DE2321461C3 - Process for the fermentative production of L-lysine - Google Patents

Description

begins.

2. The method according to claim 1, characterized in that that acetic acid is used as an assimilable source of carbon.

3. The method according to claim I, characterized in that there is an assimilable source for Carbon ethanol used.

4. The method according to any one of claims 1 to 3, characterized in that for the Substances required for growth in the medium in the form of natural ones containing them Substances used.

The second type is caused by threonine or sensitive mutants and by threoninidiicne and mutants requiring threonine, growth is inhibited by a low concentration of threonine or methionine the latter mutant requiring threonine to grow. These mutants are in the FR PS 15 33 688 described. The third type is one Mutantc'which compared to S- (2-aminoethyl) -L-cysteine (hereinafter abbreviated as AÄC) which is the sulfur maloBC of L-lysine is permanent. These mutans " "hoof in the L) S-PS 37 07 441.

The invention is based on the object of providing L-Lys.n, us with readily available starting materials to manufacture at a low cost. .-Lysine is well known for human and animal nutrition “awkward. so that door this substance one further Can also be used as a supplement for food and drink Feed can be expected.

It has now been found that cine Mulantc. which is stable through a feedback control of F vsine and Lysm analogs and the nutrient requirement for at least one substance from the group of serine. Proline. Alanine. Nicotinamide. Pantothenic acid thiamine. Guanine, hpoxanlhm! And vitamin B _n "has, in the cultivation my Beeimictcn medium containing assimilable Kohlenstoffyucllcn. Stickstoffquellcn and minerals. Produced lysine, which is accumulated in the medium in large Mermen. The thus generated i.- Lysine can easily be obtained from the medium.

~ The certain strains listed below are used, which are able to generate L-Lys.n. and selected from mutants exhibiting resistance to lysine and the lysine analog S- (2-aminoethyl) -L-cysteine! Abbreviation AAC) and one or more of the above nutritional requirements own. .

The object of the invention is therefore a Process for the production of i.-lysine by aerobic; Cultivating a lysine producing strain of Brevibi-cterium oder'Corynebacterium in a medium which contains assimilable sources of carbon unc Nitrogen: Inorganic salts, substances required for growth and organic growth-promoting substances dcrndc contains substances, at a pH value of 5 to 9, which is characterized in that one al Lysine producing strain

60

The invention relates to a method for producing L-lysine by aerobically growing a lysine-producing one Strain of Brevibacterium or Corynebacterium in a medium which is assimilable Sources of carbon and nitrogen, inorganic salts, substances necessary for growth and contains organic growth-promoting substances, at a nll value of 5 to 9.

Brevibacterium
Brevibacterium
Brevibacterium
Brevibacterium
Brevibacterium
Brevibacterium
Brevibacterium
Brevibacterium

lactofermentum ATCC 21 798, lactofermentum ATCC 21 799. laclofcrmentum ATCC 21 800, lactofermentum ATCC 21 801. lactofermcnlum FFRM P-1711. lactofermenlum FFRM P-1712, lactofermentum 1'FRM P-1857, laclofcrmenuim IERM P-1570.

Brevibacterium lactofermentum FERM P-1573, Brevibacterium laciofermentum FERM P-1574, Brevibacterium lactofermentum FERM P-1575, Brevibacterium lactofermentum FERM P-1571. Brevibacterium lactofermentum FbRM P-1572, Corynebaclerium glutamicum FERM P-1613, Corynebacterium lilium FERM P-2026. Corynebacterium acetoacidophilum FERM P-2027,

Corynebacterium glutamicum FERM P-1982. Corynebacterium glutamicum FERM P-1984 or
Corynebacterium giutamicum FERM P-1985

begins. , ₅

The fermentative production of L-lysine under Consideration of certain nutrient requirements is already known. On the other hand, however, are the nutritional requirements the listed mutants to be used according to the invention (see below) completely new requirements. where was not yet known. that this has a specific effect on the generation of exercise i.-lysine.

The strains used in the method of the invention have been selected by selection or by conventional means Induction methods for artificial coals produced.

The strains used in the method according to the invention have the following properties and the following Nutritional requirements: jo

Brevibacterium lactofermenlum ATCC 21 798 (a strain that needs a herdsman and is resistant to AÄC).

Brevibacterium lactofermentum ATCC 2! 799 strain that requires pantothenic acid and is resistant to AÄC),

Brevibacterium laciofermentum ATC C 21 XOO (a strain that requires thiamine and is resistant to AÄC).

