DE2166090C3 - 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 ^ -propionamido-lp-nitrophenyl-lS-pro-

pandiol (referred to here as substance A) and d-threo- The cultivation was taking place at 30 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-1-p-nitrophenyl-1, 3-propanediols (chloramphe-6,8 adjusted. There were the medium three times each nicol), in which the 2-amino group is added by propionamide 100 ml η-paraffin every 12 hours. The or isobutyramide is substituted. Via a process n-paraffin substrate was after fermentation was complete Almost consumed directly by the company for the production of these substances.

tation using a bacterium has so far been 35. The obtained ferment broth (2 1) 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 - the fermentation broth - were shaken 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 sources. The liquid extracted in this way (2.3 l) was cultured and then the reaction mixture was dehydrated with anhydrous Na ₂ SO ₄ and worked up at 15 ° C.: evaporated under reduced pressure

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

Corynebacterium hydrocarboclastus ATCC15 592, extracted. After centrifugation, the solution became Nocardia gloverula ATCC 13 130, 45 middle 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. After that

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. Any conical C ₁₂ -Ci ₈ -n paraffins. They are supplemented by the 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. She is in a relationship.

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

45 C. During the cultivation, the recrystallization from ethylenedipH is repeated 60 times set 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 the sodium solution. The cultivation is finished after 2 to 7 days chloramphenicol, however, they had led with regard to the. The completion of the cultivation can through the 6 ₅ antibiotic spectrum the same tendencies. Both values of the obtained chloramphenicol analogs so-substances A and B had a UV absorption maximum determined and also determined by the mum of 273 ΐημ, which was the same as that of the "paper disc method" in order to achieve a maximum . Chloramphenicols.

Example 2

Nocardia gloverula ATCC 13 130 was produced in one culture medium similar to that described in Example 1, but with the difference that n-paraffin was replaced by glucose. After 75 hours, 0.9 g / l substance A and 0.3 g / l substance B were produced.

Example 3

Arthrobacter praaffineus ATCC 15 590 was cultured in a manner similar to that described in Example 1, but with the difference that the C _u -C _is fraction (400 ml) of the η paraffin was added. After culturing for 98 hours, 1.2 g / l of vcix 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% 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. Ajoschließend were grown for 72 hours at 30 ⁰ C on (aerobic).

Sucrose 8%

K-HPO ₄ G, l%

MnSO ₄ 4H ₂ O 0.01%

(NH ₄ ) ₂ HPO ₄ 0.3%

Na ₂ HPO ₄ 0.1%

MgSO ₄ -7H ₂ O 0.1%

ZnSO ₄ 4H ₂ O 0.001%

Yeast xiract 1.0%

The pH of the medium was during the cultivation automatically set to 6.5 to 6.8 with ammonia water.

ao 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-2-isobutyramido-l-p-nitrophenyl-l, 3-pro medium grown, which has a composition of pandiol through fermentation, resulting in 1% meat extract, 1% peptone, 0.5% NaCl and 2% indicates that one of the following sorbitol, as well as a pH of 7.2 (before sterilization) Had microorganisms. With the obtained seed culture, 3 1 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 10146 KH ₂ PO ₄ 0.2% 15 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 maize water OJ% mixed up in the usual way. η-paraffin 10% (V / V) 20 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%