Escherichia coli enzymes

Among its inhibitors are methotrexate (MTX), trimethoprim, and other derivatives of pyrimidines, triazines, pteridines, and related heterocyclic compounds. Some of these inhibitors, such as MTX, bind more tightly to Escherichia coli enzyme than does the substrate dihydrofolate. This fact has been attributed to ion-pair formation between protonated MTX and a negative carboxyl, presumably Asp-27, as well as to hydrophobic interactions.33... 

Mammalian thioredoxin reductase is able to reduce many substances in addition to thioredoxin such as insulin, vitamin K, alloxan, and others, while Escherichia coli enzyme is a... 

ChIP-chop allows one to determine if the DNA that is associated with an antigen of interest is highly CpG-methylated. The Escherichia coli enzyme McrBC is used to hydrolyze ( chop ) the DNA that is methylated specifically at purine/cytosine sites. Hydrolysis within the target DNA sequence prevents detection of the DNA by PCR [24]. 

Selected entries from Methods in Enzymology [vol, page(s)] Equilibrium isotope exchange study of kinetic mechanism, 249, 466 site-directed mutagenesis of Escherichia coli enzyme, 249, 93 positional isotope exchange studies, 249, 423 product inhibition studies of three substrates three products reactions, 249, 207-208. 

Hampton, R.Y. Raetz, C.R.H. Lipid A 4-kinase from Escherichia coli enzyme assay and preparation of 4- 2p-labeled probes of high specific radioactivity. Methods Enzymol., 209, 466-475 (1992)... 

Chauvin, R Brand, L. Roseman, S. Sugar transport by the bacterial phosphotransferase system. Characterization of the Escherichia coli enzyme I monomer/dimer transition kinetics by fluorescence anisotropy. J. Biol. Chem., 269, 20270-20274 (1994)... 

Chauvin, F. Fomenkov, A. Johnson, C.R. Roseman, S. The N-terminal domain of Escherichia coli enzyme I of the phosphoenolpyruvate/glycose phosphotransferase system molecular cloning and characterization. Proc. Natl. Acad. Sci.USA, 93, 7028-7031 (1996)... 

The PDH complex contains three enzymes—pyruvate dehydrogenase (EJ, dihydrolipoyl transacetylase (E2), and dihydrolipoyl dehydrogenase (E3)—each present in multiple copies. The number of copies of each enzyme and therefore the size of the complex varies among species. The PDH complex isolated from mammals is about 50 nm in diameter—more than five times the size of an entire ribosome and big enough to be visualized with the electron microscope (Fig. 16-5a). In the bovine enzyme, 60 identical copies of E2 form a pentagonal dodecahedron (the core) with a diameter of about 25 nm (Fig. 16-5b). (The core of the Escherichia coli enzyme contains 24 copies of E2.) E2 is the point of... 

Other bacterial mutants, such as a mutant Escherichia coli enzyme tolerant to glyphosate, have been described (47). The E. coli mutant had a single amino acid change from the wild type, resulting in substitution of glycine 96 with alanine. An identical mutation was reported in glyphosate-tolerant Klebsiella pneumoniae (48). The nature of changes in the kinetic constants of the K pneumoniae enzyme is similar to that of the E. coli enzyme. 

Yoshida et al. (1997) achieved the production of vanillin by oxidation of vanillylamine using amine oxidase (AO) from Aspergillus niger and monoamine oxidase (MAO) from Escherichia coli. Enzyme kinetic studies have revealed that AO is... 

Kim, S.H. and Oriel, P. 2000. Cloning and expression of the nitrile hydratase and amidase genes from Bacillus sp. BR449 into Escherichia coli. Enzyme Microbiology and Technology, 27 492-501. 

Atkin, C. L., Thelander, L., Reichard, P., and Lang, G., 1973, Iron and free radical in ribonucleotide reductase. Exchange of iron and M sshauer spectroscopy of the protein B2 subunit of the Escherichia coli enzyme. J. Biol. Chem. 248 7464n7472. 

The archetype Fe(II)/aKG hydroxylase is taurine/ q KG dioxygenase (TauD), an Escherichia coli enzyme that catalyzes the conversion of taurine (2-aminoethanesulfonic acid) to sulfite and aminoacetaldehyde, as illustrated in Scheme 1. TauD catalyzes the hydroxylation of a C H bond on the carbon adjacent to the sulfonate group of taurine. The product of this reaction then decomposes to yield hydrogen sulfite, which serves as an important source of sulfur for many microorganisms. A catalytic mechanism that has been proposed for these enzymes is provided as Scheme 2. Prior to the activation and hydroxylation of the Ci carbon on taurine, q KG binds to the Fe(II) center as a chelate, displacing two of the coordinated waters. Taurine then binds to the enzyme in the vicinity of the Fe(II) center, displacing the remaining water. 

