SU626117A1 - Toxin producing method - Google Patents

Description

one

This invention relates to the field of medicine and microbiology.

A known method for producing a toxin by growing inoculum of a culture of Clostridium perlringens followed by inoculation of the inoculum for cultivation and release of the desired product 1.

However, this method does not provide a high yield of toxin. In addition, the method is time consuming.

The aim of the invention is to improve the method and increase the yield of toxin.

This is achieved in that the inoculum is grown for 2.5-3.5 hours and an inoculum is selected for sowing, having a specific growth rate of puffed cpp 0.7-0.8 1 / h; the specific rate of growth of the toxigenic properties of the toxin is 450-550 01 b / bn and the specific rate of increase of the antigenic properties of the toxin is 7.0-9.0 b + / billion h.

FIG. 1 shows the dependence of the specific growth rates of biomass ji on time /; in fig. 2 - dependence of toxigenic rpi toxin properties on time t; in fig. 3- dependence of antigenic gr2 toxin properties on time t.

Example. The strain Clostridium perfringens type A 28 VR-28 is cultivated in casein-napcreatic nutrient medium with

by adding 1% glucose in vials for 16-18 hours. Sow in the bottle with the same medium with a working volume of 9 L, then the culture in the bottle is used as a seed (or masterbatch - inoculum) for seeding a reactor with a working volume of 250 l. In the culturing process in bottles, samples are taken every hour, in samples the concentration of biomass X is determined by counting the microbial cells in the Highway chamber.

The activity of the culture fluid is also studied in dynamics, determining the growth of toxigenic pi (B1 / ml) and antigenic PZ (L + / Mi) properties of the toxin.

Based on the data obtained, the specific growth rates of the population and time of the toxigenic rpi and antigenic rpz properties of the toxin are calculated (see table).

Graphs of dependences (as shown in Fig. 1) of specific biomass growth rates (d, growth of toxigenic rpi and antigenic c2 properties of the toxin (ordinate axis) versus time (abscissa axis) of growth are constructed.

The period of population development is determined, in which the specific growth rate p is 0.7-0.8 1 / H, the specific growth rate of the toxigenic properties of the toxin rpi is 450-550 Dlm / bn. H, and the specific velocity is

increase in the antigenic properties of the toxin rpz 7-9 b + / billion This period in our example corresponds to 2.5-3.5 hours from the start of cultivation. It is bounded by the dotted lines in FIG. one.

In production, a 6-8 hour culture is usually used, in which, as can be seen from FIG. 1, the values of all three specific velocities approach zero. The use of iozlyum in the development period of 2.5-3.5 hours from the cultivation period for planting the reactor allows to obtain a balanced growth in it, which is characterized by soviadepy in time of maximum values of the specific growth rates of the population, an increase in the toxin and antigenic properties of toxin and an increase in activity ( output) of toxin from 7 to 8.7-9.5 b + / ml, which is compared with the average annual process, giving 7 b + / ml, 28% more.

The table below shows the formula for calculating the specific growth rates of biomass (.i, increases in toxigenic rpi and antigenic c2 toxin PROPERTIES.

The proposed method provides to novysheyn activity (output) of the toxin from 7 to 8.7-9.5 b + / ml, which is 28% more compared to the average annual process, dyush them 7 + / ml. In addition, a significant simplification of the method is noted, since the preparation time of the uterine culture (inoculum) of a Clostridium porfringens culture is reduced from 6--8 to 3 hours with 100% intensive growth of the culture and high activity of the toxin in the reactor.

Table

Designation

Formulas for calculating the index, dimension

J

dx

 1 / h

V-

di

X

2.3 ()

and, ::; (h - to)

ft J

rpt

rpi t / bn

dt x

2, 3 (jPll - AO) (- Ig XQ)

// ,. ((Xl-Xo}

dp2

rp2 dt

2.3 (/ 21 - Ptu (Ig-Y -)

(1 - o) (- Jfo) of the invention

The method of producing toxin by growing the inoculum of Clostridium perfringens culture followed by sowing the ipoculum for cultivation and isolating the target product, characterized in that, in order to simplify the process and increase the yield of the toxin, the growth of the inoculum is carried out for 2.5-3.5 hours seeding, an inoculum is selected that has a specific growth rate of 0.7 to 0.8 1 / H of a population, a specific growth rate of the toxigenic properties of the toxin 450-550 Dim / bn, and a specific growth rate of the antigenic properties of the toxin 7.0-9.0 b + billion h

ten /