Brevibacterium lactofermenlum ATCC 21 KOi (a strain that requires adenine or guanine and is resistant to AÄC). Brevibacterium laciofermentum FERM P-1711 (a strain that requires alanine and is resistant to AAC),

Brevibaclenum lactofermentum 1-'ERM P-1712 (a strain that requires alanine or valine and is resistant to AÄC), Brevibacterium lactofermentum FERM P-1857 (a strain that requires alanine plus leucine and is resistant to AÄC), Brevibacterium lactofermentum FERM P-1570 (a strain that requires alanine plus leucine) S la mm I, Brevibacterimr.lactofermentum 1-'ERM P-1573 (a vitamin B ₁ , required and AAC-resistant strain) that is required and resistant to AÄC,
Brevibacterium lactofermentum FHRM P-1574 (a strain requiring nicotinamide and resistant to AAC).

Brevibacterium lactofermentum FERM P-1575 (a strain that requires I lypoxanthin and is resistant to AÄC),

Brevibacterium lactofermentum FERM P-1571 (a strain that requires nicotinamide and leucine and is resistant to AÄC). Brevibacterium lactofermentum F'ERM P-1572 (a strain that requires serine and leucine and is resistant to AÄC),
Corynebacterium gluiamicum FERM P-1613 (a serine-requiring and AÄC-resistant strain),

Corynebacterium glutamicum FERM P-1982 {requiring a proline and compared to AÄC constant strain),

Corynebacterium glutamicum FERM P-1984 (requires a guanine and compared to AÄC constant strain),

Corynebacterium glutamicum FERM P-1985 (a _{strain that requires vitamin B 12} and leucine and is resistant to AÄC),
Corynebacterium lilium FERM P-2026
(a strain that requires Ah'.nin and is resistant to AAC),
Corynebacterium acetoacidophilum
FERM P-2027

(a strain that requires alanine and is resistant to AÄC).

The specified FERM P numbers are the registration numbers at Fermentation Research Institutes. Agency of Industrial Science of Technology, the Ministry of the Industrial Trade an Industry. Japan. These strains with the FERM P numbers are available from this depository.

An advantage of the present invention over the conventional methods is that the certain new mutants that produce significant amounts of i.-lysine are made available be able. Another advantage of the invention is that the L-lysine productivity is not very high influenced by variations in the amount of threonine and the other nutrients in the medium because the strains used according to the invention are resistant to lysine analogs.

The composition of the formulation medium and the cultivation methods correspond to the conventional ones Processes used for the fermentation of amino acids. For the purposes media commonly used for conventional amino acid fermentations are suitable for the invention can be used, as well as the known media for the production of lysine by fermentation. Thus, for the cultivation of the strains used according to the invention either a synthetic one Culture medium or a natural nutrient medium are suitable as long as it contains the essential nutrients for the growth of the strains used. Examples of this are carbon sources. Sources of nitrogen and inorganic compounds produced by the microorganisms used in recycled in the appropriate quantities.

As carbon sources z. B. carbohydrates such as glucose, fructose, maltose. Sucrose, starch. Starch hydrolysates, molasses, and the like, as well as other suitable carbon sources such as organic acids. z. B. acetic acid, propionic acid, fumaric acid etc .. organic substances, / .. B. benzoic acid, or alcohols, e.g. B. methanol and ethanol are mentioned. These substances can be used either alone or in admixture of two or more.

Depending on the assimilability of the strains used, it is possible in the l'ermcnlalionsmedium to use hydrocarbons in larger or smaller quantities as a source of carbon.

Examples of suitable nitrogen sources are various Types of inorganic or organic salts or of compounds such as urea or Ammonium salts, e.g. ammonium chloride, ammonium sulfate, ammonium nitrate, ammonium phosphai 5 etc. as well as one or more amino acids or natural nitrogen-containing substances such as corn steep liquor, yeast extract, meat extract, fish meal, Peptones, bouillon, casein hydrolyzate. ^, Soluble fish products, Rice bran extract etc. These substances can be used either on their own or in a mixture of two or more can be used.

Examples of inorganic compounds that can be added to the culture medium are Magnesium sulfate, sodium phosphate, potassium dihydrogen phosphate, potassium monohydrogen phosphate, iron sulfate and other iron salts, manganese chloride, calcium chloride and sodium chloride.

