DE448947C - Process for the production of nucleic acid from yeast - Google Patents
Process for the production of nucleic acid from yeastInfo
- Publication number
- DE448947C DE448947C DEE31387D DEE0031387D DE448947C DE 448947 C DE448947 C DE 448947C DE E31387 D DEE31387 D DE E31387D DE E0031387 D DEE0031387 D DE E0031387D DE 448947 C DE448947 C DE 448947C
- Authority
- DE
- Germany
- Prior art keywords
- nucleic acid
- yeast
- production
- alcohol
- iron
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired
Links
- 150000007523 nucleic acids Chemical class 0.000 title claims description 29
- 102000039446 nucleic acids Human genes 0.000 title claims description 29
- 108020004707 nucleic acids Proteins 0.000 title claims description 29
- 240000004808 Saccharomyces cerevisiae Species 0.000 title claims description 9
- 238000000034 method Methods 0.000 title claims description 6
- 238000004519 manufacturing process Methods 0.000 title claims description 3
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 9
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 8
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 claims description 6
- 239000001632 sodium acetate Substances 0.000 claims description 6
- 235000017281 sodium acetate Nutrition 0.000 claims description 6
- 150000002505 iron Chemical class 0.000 claims description 4
- 239000000706 filtrate Substances 0.000 claims description 2
- 229910001385 heavy metal Inorganic materials 0.000 claims 1
- 150000003839 salts Chemical class 0.000 claims 1
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 11
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 6
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 5
- 150000002506 iron compounds Chemical class 0.000 description 3
- 229960000583 acetic acid Drugs 0.000 description 2
- 239000003513 alkali Substances 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 229910052742 iron Inorganic materials 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 238000001556 precipitation Methods 0.000 description 2
- 239000011734 sodium Substances 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 1
- 229910021578 Iron(III) chloride Inorganic materials 0.000 description 1
- 150000001242 acetic acid derivatives Chemical class 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- 238000009412 basement excavation Methods 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 238000003776 cleavage reaction Methods 0.000 description 1
- 238000011109 contamination Methods 0.000 description 1
- 238000000354 decomposition reaction Methods 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 230000008021 deposition Effects 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 239000012362 glacial acetic acid Substances 0.000 description 1
- 235000014413 iron hydroxide Nutrition 0.000 description 1
- RBTARNINKXHZNM-UHFFFAOYSA-K iron trichloride Chemical compound Cl[Fe](Cl)Cl RBTARNINKXHZNM-UHFFFAOYSA-K 0.000 description 1
- NCNCGGDMXMBVIA-UHFFFAOYSA-L iron(ii) hydroxide Chemical compound [OH-].[OH-].[Fe+2] NCNCGGDMXMBVIA-UHFFFAOYSA-L 0.000 description 1
- 238000002386 leaching Methods 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 230000001376 precipitating effect Effects 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 230000007017 scission Effects 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- -1 sodium acetate Chemical class 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
- C12N15/1003—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor
Landscapes
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Biomedical Technology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Genetics & Genomics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Biotechnology (AREA)
- General Engineering & Computer Science (AREA)
- Crystallography & Structural Chemistry (AREA)
- Plant Pathology (AREA)
- Molecular Biology (AREA)
- Microbiology (AREA)
- Biophysics (AREA)
- Physics & Mathematics (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Analytical Chemistry (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Saccharide Compounds (AREA)
Description
Verfahren zur Gewinnung von Nucleinsäure aus Hefe. Die Gewinnung der Nucleinsäure aus Hefe durch Behandeln der Hefe mit Alkalien ist bekannt. Man erhält hierbei nach dem Abfiltrieren der Hefe eine alkalische Flüssigkeit, die auf Zusatz von Alkohol und Salzsäure die Nucleinsäure ausfallen läßt. Dieses Verfahren hat offenbar zwei Nachteile: einmal wird durch diese Fällungsmittel nicht nur Nucleinsäure ausgefällt, sondern auch noch andere Stoffe, die die Nucleinsäure verunreinigen; dann aber wird hierbei ein bedeutendes Vo-Iumen Alkohol benötigt. Diesem Übel abzuhelfen, wird die Nucleinsäure erst in die Eisenverbindung übergeführt, aus der dann mit einem besonderen Lösungsmittel für Nucleinsäure, nämlich Natriumacetat, die Nucleinsäure ausgelaugt wird.Process for the recovery of nucleic acid from yeast. The extraction of the Nucleic acid from yeast by treating the yeast with alkalis is known. You get here after filtering off the yeast an alkaline liquid, which on addition from alcohol and hydrochloric acid causes the nucleic acid to precipitate. This procedure has apparently two disadvantages: on the one hand, these precipitating agents not only produce nucleic acid precipitated, but also other substances that contaminate the nucleic acid; but then a significant volume of alcohol is required. To remedy this evil, the nucleic acid is first converted into the iron compound, from which then with a special solvent for nucleic acid, namely sodium acetate, the nucleic acid is leached out.
Ferner ist die Herstellung von nucleinsaurem Eisen aus Hefe bekannt. Bei der Erfindung dagegen handelt es sich um die Darstellung freier Nucleinsäure aus dem Eisensalz. Das Neue des vorliegenden Verfahrens besteht einmal in der Auslaugung des Eisensalzes mit Natriumacetatlösung und zweitens in der Fällung der freien Nucleinsäure aus dieser Acetatlösung mit Hilfe von Salzsäure und Alkohol. Auf diese Weise gelingt es, die Nucleinsäure in guter Ausbeute und in reinem Zustande zu gewinnen. Die Zersetzung der Eisenverbindung mit Alkali ist sehr unvorteilhaft, da freies Alkali die Nucleinsä.ure teilweise zerstört und so die Ausbeuten sehr herabsetzt. Außerdem erhält man auf diese ZVeise eine durch Spaltprodukte verunreinigte Nucleinsäure.The production of nucleic acid iron from yeast is also known. In contrast, the invention concerns the preparation of free nucleic acid from the iron salt. The novelty of the present process consists on the one hand in the leaching of the iron salt with sodium acetate solution and secondly in the precipitation of the free nucleic acid from this acetate solution with the help of hydrochloric acid and alcohol. In this way it succeeds it is necessary to obtain the nucleic acid in good yield and in a pure state. The decomposition the iron compound with alkali is very unfavorable, since free alkali the nucleic acid partially destroyed and so greatly reduces the yields. You also get on this ZVeise a nucleic acid contaminated by cleavage products.
