CS240342B1 - The method of fermentation production of tylosin - Google Patents

Abstract

The solution concerns a new method of controlling the fermentation production of tylosin according to the theological characteristics of the fermentation medium during aerobic cultivation of the high-yielding microorganism Streptomyces fradiae, with intensive mixing in a nutrient medium containing sources of assimilable nitrogen, carbon and phosphorus and with continuous or semi-continuous dosing of nutrients and a diluent. In the production phase, NH4+ and NO3- ions are added continuously or semi-continuously in such an amount as to maintain the concentration of ammonia nitrogen in the range of 0.02—0.3% and, in order to maintain optimal viscosity, sterile water is added continuously or semi-continuously and the pH is maintained in the range of 6.7—7.1 throughout the cultivation.

Description

240342

BACKGROUND OF THE INVENTION 1. Field of the Invention The invention relates to a process for controlling the production of tyrosine according to the rheological characteristics of an aerobic cultivation of a high-yielding microorganism Strepytesces fradiae, with intensive mixing of nutrient nitrogen, carbon and phosphorous, mineral or nutrient salts under continuous or semi-continuous dosing. nutrients and dilution solution in the production phase-fermentation.

Increasing the industrial production of tylosin can be achieved by increasing production capacity or by increasing production in its bio-synthesis. The second method does not require a major investment cost for the difference. Incorporated on a better production capability used for biosynthesis, or better fermentation conditions, which ensure better management of fermentation. From these sources, new mutants of the microorganism Streptomyces fradiae are obtained by mutagenesis and developed! new methods of blosynthesis of thylogen. The basic conditions for high-production fermentation include - the amount of nutrients in the fermentation broth, the maintenance of some physical, chemical and biochemical quantities at the optimum level, aeration and mixing of the fermentation medium, and the like. In practice, it is often the case that for technical or other reasons it is not possible to change some of the conditions for the optimal course of biosynthesis. Such a variable factor may be, for example, the intensity of the fermentation broth mixing. Recently, as a result of the energy crisis, efforts have been made to prolong the cultivation time in order to reduce the amount of fermentation deployments, thereby saving the energy of the asuria in biosynthesis and chemical isolation into the final product. For this reason, in the development of long-term fermentation, following the increase in yield, the development led to an increasing concentration of the main soil components. However, increasing the nutrient concentration is limited by the fact that excessive growth of the biomass causes disruption of the metabolism optimum. In this case, due to insufficient mixing, oxygen levels are limited. The limiting factor in some cases may be the adverse effect of increased osmotic pressure.

By increasing the concentration of nutrients, the concentration limit of the harmful factors may be exceeded in one or more components of the culture medium. This fact has also been demonstrated in the tyrosine biosynthesis, namely the inhibition of antibiotic-NH4 + production by ions in fast nitrogen consumption (S. Omura: Control of Ammonium Ion Level in Antibiotic Fermentation, Fifth Internattional Symposium on Actin- mycetes Biology, Oaxtepec, Morelos, Mexi-co, August 16—19, 1982, Abstracts pp. 71 to 72).

If, at the start of the culture, the concentration of -NH4 + ions was increased, the fermentation of tylosin by Streptomycesfradiae resulted in excessive growth of biomass at the production phase and thus by oxygen limits. At the same time, the production of tylosin in nutrient medium was reduced,

It has been found that excessive growth of bio-mass, and thus high viscosity, can be prevented by the sterile water being continuously or semi-continuously dispensed at the end of the growth phase. This maintains the viscosity at which no oxygen climates occur. At the same time, conditions for continuous or semicontinuous addition of NH4 + ions and —NCb-ions in the production phase of the fermentation are formed. By dosing —NH4 + and —NOS “ions that use at. the metabolism and continuous or semi-continuous maintenance of pH in the 6.8-7.1 range maintains a good physiological state of the culture.

Maintaining optimum viscosity and optimum pH and dosing —NHi + and —NO3 “ions in the production portion of the fermentation resulted in a high yield of tylosin, but also in the suppression of desmicosine and other decomposition products of tylosin.

Fermentation in laboratory fermentation tanks.

Vegetatively inoculum was prepared in 500 ml boiling flasks filled with 60 ml of inoculum (corn extract, yeast extract, soybean oil, soybean oil and CaCCU). The nutrient broth after inoculation was inoculated with a spherical agar spore and incubated on a rotary shaker for 48 hours. The vegetative inoculum thus prepared was inoculated with a laboratory fermenter with sterilized soil of the same composition as used in 500 ml boiling flasks. Labo-rat fermentor with a total volume of 20 lbol filled with 10 l of nutrient broth. Cultivation was run at 29 ° C, with 6 lsterile air / min. The growth of the vegetative inoculum in the laboratory fermentor took 28 hours. The prepared inoculum was seeded with 10 L of production soil in a laboratory fermentor with a total volume of 20 L. The production soil contained beet molasses, soybean oil, fish crust, corn torture, corn gluten, inorganic salts and CaCO 3. The fermentation was carried out for 9 days with a culture temperature of 28 ° C and with an additional air of 40 hours of culture of 6 1 / min. and from 40 hour of culture 8 1 / min.

After 9 days, a production of 5,500 mcg milliliter was achieved. Example 2

Fermentation in laboratory fermentation tanks with regulation of fermentation conditions

Claims (1)

Process for the fermentative production of tylosin in aerobic cultivation of high-production mutants of Streptomyces fradiae microorganism under intensive stirring, in a culture medium containing sources of assimilable carbon, nitrogen and phosphorus and mineral nutrient, characterized in that the pH is maintained continuously or semi-continuously throughout the cultivation. 7-7.1, preferably 6.8-7.0, in the production phase, the —NH4 ⁺ and —NO3- ions are added continuously or semi-continuously in an amount to maintain an ammonia nitrogen concentration of 0.02-0. 30%, preferably 0.03-0.20%, and to maintain optimal viscosity, continuously or semi-continuously sterile water is added from the end of the growth phase.