# US 2008/0187967 A1 · Device and method for the production of biologically active compounds by fermentation > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 2008/0187967 A1 - Document ID: US20080187967A1 - Status: Published application - Kind: A1 - Application number: 12/001,959 - Filed: 2007-12-13 - Published: 2008-08-07 - Earliest claimed: 2005-06-17 - Assignees: MERZ PHARMA GmbH & CO. KGaA - Inventors: Bernd Doelle; Michael Pfeil - Web page: https://patentgrep.com/patent/US20080187967A1 ## CPC - C12M 41/14: {Incubators; Climatic chambers (per se B01L1/00)} - C07K 14/33: from Clostridium (G) - C12M 37/02: {Filters} - C12M 41/40: {of pressure} - C12P 21/02: having a known sequence of two or more amino acids, e.g. glutathione - Y10S 435/809: Incubators or racks or holders for culture plates or containers - C12M: APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; {APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS} - C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00) - C12P: FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE {(brewing of beer C12C; producing vinegar C12J; producing specific peptides or proteins C07K; producing enzymes C12N9/00; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification C12N15/00; measuring or testing processes involving enzymes or microorganisms C12Q; measuring or testing processes involving nucleic acid amplification reactions C12Q1/6844; fermentation processes to form a food composition, A21 or A23; compounds in general, see the relevant compound class, e.g. C01, C07)} - Y10S: TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS ## Abstract The present invention relates to a process and an apparatus for the fermentational production of biologically active materials, wherein a fermenter is located in an insulator which, in turn, is located within a working chamber or is adjacent to it. A pressure gradient in relation to ambient pressure prevails in both the insulator and in the working chamber. ## Claims ### Claim 1 An apparatus for the fermentative production of a biologically active compound, comprising at least a first insulator containing a fermenter, the first insulator being surrounded or adjacent to a working chamber, wherein the working chamber is connected to the environment via a pressure sluice, wherein a low pressure prevails in the insulator and in the working chamber, wherein the pressure (in relation to ambient pressure) in the insulator is lower than the pressure (in relation to ambient pressure) in the working chamber, and wherein an over pressure prevails in the pressure sluice in relation to ambient pressure. ### Claim 2 (depends on 1) The apparatus according to claim 1, wherein the pressure in the insulator is 20 to 200 Pa lower than ambient pressure. ### Claim 3 (depends on 1) The apparatus according to claim 1, wherein the pressure in the working chamber of 5 to 50 Pa is lower than ambient pressure. ### Claim 4 (depends on 1) The apparatus according to claim 1, wherein the pressure in the pressure sluice of 10 to 100 Pa is higher than ambient pressure. ### Claim 5 (depends on 1) The apparatus according to claim 1, wherein the apparatus comprises at least a second insulator, and wherein the second insulator does not contain a fermenter. ### Claim 6 (depends on 1) The apparatus according to claim 1, wherein the apparatus and/or the first insulator and/or the second insulator comprise at least a supply air duct and an exhaust air duct, wherein the supply air duct and the exhaust air duct comprise a filter, selected from a HEPA filter. ### Claim 7 (depends on 5) The apparatus according to claim 5, wherein the internal pressure of the first insulator is equal to the internal pressure of the second insulator. ### Claim 8 (depends on 5) The apparatus according to claim 5, wherein the internal pressure of the first insulator is different than the internal pressure of the second insulator. ### Claim 9 (depends on 5) The apparatus according to claim 5, wherein the first and the second insulator are connected with each other via a passage, wherein the passage allows for material transport from the first insulator to the second insulator and also allows for material transport from the second insulator to the first insulator. ### Claim 10 (depends on 9) The apparatus according to claim 9, wherein the passage is a sluice. ### Claim 11 (depends on 1) The apparatus according claim 1, wherein the apparatus comprises a sterilizing facility and/or a disinfection facility. ### Claim 12 (depends on 1) The apparatus according to claim 1, wherein the first insulator comprises an anaerobic working fermenter. ### Claim 13 (depends on 1) The apparatus according to claim 1, wherein the first insulator comprises a precipitation facility. ### Claim 14 (depends on 1) The apparatus according to claim 1, wherein the first insulator comprises a filtration facility. ### Claim 15 (depends on 5) The apparatus according to claim 5, wherein the second insulator comprises an extraction facility. ### Claim 16 (depends on 5) The apparatus according to claims 5, wherein the second insulator comprises a precipitation facility. ### Claim 17 (depends on 5) The apparatus according to claim 5, wherein the second insulator comprises at least a chromatography facility. ### Claim 18 (depends on 1) A process for the fermentational production of a biologically active compound comprising: a fermentation step for the production of the biologically active compound, a purification step for the biologically active compound, wherein the process is conducted in the apparatus according to claim 1. ### Claim 19 (depends on 18) The process according to claim 18, wherein the biologically active compound is a toxin or another protein obtained from fermentation, selected from a botulinum toxin and a botulinum neurotoxin. ### Claim 20 (depends on 19) The process according to claim 19, wherein the botulinum toxin is a botulinum toxin belonging to Clostridium botulinum of types A, B, C, D, E, F, and G, or is a mixture of two or more of these types. ### Claim 21 (depends on 20) The process according to claim 20, wherein the botulinum toxin or the mixture of botulinum toxins is/are a botulinum neurotoxin or a composition of botulinum neurotoxins. ### Claim 22 (depends on 18) The process according to claim 18, wherein the fermentation step occurs in the first insulator and the purification step occurs totally or partially in a second insulator. ### Claim 23 (depends on 18) The process according to claim 18, wherein the fermentation step and a part of the purification step occur in the first insulator and another part of the purification step occurs in the second insulator. ### Claim 24 (depends on 18) The process according to claim 18, wherein the fermentation step, a precipitation and filtration of a product of the fermentation step, occur in the first insulator as a part of the purification step. ### Claim 25 (depends on 18) The process according to claim 18, wherein the first insulator and the second insulator are operated at the same temperature or at different temperatures. ### Claim 26 (depends on 18) The process according to claim 18, wherein the temperature in the first and/or the second insulator are changed according to each process step. ### Claim 27 (depends on 26) The process according to claim 26, wherein the temperature in the first insulator is in a range of approximately 20° to approximately 50° C. and the temperature in the second insulator is in a range of approximately −5° to approximately +25° C. ### Claim 28 (depends on 18) The process according to claim 18, wherein the process comprises the following steps: a) in the first insulator inoculating fermentation media with a producing strain which produces a biologically active compound; fermenting the producing strain; separating supernatant from cells of the producing strain; and precipitating the supernatant; b) in the second insulator centrifuging the precipitated supernatant to obtain a pellet; extracting the pellet, centrifuging to obtain a supernatant; precipitating the supernatant with subsequent centrifuging to obtain a supernatant; c) in the first or the second insulator precipitating the supernatant; centrifuging the precipitate; solubilizing the pellet obtained by centrifuging the precipitate; dialysing the solubilized pellet and centrifuging the dialysate; performing chromatography of the dialysate; filtration of the dialysate to obtain an eluate.