Purification and Properties of the Phosphotriesterase from Pseudomonas diminuta

Dumas, David P.; Caldwell, Steven R.; Wild, James R.; Raushel, Frank M.

doi:10.1016/s0021-9258(19)47164-0

Cited by 461 publications

(195 citation statements)

References 22 publications

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“…Accessibility to the metal-containing sites in the protein is governed among other factors by the polarity of the chelators. Both 1,10-phenanthroline and N,N,N h,N h-tetrakis-(2-pyridylmethyl)ethylenediamine are less polar than EDTA and therefore they are more permeant to the hydrophobic sites [35,36]. In addition to the Zn# + -binding sequence HXXE … H and the cysteine residue already discussed, two co-catalytic sequences GDV and GDY [34] are also found at residues 390-392 and 394-396 in human serum BChE.…”

Section: Discussionmentioning

confidence: 89%

Evidence for a Zn2+-binding site in human serum butyrylcholinesterase

Bhanumathy

Balasubramanian

1996

Biochemical Journal

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Purified human serum butyrylcholinesterase after treatment with either of the metal chelators EDTA or NaCN was able to bind to a Zn(2+)-chelate-Sepharose affinity column and was eluted from the column by EDTA or imidazole. Prior EDTA treatment of the enzyme was essential for binding to this affinity column. The enzyme could be labelled with (65)Zn(2+) after EDTA treatment of the enzyme. Diethylpyrocarbonate modification of histidine residues in the EDTA-treated enzyme resulted in the abolition of both binding to the Zn(2+)-chelate-Sepharose column and labelling by (65)Zn(2+). Stoicheiometry of (65)Zn(2+) binding indicated approximately 0.85 mol of Zn(2+)/mol of subunit of the EDTA-treated enzyme. EDTA or NaCN treatment resulted in the loss of thermal stability of the enzyme at 37 degrees C which could not be reversed by Zn(2+). Whereas the cholinesterase activity of butyrlcholinesterase was not affected by EDTA, there was significant loss of its carboxypeptidase activity in the presence of EDTA, and the loss could be reversed by added ZnCl2. These results suggest the presence of a Zn(2+)-binding site on human serum butyrylcholinesterase and the involvement of histidine residues in the metal binding. The presence in human serum butyrylcholinesterase of a sequence HXXE...H found in many known Zn(2+)-containing enzymes supports these findings.

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Section: Discussionmentioning

confidence: 89%

Evidence for a Zn2+-binding site in human serum butyrylcholinesterase

Bhanumathy

Balasubramanian

1996

Biochemical Journal

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“…The phosphotriesterase enzyme from Pseudomonas diminuta catalyses the hydrolysis of a range of synthetic insecticides and chemical warfare agents, none of which were synthesized until the 1940s [19]. Remarkably, phosphotriesterase hydrolyses the insecticide parathion with an efficiency that approaches catalytic perfection (k cat /K M ¼ 4 Â 10 7 s 21 M 21 ) [20]. The enzyme appears to have evolved from a lactonase ancestor with a very weak promiscuous phosphotriesterase activity (k cat /K M , 10 s 21 M 21 ), to near perfection, in approximately 50 years [21].…”

Section: Punctuated Equilibria In Enzyme Evolutionmentioning

confidence: 99%

Rapid bursts and slow declines: on the possible evolutionary trajectories of enzymes

Newton

Arcus

Patrick

2015

J. R. Soc. Interface.

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The evolution of enzymes is often viewed as following a smooth and steady trajectory, from barely functional primordial catalysts to the highly active and specific enzymes that we observe today. In this review, we summarize experimental data that suggest a different reality. Modern examples, such as the emergence of enzymes that hydrolyse human-made pesticides, demonstrate that evolution can be extraordinarily rapid. Experiments to infer and resurrect ancient sequences suggest that some of the first organisms present on the Earth are likely to have possessed highly active enzymes. Reconciling these observations, we argue that rapid bursts of strong selection for increased catalytic efficiency are interspersed with much longer periods in which the catalytic power of an enzyme erodes, through neutral drift and selection for other properties such as cellular energy efficiency or regulation. Thus, many enzymes may have already passed their catalytic peaks.

