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{{Orphan|date=February 2009}}
{{chembox {{chembox
| Watchedfields = changed
| verifiedrevid = 414422485 | verifiedrevid = 443637574
| ImageFile = Diethyl pyrocarbonate.svg | ImageFile = Diethyl pyrocarbonate.svg
| ImageSize = 250px | ImageSize =
| ImageFile2 = Diethyl pyrocarbonate3d.png | ImageFile2 = Diethyl pyrocarbonate3d.png
| ImageSize2 = 250px | ImageSize2 =
| IUPACName = diethyl dicarbonate | PIN = Diethyl dicarbonate
| OtherNames = {{bulletedlist|Diethylpyrocarbonate|Diethyl oxydiformate|Ethoxyformic anhydride|Pyrocarbonic acid diethyl ester|DEPC}}
| OtherNames =
| Section1 = {{Chembox Identifiers |Section1={{Chembox Identifiers
| ChemSpiderID_Ref = {{chemspidercite|correct|chemspider}} | ChemSpiderID_Ref = {{chemspidercite|correct|chemspider}}
| ChemSpiderID = 2943 | ChemSpiderID = 2943
| KEGG_Ref = {{keggcite|correct|kegg}} | KEGG_Ref = {{keggcite|correct|kegg}}
Line 17: Line 17:
| ChEMBL_Ref = {{ebicite|correct|EBI}} | ChEMBL_Ref = {{ebicite|correct|EBI}}
| ChEMBL = 55517 | ChEMBL = 55517
| EC_number = 216-542-8
| StdInChI_Ref = {{stdinchicite|correct|chemspider}} | StdInChI_Ref = {{stdinchicite|correct|chemspider}}
| StdInChI = 1S/C6H10O5/c1-3-9-5(7)11-6(8)10-4-2/h3-4H2,1-2H3 | StdInChI = 1S/C6H10O5/c1-3-9-5(7)11-6(8)10-4-2/h3-4H2,1-2H3
Line 23: Line 24:
| CASNo_Ref = {{cascite|correct|CAS}} | CASNo_Ref = {{cascite|correct|CAS}}
| CASNo = 1609-47-8 | CASNo = 1609-47-8
| UNII_Ref = {{fdacite|correct|FDA}}
⚫ | PubChem = 3051
| ChEBI = 59051 | UNII = LMR3LZG146
⚫ | PubChem = 3051
| ChEBI_Ref = {{ebicite|correct|EBI}}
| ChEBI = 59051
| SMILES = O=C(OCC)OC(=O)OCC | SMILES = O=C(OCC)OC(=O)OCC
| MeSHName = Diethylpyrocarbonate | MeSHName = Diethylpyrocarbonate
}} }}
| Section2 = {{Chembox Properties |Section2={{Chembox Properties
| C=6 | H=10 | O=5
| Formula = C<sub>6</sub>H<sub>10</sub>O<sub>5</sub>
| Appearance = Clear, colorless liquid
| MolarMass = 162.141 g/mol
| Density = 1.101 g/mL at 25 °C<br />1.121 g/mL at 20 °C
| Appearance =
| Density = | MeltingPt =
| BoilingPtC = 93 to 94
| MeltingPt =
| BoilingPt_notes = at 24 hPa
| BoilingPt =
}} }}
| Section3 = {{Chembox Hazards |Section3={{Chembox Hazards
| Solubility = | MainHazards = Harmful
| MainHazards = | FlashPtC = 69
| FlashPt_notes = closed cup
| FlashPt =
| Autoignition = | AutoignitionPt =
| LD50 = Oral - rat - 850 mg/kg
| GHSPictograms = {{GHS07}}
| GHSSignalWord = Warning
| HPhrases = {{H-phrases|302|315|319|332|335}}
| PPhrases = {{P-phrases|261|264|270|271|280|301+312|302+352|304+312|304+340|305+351+338|312|321|330|332+313|337+313|362|403+233|405|501}}
}}
|Section8={{Chembox Related
| OtherCompounds = ]<br />]
}} }}
}} }}
'''Diethylpyrocarbonate''' (DEPC), also called diethyl dicarbonate (] name), diethyl oxydiformate, ethoxyformic anhydride, or pyrocarbonic acid diethyl ester, is used in the ] to inactivate the ] ] from water and other laboratory utensils. It inactivates the RNases by the ]s of the ] residues. DEPC cannot be used with ] buffer or ] since they inactivate DEPC by reacting with it. In contrast it can be used with PBS or ]. 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 ].