The culture medium must contain nutrients and other substances that are necessary for the growth of the strain. The growth promoting agents and minor nutrients which improve the yield and the rate of production of i.-lysine include amino acids such as serine. Proline, alanine. Leucine etc., various vitamins 2s like vitamin B ₁₂ . Thiamine. Pantothenic acid. Nicotinamide (or nicotinic acid) etc. Purine bases such as guanine, adenine and hypoxanthine. Soybean protein hydrolysates, yeast extract, corn steep liquor. Peptones. Casein hydrolyzate and the like

It is an important factor. The amount of amino acid and other nutritional j. substances to less than the optimal concentration for the growth of the strain of microorganisms too limit what is a technique used in amino acid fermentations is applied fairly generally.

The fermentation is carried out at temperatures between 24 and 37 ¹ C for periods of about 24 to 96 hours and under aerobic conditions with shaking or aeration and stirring. The pH of the medium is adjusted between 5 and 9. If the pH of the medium falls below 5.0, then it is adjusted with neutralizing agents such as calcium carbonate and aqueous ammonia. When organic acids are used as carbon sources, the pH of the medium tends to rise. He is then by the addition of acids such. B. those organic acids that serve as a carbon source or adjusted by hydrogen chloride or sulfuric acid to a value within the abovementioned range. The L-lysine can be isolated from the culture broth by known methods. The bacterial cells can be removed by filtration or centrifugation. The i.-lysine can be isolated using cation exchange resins.

The amount of lysine produced in the culture broth was determined using lysine decarboxylasc definitely.

5 g / dl calcium carbonate and having a pH of 7.2 was prepared. 20 ml batches of the Medium were placed in 500 ml shake flasks and sterilized. The strains given in Table 1 were inoculated into the medium and cultured with shaking for 72 hours at 3PC. The amount of L-lysine formed (as hydrochloride) is shown in Table 1.

The culture media door the strains ATCC 2i 799 and ATCC 21 801 contained 5 mg / ml calcium pantothenate and 100 mg / l adenine and guanine, respectively.

The bacterial cells and solid calcium salts were obtained from the culture broth of the ATCC 21 798 strain Centrifugation removed. 1 1 of the supernatant liquid was placed in a column which with a strong cation exchange resin (crosslinked polystyrene with —SO, H groups) in the Hydrogen form was filled. The L-lysine adsorbed on the resin was washed with 3% aqueous Ammonia elutes. The eluate was concentrated under reduced pressure. To the concentrated Solution was given hydrochloric acid. It was chilled in a refrigerator to precipitate the i.-lysine. There was thus obtained crude crystalline L-lysine hydrochloride dihydrate in an amount of 30.2 g receive.

Example 2

Brevibacterium laclol'ermentum ATCC 21 798, ATCC 21 799, ATCC 21 8 (X) and ATCC 21 801 were each inoculated into a germination medium which contained 1.5 g / dl glucose. 0.3 g / dl ammonium acetate. 0.1 g / dl KH ₂ PO ₄ . 0.04 u / dl MgSO ₄ 7H ₂ O, 2 ppm Fe and Mn ions. 50! Ig / 1 biotin, 200 ag fThiamin HCl. 1.5 ml / dl soybean proteinhydrol) sat (with a total nitrogen content of 7%) and 0.5 g / dl yeast extract and which had a pH of 8.0. It was cultivated for 16 hours at 31 ° C. with stirring and aeration.

For inoculation, 15 ml of the individual germ cultures were added to 300 ml of a main culture medium which contained 3 g / dl glucose, 0.5 gdl ammonium acetate, 0.2 g / dl urea, 0.1 g / dl KH ₁ PO ₄ . 0.04 g / dl MgSO ₄ ■ 7H ₂ O, 2 ppm Fe and Mn ions, 50 ;;. G / l biotin, 15 [xg / l thiamine · HCl, 3 ml / dl soybean protein hydrolyzate and those in Table 2 contained nutrients shown and that had a pH of 7.5. Cultivation took place at 31.5 ° C. with stirring at 1500 rpm, an equal volume of air being introduced per minute. During the cultivation, the pH of the medium was kept between 7.2 and 8.0 by automatically adding a mixed solution of acetic acid and ammonium acetate. This contained 0.25 mol of ammonium acetate per mol of acetic acid. Its acetic acid concentration was 60%.

