SU767191A1 - Method of culturing microorganisms - Google Patents

Description

(54) METHOD FOR CULTIVATING MICROORGANISMS

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This invention relates to the microbiological industry, namely to a method for cultivating microorganisms.

There is a known method of cultivating microorganisms on acidic nutrient media containing sources of carbon, nitrogen, phosphorus, and mineral salts, which involves seeding into a fermenter from the inoculant of the same tlJ culture.

The disadvantages of this method are the relatively high values of the yield of the target products and the productivity of the fermenters.

The aim of the invention is to increase the yield of the target products and increase the productivity of farmers.

 This goal is achieved by the fact that according to the proposed method, the seeding is carried out by a culture adapted to one or several carbon-containing components of the culture medium.

Depending on the ultimate goal of cultivation, adaptation is carried out in various ways by growing the culture in the inoculum on the original nutrient medium in

periodic mode to the state in which it proceeds to the disposal of metabolic products; by growing the culture in inoculum 5 torus on a nutrient medium containing one or more products of metabolism as the main carbon source; by grown. inoculum culture on

to a nutrient medium containing carbon in an e-source and one of the mixtures of carbon sources in the nutrient medium used in fermentor 1,

) 5 Seed in culture fermenter,

adapted to one or several carbon-containing components of the culture medium, leads to the fact that in the fermenter simultaneously develop two or more fractions of the same culture, different in relation to substrate i of the nutrient medium. This leads to a more complete utilization of the carbon-containing components of the environment, and, consequently, to an increase in the economic coefficient or yield of the target products.

The proposed method is as follows:

30 following examples.

Approx. 1. The culture of Candida tropicaEis YeBFM-303 yeast is grown in a AK-10 fermenter per 6 liters of modified medium Reader of the following composition, (g / l):

Glucose, 20

LN4NgP048,0

(NH4) 2HPO.8,0

KSe. . 0.4

MDEA04.7 HgO1.0

PeS04-7 HgO0.005

SLOT -7 HjO0,0006

ZnS04 -7 HjO0,0008

CuS04 -5 NgO0.00008

Yeast water 20 ml

The cultivation is carried out in periodic mode. After 12 h, no glucose is detected in the medium. The yield of biomass cultivation is 8.5 g / l. During the subsequent days, the biomass concentration does not change significantly. The economic rate of 44%.

According to the proposed method, a culture of yeast in an inoculum, i.e., is harvested on a medium above for 36 hours in a batch mode. During this time, the culture goes to the consumption of metabolic products. The main fermenter of AK-10 is grown in the same mode. After 12 h, culture is sown in this fermenter. From an inoculum in the amount of 600 ml (10 of the volume of the culture medium). Through the final biomass, the main fermenter is 11.5 g / l (minus the sub-biomass), and the economic coefficient is 57.5%.

Example 2. Culture. Aerobacter aerogenes tank is grown in order to obtain acetoin at a flow rate of 0.2 h in Lermentera AK-Yu per 5 l of medium containing composition, g / l: glucose 10.0, peptone 5fco 5.0, CrNRO-MH O 2.0, 2.0, СаСЕ2 0.2, Na2SiO3 0.05, FeS047H O 0.005, ZnSO4-7H2O, MnSO4 5H.jO, CoCC2 ertjO and NajMob 2H2P no 0 ,. Reduction pressure: pH 6.0, temperature L l / h. Neshalki speed 800 rev / min. At the exit of the fermenter, the culture fluid contains 20.4-10 tank

teriapnyh cells / ml. Acetone yield

  -., ... ,,, .... ... ,, ,,,. ,,,.

In order to increase the yield of acetone, a system of two successively connected inoculators is used, each of which is combined with the main fermentor. The working volume of the primary inoculant is 1 liter, the second one is 1.5 liter. In both inoculators, they grow in AeroBiactef aerogehes periodically for 12 hours on the above medium, except that 2% ethanol is used as a source of carbon in the second inoculant. .