'''Diethyl pyrocarbonate''' ('''DEPC'''), also called '''diethyl dicarbonate''' (] name), is used in the ] to inactivate ] ]s in water and on laboratory utensils. It does so by the ] modification of ] (most strongly), ], ], and ] residues.<ref>{{cite journal|author=Chirgwin, John M|year=1979|title=Isolation of biologically active ribonucleic acid from sourcesenriched in ribonuclease|journal=Biochemistry|volume=18|issue=24|pages=5294–5299|doi=10.1021/bi00591a005|pmid=518835|display-authors=etal}}</ref><ref>{{cite journal|doi=10.1111/j.1432-1033.1970.tb00955.x|title=A Mechanism of the Irreversible Inactivation of Bovine Pancreatic Ribonuclease by Diethylpyrocarbonate. A General Reaction of Diethylpyrocarbonate with Proteins|year=1970|last1=Wolf|first1=Barry|last2=Lesnaw|first2=Judith A.|last3=Reichmann|first3=Manfred E.|journal=European Journal of Biochemistry|volume=13|issue=3|pages=519–25|pmid=5444158|doi-access=free}}</ref>
⚫ Water is usually treated with 0.1% v/v diethylpyrocarbonate for at least 1hour at 37°C and then autoclaved (at least 15 min) to inactivate traces of DEPC. Inactivation of DEPC in this manner yields CO<sub>2</sub>, H<sub>2</sub>O and EtOH. Higher concentrations of DEPC are competent of deactivating larger amounts of RNase but remaining traces or byproducts may inhibit further biochemical reactions such as in vitro translation. Further on, chemical modification of RNA such as carboxymethylation is possible when traces of DEPC or its byproducts are present, resulting to reduced usage of RNA even after buffer exchange (after precipitation).

⚫ DEPC-treated (and therefore RNase-free) water is used in handling of ] 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&nbsp;°C and then ]d (at least 15 min) to inactivate traces of DEPC. Inactivation of DEPC in this manner yields CO<sub>2</sub> and ]. 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 transcription. 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 (and therefore RNase-free) water is used in handling of RNA in the laboratory, to reduce the risk of RNA being degraded by RNases.


DEPC is unstable in water and susceptible to hydrolysis to carbon dioxide and ethanol, especially in the presence of a ]. For this reason, DEPC cannot be used with ] or ] buffers. In contrast, it can be used with ] or ].<ref name=sigma>{{cite web|title=FAQ about DEPC|url=http://sigma-aldrich.custhelp.com/app/answers/detail/a_id/268/p/7,117/session/L3RpbWUvMTM0NDc1NzUxNC9zaWQvbzh6M3d1M2w%3D|publisher=Sigma-Aldrich|accessdate=12 August 2012}}</ref> 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 ] by DEPC results in carbethoxylate derivates at the N-omega-2 nitrogen of the ] ring. DEPC modification of histidines can be reversed by treatment with 0.5M ] at neutral pH.


⚫ DEPC ] of histidines is also used to study the importance of histidyl residues in enzymes. Modification of ] by DEPC results in carbethoxylated derivates at the N-omega-2 nitrogen of the ] ring. DEPC modification of histidines can be reversed by treatment with 0.5 M ] at neutral pH.
⚫ == External links ==
⚫ *
⚫ *


DEPC can also be used for probing the structure of double-stranded DNA.{{citation needed|date=March 2021}}


==See also==
{{biochem-stub}}
* ]


==References==
⚫ ]
{{Reflist}}

⚫ == External links ==
⚫ *
⚫ *


⚫ ]
]
]
]
]
]
]

Latest revision as of 13:27, 21 November 2024

Diethyl pyrocarbonate
Names
Preferred IUPAC name Diethyl dicarbonate
Other names
  • Diethylpyrocarbonate
  • Diethyl oxydiformate
  • Ethoxyformic anhydride
  • Pyrocarbonic acid diethyl ester
  • DEPC
Identifiers
CAS Number
3D model (JSmol)
ChEBI
ChEMBL
ChemSpider
ECHA InfoCard 100.015.039 Edit this at Wikidata
EC Number
  • 216-542-8
KEGG
MeSH Diethylpyrocarbonate
PubChem CID
UNII
CompTox Dashboard (EPA)
InChI
  • InChI=1S/C6H10O5/c1-3-9-5(7)11-6(8)10-4-2/h3-4H2,1-2H3Key: FFYPMLJYZAEMQB-UHFFFAOYSA-N
  • InChI=1/C6H10O5/c1-3-9-5(7)11-6(8)10-4-2/h3-4H2,1-2H3Key: FFYPMLJYZAEMQB-UHFFFAOYAU
SMILES
  • O=C(OCC)OC(=O)OCC
Properties
Chemical formula C6H10O5
Molar mass 162.141 g·mol
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
Occupational safety and health (OHS/OSH):
Main hazards Harmful
GHS labelling:
Pictograms GHS07: Exclamation mark
Signal word Warning
Hazard statements H302, H315, H319, H332, H335
Precautionary statements P261, P264, P270, P271, P280, P301+P312, P302+P352, P304+P312, P304+P340, P305+P351+P338, P312, P321, P330, P332+P313, P337+P313, P362, P403+P233, P405, P501
Flash point 69 °C (156 °F; 342 K) closed cup
Lethal dose or concentration (LD, LC):
LD50 (median dose) Oral - rat - 850 mg/kg
Related compounds
Related compounds Di-tert-butyl dicarbonate
Dimethyl dicarbonate
Except where otherwise noted, data are given for materials in their standard state (at 25 °C , 100 kPa). checkY verify (what is  ?) Infobox references
Chemical compound

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.

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 transcription. 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. 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.

See also

References

  1. Chirgwin, John M; et al. (1979). "Isolation of biologically active ribonucleic acid from sourcesenriched in ribonuclease". Biochemistry. 18 (24): 5294–5299. doi:10.1021/bi00591a005. PMID 518835.
  2. 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. doi:10.1111/j.1432-1033.1970.tb00955.x. PMID 5444158.
  3. "FAQ about DEPC". Sigma-Aldrich. Retrieved 12 August 2012.

External links

Categories: