DE2308059C3 - Process for the biosynthetic production of griseofulvin - Google Patents

Description

Hs are already numerous processes for biosynthesis: of griseofulvin by means of various species of the genus Penicillium. how /. B. by means of the P.patulum-, P.griseofulvum and P.urticae strains known (US-V patents 3 095 360. 3 069 329 and 3 069 328).

In these known methods, culture media are used, their composition in terms of the nitrogen, carbon and other component sources depending on the metabolic peculiarities the species or the strain used is varied. The physico-chemical growing conditions (Temperature, ventilation, stirring process, duration, etc.) are both the trunks in the known methods as well as the media used.

The known methods (U.S. Patents 3,095,360, 3,069,329 and 3,069,328). at which Siamese of P.griseofulviim and P.urticae used have the disadvantage that low activities (antibiotic concentration) are achieved. In the known processes according to the above-mentioned USA patents, inoculation (Inoculation) of the medium / ur production of the preculture (the vegetative inoculum) in agar-agar-containing Spore suspensions generated by culture media are used. These methods have in particular the The disadvantage that the biosynthesis potential becomes unstable over time and that constant laboratory processing of cultures is necessary.

In some of the known processes (U.S. Patents 3,069,329, 3,069,328) in which Penicillium patulum strains, methyl group donors are used as stimulators, and the Carbon source is added continuously during the biosynthesis process, which has disadvantages in terms of additional expenses for the materials and plant costs required for continuous addition are connected.

According to the present invention, a method / for the biosynthetic production of Griseofulvin by growing Penicillium griseofulvum or Penicillium urticae in a culture medium that is characterized by. that the culture medium from 5 to 15 g / l corn germs. 5 to 8 g / l corn extract, HM) to 140 g / l lactose 7 to 15 g'l glucose. IO to 14 g I (calcium carbonate, I to 2g I potassium chloride, 6 to 10 g / l monopotassium phosphate, 0.05 to 0.1 gl magnesium sulfate. 1 to 2 g / l urea and 2 to 10 g'l hydricitem sunflower oil (Sonit) consists.

g / l

G/

G/

G/

g / gi

G/

g / l

The invention is based on the following exemplary embodiment explains in which strain Penicillium urticae was used:

a) Preparation of the spore-bearing inoculum l-jHsehalle. soaked with water and sterilized

Millet or rice grains were treated with a spore

suspensiiin inoculated. those from one by selection

new and pre-tested pure culture with!.: '. ^ m

Potential has been prepared.

The licbrühing was up to the welcome spore formation led, the culture then dried under vacuum urrJ! stored at 4 C for up to 2 years, being the survivable wedge of the spores and the biosynthetic potential during this time good words showed.

b) Preparation of the I. vegetative inoculum 1 of a medium of the following composition! 2Ci were prepared:

Corn extract (50 ".,) 5

Sodium acetate 2

Monopotassium phosphate 4

Potassium chloride 0.5

Magnesium sulfate 0.5

Hiseiui! »-Sulphate 0.01

Glucose (100 "ig calculated) 28

Hydrogenated sunflower oil (Sonit) 1 pH value 5.95 to 6.0

After sterilization, spore-bearing material was used nipft and at 27 to 28 C with stirring with a Belüf'ungsquote of 1 1 air / l / minute, at a gauge pressure of 0.3 to 0.5 atm for 25 to .55 30 hours, with under optimal transferring conditions a biomass of 20 to 30 ". ', characterized by the II. to Ml. degree metachromatic Corpuscles.

c) Preparation of the II. vegetative inoculum 25 (X) 1 of a medium of the following composition were prepared:

Soybean meal 4

Calcium carbonate 8

Monopotassium phosphate 4

Glucose (100% calculated) 20

Lactose 20

Sodium chloride 3

Ammonium sulfate 2

Magnesium sulfate 0.01 g / l

Hydrogenated sunflower oil (Sonit) 1 g / l pH value 5.95 to 6.0

After sterilization, 5 to 10% vol / vol des I. Inoculated vegetative inoculum and 18 to 25 hours among those used in the 1st vegetative inoculum Conditions bred.

g / l

g / l

g / i

g / l g / l

g / l

g / l

d) The actual biosynthesis 30 tons of the medium of the following composition were prepared:

Corn germ 5 to 15 g / l

Corn extract 5 to 8 g / l

^fi 5 lactose KX) up to 140 g / l

Glucose 7 to 15 g / l

(. "aleium carbonal 10 to 14 g / l

Potassium chloride I to 2 g / l

3 4

Mortokaliumpho-, phat ft to lüg I regulated the whole sterilization system with water

Magnesium sulfate 0.05 to (J. I g 1. One must note that the sterilization time

Urea 1 to 2 u I for a batch (30 t) 4 to 5 hours not

Hydrogenated sunflower oil strides. Both the extension of the sterilization

(Sonit) 2 to IO g I 5 duration as well as increasing the sterilization temperature

pH value 5.S to 7.5 door leads to a partial degradation of those present Substances, especially the sugar, which were used in temples in a 15 to 20 ton container Tons of raturcn over! 20 C caramelized, which is negative Introduced water, the maize flour and the maize impact.

Extract added, and 15 to 60 minutes at 100 C io. After inoculation with mild II. vegetative inoculum

cooked. In another identical vessel were in a ratio of 5 to 10 ", vol / vol was the

6 tons of water the lactose and glucose at Üiosynthesepro / .eß with the following parameters

55 to 65 C dissolved, then passed through the thermally treated:
ten corn cimmihl and extract. In a / ur

For the preparation of the salts, a 500-liter jar was set at 15 temperature: 26 to 30 C.

the monopotassium phosphate. the potassium chloride, the Bclühunt - during the first 12 hours between

Magnesium sulfate and the urea dissolved and then - _{schcn (! S uiul} | ₂ 1 air / 1, minute,

the solution obtained the other components / u _nadl , ₂ 'hours and until the end

given. In a 5-ton vessel, the CaI- _der (.ermentation between 1.2 and

cium carbonate slurry prepared, which is then 20 "> 1 air minute
was also given about the other components.

The volume calculated for the mixing formula was pH value: the natural pH value between

de filled with water, the pH value is 10 to see 5.8 and 7.0.
I5 "igcr sodium hydride solution to 5.7 to 6.3

and the hydrogenated sunflower oil as an anti-25 The fermentation process lasted 300 to 350

foaming agent added. Hours and an activity of 12,000 gamma / inl

The medium thus prepared was obtained for sterilization.

carried out according to a continuous process. The method according to the invention has the advantage

Sterilization was carried out at 120 to 125 C. on. that yields of 12,000 gamma / ml are achieved

In order to avoid the breakdown of the sugars, the cleaning process can be carried out under convenient conditions.

this takes place separately at a lower temperature, since there are no intermediate products of the

(115 to 120 C), but only after biosynthesis are formed.

Claims (1)

Claim: Process / ur biosynthetic production of Griseofulvin by growing Pcnic.llium griseofulvum or Penicillium urtieae in a κι. turmedmm. d: i d u r c h g e k c η η / c i η et. that the culture medium from 5 to 15 g-'l Ma.skemun. 5 to 8 g / l corn extract. IÜÜ up to 140 g I lactose, 7 to 15 g / l glucose. 10 to 14 g / l calcium carbonate. 1 to "» g / l potassium chloride, 6 to 10 g / l monopotassium phosphate, 0.05 to 0.1 g / l magnesium sulfate. 1 his 2 g-l urine peat and 2 to 10 g-l hydrogenated sunflower oil (Sonit) considered.