High-throughput screening method of guanosine production strains

Abstract

The invention discloses a high-throughput screening method of guanosine production strains, comprising the following steps of: (1) inoculating the guanosine production strains to be screened into a microtiter plate I containing a culture medium, and carrying out shaking culture; (2) centrifugalizing the microtiter plate I, and removing thalli to obtain supernate; and (3) diluting the supernatant supernate of the microtiter plate I, then sequentially transferring to a microtiter plate II, measuring a light absorption value by using a full-waveband enzyme-labeling instrument, and screening out the guanosine production strains. The high-throughput screening method integrates the effects of microtiter plate culture, fast enzyme-labeling instrument detection, a continuous sample injector and a volley rifle, achieves the handling capacity of more than 200 samples / day everyone and provides a simple, convenient and efficient screening method for the screening of the guanosine production strains.

Description

technical field [0001] The invention belongs to the technical field of fermentation engineering, and in particular relates to a breeding method for a microbial strain producing guanosine. Background technique [0002] Guanosine, also known as guanosine, its trade name is GR, also known as 9-13-D-ribofuranosylguanine. It has a wide range of uses and is an important intermediate of food and pharmaceutical products. It can be used to synthesize food flavor enhancer disodium 5'-guanylate, taste nucleotide disodium and nucleoside antiviral drugs such as ribavirin , Acyclovir, etc., are also the main raw materials for the manufacture of drugs such as sodium guanosine triphosphate. Someone selected mutant strains resistant to guanine or guanine structural analogs when breeding guanosine-producing bacteria, and found that the strains that lost GMP reductase mainly accumulated guanosine. Watanabe et al. in Japan cloned the key enzyme PRPP transamidase gene in the guanosine biosynth...

AI Technical Summary

Problems solved by technology

However, the screening method of traditional microbial strains is a heavy work, the workload is large, the screening period is long, and it is often useless, the target strains cannot be obtained, and the blindness of screening is very high.