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“…Like other organophosphorus pesticides, MP kills insects by inhibiting their acetyl cholinesterase, in an irreversible manner (Karalliedde & Senanayake, 1989). Unfortunately, as acetyl cholinesterase occurs in all vertebrates, the potential for organophosphorus pesticides causing damage to humans and other vertebrates is very high (Dumas et al , 1989; Sogorb et al , 2004). Soil micro-organisms can play a significant role in the bioremediation of pesticide-contaminated soil, and various MP-degrading soil bacteria have already been isolated and characterized (Chaudhry et al , 1988; Hayatsu et al , 2000; Liu et al , 2005; Zhang et al , 2006; Shen et al , 2010).…”

mentioning

confidence: 99%

Burkholderia zhejiangensis sp. nov., a methyl-parathion-degrading bacterium isolated from a wastewater-treatment system

Zheng

Sun

et al. 2012

International Journal of Systematic and Evolutionary Microbiology

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The taxonomic status of a methyl-parathion-degrading strain, OP-1 T , isolated from a wastewatertreatment system in China, was determined using a polyphasic approach. The rod-shaped cells were Gram-staining-negative, non-spore-forming and non-motile. Phylogenetic analysis based on 16S rRNA gene sequences indicated that the novel strain belonged to the genus Burkholderia, as it appeared closely related to Burkholderia glathei ATCC 29195 T (97.4 % sequence similarity), Burkholderia sordidicola KCTC 12081 T (96.5 %) and Burkholderia bryophila LMG 23644 T (96.3 %). The major cellular fatty acids, C 16 : 0 , C 17 : 0 cyclo and C 18 : 1 v7c, were also similar to those found in established members of the genus Burkholderia. The genomic DNA G+C content of strain OP-1 T was 59.4 mol%. The level of DNA-DNA relatedness between the novel strain and the closest recognized species, Burkholderia glathei ATCC 29195 T , was only 30 %. Based on the phenotypic, genotypic and phylogenetic evidence, strain OP-1 T represents a novel species of the genus Burkholderia, for which the name Burkholderia zhejiangensis sp. nov. is proposed. The type strain is OP-1 T (5CCTCC AB 2010354 T 5KCTC 23300 T ).Methyl parathion (O,O-dimethyl O-p-nitrophenylphosphorothioate; MP) is widely used to control a large range of insect pests. Like other organophosphorus pesticides, MP kills insects by inhibiting their acetyl cholinesterase, in an irreversible manner (Karalliedde & Senanayake, 1989). Unfortunately, as acetyl cholinesterase occurs in all vertebrates, the potential for organophosphorus pesticides causing damage to humans and other vertebrates is very high (Dumas et al., 1989;Sogorb et al., 2004). Soil microorganisms can play a significant role in the bioremediation of pesticide-contaminated soil, and various MP-degrading soil bacteria have already been isolated and characterized (Chaudhry et al., 1988;Hayatsu et al., 2000;Liu et al., 2005;Zhang et al., 2006;Shen et al., 2010).Burkholderia is a genus of metabolically versatile Gramstaining-negative bacteria. The genus was first proposed by Yabuuchi et al. (1992), who transferred seven species from rRNA group II of the genus Pseudomonas to the novel genus. Many species of the genus Burkholderia play an important role in the biodegradation of pollutants (Coenye & Vandamme, 2003;Lehning et al., 1997;Hayatsu et al., 2000). In this paper, the taxonomy of an MP-degrading bacterium, designated strain OP-1 T , was investigated using a polyphasic approach, The new strain was eventually identified as a novel species in the genus Burkholderia.In 2008, a conventional enrichment method was employed in an attempt to isolate MP-degrading bacteria from sludge collected from the wastewater-treatment system of a pesticidemanufacturing company in the Chinese province of Zhejiang. A subsample (1 g) of the collected sludge was inoculated into an MP-supplemented mineral salts medium (MP-MSM) which contained (l 21 ): 1.5 g K 2 HPO 4 , 0.5 g KH 2 PO 4 , 1.0 g NH 4 NO 3 , 0.2 g MgSO 4 . 7H 2 O, 0.5 g NaCl and, as the so...

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Purification and Properties of the Phosphotriesterase from Pseudomonas diminuta

Cited by 461 publications

References 22 publications

Evidence for a Zn2+-binding site in human serum butyrylcholinesterase

Evidence for a Zn2+-binding site in human serum butyrylcholinesterase

Rapid bursts and slow declines: on the possible evolutionary trajectories of enzymes

Burkholderia zhejiangensis sp. nov., a methyl-parathion-degrading bacterium isolated from a wastewater-treatment system

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