Abstract:
:The 2-aminoethylphosphonate transaminase (AEPT; the phnW gene product) of the Salmonella enterica serovar Typhimurium 2-aminoethylphosphonate (AEP) degradation pathway catalyzes the reversible reaction of AEP and pyruvate to form phosphonoacetaldehyde (P-Ald) and L-alanine (L-Ala). Here, we describe the purification and characterization of recombinant AEPT. pH rate profiles (log V(m) and log V(m)/K(m) versus pH) revealed a pH optimum of 8.5. At pH 8.5, K(eq) is equal to 0.5 and the k(cat) values of the forward and reverse reactions are 7 and 9 s(-1), respectively. The K(m) for AEP is 1.11 +/- 0.03 mM; for pyruvate it is 0.15 +/- 0.02 mM, for P-Ald it is 0.09 +/- 0.01 mM, and for L-Ala it is 1.4 +/- 0.03 mM. Substrate specificity tests revealed a high degree of discrimination, indicating a singular physiological role for the transaminase in AEP degradation. The 40-kDa subunit of the homodimeric enzyme is homologous to other members of the pyridoxalphosphate-dependent amino acid transaminase superfamily. Catalytic residues conserved within well-characterized members are also conserved within the seven known AEPT sequences. Site-directed mutagenesis demonstrated the importance of three selected residues (Asp168, Lys194, and Arg340) in AEPT catalysis.
journal_name
J Bacterioljournal_title
Journal of bacteriologyauthors
Kim AD,Baker AS,Dunaway-Mariano D,Metcalf WW,Wanner BL,Martin BMdoi
10.1128/jb.184.15.4134-4140.2002keywords:
subject
Has Abstractpub_date
2002-08-01 00:00:00pages
4134-40issue
15eissn
0021-9193issn
1098-5530journal_volume
184pub_type
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doi:10.1128/jb.174.11.3789-3799.1992
更新日期:1992-06-01 00:00:00
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更新日期:1970-10-01 00:00:00
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更新日期:1962-12-01 00:00:00
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更新日期:1976-06-01 00:00:00
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更新日期:1970-09-01 00:00:00
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更新日期:1970-04-01 00:00:00
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更新日期:1981-03-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
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更新日期:2012-11-01 00:00:00
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更新日期:1981-11-01 00:00:00
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pub_type: 杂志文章
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更新日期:2008-08-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.00468-16
更新日期:2016-09-22 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/jb.182.23.6824-6826.2000
更新日期:2000-12-01 00:00:00
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更新日期:1967-11-01 00:00:00
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更新日期:1982-10-01 00:00:00
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更新日期:2000-03-01 00:00:00
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pub_type: 杂志文章
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更新日期:2000-10-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.84.2.295-301.1962
更新日期:1962-08-01 00:00:00
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pub_type: 杂志文章
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更新日期:1979-10-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
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更新日期:1976-11-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.124.3.1424-1428.1975
更新日期:1975-12-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.94.3.557-561.1967
更新日期:1967-09-01 00:00:00
abstract::The Yersinia pestis proteome was studied as a function of temperature and calcium by two-dimensional differential gel electrophoresis. Over 4,100 individual protein spots were detected, of which hundreds were differentially expressed. A total of 43 differentially expressed protein spots, representing 24 unique protein...
journal_title:Journal of bacteriology
pub_type: 杂志文章
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更新日期:2005-12-01 00:00:00