SU339191A1 - METHOD FOR CULTIVATING MICROORGANISMS - Google Patents

Description

The invention relates to the microbiological industry, more specifically to methods for the cultivation of microorganisms.

A known method of cultivation of microorganisms is involved, including preparing a seed culture, sowing it on a nutrient medium and illuminating the visible area with light.

The proposed method allows to intensify the process of biosynthesis, to increase the yield of substances produced by microorganisms and to accelerate certain stages of the development cycle. This is achieved by subjecting the seed culture to light before sowing it on a nutrient medium, and the light is transmitted with a spectral range of 300 to 650 nm and an intensity of no more than 5 to 10 erg / cm-sec for 1 to 30 minutes.

The essence of the invention is as follows.

The seed of microorganisms (inoculum) immediately before sowing is illuminated with the light of the spectral composition of 300-650 for 1 to 30 minutes. The intensity of light does not exceed 5-10 erg / cm-sec.

The pre-seeding illumination causes a significant intensification of the vital processes of microorganisms, for example, protein and vitamin biosynthesis, and acceleration of certain stages of the development cycle during

sowing of microorganisms on the culture medium.

For a number of microorganisms, for example, Pseudomonas fluorescens, Clostridium butiricum, Schizosaccharomyces pombe, a stable effect was found to increase the yield of protein products up to 75%, increase the speed of cell division to 170%, reduce the duration of lag phase by half (from 6 hours to 3.5 hours). ), increasing the yield of vitamin B to 130%. The light of the spectral composition of 300-390 plays the main role in the integration of the processes of biosynthesis in microorganisms.

One of the main features of the proposed method is the use of microorganism cultures in experiments that are in a strictly defined physiological age.

Example 1. Pseudomonas fluorescens culture was subcultured twice in m septon broth with 0.1% saltpeter. The third reseeding was done in such a way that by the beginning of the harvest, the seed had 18 hours of age.

The seed obtained is poured into quartz test tubes and illuminated for 1-30 minutes with monochromatic light from a DRSh1000 lamp. After that, irradiated and non-irradiated samples are sown on Wednesday

test tubes. A paraffin layer is applied to the surface of the medium to create strictly anaerobic conditions. After seeding, all specimens with P. lluorescens osteppepym cultivars are put into a thermostat at. At the time of entering the stationary phase of development in culture, the total protein content is determined from and the number of cells is counted.

PRI mme R 2. Use the spore material Clostridium butiricum, which is obtained as follows. A 25% potato mash is poured in a high layer into microbeads, with a little chalk at the bottom. Before the nasal medium, they are heated 40–60 min by a boiling water bath. After cooling the tubes to 30-40 ° C, 0.2 ml of the spore material is introduced into each of them. Fermentation lasts 7-10 dpi at. By filling the tube, the tubes are sealed and stored at a tempo. Before setting the experience, spores are germinated in a glucose-peptone medium; glucose 2%, peptone 1%, KH2PO4 0.1%, chalk pas dpe tubes, tap water. Before sowing, the medium is heated as in example 1. The seed in the amount of 1% of the volume of the medium is added to the bottom of the quartz tube and grown for 24 hours at a tempo at. Quartz test tubes with seed culture will be inspected with light from the BUV-30 lamp for 1-30 minutes or with monochromatic light from the DRSh-1000 lamp. From irradiated and non-irradiated samples of C1, Buliricum were sown on synthetic medium with 1% glucose, 0.1% fumaric acid, 0.3% (NH4) 2HPO4, potassium in the composition of the CPC 140 mg / ml, biotinol - 0.001 mg / ml , RP environment 6.7-6.8. Then all are imaged with svezhezase other cultures C1. bntiricum is placed in a single thermostat at 37 ° C. After 21 hours of growth at the end of the exothermal phase of development in culture, the total protein content is determined and the number of cells is counted.

Example 3. Schizosaccharomyses pombe inoculum is used, which is seeded twice before the experiment on Gaisep medium: 10 g peptone, 3 g KH2PO4; 4 g MgSO47H2O, 20 g

GLUCOSES in 1000 ml of distilled water. The third reseeding was done in such a way that by the beginning of the experiment the inoculum was at the age of 24 hours. After that, the ipocool is illuminated for 1–30 min with monochromatic light from the DRSh-1000 lamina. Illuminated and unlighted samples Sch. The pombe is subcultured into Reader medium flasks, poured with a Toiki layer for good aeration.

Then all samples with fresh

cultures placed in a temp thermostat at 30 ° C. After 16 hours of growth (nasledp one-third of the expo-phase phase), the total protein content in the culture is determined and the number of cells is counted.

In examples, the total protein content is determined according to Lowry, and the number of cells is calculated according to Vipogradsky. The error in determining the percentage of stimulation of the formation of total protein in the experiment compared with the control does not exceed 4%, and when counting cells. The determination of the amount of vitamin B2 is carried out on a fluorometer.

Subject invention

Claims (2)

1. A method of cultivating microorganisms, including preparing a seed culture, sowing it on a nutrient medium and refreshing the visible area with light, characterized in that, in order to increase the yield of substances produced by microorganisms and speed up individual stages of the development cycle, the seed culture is subjected to illumination before sowing on a nutrient medium. 2. A method according to claim 1, characterized in that the illumination is carried out with a light of a spectral range of 300-650 with an intensity of no more than 510 erg / cm-sec for 1-30 minutes.