ジエチルピロカルボネート
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- DEPC、diethyl pyrocarbonate
Wikipedia preview
出典(authority):フリー百科事典『ウィキペディア（Wikipedia）』「2017/09/22 00:12:50」(JST)
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Diethyl pyrocarbonate
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| Names |
| IUPAC name
Diethyl dicarbonate
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Other names
- Diethylpyrocarbonate
- Diethyl oxydiformate
- Ethoxyformic anhydride
- Pyrocarbonic acid diethyl ester
- DEPC
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| Identifiers |
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CAS Number
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3D model (JSmol)
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| ChEBI |
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| ChemSpider |
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| ECHA InfoCard |
100.015.039 |
| KEGG |
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| MeSH |
Diethylpyrocarbonate |
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InChI
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InChI=1S/C6H10O5/c1-3-9-5(7)11-6(8)10-4-2/h3-4H2,1-2H3 Y
Key: FFYPMLJYZAEMQB-UHFFFAOYSA-N Y
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InChI=1/C6H10O5/c1-3-9-5(7)11-6(8)10-4-2/h3-4H2,1-2H3
Key: FFYPMLJYZAEMQB-UHFFFAOYAU
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| Properties |
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Chemical formula
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C6H10O5 |
| Molar mass |
162.14 g·mol−1 |
| Appearance |
Clear, colorless liquid |
| Density |
1.101 g/mL at 25 °C
1.121 g/mL at 20 °C |
| Boiling point |
93 to 94 °C (199 to 201 °F; 366 to 367 K) at 24 hPa |
| Hazards |
| Main hazards |
Harmful |
| R-phrases (outdated) |
R22 R36/37/38 |
| Flash point |
69 °C (156 °F; 342 K) closed cup |
| Lethal dose or concentration (LD, LC): |
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LD50 (median dose)
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Oral - rat - 850 mg/kg |
| Related compounds |
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Related compounds
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Di-tert-butyl dicarbonate
Dimethyl dicarbonate |
Except where otherwise noted, data are given for materials in their standard state (at 25 °C [77 °F], 100 kPa).
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| Y verify (what is YN ?) |
| Infobox references |
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Diethyl pyrocarbonate (DEPC), also called diethyl dicarbonate (IUPAC name), is used in the laboratory to inactivate RNase enzymes in water and on laboratory utensils. It does so by the covalent modification of histidine (most strongly), lysine, cysteine, and tyrosine residues.[1][2]
DEPC-treated (and therefore RNase-free) water is used in handling of RNA in the laboratory to reduce the risk of RNA being degraded by RNases.
Water is usually treated with 0.1% v/v DEPC for at least 2 hours at 37 °C and then autoclaved (at least 15 min) to inactivate traces of DEPC. Inactivation of DEPC in this manner yields CO2 and ethanol. Higher concentrations of DEPC are capable of deactivating larger amounts of RNase, but remaining traces or byproducts may inhibit further biochemical reactions such as in vitro translation. Furthermore, chemical modification of RNA such as carboxymethylation is possible when traces of DEPC or its byproducts are present, resulting in impaired recovery of intact RNA even after buffer exchange (after precipitation).
DEPC treated water for use in a laboratory
DEPC is unstable in water and susceptible to hydrolysis to carbon dioxide and ethanol, especially in the presence of a nucleophile. For this reason, DEPC cannot be used with Tris or HEPES buffers. In contrast, it can be used with phosphate-buffered saline or MOPS.[3] A handy rule is that enzymes or chemicals which have active -O:, -N: or -S: cannot be treated with DEPC to become RNase-free, as DEPC reacts with these species. Furthermore, DEPC degradation products can inhibit in vitro transcription.
DEPC derivatization of histidines is also used to study the importance of histidyl residues in enzymes. Modification of histidine by DEPC results in carbethoxylated derivates at the N-omega-2 nitrogen of the imidazole ring. DEPC modification of histidines can be reversed by treatment with 0.5 M hydroxylamine at neutral pH.
DEPC can also be used for probing the structure of double-stranded DNA.<needs reference />
References
- ^ Chirgwin, John M; et al. (1979). "Isolation of biologically active ribonucleic acid from sourcesenriched in ribonuclease". Biochemistry. 18 (24): 5294–5299. PMID 518835. doi:10.1021/bi00591a005.