Durch die Überführung der -Nucleinsäure in das unlösliche Eisensalz wird der große Ballast an Wasser aus dem Verfahren entfernt. Die darauffolgende Aushub ng mit Natriumacetat erfordert nur ein kleines Volumen, weshalb dann -die Fällung der freien Nucleinsäure mit wenig Salzsäure und wenig Alkohol vor sich gehen kann.By converting the nucleic acid into the insoluble iron salt the large ballast of water is removed from the process. The following Excavation ng with sodium acetate requires only a small volume, which is why then -the Precipitation of the free nucleic acid with a little hydrochloric acid and a little alcohol proceed can.
Beispiel. 5oo g abgepreßte Hefe werden mit starker Natronlauge übergossen und verrührt. Man kocht und säuert .alsdann mit Eisessig an. Da Nucleinsäure durch starke Essigsäure gefällt wird, so gibt man nur so viel Säure hinzu, daß eine ganz schwach saure Reaktion eintritt, also ein pH von etwa 5 bis 6. Die von der Hefe abzentrifugierte Flüssigkeit wird mit der genügenden Menge einprozentiger Eisenchloridlösung versetzt, worauf sich das nucleinsaure Eisen abscheidet. Man saugt dieses ab und laugt es mit ioprozentiger Natriumacetatlösung aus. Der Umsatz erfolgt, da Natriumacetat schwach alkalisch reagiert, wahrscheinlich nach folgendem Schema: (Nu)3Fe -;- 3 NaOH = Fe(OH)3 -E- 3 (Nu)Na (Nu = Nucleinsäurerest). Es entsteht das leichtlösliche nucleinsaure Natrium, während sich Eisenhydroxyd abscheidet. Es wird wieder abgenutscht und des Eisenrückstand auf dem Filter mit wenig Wasser gewaschen. Das klare Filtrat wird mit der gleichen Menge Alkohol und verdünnter Salzsäure versetzt. Die Nucleinsäure setzt sich zu Boden und kann nun abgesaugt werden. Zur besseren Trocknung wird der Niederschlag nochmals mit Alkohol nachbehandelt und nach Abtrennung # des Alkohols getrocknet. . . .Example. 500 g of squeezed yeast are poured over with strong sodium hydroxide solution and stirred. It is boiled and then acidified with glacial acetic acid. Because nucleic acid through If strong acetic acid is precipitated, only enough acid is added to make one whole weakly acidic reaction occurs, i.e. a pH of about 5 to 6. That of the yeast centrifuged liquid is mixed with a sufficient amount of one percent ferric chloride solution added, whereupon the nucleic acid iron is deposited. One sucks this off and leaches it with 10 percent sodium acetate solution. The conversion takes place because sodium acetate reacts weakly alkaline, probably according to the following scheme: (Nu) 3Fe -; - 3 NaOH = Fe (OH) 3 -E- 3 (Nu) Na (Nu = nucleic acid residue). The easily soluble one emerges nucleic acid sodium, while iron hydroxide is deposited. It is sucked off again and Washed the iron residue on the filter with a little water. The same amount of alcohol and dilute hydrochloric acid are added to the clear filtrate. The nucleic acid settles on the ground and can now be suctioned off. For better Drying, the precipitate is treated again with alcohol and after separation # of the alcohol dried. . . .
Wenn oben betont wurde, ;daß bei-der.Abscheidung der Nucleinsäure aus der Eisenverbindung keine Natronlauge verwendet werder, darf, da diese die NucIeinsäure teilweise zerstört, so steht dies nicht in unlösbarem Widerspruch dazu, daß. die Loslösung der Nucleinsäure aus dem Zellverbande mit Na-. tronlauge bewirkt werden soll. Denn bei dieser Maßnahme ist man auf Lauge angewiesen und muß daher auch die hierdurch bedingten Verluste und Verunreinigungen der Nucl.einsäure in Kauf nehmen.If it was emphasized above that during the deposition of the nucleic acid No sodium hydroxide solution may be used from the iron compound, as this is the nucleic acid partially destroyed, this is not in insoluble contradiction to the fact that. the Detachment of the nucleic acid from the cell structure with Na-. tronlauge be effected target. Because with this measure one is dependent on lye and must therefore also the accept the losses and contamination of the nucleic acid caused by this.
Claims (1)
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| DEE31387D DE448947C (en) | 1924-10-12 | 1924-10-12 | Process for the production of nucleic acid from yeast |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| DEE31387D DE448947C (en) | 1924-10-12 | 1924-10-12 | Process for the production of nucleic acid from yeast |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| DE448947C true DE448947C (en) | 1927-09-02 |
Family
ID=7075028
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| DEE31387D Expired DE448947C (en) | 1924-10-12 | 1924-10-12 | Process for the production of nucleic acid from yeast |
Country Status (1)
| Country | Link |
|---|---|
| DE (1) | DE448947C (en) |
-
1924
- 1924-10-12 DE DEE31387D patent/DE448947C/en not_active Expired
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