10.0; Yeast powder: 2.0; Casein: 2.0; K 2 HPO 4 : 0.2; MgSO 4 : 0.3.

[0038] 3. Secondary Seed Liquid Preparation of Sacc...

Embodiment 2

[0046] The composition and inoculum size of slant medium, primary seed medium, secondary seed medium, and fermentation medium are the same as in Example 1.

[0047] The control conditions of the fermentation process: at the beginning of the fermentation, the ventilation volume is controlled at 0.5vvm, and the rotation speed is controlled at 200rpm, so that the initial k L a reached 115.5h -1 ; Fermentation 96 hours thalline growth enters logarithmic growth terminal stage, from 96 hours to the end of fermentation, the ventilation volume is controlled at 0.3vvm, and the rotating speed is controlled at 150rpm, so that the initial k of 96 hours L a reached 91.4h -1 , the two-stage dissolved oxygen control made the production of spinosyn reach 160.9mg / L in 196 hours, and the DCW reached 24.1g / L.

Embodiment 3

[0049] The composition and inoculum size of slant medium, primary seed medium, secondary seed medium, and fermentation medium are the same as in Example 1.

[0050] The control conditions of the fermentation process: at the beginning of the fermentation, the ventilation volume is controlled at 0.7vvm, and the rotation speed is controlled at 150rpm, so that the initial k L a reached 110.1h -1 ; Fermentation 98 hours thalline growth enters logarithmic growth terminal stage, from 98 hours to the end of fermentation, the ventilation volume is controlled at 0.3vvm, and the rotating speed is controlled at 150rpm, so that the initial k of 98 hours L a reached 91.4h -1 , the two-stage dissolved oxygen control made spinosad production reach 70.1mg / L in 215 hours, and DCW reached 21.9g / L.

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Abstract

The invention provides a two-stage dissolved oxygen control method for increasing pleocidin output. The method is characterized in that the volumetric mass transfer coefficient kLa of oxygen is adjusted and controlled by adjusting throughput and stirring speed at the beginning of fermentation of saccharopolyspora spinosa in the process of preparing saccharopolyspora spinosa fermentation liquor, thus the initial oxygen volumetric mass transfer coefficient kLa can be controlled to be 100-130h<-1> and the accumulation of biomass of saccharopolyspora spinosa is promoted; then the volumetric mass transfer coefficient kLa of oxygen is adjusted and controlled by adjusting throughput and stirring speed when the fermentation enters the final cell growth exponential phase, and thus the initial oxygen volumetric mass transfer coefficient kLa can be controlled to be 60-92h<-1> and the synthesis and accumulation of pleocidin are promoted. According to the method, the two-stage dissolved oxygen control is performed, which obviously increases the pleocidin output and shortened the fermentation cycle. The two-stage dissolved oxygen control and fermentation method is easily realized and has a high application value.

Description

technical field [0001] The invention relates to the fields of fermentation engineering and biopharmaceuticals, in particular to the application of dissolved oxygen control technology in the optimization of fermentation process, specifically a two-stage dissolved oxygen control method for increasing spinosad production. Background technique [0002] Spinosyn (spinosad), also known as spinosad or spinosad, is an intracellular secondary metabolite produced by the soil actinomycete Saccharopolyspora spinosa after aerobic fermentation. The active ingredients are spinosyn component A (spinosyn A) component D (spinosyn D). Spinosyn is a naturally occurring antibiotic that has good control effects on pests such as Lepidoptera and Diptera. Spinosyn has rapid contact and ingestion toxicity to insects, and stimulates the nervous system of insects, resulting in non-functional muscles. Contraction, collapse, with tremors and paralysis. Compared with common insecticides, spinosad has bo...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P19/62C12R1/01
Inventor 余龙江白云周蓬蓬
Owner HUAZHONG UNIV OF SCI & TECH
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