Aldose-1-epimerase), quite widespread in animal tissue and bacteria, which catalyses mutarotation. The Escherichia coli enzyme has a maximum activity close to neutral pH. The activation energy 4G =11.9 kcal mol" is greatly lowered, as usual with respect to that of the non-enzymically catalysed reaction, close to 17 kcal mol". o-Glucose, D-galactose, and D-fucose are substrates but not D-mannose (Hucho and Wallenfels 1971). 

The enzyme 2-C-methyl-D-erythritol-4-phosphate synthetase appears to catalyse a Bilik reaction (Figure 6.10) the substrate l-deoxyxylulose-5-phosphate is converted to the title compound via an intermediate aldehyde, whose carbonyl derives from C3 of the substrate. The first step is thus a Bilik reaction and the aldehyde is subsequently reduced by the enzyme using NADPH as reductant, The X-ray crystal structure of the Escherichia coli enzyme in complex with the promising antimalarial Fosmidomycin (a hydroxamic acid) reveals a bound Mn " coordinated to oxygens equivalent to the substrate carbonyl and 03. The stereochemistry and regiochemistry follow the normal Bilik course, although the crystallographers favour an alkyl shift rather than a reverse aldol-aldol mechanism. The intermediate aldehyde has been shown to be a catalytically competent intermediate. 

The X-ray structure of the transferase MurA from Enterobacter cloacae highly homologous to the Escherichia coli enzyme, is of a two-domain enzyme in an open conformation it was reasonably surmised that when the positive charges of an arginine and a lysine, whose repulsion were keeping the protein in the open conformation, were neutralised by the binding of UDPGlcNAc, the active site would close and eliminate water. 

D. Ridgway, R. C. Neerhout, and A. Bleyer. Attenuation of asparaginase-induced hyperglycemia after substitution of the Erwinia carotovora for the Escherichia coli enzyme preparation. Cancer 63 561 (1989). 

NAD and NADP co factor biosynthesis NadD - nicotinate mononucleotide [B] 27 Cloned from Escherichia coli, enzyme assay 28... 

Figure 6 A model of the transition state interactions in the reaction catalyzed by APs, based on X-ray structures7 ° The residue numbering is that for the Escherichia coli enzyme, but the critical Arg and Ser are conserved among all known APs.

Park S J, Park JP, Lee SY, Doi Y (2003) Enrichment of specific monomer in medium-chain-length poly(3-hydroxyalkanoates) by amplification of fadD and fadE genes in recombinant Escherichia coli. Enzyme Microb Technol 33 62-70 Park SJ, Choi JI, Lee SY (2004) Engineering of Escherichia coli fatty acid metabolism for the production of polyhydroxyalkanoates. Enzyme Microb Technol 36 579-588 Park JH, Lee KH, Kim TY, Lee SY (2007) Metabolic engineering of Escherichia coli for the production of L-vafine based on transcriptome analysis and in silico gene knockout simulation. Proc Natl Acad Sci U S A 104 7797-7802... 

Takahashi C, Shirakawa J, Tsuchidate T, Okai N, Hatada K, Nakayama H, Tateno T, Ogino C, Kondo A. (2012). Robust production of gamma-amino butyric acid using recombinant Corynebacterium glutamicum expressing glutamate decarboxylase from Escherichia coli. Enzyme Microb Technol, 51,171-176. 

Alkaline Phosphatase. Alkaline phosphatase (AP) is probably the most extensively studied phosphatase (7-9). This ubiquitous enzyme is a nonspecific phosphatase found in prokaryotes and eukaryotes. The AP reaction proceeds via an intermediate in which a serine residue (Serl02 in the Escherichia coli enzyme) is phosphorylated (mechanism b in Fig. 1). The hydrolysis of the intermediate to produce inorganic phosphate competes with phosphoryl transfer to other acceptors such as alcohols or amines, if present in solution. 

Chen, G.S., Siddiqui, M.A. Biosynthesis of transfer RNA in vitro conversion of transfer RNA precursors from Bombyx mori to 4S RNA by Escherichia coli enzymes. Proc. nat. Acad. Sci. (Wash.) 70, 2610-2613 (1973)... 

Aminoalkanes Amphiphilic gels for the purification of Escherichia coli enzymes 59... 

Pugh, E.L., Sauer, F., Waite, M, Toomey, R.E., and Wakil S.J., 1966. Studies on the mechanism of fatty add synthesis. 13. The role of beta-hydroxy adds in the synthesis of pahnitate and ds vaccenate by the Escherichia coli enzyme system. Journal of Biological Chemistry. 241 2635 2643. 

Wang, D., Li, Q., Li, W, Xing, J., and Su, Z. (2009) Improvement of succinate production by overexpression of a cyanobacterial carbonic anhydrase in Escherichia coli. Enzyme Microb. Technol, 45, 491-497. 

---