Properties of a thermostable nitrate reductase from the hyperthermophilic archaeon Pyrobaculum aerophilum

The nitrate reductase of the hyperthermophilic archaeon Pyrobaculum aerophilum was purified 137-fold from the cytoplasmic membrane. Based on sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis, the enzyme complex consists of three subunits with apparent molecular weights of 130,000, 5...

Full description

Saved in:
Bibliographic Details
Published in:Journal of bacteriology Vol. 183; no. 19; p. 5491
Main Authors: Afshar, S, Johnson, E, de Vries, S, Schröder, I
Format: Journal Article
Language:English
Published: United States 01.10.2001
Subjects:
Enzyme Stability
Hydrogen-Ion Concentration
Kinetics
Molecular Weight
Molybdoferredoxin
Nitrate Reductase
Nitrate Reductases - chemistry
Nitrate Reductases - isolation & purification
Nitrate Reductases - metabolism
Protein Subunits
Temperature
Thermoproteaceae - enzymology
ISSN:0021-9193
Online Access:Get more information
Tags: Add Tag
No Tags, Be the first to tag this record!
Abstract The nitrate reductase of the hyperthermophilic archaeon Pyrobaculum aerophilum was purified 137-fold from the cytoplasmic membrane. Based on sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis, the enzyme complex consists of three subunits with apparent molecular weights of 130,000, 52,000, and 32,000. The enzyme contained molybdenum (0.8-mol/mol complex), iron (15.4-mol/mol complex) and cytochrome b (0.49-mol/mol complex) as cofactors. The P. aerophilum nitrate reductase distinguishes itself from nitrate reductases of mesophilic bacteria and archaea by its very high specific activity using reduced benzyl viologen as the electron donor (V(max) with nitrate, 1,162 s(-1) (326 U/mg); V(max) with chlorate, 1,348 s(-1) (378 U/mg) [assayed at 75 degrees C]). The K(m) values for nitrate and chlorate were 58 and 140 microM, respectively. Azide was a competitive inhibitor and cyanide was a noncompetitive inhibitor of the nitrate reductase activity. The temperature optimum for activity was > 95 degrees C. When incubated at 100 degrees C, the purified nitrate reductase had a half-life of 1.5 h. This study constitutes the first description of a nitrate reductase from a hyperthermophilic archaeon.
AbstractList The nitrate reductase of the hyperthermophilic archaeon Pyrobaculum aerophilum was purified 137-fold from the cytoplasmic membrane. Based on sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis, the enzyme complex consists of three subunits with apparent molecular weights of 130,000, 52,000, and 32,000. The enzyme contained molybdenum (0.8-mol/mol complex), iron (15.4-mol/mol complex) and cytochrome b (0.49-mol/mol complex) as cofactors. The P. aerophilum nitrate reductase distinguishes itself from nitrate reductases of mesophilic bacteria and archaea by its very high specific activity using reduced benzyl viologen as the electron donor (V(max) with nitrate, 1,162 s(-1) (326 U/mg); V(max) with chlorate, 1,348 s(-1) (378 U/mg) [assayed at 75 degrees C]). The K(m) values for nitrate and chlorate were 58 and 140 microM, respectively. Azide was a competitive inhibitor and cyanide was a noncompetitive inhibitor of the nitrate reductase activity. The temperature optimum for activity was > 95 degrees C. When incubated at 100 degrees C, the purified nitrate reductase had a half-life of 1.5 h. This study constitutes the first description of a nitrate reductase from a hyperthermophilic archaeon.