After 48 hours of cultivation, 4.8 to 6.4 g / dl were found in the culture broth of each strain I. Lysine hydrochloride found as shown in Table 2 is approved.

example 1

A culture medium containing 10gdl glucose. 5 g, -dl (NH ₄ I ₂ SO _4. 0.1 g / dl KH ₂ PO _4. 0.04 g dl MgSO ₄ · H ₂ O. 2 ppm Fe and Mn ions. 50 µg I thiamine-HCl. 50 ; ig / i biotin, 1.5 ml dl soybean protein indrolysis (with 7% total nitrogen) and

Example 3

Brevibacterium lactofcrmentum ATCC 21 798.

(.5 ATCC 21 799, ATCC 2180 and ATCC 21 801 were each cultivated for 16 hours at 31.5 C with shaking in a germ culture, which contained 1.5 u dl glucose, 0.3 g dl urea, 0.1 g / dl KH ₂ PO ₄ 0.04 g dl MuSO ₄ · 7H ₂ O.

I ppm Fe and Mn ions, 50 ag / 1 biotin, 200 ag I rhiamine hydrochloride, 1.5 ml / dl soybean protcinyl hydrolyzate and 0.5 g / dl kefe extract and which had a pH of 8.0.

15 ml of each Kcim culture was used da / u, .inoculate in 300 ml of a main culture medium / u, which contained the following components:

Glucose 1 i: dl

Ethanol 1.5 g dl

(NH ₄ ), SÜ ₄ 0.5 g / dl

Urea 0.2 g / dl

KH ₁ PO ₄ 0.1 g / dl

MgSO ₄ · 7 H ₂ O 0.04 g / dL

Fe ⁺ ^ 2 ppm

Mn ^{4 +} 2 ppm

Biotin 50 ag / 1

Vitamin B ₁ HC! 5 () '; xg / l

Soybean proline hydrolyzate

(Total nitrogen: 7%) 3 ml / dl

Nutrients according to Table 3
(pH: 7.5)

The cultivation took place at 31.5 ° C. with stirring at 1500 rpm, one volume of the medium being used the same volume of air was introduced.

During the fermentation, the pH of the medium was adjusted between 7.0 and 7.5 by introducing it of gaseous ammonia echoed into the medium.

The amount of remaining ethanol was determined by gas chromatography. Ethanol was added to the Entered medium when the remaining ethanol had decreased to about 0.3 g / dL.

After 48 hours, the culture broth of each strain was examined. It was found that she i.-lysine hydrochloride in the amounts shown in Table 3.

Example 4

Brevibacterium iaclofermentum FERM P-1570. FERM P-1571, FERM P-1572, FERM P-1573, FERM P-1574 and FERM P-1575 were used as seed strains used and cultivated in the same manner as in Example 1, except that in the in Table 4 specified amounts of soybean protein hydrolyzate was added and that to the medium for the Strain FERM P-1575 5 mg / ml hypoxanthine were given.

Table 4 shows the amounts of i.-lysine (as hydrochloride which after 72 hours of incubation formed in the resulting broth.

Example 5

Brevibacterium lactofermentum FERM P-1570. FERM P-1571. FERM P-1572. FERM P-1573. FERM P-1574 and FERM P-1575 were each used as seed strains and using acetic acid cultivated as a main carbon source as in Example 2, with the exception that 5 mg / ml hypoxanthine strain FERM P-1572 was added to the medium door.

In table 5 the amount of L-lysine (as hydrochloride) indicated which after 48 hours of incubation in the resulting broth of each strain had been formed.

After culturing the strain FERM P-1571 for 48 hours in the manner described above 30 percent by volume of acetic acid per initial volume of medium had been consumed. It was determined, that the Kullur broth 7.1 g / dl i.-lysine hydrochloride contained. One liter of culture broth gave 56.3 g of i.-lysine hydrochloride dihydrate according to the procedure of Example 1 obtained.

Example 6

ίο Brevibacterium lactofermentum FERM P 1570. IERM P-1571, FERM P-1572, FERM P-1573, FERM P-1574 and FERM P-1575 were each used as seed strains and using ethanol as the main carbon source in the same manner as in Example 3 except that the strain FERM P-1575 5 mg / ml hypoxanthine was added to the medium for this.

Table 6 shows that in the obtained broth of each strain after 48 hours of incubation amount of L-lysine obtained (as hydrochloride).