After 12 hours, the duct from the first inoculum is turned on to the main fermenter at a rate of 0.2 hours and seeded for 5 hours. At the same time, fresh nutrient medium is fed to the first inoculum at the same rate. Then, the duct from the first inoculum is switched to the second inoculum, from which the continuous continuous sowing of the adapted culture to ethyl alcohol is fed into the main fermenter. Fresh fresh medium is fed at a rate of 0.2 hours. At the exit of the fermenter, the culture fluid contains 24.3.10 bacterial cells / ml. The output of acetoin 1.3 g / l

EXAMPLE 3. A culture of Candida troDicaCis IBFM-303 is grown in 5 liters of a modified Reader medium containing, g / l: NHiHyPOj 8.0, GNH4) 2IR041-8.0, KCE 0.6 NaCf MgS047Y2 J If O GO 0.004, MnSO4-7H, jO 0.0004,. O, O 00 4, Syzod 0.0002, yeast water 20 ml, glucose 10.0, maltoza 10.0 in fermenter AK-10 at a flow rate of 0.25 h. The concentration of substances at the exit of the fermenter, glucose 0.01%, maltose 0.90%, biomass 6.94 g / l, ekontichesky coefficient 34.7%.

According to the proposed method, the processes are carried out using a bakterrpo of three inoculums; oors (each 1.5 liter in volume) connected in series. The second and third inoculators are connected to the main fermenter with a volume of 6 liters.

The yeast is seeded in the first inoculum and cultured periodically on the above medium for 12 hours. Then the fresh nutrient is started to flow at a flow rate of 0.2 at the same rate as the culture fluid from the first inoculant to the second. The filling of the second inoculum, tO15E1Pr6, occurs in 7.5 hours after which cultivation is continued in batch mode for another 7 hours. Since the first inoculant is almost completely consumed with glucose, the source of carbon in the second inoculant is mainly maltose. The yeast leaving the second inoculum is adapted to maltose. From the second inoculum, the culture medium is fed 150 ml each to the main fermenter and the third inoculum. In the third inoculum, cultivation is carried out in an intermittent mode of 36 hours. In this case, the culture is transferred to the product of the metabolism of maltose. Then third

the inoculum is connected to OSIOVNO1 U

the fermenter with Ckopost feed culture medium 150 ml / h.

Thus, a mixture of the initial and maltose-adapted cultures from the second inoculum of the trr and adapted from the products of the metabolism of maltose from the third inoculum is sown into the main fermenter. In addition, in the main fermenter serves a fresh nutritious chapters with a flow rate of 0.25 H-.

Upon reaching a steady flow regime of the fermenter, the concentration of substances is determined at the outlet: glucose 0.03%, maltose 0.01%, biomass 14.87 g / l. Economic coefficient is 74.35%. Example 4, the Culture of Lactobacteriim deBbrucku is grown periodically with aeration

1 l / l per minute at, at the barley wort 3 ° B, at an absolute value of 7, and at the initial biomass concentration taken from the exponential growth phase. About 34-10 ° cells / ml. After 12 hours of cultivation, the yield of lactic acid is 49.8%. The process ends after 26 hours. The yield of lactic acid is 90% (see table);

According to the proposed method, cultivation of the Lactobacteriun deCbriJcku culture is carried out in a fermenter in an analogous manner for 3 hours, after which the culture is seeded from an inoculant, in which the cultivation is carried out on the initial medium for 26 hours. The process in the fruit fermentor ends after 18 hours.

Claims (4)

Claim 1. A method of cultivating microorganisms in liquid nutrient media 45 containing sources of carbon, nitrogen, phosphorus and mineral salts, which involves inoculating the fermenter from an inoculator of the same culture, as described above. In order to increase the yield of target products, seedlings are carried out by a culture adapted to one or more carbon-containing components of the culture medium. 2. The method according to claim 1, characterized in that the adaptation is carried out by growing the culture in an inoculator on the initial nutrient medium in a batch mode until the state in which it proceeds to the utilization of metabolic products. 3. The method according to π. 1, with the fact that the adaptation is carried out by growing the culture in an inoculator on a nutrient medium containing, as the main carbon source, one willow of metabolic products, 4. The method according to π. 1, characterized in that the adaptation is carried out by growing the culture in an inoculator on a nutrient medium containing, as a carbon source, one of a mixture of carbon sources of the initial nutrient medium.