KR830001444B1 - Method for preparing lysine and glutamic acid using immobilized cells - Google Patents

Abstract

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Description

Method for preparing lysine and glutamic acid using immobilized cells

The present invention relates to a method for producing lysine and glutamic acid by batch or continuous fermentation using immobilized cells. This method not only saves raw materials and energy than conventional batch fermentation, but also improves productivity in terms of productivity. In the present invention, the selection of the carrier necessary for immobilization is very important, but the polyacrylamide gel previously used for immobilization of enzymes is very disadvantageous because it is difficult to immobilize and also harms the cells during immobilization. .

However, in the present invention, the immobilized carrier was selected as a carrageenan and researched and developed, and thus, the gel can be efficiently used to immobilize the cells because the gel has good water transfer properties, high strength, and simple immobilization. This was confirmed.

In order to achieve the present invention, the first cell immobilized on the carrier is low in both concentration and titer, so it is incubated in the seed medium for 48 hours to activate the titer to the maximum. The purification method heats the carrageenan solution to 80 ° C. After melting, the mixture was cooled to 45 ° C., and then mixed with a certain amount of the bacterial culture solution. The mixture was hardened at room temperature and cured for about 1 hour in 2% potassium chloride solution.

Optimization of the cell immobilization method and optimization of the physicochemical properties of the immobilized gel are one of the important features of the present invention.

The maximum yield reached 25% when the gel of activated immobilized cells in the air stirred fermenter was added and batch fermented. In the case of continuous, it was found to be four times better than the conventional batch fermentation method.

Investigation of the effect on operating conditions was also carried out to find and confirm the optimal conditions such as the aeration rate for lysine and glutamic acid.

The productivity and yield were also investigated continuously under the optimum conditions obtained above. As a result, the yield increased as the average residence time increased, while the productivity showed the maximum value. The maximum productivity was found to be 0.265 kg glutamic acid / ton / hr. As a result of continuous operation to investigate the stability of the carrier at the average residence time showing the maximum productivity, the gel was not broken and the initial form was maintained even after more than one month of operation, and the titer was also maintained without decreasing. It is another feature of the present invention to adjust the immobilized cells well to maintain a certain growth rate. As a result, when the production of lysine and glutamic acid using immobilized cells, the raw material and energy can be greatly saved than the conventional batch fermentation method, and at the same time, the invention of the production process technology that can produce lysine and glutamic acid by continuous fermentation This is another feature.

Hereinafter, the present invention will be described in detail as follows.

Example 1

Immobilization Method and Activation Method

The 4% (w / v) carrageenan concentration is dissolved by heating to 85 ° C and then slowly cooled to 45 ° C. At this time, 10% (v / v) of the culture medium incubated to 45 ℃ in advance. Mix the carrageenan solution and the bacterial culture well, and pour into a flat plate of constant thickness and allow to cool at room temperature for 30 minutes. The gel was cut into squares of constant size and then cured with stirring for 1 hour in 2% potassium chloride solution. The gel thus cured was activated for 48 hours in the spawn medium to maximize the titer of the gel.

Example 2

Batch Fermentation of Glutamic Acid Using Immobilized Cells

The immobilized cells activated in Example 1 were fermented batchwise in an air agitating reactor. At this time, the gel volume 50mι, the medium volume 280mι, the ventilation rate was 6.0VVM and the pH was adjusted with 1N NaOH solution. As a result, a maximum yield of about 25% was obtained based on the carbon source.

Example 3

Continuous Fermentation of Glutamic Acid Using Immobilized Cells

The immobilized cells activated in Example 1 were operated continuously in an air stirred reactor with a gel volume of 30 mι, a medium volume of 270 mι, and a ventilation rate of 6.0 VVM. As a result of examining the production rate according to the average residence time, the maximum was obtained in about 10 hours. Continuous operation at the maximum value obtained above allowed operation for more than a month without decreasing the potency.

Example 4

Batch Fermentation of Lysine Using Immobilized Cells

Fermentation was carried out batchwise in an air agitating reactor using the immobilized cells activated in Example 1. At this time, the gel volume was 23mι, medium volume 280mι, ventilation rate 6.0VVM, and adjusted with PH 1N NaOH solution.

As a result, a maximum yield of about 30% was obtained based on the carbon source.

Example 5

Continuous Fermentation of Lysine Using Immobilized Cells

Fermentation was carried out continuously in an air agitating reactor using the activated immobilized cells as in Example 1. Conditions at this time were 40mι of gel volume, 260mι of medium volume, 6.0VVM of aeration rate, and the pH was adjusted to 1N NaOH solution. As a result of continuous operation, the maximum yield was about 25% and it could be operated continuously for more than 15 days.

Claims (1)

Using carrageenan, the corynebacterium and glutamicum (ATCC 13058) cells are immobilized, and then optimized in a continuous or batchwise manner, while maintaining the activity of the immobilized cells. Lysine and glutamic acid using immobilized cells characterized in that glutamic acid (L-Glutamic aci) or lysine (L-Lysine) is produced under fermentation conditions (PH, 7.0; 30 ° C; exhaust rate, 6 times per liquid volume per minute). Manufacturing method.