Method for improving saccharopolyspora spinosa spinosad fermentation yield

A technology of Saccharopolyspora spinosa and spinosad, which is applied in the field of improving the yield of spinosad fermentation of Saccharopolyspora spinosa, can solve the problems of increasing waste of resources, residues, and low utilization rate of starch, and achieve the reduction of total sugar and Non-reducing sugar content, avoiding very sticky, excellent high-yield performance

Inactive Publication Date: 2016-06-01
牡丹江佰佳信生物科技有限公司
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Problems solved by technology

In the process of using Saccharopolyspora spinosa to ferment spinosad, starch is often added to the medium as a late-acting carbon source because of its low price, wide source, and ability to relieve the glucose effect. In production practice , the existing Saccharopolyspora spinosa used to produce spinosad generally has the defect of low utilization of starch in the fermentation ...

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  • Method for improving saccharopolyspora spinosa spinosad fermentation yield

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Embodiment approach

[0036] According to one embodiment of the present invention, the method is carried out according to the following steps:

[0037] Step 1: performing ultraviolet compound streptomycin mutagenesis on Saccharopolyspora spinosa for the first starch utilization ability screening to obtain the first mutant strain;

[0038] Step 2: performing a second starch utilization ability screening on the first mutant strain after mutagenesis with 5-fluorouracil to obtain a second mutant strain;

[0039] Step 3: performing the third starch utilization ability screening on the second mutant strain after microwave-complexed streptomycin mutagenesis to obtain the third mutant strain;

[0040] Step 4: Perform the fourth starch utilization ability screening on the third mutant strain after mutagenesis with nitrosoguanidine and streptomycin to obtain the fourth mutant strain.

[0041] According to the method provided by the present invention, the pH value of the starch hydrolysis medium is 7.0-7.4, ...

Embodiment 1

[0048] Utilize 0.1mol / L, the phosphate buffer solution of pH6.5 is formulated as the concentration of 10 8 spore suspension per ml.

[0049] Preparation of separation medium: glucose 10g / L, soluble starch 20g / L, yeast extract powder 5g / L, enzyme hydrolyzed casein 5g / L, calcium carbonate 1g / L and agar 20g / L. Starvation medium: glucose 10g / L, magnesium sulfate 0.5g / L, potassium dihydrogen phosphate 1g / L, potassium chloride 0.5g / L. Starch hydrolysis medium: soluble starch 10g / L, dipotassium hydrogen phosphate 0.3g / L, magnesium carbonate 1g / L, sodium chloride 0.5g / L, potassium nitrate 1g / L and agar 20g / L, pH 7.2. The iodine solution is composed of iodine tablet 3g / L and potassium iodide 6g / L, prepared with water.

[0050] Slant medium composition: cornstarch 25g / L, corn steep liquor 20g / L, yeast extract 10g / L and agar 20g / L, pH value 7.0, prepared with water; sterilized at 115°C for 20 minutes.

[0051] (1) Ultraviolet compound streptomycin mutagenesis:

[0052] 1) Ultraviolet...

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Abstract

The invention relates to a method for improving a saccharopolyspora spinosa spinosad fermentation yield. The method utilizes mutagenesis treatment to screen a saccharopolyspora spinosa spinosad high-yield mutant strain with a good starch utilization capability. The mutagenesis treatment comprises at least two of UV-streptomycin combined mutagenesis, 5-fluorouracil mutagenesis, microwave-streptomycin combined mutagenesis and nitrosoguanidine-streptomycin combined mutagenesis. Preferably, the mutagenesis treatment comprises the above four mutagenesis methods. Preferably, the UV-streptomycin combined mutagenesis, 5-fluorouracil mutagenesis, microwave-streptomycin combined mutagenesis and nitrosoguanidine-streptomycin combined mutagenesis processes are orderly carried out. The broth of the obtained mutant strain has spinosad titer substantially higher than that of an original strain screened through mutagenesis and thus the method can improve a saccharopolyspora spinosa spinosad fermentation yield.

Description

technical field [0001] The invention relates to the field of biological fermentation, in particular to a method for increasing the spinosad fermentation yield of Saccharopolyspora spinosa. Background technique [0002] Spinosyn, a secondary metabolite produced by the aerobic fermentation of Saccharopolyspora spinosa, belongs to macrolide compounds, and the effective active ingredient in its commercial products is spinosad A (85%-90%) and spinosyn D (10%-15%). Spinosad has no antibacterial activity, but it has good insecticidal activity. It has a wide insecticidal spectrum against pests of Lepidoptera and Thysanoptera. Lepidopteran pests have a good control effect. Spinosad is one of the most selective compounds among existing insecticides in the control of Lepidoptera pests, and its activity is equivalent to cypermethrin. While spinosyn has high insecticidal activity, it exhibits low toxicity to non-target organisms. It has low toxicity to mammals, birds and beneficial ins...

Claims

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Application Information

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IPC IPC(8): C12N13/00C12N15/01C12P19/62C12R1/645
Inventor 王书睿周贤龙刘静
Owner 牡丹江佰佳信生物科技有限公司
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