SU1082815A1 - Process for preparing l-lysine - Google Patents

Abstract

A method for producing oL-lysine by deep cultivating its microorganisms / catches on a nutrient medium containing molasses as a carbon source, a nitrogen source, essential mineral salts and a lysine biosynthesis stimulator, under conditions of aeration of the medium, followed by release of the target product, characterized by that, in order to increase the yield, 1 5% aqueous extract of seaweed of the genus FUCUS .vesiculosus in an amount of 0.02-0.8 g / l is used as a stimulator of the biosynthesis of cC-lysine in the amount of 0.02-0.8 g / l (Lredy.

Description

The invention relates to the microbiological industry, and specifically to methods for producing L-lion by fermentation.

  IfSfeecTeH is a method for producing 1-lysine by submerged cultivation of microorganisms producing it in a nutrient medium containing mineral salts, with the introduction of carbon sources necessary for the growth of amino acids into the medium or continuously, followed by release of L, -lysine from culture fluid one of the known methods. Corn extract, potato clutch juice, yeast hydrolyzate, or yeast extract are used as growth stimulants. Fermentation is carried out at 3032 ° C, pH 7.0-7.2, continuous aeration and stirring l.

It is known to use extracts of various algae as stimulants for the growth of microorganisms, such as Chlorella 2, green algae S.acuminatus NW, as well as extracts of seaweed of the genus FUC.US veciculosus.

However, the above algae extracts, in particular, aqueous or alcoholic extracts of seaweed of the genus F. Vesiculosiis are used as stimulators of biomass growth, i.e. their use in the cultivation of yeasts, mushrooms Aapergillus niger and p ski.Lemna minor aimed at increasing the yield of biomass.

Closest to the proposed method for producing L-lysine by submerged cultivation of its producers on a nutrient medium containing molasses, as a carbon source, corncurrant extract or enzymatic hydrolyzate of yeast cells as a biosynthesis stimulator and nitrogen source and mineral salts, with aeration of the medium followed isolation of L-lysine by a known method 51.

The disadvantage of this method is that it does not provide a sufficiently high yield of L-lysine (33.4 39, 0 g / l), and also involves the use of expensive and scarce components - a source of nitrogen, which is corn extract.

The purpose of the invention is to increase the yield of L-lysine.

The goal is achieved by the fact that according to the method of obtaining C-lysin, which involves the submerged cultivation of microorganisms producing it on a nutrient medium containing molasses as a carbon source, a nitrogen source, essential mineral salts and a lysine biosynthesis stimulator, under conditions of aeration of the medium with subsequent release the target product, as a stimulator of biosynthesis of L, α-lysine, use 1–5% aqueous extract of algae of the genus Fucus vesicu losus in an amount of 0.2–0.8 g / l of medium.

 The extract is prepared by infusing dry algae for 2-4 hours at 25-30 ° C, after which it is centrifuged, sterilized and injected 100 16 hours after the seed is introduced into the nutrient medium. ,

The cultivation of the producers of I, α-lysine Corynebacterium glutamicum T-3 or the VNIIgenetika strain 4995, or 5 Brevibacterium Sp 221, or 22 L, D is carried out on a nutrient medium of the following composition, wt.%;

Molasses16-21 (sugar

48%)

- Acid hydroli BVK3,5-4,1

Corn

extract1.0-2.0

1-5% aqueous 5 sea extract

algae0.002-0.008

NH.C11.5 - 2.5

Mel1.5 - 2.5

The tap water has a pH of 7.4. The rest

EXAMPLE 1. To obtain C lysine, a culture of the genus Cor.glutamicum T-3 is used, which is grown by Growing on a nutrient medium of the following composition, wt.%:

Molasses16 (sugar

48%)

Acidic hydrolyzate BVK 3,5 Corn extract 1,0 0 NH.Cl1,5

Mel1,5

The water is tap The remaining pH of the medium is 7.4. 10 h after the application of the inoculum grown in test tubes on MPA agarized media, sterile 1% centrate of an aqueous extract of seaweed of the genus Fucus.vesicuQ losus is added at the rate of 0.2 g / l (0.002 wt.%)

The aqueous extract is prepared from dry seaweed of the genus Fucus vesiculosus by insisting for 2 hours at 25 ° C. At the end of the extraction, the suspension is centrifuged

at 6000 rpm, sterilized for 0.5 h at 1.0 ati and the resulting crescent is used as a stimulator in the biosynthesis of L-lysine.

Fermentation is carried out in Erpenmeyer flasks with a capacity of 250 ml under laboratory conditions on a shaker (240 rpm). The process takes place at 30 + and pH 7.4. The volume of fermentation medium 10 ml. At the same time, at the output of I, lysine (determined by the method of paper electrophoresis in culture liquid is 30 g / l, EXAMPLE 2. The mode of cultivation of the producer of t-lysine Cor. Gl tamicum VNIIgenetika 4995 analogue to example 1 . A nutrient medium of the following composition is used, wt.%: Molasses19 (sugar 48 Acid hydroli3, 8 bw BVK 1.5 Maize extract 2.0 2.0 tap water The balance of the medium is 7.4, 13 h after the inoculation the material is added to the ptstelyuyu environment sterile fugat, 3% aqueous extract of sea water of the genus FU CUS vesiculosus in an amount of 0.5 G / L. The infusion time of dry algae is 3 parts at 30 ° C. After 72 fermentations, the yield of L-lysine in the cultured liquid is 45 g / l Froze. , -lysin Brevibacterium Sp.2L. is similar to examples 1 and 2. A nutrient medium of the following composition is used, wt .-%: 21 (sugar Molasses 48%) Acid hydrol4, 1 bVC 1.5 Corn extract 2.5 2.5 The water in the water pH of the medium is 7.4. After 16 parts after adding the inoculum, sterile centrate of 5% aqueous extract of algae of the genus Fucus vesiculosus is added to the nutrient medium. 0.8 g / l. Dry algae infusion time is 4 hours at 35 ° C. The yield of U-lysine in the culture liquid after 72 hours of fermentation is 39.1 g / l. EXAMPLE 4 The mode of cultivation of the producer of C-lysine B.Sp 22LD is similar to Example 3. After 72 hours of fermentation, the activity of lysine is 42, 8 g / l. Thus, the proposed method allows to increase the yield of lysine from 30 to 45 g / l as compared with the known (33.4 - 39.6 g / l).

Claims (1)

METHOD FOR PRODUCING cL-LYSINE by deep cultivation of microorganisms producing it on a nutrient medium containing molasses as a carbon source, nitrogen source, necessary mineral salts and a lysine biosynthesis stimulator, under conditions of aeration of the medium with subsequent isolation of the target product, characterized in that, in order to increase the yield, 1 5% aqueous extract of seaweeds of the genus Fucus, vesi culosus in the amount of 0.02-0.8 g / l dreadlocks is used as a stimulator of d · lysine biosynthesis. containing misoli, with the introduction into it or continuously of the sources of growth stimulators necessary for the growth of the amy; losus in an amount of 0.2-0.8 g / l of medium. * The extract is prepared by infusion of dry algae for 2-4 hours at 25-30 ° C, after which it is centrifuged, sterilized and introduced 1016 hours after the inoculation of the seed material.