Method for increasing yield of riemerella anatipestifer gelatin liquefying enzyme through batch fermentation

Abstract

The invention belongs to the technical field of microbial fermentation culture, and discloses a method for increasing the yield of riemerella anatipestifer RA1 gelatin liquefying enzyme by batch fermentation, which comprises the following steps: performing fermentation culture on serum type 1 riemerella anatipestifer by a batch fermentation method to obtain fermentation liquor; centrifuging, precipitating and dialyzing the fermentation liquid, repeatedly washing the recovered target product with a PBS buffer solution to obtain a gelatinase pure product, and verifying that the gelatinase pure product is positive through a gelatin liquefying enzyme liquefaction experiment, so as to obtain a gelatin liquefying enzyme final product; and carrying out molecular weight determination on the gelatin liquefying enzyme end product by adopting an SDS-PAGE method. The fermentation liquor culture specifically comprises the following steps: strain recovery: melting serum type 1 riemerella anatipestifer RA1 stored in a refrigerator, taking bacterial sludge by using an inoculating loop through a sterile operation method, coating a freshly prepared blood plate with the bacterial sludge, and carrying out streak culture to obtain a single colony; rA1 amplification culture: coating a blood plate with RA1 single colonies, and culturing overnight after a large number of bacterial clusters grow on the plate; and culturing RA1 by a batch fermentation method.

Description

technical field [0001] The invention belongs to the technical field of microbial fermentation and culture, in particular to a method for increasing the yield of Riemerella anatipestifer gelatin liquefaction enzyme by batch fermentation. Background technique [0002] At present, Rimerella-anatipestifer (RA) is the causative agent of infectious serositis in ducks. It invades the body and can cause systemic diseases characterized by fibrinous polyserositis and meningitis. Riemerella anatipestifer is a non-hemolytic, non-motile, gram-negative bacterium. With the development of duck farming in the world, the disease has become an important bacterial disease affecting duck farming worldwide. Studies have found that RA (serotype 1, RA1) can secrete a protease (gelatin liquefaction enzyme) that liquefies gelatin, which is an important cause of its pathogenesis. At present, the enzyme is mainly obtained in the laboratory by culturing in an Erlenmeyer flask, and the yield obtained i...

AI Technical Summary

Problems solved by technology

[0004] (1) adopt Erlenmeyer flask small batch to cultivate output very low;

[0005] (2) Cannot automatically monitor the fermentation process

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