SU258845A1 - METHOD OF MANUFACTURE 1-LYSINE - Google Patents

Description

The present invention relates to the microbiological field, in particular the production of L-lysine.

There are known methods for producing /. Lysin and by preparing sowing bacterial cultures, 1 for example, Brevibacterium 22, the producer of 1t / .lizi a, and a sterile nutrient medium containing; sources of carbon, nitrogen and a number of physiologically active veU; ecTB, followed by L-lysine biosynthesis (main fermentation) and its release from the resulting culture fluid.

However, with these methods of L-lysine production, the biosynthesis rate is reduced with a sufficiently high yield of L-lys iia.

To speed up the production process of Lipipa and increase its yield, a nutrient medium is proposed for the preparation of CeBHoii bacterially; Cultivators and fermentations should be supplied in a continuous flow in an amount that ensures at these stages the maintenance of a dilution factor of 0.03-0.2, while taking a bacteriological solution and producing a per-stream flow at the main fermentation.

The first strain No. 22 of the genus Vévibacteriuni, which grows on a dry feeder for 24 hours at ambient temperature, was taken from the surrounding environment of the amphosplotome, aprnmer – oleserindef cyt 1 strain No. 22 of the genus Веvibacteriuni. Your diet is salted with its physiological solution i in the amount of 5% in {alb. In flasks, irosteril 3opp 1a is in advance a medium that contains waste; in case of nitrogen, nitrogen, and a number of physiologically active substances. In flasks of seed about klyzru, 24 hours at. Pz flasks of seme1 1 st material are fed into the inoculum. Further 1st process IN eIRERYV OM MIOTOK.

Steril in O P tatelia on the medium in the reservoir, from which the ip of the dosing iiacoca is transferred to the O ocul tor. .i, a levels in the inoculum reaches 1 / 3-1 / 2 (from the working volume), culture is added from the spit (sludge from flasks). 6–12 hours after seeding, the incubation period H1b and the end period begin and the inflow with dilution coefficient (D) ranges from 0.03 to 0.2.

ACHIEVING 1 AND 10Ku.1HATOR volume and sowing material ipper :) 1 current is fed to the fermenter for wastes 1; 51 biosynthesis of L-lizi ia. At one oravre: flaxcoat, a fermeitaciophenium flask is added to the fermentation tank from the reservoir.

The dilution factor is maintained, as necessary, from 0.03 to 0.2. The fermentation temperature is 31 ± 1 C, the pH of the medium in the inoculum is 7.0-7.2, in the fermenters 6.2-7.0, the intensity of aeration in the inoculum

50-65 mg, and in the fermenters 35-

l / man

40 (but sulfite method), l / min.

The culture liquid after fermentation is collected in a separate container for further processing by one of the known methods, for example by evaporation and drying in a spray dryer, in order to obtain L-Lysine feed concentrate.

The seed is prepared by the homogeneous-continuous method in the inoculum with a useful capacity of 1.5 liters. At a flow rate of 07 (100 ml / hour), the highest biomass concentration is achieved. As the flow rate increases, the growth rate of the culture increases, and the concentration of biomass in the culture fluid decreases. At a flow rate of 2 (300 ml / hour), the cell tightness decreases, and leaching begins. With an increase in the flow rate, the residual sugar and pH increase, the L-lysine concentration decreases.

For the manufacture of seed material, any variation of the flow rate in the range of 03-0.2 can be used if necessary.

From the point of view of biomass production, the most profitable option is to cultivate the N-producer of L-lysine at a flow rate of 05 (75 ml / hour). The highest productivity (see table) is achieved in the cultivation mode with the flow rate of the medium, 17 (250 ml / hour). Under these conditions, the growth rate of the culture itself is also very high, with a 25% less biomass concentration than the dilution ratio of 0.03.

A continuous inoculum for 24 hours gives, at D 0.17, 250 ml / hour X24, 6000 ml / hour. For 24 hours in the same apparatus with a periodic method you can get 1.5 l of inoculated material.

Productivity of inoculum at different flow rates

The main L-lysine fermentation process in an uninterrupted stream is carried out in several interconnected fermenters. The dilution rate is 0.03-0.2, temperature 31 ±; 1 ° C, aeration intensity 35-40 mg Oa

L; min

Subject invention

A method for producing L-lysine by preparing a seed bacterial culture, for example, Brevibacterium 22, producer Lipipa, on sterilized nutrient medium containing sources of carbon, nitrogen and a number of physiologically active substances, followed by L-lysine biosynthesis (main fermentation) and isolating it from the resulting culture liquid, characterized in that, in order to accelerate the process of producing L-lysis, and increase its yield, nutrient medium is applied to prepare the seed bacterial culture and the main fermentation in a continuous flow in an amount that will ensure at these stages the maintenance of a dilution coefficient of the medium equal to 0.03-0.2, while preparing the bacterial culture is added continuously to the main fermentation stage.