The nitrate reductase of the hyperthermophilic archaeon Pyrobaculum aerophilum was purified 137-fold from the cytoplasmic membrane. Based on sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis, the enzyme complex consists of three subunits with apparent molecular weights of 130,000, 52,000, and 32,000. The enzyme contained molybdenum (0.8-mol/mol complex), iron (15.4-mol/mol complex) and cytochrome b (0.49-mol/mol complex) as cofactors. The P. aerophilum nitrate reductase distinguishes itself from nitrate reductases of mesophilic bacteria and archaea by its very high specific activity using reduced benzyl viologen as the electron donor (V(max) with nitrate, 1,162 s(-1) (326 U/mg); V(max) with chlorate, 1,348 s(-1) (378 U/mg) [assayed at 75 degrees C]). The K(m) values for nitrate and chlorate were 58 and 140 microM, respectively. Azide was a competitive inhibitor and cyanide was a noncompetitive inhibitor of the nitrate reductase activity. The temperature optimum for activity was > 95 degrees C. When incubated at 100 degrees C, the purified nitrate reductase had a half-life of 1.5 h. This study constitutes the first description of a nitrate reductase from a hyperthermophilic archaeon.The nitrate reductase of the hyperthermophilic archaeon Pyrobaculum aerophilum was purified 137-fold from the cytoplasmic membrane. Based on sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis, the enzyme complex consists of three subunits with apparent molecular weights of 130,000, 52,000, and 32,000. The enzyme contained molybdenum (0.8-mol/mol complex), iron (15.4-mol/mol complex) and cytochrome b (0.49-mol/mol complex) as cofactors. The P. aerophilum nitrate reductase distinguishes itself from nitrate reductases of mesophilic bacteria and archaea by its very high specific activity using reduced benzyl viologen as the electron donor (V(max) with nitrate, 1,162 s(-1) (326 U/mg); V(max) with chlorate, 1,348 s(-1) (378 U/mg) [assayed at 75 degrees C]). The K(m) values for nitrate and chlorate were 58 and 140 microM, respectively. Azide was a competitive inhibitor and cyanide was a noncompetitive inhibitor of the nitrate reductase activity. The temperature optimum for activity was > 95 degrees C. When incubated at 100 degrees C, the purified nitrate reductase had a half-life of 1.5 h. This study constitutes the first description of a nitrate reductase from a hyperthermophilic archaeon.
Author Johnson, E
de Vries, S
Afshar, S
Schröder, I
Author_xml – sequence: 1
  givenname: S
  surname: Afshar
  fullname: Afshar, S
  organization: Department of Microbiology and Molecular Genetics, University of California, Los Angeles, California 90095-1489, USA
– sequence: 2
  givenname: E
  surname: Johnson
  fullname: Johnson, E
– sequence: 3
  givenname: S
  surname: de Vries
  fullname: de Vries, S
– sequence: 4
  givenname: I
  surname: Schröder
  fullname: Schröder, I
BackLink https://www.ncbi.nlm.nih.gov/pubmed/11544209$$D View this record in MEDLINE/PubMed
BookMark eNo1kMtOwzAQRb0oog_4BeQVuwTbcR5eQsVTlegC1tHEHitBSVxsZ9G_J5SymSvNPTOauWuyGN2IhFDOUs5Fdff2kPIqS7lKc6l4Mpc8FYzxBVkxJniiuMqWZB3C19yUMheXZMl5LqVgakW6vXcH9LHDQJ2lQGOLfnAhQtMjHbvoISL1aCYdISC13g2_DG2Pv2Mn-NB2facpeN0CupHuj941oKd-GiigP_nTcEUuLPQBr8-6IZ9Pjx_bl2T3_vy6vd8lWioVE1UVYJGBgKpoLNr5fF1UaHK0RdlkpTHKiqowJfDKSm4yo1GzEnKDuSyMFBty-7f34N33hCHWQxc09j2M6KZQl_PzqiyqGbw5g1MzoKkPvhvAH-v_cMQPSNhr0g
CitedBy_id crossref_primary_10_1089_cmb_2007_R018
crossref_primary_10_1007_s00792_019_01112_9
crossref_primary_10_1038_ismej_2014_216
crossref_primary_10_1089_ast_2008_0248
crossref_primary_10_1007_s10295_010_0717_6
crossref_primary_10_1038_s41598_020_67892_9
crossref_primary_10_1111_1758_2229_12335
crossref_primary_10_1146_annurev_micro_102215_095406
crossref_primary_10_1007_s10534_011_9442_5
crossref_primary_10_1016_j_bbagen_2005_02_008
crossref_primary_10_1016_j_biochi_2010_06_003
crossref_primary_10_1016_j_jbiotec_2010_12_025
crossref_primary_10_1016_j_pep_2005_02_002
crossref_primary_10_1111_j_1742_4658_2010_07725_x
crossref_primary_10_1016_j_ccr_2019_05_005