From Table 6 it can be seen that in the culture broth after 48 hours of cultivation 6.3 g of L-lysine hydrochloride (27.7% yield based on the ethanol used) were found. From one liter of the culture broth, following the procedure of Example 1, 50.4 g of i.-lysine hydrochloride dihydrate were obtained.

B e i s ρ i e 1 7

Brevibacterium lactofermentum FERM P-1711. FERM P-1712 and No. 872 (control sample) were used as the germ strains and cultivated in the same manner as in Example 1 except that the medium for the strain FERM P-1711 was added with 200 µg / 1 thiamine hydrochloride and 3% soybean protein hydrolyzate were given.
Table 7 shows the amount of formed lysine (as hydrochloride) in the resulting broth each; Strain after 72 hours of incubation.

Example 8

Brevibacterium lactofermentum FERM P-1711 FERM P-1712 and No. 872 (control sample) were each grown in a seed culture medium of the following Composition inoculated and cultivated at 3Γ C with stirring and ventilation.

Germ culture medium

glucose

Ammonium acetate

urea

KH ₂ PO ₄

1.5%

0.3%

0.1%

0.1%

MgSO ₄ 7H, O 0.04%

Fe ^{+ +} 2 ppm

Mn ^{+ +} 2 ppm

Biotin 50 ag / 1

Thiamine hydrochloride 200 ag /!

Soybean protein hydrolyzate

(with 7% total nitrogen) 2%

(pH: 7.5)

300 ml batches of a main culture medium with de The following compositions were placed in 1 -! - GIa: fermentation flasks.

509 685/31

/ LD 6 9

IlauptkulUirmcdium

Glucose - "'<>

Ammonium acetate 0.5%

Urea 0.2 "/ ,,

KH ₂ PO ₄ 0.1%

MgSO ₄ - 71I ₂ O 0.04%

Fe ^f '". 2 ppm

Mn ^M - ppm

Biotin 50 ag I.

Thiamine hydrochloride 50 ag 1

Soybean protein hydrolyzate

(with 7% total fuel) 2.5%

(pH: 7.5)

After sterilization, each medium was inoculated with 15 ml of the above seed culture broth of each strain. With stirring at 1500 rpm C was cultured at 31 to 33 ^1, wherein an equal Luflvolunicn per minute was initiated. During the cultivation, the pH of the medium was kept between 7.2 and 8.0 by automatically adding a mixed solution of acetic acid and ammonium acetal which contained 0.25 mol of ammonium acetal per mol of acetic acid and at which the acetic acid concentration was 60%.

After culturing for 55 hours, i.-lysine hydrochloride formed in the amounts shown in Table S.

Example 9

Brcvibaclcrium laetofermentum FERM P-17II. PERM P-1712 and Nr, 872 (control sample) were placed in a seed culture medium of the same composition inoculated as in Example 8, with the exception that for this 0.5% ethanol instead of ammonium acetate and 0.3% urea were given. It was stirred and aerated at 31 C for 18 hours cultivated.

Fermentation was carried out in the same manner as in Example 8 using the obtained ones Germ culture broth, with the exception that 1% glucose, 1% ethanol and 0.5% ammonium sulfate was added instead of ammonium acetal.

During the fermentation the pH of the medium was increased between 7.2 and 8.2 by introduction held in the medium by gaseous ammonia.

The amount of residual ethanol was determined by gas chromatography. Was in the medium Entered ethanol when the remaining ethanol content dropped to about 0.3%.

After 48 hours, the culture broth of each strain contained L-lysine hydrochloride in those shown in Table 9 specified quantities

Example 10

A culture medium of the following composition was prepared. 20 ml batches of this medium were placed in 500 ml shake flasks and 5 minutes Sterilized at 110 C.

Composition of the medium

Glucose 10%

(NH ₄ I ₁ SO ₄ 4.5%

KH ₁ PO ₄ 0.1%

MgSO ₄ - 7H ₂ O 0.04%

Fe ^{+ T} 2 ppm

Mn ⁺ "" 2 ppm

1 VV

Biotin 50 iig / 1

Thiamine hydrochloride 200 [ig / 1

Soybean prolein hydrolyzal

(with 7% total stickslol'O 1%

CaCO, 5%

(pH 7.0)

Brevibacterium lactofermentuin FERM P-1857, which has previously been cultivated on a bouillon gelatinous plant had been. was inoculated into the above culture medium. It was shaking for 72 hours cultivated at 31 C.

The result was 5.1 g dl i.-l.ysine (as hydrochloride) accumulated in the Kullurbrühc (51% yield, based on the glucose used).

Example 11

Corynchactcrium glutamicum FERM P-HiI3, FERM P-19H2, FERM P-1984, FERM P-1985, Corynebacterium lilium FERM P-2026 and C'orynebacterium aceloacidophilum FERM P-2027 were used as seed strains and, as in Example 10 described, cultured, with the exception that continued to the medium for the strain FERM P-1984 another 0.5% lleefy extract was given.

In Table 10 are the amounts of formed ! .- Lysine (as Ilydrochlorid) after a 72 hour Cultivation in the resulting broth of the individual strains put together.

Example 12

Coryncbactcrium glutamicum FERM P-1613, FERM P-1982. FERM P-1984, FERM P-1985. Corynebacterium lilium IERM P-2026 and Corynebacterium aeetoacidophilum 1 ERM P-2027 were identified as Germ strains used and, as described in Example 2, using acetic acid as Main carbon sources cultivated, with the exception that 0.5% yeast extract is added to the germ culture medium were given that the pH of the Kcimkullurmcdium was adjusted to 7.0 and that to the ilauplkuHurrnedium 2.5% soybean protein hydrolyzate were given.

The table shows the amounts of!, - lysine (as

Hydrochloride), which after 55 hours Cultivation in the resulting height of each strain had been formed

Example 13

Coryncbactcrium glutamicum PERM P-1613, Coryncbactcrium lilium FERM P-2026 and Coryncbactcrium aeetoacidophilum FERM P-2027 inoculated into the culture medium given below and inoculated for IX hours with stirring and aeration cultivated at 31 C.

Culture medium

Glucose 1.5%

Urea ... 0.3%.

KH ₁ PO ₄ 0.1%

MgSO ₄ - 7H ₁ O 0.04%

Fe ⁺ ^ ". 2 ppm

Mn ⁺ 42 ppm

Biotin 50; ig 1

Thiamine hydrochloride 2 (K); ig 1

Soybean protein hydrolyzate

(with 7%, total nitrogen) 1.5%

(pH 7.5)

23

300 ml batch of a main battery. Mediums of ¹ , the composition below, were placed in II glass mixing flasks.

Main culinary medium

(jlucose 1 "»

Ethanol 1%

(Nl I ₄ I ₁ SO ₄ 0.5 "ο

Urea 0.2%

KH ₁ PO ₄ 0.1 ",,

MgSO ₄ - 7 II, O 0.04 ",,

Fc "" '2 ppm

Mn * '2 ppm

Biotin 50; j.g 1

1 amine hydrochloride 50; jg 1 ' ⁵

Soy bean protein hydrolyzate

(with 7 "" Ciesaml nitrogen) 2.5 "/ ,,

IpH 7.5)

After sterilization, each medium was mixed with 15 ml of the cultured germ broths inoculated into various mutes. It was cultivated at up to 33 C with stirring, with the same Volume of air per minute was introduced. During the cultivation, the pH of the 2s Medium / between 7.2 and 7.H held by introducing gaseous ammonia into the medium.

The amount of residual ethanol in the medium was determined by gas chromatography. In the Ethanol was added to the medium as the concentration of the remaining ethanol had dropped to about 0.3%.

In Table 12 the amount of i.-lysine (as H vdrochloridl indicated which in the resulting Broth of the individual strains had been formed after culturing for 48 hours.

Table 1

461

Table 3

Table 4

Sla mm

FERM P-1570 FERM P-1571 FERM P-1572 FERM P-1573 FERM P-1574 FERM P-1575

Table 5

Amount of L> sin Ig dl)

12th

Addition of Hypoxanlhin

Img.'dl)

Mute 21 79 H. Admitted nutrient lOOmii / 1 and
100 mg / 1 Amount of
educated
Lysine 21 799 (g dl) ATCC 21 KOO 5.5 ATCC 21 Sol 4.4 ATCC 5.5 ATCC Ca pantothenate:
5 mg ml 5.7 Adenine:
Guanine:

Amount of
Soybean
proiein-
hydrolysal Amount of
educated
l.-lysine (%) Ig dll 3 2.9 1.5 5.0 1.5 4.6 3 3.5 2 3.1 3 3.0

Amount of 1-lysine formed

(g dl)

Brevibacterium laetofermentum ATCC 21 798

Brevibacterium lactofermcnUim ATCC 21 799

Brevibacterium laetofermentum ATCC 21 800

Brevibuclerium laetofermentum ATCC 21 801

Table 2

ATCC 21 798
ATCC 21 799

ATCC 21 800
ATCC 21 801

Additional nutrient

Ca pantothenate: 5 mg'ml

Adenine: 100mglund Guanine: 100 mg / 1

4.1 2.5 3.0 2.8

Amount of lysine formed

lg dl)

6.4

4.8

5.7 6.0

45

55 IERM P-1570 FERM P-1571 FERM P-1572 FERM P-1573 FERM P-1574 FERM P-1575

Table 6
tribe

60 FERM P-1570 FERM P-1571 FERM P-1572 FERM P-1573 FERM P-1574 FERM P-1575

5.4 7.1 7.0 6.5 5.8 6.1

adding
Hypoxanthine Amount of
educated
i -Lysine (mg / dl) tg-dll 5.2 - 6.3 - 6.5 6.1 5.0 5 5.5

. MJ ^ im &?

13th

Table 7

Sl LI 111 111

IHRM P-171 I.
FFRM P-1712

No. K72

Table X

Si am m

FERM P-17II
FERM P-1712

No. 872

Table 9

Amount of l-lysine formed

Ig dl)

4.4 3.2

1.8

Amount of l-lysine formed

(gdl)

7.2 6.1 4.0

Δ0 <L 1 ^ D 1 IO table tribe Today. related to
the cinucsel / le (ilueose
I "'., I P-1613 44 IO 1 ' ¹ IiRM P-1982 32 I FR M P-198 5 18th FIiRM P-1984 1 HRM P-2026 15th I HRM P-2027 1 "HRM Il table

14th

_{Consumes 20} Slaiiuii Lssigsaurc

T)

24

32

FHRM P-1613 FHRM P-1982 FFRM P-1984 FERM P-1985 IFRM P-2026 FFRM P-2027

Table 12

Amount of Howl. formed related to 1 -lysine the A set lilUL'ose Ig dl) ("nl 3.0 30th 3.0 30th 4.1 41 3.1 31 1 0 ^Ί 9

Amount of
educated
1 -lysine Yeihiauchli
Acid Acid Ig dlI ("111 5.0 T) 5.2 23.5 4.7 25th 6.4 23 5.5 24 5.8 T)

tribe

FERM P-1711
FERM P-1712

No. 872

Amount of 1-lysine formed

Igdll

6.6 5.6 3.6

Howling, based on the ethanol used

t "I ₁ )

26 20 17

Slamin

1 ERM P-1613 FERM P-2026 IFRM P-2027

Amount of
educated
I -lysine Howl,
be / ogen in
to be there
set
Ethanol (g dll 1 ".. 1 4.S 23 4.1 20.5 4 6 24

Claims (1)

Patent claims: 1. Method for producing L-lysine by aerobically growing a lysine producing strain of Brevibacterium or Corynebacterium in a medium containing assimilable sources for carbon and nitrogen, inorganic salts, substances necessary for growth and Contains organic growth-promoting substances at a pH value of 5 to 9, d a d u r c h characterized in that one is called an L-lysine-producing strain Brevibacterium lactofermentum ATCC 21 798, Brevibacterium lactofermentum ATCC 21 799, ' ⁵ Brevibacterium lactofermentum ATCC 21 800, Brevibacterium lactofermentum ATCC 21 801, Brevibacterium lactofermentum FERMP-1711.
Brevibacterium lactofermentum FERM P-1712,
Brevibacterium lactofermentum FERM P-1857.
Brevibacterium lactofermentum FERM P-1570,
Brevibai erium lactofermentum FERM P-1573.
Brevibacterium lactofermentum FERM P-1574,
Brevibacterium! Actofermenium FERM P-1575.
Brevibacterium lactofermentum FERM P-1571.
Brevibacterium laclofermentum FERM P-1572. ^ Corynebacterium glutamicum FERM P-1613. Corynebacterium Hlium FERM P-2026. Corynebacterium acetoaeidophiluin FERM P-2027. Corynebacterium glutamicum FERM P-1982, Corynebacterium glutamicum FERM P-1984 or
Corynebacterium glutamicum FERM-P 1985 The formation of L-lysine from _ fermentable carbohydrates by microorganisms is bere.ts Sannt Among the L-lysine-producing microorganisms are three types of bacteria representative be _ ^m. rw,. «te Tvp represents a mutant that belongs to the