- ^ Wolf, Barry; Lesnaw, Judith A.; Reichmann, Manfred E. (1970). "A Mechanism of the Irreversible Inactivation of Bovine Pancreatic Ribonuclease by Diethylpyrocarbonate. A General Reaction of Diethylpyrocarbonate with Proteins". European Journal of Biochemistry. 13 (3): 519–25. PMID 5444158. doi:10.1111/j.1432-1033.1970.tb00955.x.
- ^ "FAQ about DEPC". Sigma-Aldrich. Retrieved 12 August 2012.
External links
- DEPC at OpenWetWare life scientist wiki
- RNase and DEPC Treatment
English Journal
- Purification and biochemical characterization of a moderately halotolerant NADPH dependent xylose reductase from Debaryomyces nepalensis NCYC 3413.
- Kumar S, Gummadi SN.SourceApplied and Industrial Microbiology Laboratory, Department of Biotechnology, Indian Institute of Technology-Madras, Chennai 600 036, India.
- Bioresource technology.Bioresour Technol.2011 Oct;102(20):9710-7. Epub 2011 Jul 22.
- A Xylose reductase (XR) from the halotolerant yeast, Debaryomyces nepalensis NCYC 3413 was purified to apparent homogeneity. The enzyme has a molecular mass of 74kDa with monomeric subunit of 36.4kDa (MALDI-TOF/MS) and pI of 6.0. The enzyme exhibited its maximum activity at pH 7.0 and 45°C (21.2U/m
- PMID 21855330
- Nucleosome exclusion from the interspecies conserved central AT-rich region of the Ars insulator.
- Takagi H, Inai Y, Watanabe SI, Tatemoto S, Yajima M, Akasaka K, Yamamoto T, Sakamoto N.SourceDepartment of Mathematical and Life Sciences, Graduate School of Science, Hiroshima University, Higashi-Hiroshima 739-8526, Japan.
- Journal of biochemistry.J Biochem.2011 Sep 19. [Epub ahead of print]
- The Ars insulator is a boundary element identified in the upstream region of the arylsulfatase (HpArs) gene in the sea urchin, Hemicentrotus pulcherrimus, and possesses the ability to both block enhancer-promoter communications and protect transgenes from silent chromatin. To understand the molecula
- PMID 21930654
Japanese Journal
- Characterization of Heme-Coordinating Histidyl Residues of an Engineered Six-Coordinated Myoglobin Mutant Based on the Reactivity with Diethylpyrocarbonate, Mass Spectrometry, and Electron Paramagnetic Resonance Spectroscopy(ENZYMOLOGY, PROTEIN ENGINEERING, AND ENZYME TECHNOLOGY)
- Nakanishi Nobuyuki,Takeuchi Fusako,Park Sam-Yong,Hori Hiroshi,Kiyota Kohei,Uno Tadayuki,Tsubaki Motonari
- Journal of bioscience and bioengineering 105(6), 604-613, 2008-06-25
- … We further investigated the reactivity of these possible His ligands with diethylpyrocarbonate(DEPC) to clarify the coordination structure and their protonation states in ferric form. …
- NAID 110006792536
- Purification and Characterization of A Novel (R)-Hydroxynitrile Lyase from Eriobotrya japonica (Loquat)
- UEATRONGCHIT Techawaree,KAYO Ai,KOMEDA Hidenobu,ASANO Yasuhisa,H-KITTIKUN Aran
- Bioscience, biotechnology, and biochemistry 72(6), 1513-1522, 2008-06-23
- … Metal ions were not required for its activity, while activity was significantly inhibited by CuSO4, HgCl2, AgNO3, FeCl3, β-mercaptoethanol, iodoacetic acid, phenylmethylsulfonylfluoride, and diethylpyrocarbonate. …
- NAID 10027528703
Related Links
- Diethylpyrocarbonate (DEPC), also called diethyl dicarbonate (IUPAC name), diethyl oxydiformate, ethoxyformic anhydride, or pyrocarbonic acid diethyl ester, is used in the laboratory to inactivate the RNase enzymes from water and other ...
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- diethyl pyrocarbonate、diethylpyrocarbonate
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- diethylpyrocarbonate、DEPC
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ジエチルピロカーボネート、二炭酸ジエチル
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- DEPC、diethylpyrocarbonate