DE2166090B2 - Process for the preparation of d-threo-2-propionamido-1-p-nitrophenyl-1, 3-propanediol and d-threo-2-isobutyramido-1-p-nitrophenyl-1, 3-propanediol. Eliminated from: 2120153 - Google Patents

Description

The invention relates to a method for producing 25 (mixture of C ₁₂ - C ₁₅ )
of d-Threo-2-propionamido-lp-nitrophenyl-l, 3-pro-

pandiol (referred to here as substance A) and d-threo- The cultivation was taking place at 3O ⁰ C in 65 hours

2-isobutyramido-1-p-nitrophenyl-1, 3-propanediol (here stirring (650 r. P. M.) And aeration (11/1 / min.) With

referred to as substance B) by fermentation. The sterilized air running. The pH of the medium

substances mentioned are derivatives of d-Threo-2- 30 was automatically added to 6.5 to 6.5 with ammonia water

amino-l-p-nitrophenyl-l, 3-propanediols (chloramphe-6,8 adjusted to the medium three times each

nicol), in which the 2-amino group is added by propionamide 100 ml η-paraffin every 12 hours. That

or isobulyramide is substituted. Via a process n-paraffin substrate was after fermentation was complete

for the production of these substances directly by Fermen- almost consumed.

The fermentation broth obtained (21) has not yet been reported anywhere by Entin the art. removal of the microbial cells to a pH of 4.0. The method according to the invention is thereby established. After the same amount of ethyl acetate had been added, it was indicated that the following microorganisms were shaken the ferment broth for 24 hours in order to extract the analogs under aerobic conditions in a culture medium which contains the assimilable gene of chloramphenicol in the ethyl acetate layer to form carbon. The liquid thus extracted (2.3 1) was cultured and then dewatered, the reaction mixture in usual with anhydrous Na _s SO ₄ and Licher at 35 ° C worked up manner: evaporated under reduced pressure. The received

The residue was further washed with ethyl acetate (100 ml)

Corynebacterium hydrocarboclastus ATCC15 592, extracted. After centrifugation, the solution was

Nocardia gloverula ATCC 13 130, 45 medium-layer through a silica gel column (through

Arthrobacter parafineus ATCC 15 590, knife: 4 cm, height: 20 cm) sent, which then

Corynebacterium equi ATCC 10 146. was washed thoroughly with chloroform. Thereafter

a mixed solution of ChIo-

When carrying out the cultivation, co- form and methanol (95: 5) are sent,
Hydrogen or carbohydrates as the main carbon source. In the course of the elution, substance B was used in the oil. It is preferable to use different fractions from 200 to 500 ml and the substance A in the hydrocarbon fractions, which fraction _{elutes from 600 to 1000 ml, each of which contains η-paraffins, especially C 10} -C ₂₂ , preference - was collected and concentrated separately. Each conical C ₁₂ -C ₁₈ n-paraffins. They are supplemented by centered fraction was dissolved in ethanol, and the organic N sources, such as corn water, yeast extract, substances A and B were then crystallized separately by distilling off the inorganic metal salts of iron, manganese, of the solvent. On these magnesium, potassium, sodium. Substances A (4 g) and B (24 g) were isosterilized in the medium and inoculated with the microorganism. The loosing.

Cultivation is carried out under aerobic conditions at 20 to The crystals thus isolated have each been through

45 ° C carried out. During the cultivation, the 60 repetition of the recrystallization from ethylene di-

pH adjusted to 4 to 10, preferably 6 to 8, e.g. B. Chloride cleaned.

by adding a urea solution of ammonia- The isolated crystals of substances A and B have

water, ammonia or an ammonium carbide have lower antibiotic activities than those of des

natural solution. The cultivation is over after 2 to 7 days chloramphenicol, however, regarding des

guided. The completion of the breeding can through the 65 arstibiotic spectrum the same tendencies. Both

Values of the chloramphenicol analogs obtained so- substances A and B had a UV absorption maxi-

can probably be determined as well as determined by the mum of 273 πιμ, which was the same as that of the

"Paper disc method" to achieve a maximum. Chloramphenicols.

Example 2

Nocardia gloverula ATCC13 130 was grown in a culture medium similar to that described in Example 1 cultured, but with the difference that n-paraffin was replaced by glucose. After 75 hours were 0.9 g / l substance A and 0.3 g / l substance B are produced.

Example 3

IO

Arthrobacter praaffineus ATCC 15 590 was described in a manner similar to that in Example 1, however grown with the difference that the da-Qs fraction (400 ml) of the η-paraffin was added. To After culturing for 98 hours, 1.2 g / l of substance A and 1.7 g / l of substance B were obtained.

Example 4

Corynebacterium equi ATCC10146 was grown in 24 hours in a culture medium containing 2% ao glucose, 1% polypeptone, 1% meat extract and 0.3% table salt. The obtained seeds were inoculated into a medium having the following composition in a 5-1 breeding flask at a ratio of 10% by volume. Cultivation was then continued for 72 hours at 30 ^° C. (aerobic).

Sucrose 8%

K ₂ HPO ₁ 0.1%

MnSO ₄ 4H ₂ O 0.01%

(NH ₄ ) _S HPO ₄ 0.3%

Na ₂ HPO ₄ 0.1%

MgSO ₄ -7H ₂ O 0.1%

ZnSO ₄ 4H ₈ O 0.001%

Yeast extract 1.0%

The pH of the medium was automatically adjusted to 6.5 bis during the cultivation with ammonia water

6.8 set.

After working up in the usual way, 2.5 g / l substance A and 0.8 / l substance B were isolated.

Claims (1)

The following examples explain the invention: Claim: Example 1 Method for the preparation of d-Threo-2-pro- It was with shaking in 24 hours Coryne- pionamido-l-p-nitrophenyl-l, 3-propanediol and 5 bacterium hydrocarboclastus ATCC 15 592 in one d-Threo ^ -isobutyramido-i-p-nitrophenyl-l ^ -pro- medium grown, the one composition of pandiol through fermentation, resulting in 1% meat extract, 1% peptone, 0.5% NaCl and 2% indicates that one of the following sorbitol and a pH of 7.2 (before sterilization) Had microorganisms. With the obtained seed culture, 31 became one ίο inoculates culture medium, which the subsequent feed ATCC 15 592, composition in a 5-1 glass fermenter at a ATCC 13 130, ratio of 10 percent by volume had:
ATCC 15 590, ATCC 10 146 KH ₂ PO ₄ 0.2% I ₅ MgSO ₄ -7H ₂ O 0.1% in a culture medium which contains assimilable FeSO ₄ ■ 7H ₂ O 0.02% Contains carbon sources, under aerobic conditions- (NH ₄ ) ₂ SO ₄ 0.5% breeds and then the reaction corn water 0.1 / _o mixed up in the usual way. η-paraffin 10% (V / V) ac Na ₂ HPO ₄ 0.2% MnSO ₄ 4H ₂ O 0.002% ZnSO ₄ -7H ₂ O 0.001% CuCI ₂ · 2H ₂ O 0.0003% Yeast extract 0.5%