A high-throughput screening method for high-yield strains of phenazine-1-carboxamide

Abstract

The invention discloses a high-throughput screening method for phenazine-1-formamide high-yield strains. The method comprises the steps that phenazine-1-formamide high-yield mutant strains to be screened are cultured till the PCN yield is stable; the obtained fermentation broth is mixed with ethyl alcohol, copper ions are added, and a pretreated mixed solution is obtained; the pretreated fermentation broth is placed in a 96-hole plate, and the absorbancy is detected through a multiskan spectrum microplate spectrophotometer under 365 nm; phenazine-1-formamide high-yield mutant strains with the high light absorption value are selected. By means of the high-throughput screening method, the yield of multiple mutant strains can be roughly and rapidly judged, negative mutant strains can be rapidly abandoned, positive mutant strains with relatively high yield are selected, primary screening time is shortened, the workload is reduced, and a beneficial reference is provided for follow-up fermentation secondary screening work.

Description

technical field [0001] The invention relates to a high-throughput screening method for bacterial strains, in particular to a high-throughput screening method for high-yielding phenazine-1-carboxamide strains. Background technique [0002] Phenazine compounds can be used as biological pesticides. Currently, there is shenazinemycin (publication number CN1802927), the active ingredient of which is phenazine-1-carboxylic acid (PCA). Phenazine-1-carboxamide (PCN) has the function of antagonizing plant pathogenic fungi and has the potential to be developed into a biopesticide, but the main factor limiting its industrialization is low yield. For small molecule products from microbial sources, the main ways to increase yields are mutation breeding and genetic engineering breeding. Mutation breeding is a relatively simple and rapid method. Efficient primary screening means is the key factor limiting the efficiency of mutagenesis breeding. [0003] The original screening method: H...

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Problems solved by technology

However, as the yield of the strain increases, the efficiency of the method decreases

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