crossref_primary_10_1016_S0378_1097_03_00122_8
crossref_primary_10_3390_ijms20133349
crossref_primary_10_1128_JB_00965_08
crossref_primary_10_1016_j_ab_2004_01_026
crossref_primary_10_1016_j_ijbiomac_2017_04_009
crossref_primary_10_1016_j_jtice_2015_09_014
crossref_primary_10_1111_1462_2920_13068
crossref_primary_10_1134_S0006297907100124
crossref_primary_10_1007_s10541_005_0186_0
crossref_primary_10_1016_j_bbabio_2013_01_011
crossref_primary_10_3389_fmicb_2016_02123
crossref_primary_10_1128_JB_00980_13
crossref_primary_10_1007_s00775_020_01784_1
crossref_primary_10_1016_j_ccr_2008_01_017
crossref_primary_10_1042_BST0340118
crossref_primary_10_1016_j_ijbiomac_2025_145967
crossref_primary_10_1016_S0014_5793_02_02524_3
crossref_primary_10_1128_JB_188_2_525_531_2006
crossref_primary_10_1074_jbc_M300857200
crossref_primary_10_1111_j_1574_6968_2007_00887_x
crossref_primary_10_1016_j_bbagen_2004_05_007
crossref_primary_10_1080_10934529_2012_695572
crossref_primary_10_1128_spectrum_04145_22
crossref_primary_10_1042_BST0330200
crossref_primary_10_1016_j_cell_2023_05_012
crossref_primary_10_1007_s11101_009_9137_5
crossref_primary_10_1016_j_syapm_2024_126507
crossref_primary_10_1128_AEM_69_9_5585_5592_2003
crossref_primary_10_1039_C3CS60249D
ContentType Journal Article
DBID CGR
CUY
CVF
ECM
EIF
NPM
7X8
DOI 10.1128/JB.183.19.5491-5495.2001
DatabaseName Medline
MEDLINE
MEDLINE (Ovid)
MEDLINE
MEDLINE
PubMed
MEDLINE - Academic
DatabaseTitle MEDLINE
Medline Complete
MEDLINE with Full Text
PubMed
MEDLINE (Ovid)
MEDLINE - Academic
DatabaseTitleList MEDLINE
MEDLINE - Academic
Database_xml – sequence: 1
  dbid: NPM
  name: PubMed
  url: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed
  sourceTypes: Index Database
– sequence: 2
  dbid: 7X8
  name: MEDLINE - Academic
  url: https://search.proquest.com/medline
  sourceTypes: Aggregation Database
DeliveryMethod no_fulltext_linktorsrc
Discipline Biology
ExternalDocumentID 11544209
Genre Research Support, U.S. Gov't, Non-P.H.S
Journal Article
GroupedDBID ---
-DZ
-~X
.55
.GJ
0R~
186
18M
1VV
29J
2WC
39C
3O-
4.4
53G
5GY
5RE
5VS
79B
85S
8WZ
9M8
A6W
ABPPZ
ABTAH
ACGFO
ACGOD
ACNCT
ACPRK
ADBBV
AENEX
AFFDN
AFFNX
AFRAH
AGCDD
AGVNZ
AI.
AIDAL
AJUXI
ALMA_UNASSIGNED_HOLDINGS
AOIJS
BAWUL
BKOMP
BTFSW
C1A
CGR
CJ0
CS3
CUY
CVF
DIK
DU5
E3Z
EBS
ECM
EIF
EJD
F5P
FRP
GX1
H13
HYE
HZ~
IH2
KQ8
L7B
MVM
NHB
NPM
O9-
OHT
OK1
P-O
P-S
P2P
PKN
PQQKQ
QZG
RHF
RHI
RNS
RPM
RSF
RXW
TAE
TR2
UCJ
UHB
UKR
UPT
VH1
VQA
W8F
WH7
WHG
WOQ
X7M
Y6R
YIN
YQT
YR2
YZZ
ZCA
ZCG
ZGI
ZXP
ZY4
~02
~KM
7X8
AAGFI
ABUFD
ADXHL
ID FETCH-LOGICAL-c499t-986afe0a2a86bfef919c68ed5ef67b37dd9f286d7a18f41d3dcec07a5de546d42
IEDL.DBID 7X8
ISICitedReferencesCount 69
ISICitedReferencesURI http://www.webofscience.com/api/gateway?GWVersion=2&SrcApp=Summon&SrcAuth=ProQuest&DestLinkType=CitingArticles&DestApp=WOS_CPL&KeyUT=000170958900004&url=https%3A%2F%2Fcvtisr.summon.serialssolutions.com%2F%23%21%2Fsearch%3Fho%3Df%26include.ft.matches%3Dt%26l%3Dnull%26q%3D
ISSN 0021-9193
IngestDate Tue Oct 07 10:48:11 EDT 2025
Wed Feb 19 02:08:38 EST 2025
IsDoiOpenAccess false
IsOpenAccess true
IsPeerReviewed true
IsScholarly true
Issue 19
Language English
LinkModel DirectLink
MergedId FETCHMERGED-LOGICAL-c499t-986afe0a2a86bfef919c68ed5ef67b37dd9f286d7a18f41d3dcec07a5de546d42
Notes ObjectType-Article-1
SourceType-Scholarly Journals-1
ObjectType-Feature-2
content type line 23
OpenAccessLink https://www.ncbi.nlm.nih.gov/pmc/articles/95438
PMID 11544209
PQID 71159768
PQPubID 23479
ParticipantIDs proquest_miscellaneous_71159768
pubmed_primary_11544209
PublicationCentury 2000
PublicationDate 2001-10-01
PublicationDateYYYYMMDD 2001-10-01
PublicationDate_xml – month: 10
  year: 2001
  text: 2001-10-01
  day: 01
PublicationDecade 2000
PublicationPlace United States
PublicationPlace_xml – name: United States
PublicationTitle Journal of bacteriology
PublicationTitleAlternate J Bacteriol
PublicationYear 2001
SSID ssj0014452
Score 1.9559639
Snippet The nitrate reductase of the hyperthermophilic archaeon Pyrobaculum aerophilum was purified 137-fold from the cytoplasmic membrane. Based on sodium dodecyl...
SourceID proquest
pubmed
SourceType Aggregation Database
Index Database
StartPage 5491
SubjectTerms Enzyme Stability
Hydrogen-Ion Concentration
Kinetics
Molecular Weight
Molybdoferredoxin
Nitrate Reductase
Nitrate Reductases - chemistry
Nitrate Reductases - isolation & purification
Nitrate Reductases - metabolism
Protein Subunits
Temperature
Thermoproteaceae - enzymology
Title Properties of a thermostable nitrate reductase from the hyperthermophilic archaeon Pyrobaculum aerophilum
URI https://www.ncbi.nlm.nih.gov/pubmed/11544209
https://www.proquest.com/docview/71159768
Volume 183
WOSCitedRecordID wos000170958900004&url=https%3A%2F%2Fcvtisr.summon.serialssolutions.com%2F%23%21%2Fsearch%3Fho%3Df%26include.ft.matches%3Dt%26l%3Dnull%26q%3D
hasFullText
inHoldings 1
isFullTextHit
isPrint
link http://cvtisr.summon.serialssolutions.com/2.0.0/link/0/eLvHCXMwpV1LTxsxELZ4Slxa3k1pwQeuDvHGXtsSUkWrIoQgygFQbpF3PVaRym5Ykkr5951xsuKEOHDxZcfSajwez_Mbxk7xjH3QwQrIPAgaaSwK47RQsecNaj8jU2_Vw40ZDOxo5IYr7LzthaGyylYnJkUd6pJi5GcGTRd8Ou2PybOgmVGUW10O0Fhl6300ZEimzeg1h6CUXmKFS7zSrt_W8WT27PpnF0W5K10X3SMpcNHkKsq3zcz03Fx-_tiPbrNPSzOTXyzkYoetQLXLNheDJ-d77HFIMfiGwFR5HbnnZAY-1WgpFn-B4y0nAAneEKzrFJ85Tk0oRMP_zGlbIp5QKKbkCWwJ6ooP5w3qhhRN5B6a9H32tM_uL3_f_boSy6ELokTnZyqczX2Ens-8zYsIETlX5haChpibom9CcDGzeTBe2qhk6IcSyp7xOoBWeVDZAVur6gq-MB68kzbovCgdKCQolCu8tg6ClypG02EnLQfHKNSUqfAV1LOXccvDDjtcHMJ4ssDeGCf0oKznvr6794htpXqxVHj3ja1HvM7wnW2U_6aPL81xkhVcB8Pb_5BYy6Y
linkProvider ProQuest
openUrl ctx_ver=Z39.88-2004&ctx_enc=info%3Aofi%2Fenc%3AUTF-8&rfr_id=info%3Asid%2Fsummon.serialssolutions.com&rft_val_fmt=info%3Aofi%2Ffmt%3Akev%3Amtx%3Ajournal&rft.genre=article&rft.atitle=Properties+of+a+thermostable+nitrate+reductase+from+the+hyperthermophilic+archaeon+Pyrobaculum+aerophilum&rft.jtitle=Journal+of+bacteriology&rft.au=Afshar%2C+S&rft.au=Johnson%2C+E&rft.au=de+Vries%2C+S&rft.au=Schr%C3%B6der%2C+I&rft.date=2001-10-01&rft.issn=0021-9193&rft.volume=183&rft.issue=19&rft.spage=5491&rft_id=info:doi/10.1128%2FJB.183.19.5491-5495.2001&rft.externalDBID=NO_FULL_TEXT
thumbnail_l http://covers-cdn.summon.serialssolutions.com/index.aspx?isbn=/lc.gif&issn=0021-9193&client=summon
thumbnail_m http://covers-cdn.summon.serialssolutions.com/index.aspx?isbn=/mc.gif&issn=0021-9193&client=summon
thumbnail_s http://covers-cdn.summon.serialssolutions.com/index.aspx?isbn=/sc.gif&issn=0021-9193&client=summon