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PDBsum entry 4j5n
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References listed in PDB file
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Key reference
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Title
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Molecular engineering of organophosphate hydrolysis activity from a weak promiscuous lactonase template.
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Authors
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M.M.Meier,
C.Rajendran,
C.Malisi,
N.G.Fox,
C.Xu,
S.Schlee,
D.P.Barondeau,
B.Höcker,
R.Sterner,
F.M.Raushel.
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Ref.
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J Am Chem Soc, 2013,
135,
11670-11677.
[DOI no: ]
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PubMed id
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Abstract
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Rapid evolution of enzymes provides unique molecular insights into the
remarkable adaptability of proteins and helps to elucidate the relationship
between amino acid sequence, structure, and function. We interrogated the
evolution of the phosphotriesterase from Pseudomonas diminuta (PdPTE), which
hydrolyzes synthetic organophosphates with remarkable catalytic efficiency. PTE
is thought to be an evolutionarily "young" enzyme, and it has been
postulated that it has evolved from members of the phosphotriesterase-like
lactonase (PLL) family that show promiscuous organophosphate-degrading activity.
Starting from a weakly promiscuous PLL scaffold (Dr0930 from Deinococcus
radiodurans ), we designed an extremely efficient organophosphate hydrolase
(OPH) with broad substrate specificity using rational and random mutagenesis in
combination with in vitro activity screening. The OPH activity for seven
organophosphate substrates was simultaneously enhanced by up to 5 orders of
magnitude, achieving absolute values of catalytic efficiencies up to 10(6) M(-1)
s(-1). Structural and computational analyses identified the molecular basis for
the enhanced OPH activity of the engineered PLL variants and demonstrated that
OPH catalysis in PdPTE and the engineered PLL differ significantly in the mode
of substrate binding.
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