Reduction of quinones and nitroaromatic compounds by Escherichia coli nitroreductase A (NfsA): Characterization of kinetics and substrate specificity

2017 ◽  
Vol 614 ◽  
pp. 14-22 ◽  
Author(s):  
Benjaminas Valiauga ◽  
Elsie M. Williams ◽  
David F. Ackerley ◽  
Narimantas Čėnas
Keyword(s):  
Escherichia Coli ◽  
Substrate Specificity ◽  
Nitroaromatic Compounds

scholarly journals Characterization of GDP-mannose pyrophosphorylase from Escherichia coli O157:H7 EDL933 and its broad substrate specificity

2005 ◽  
Vol 37 (1-6) ◽  
pp. 1-8 ◽  
Author(s):  
Yung-Hun Yang ◽  
Young-Bok Kang ◽  
Kwang-Won Lee ◽  
Tek-Hyung Lee ◽  
Sung-Soo Park ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Substrate Specificity ◽  
Escherichia Coli O157 ◽  
Broad Substrate Specificity

scholarly journals Characterization of a Thermostable Short-Chain Alcohol Dehydrogenase from the Hyperthermophilic Archaeon Thermococcus sibiricus

2010 ◽  
Vol 76 (12) ◽  
pp. 4096-4098 ◽  
Author(s):  
Tatiana N. Stekhanova ◽  
Andrey V. Mardanov ◽  
Ekaterina Y. Bezsudnova ◽  
Vadim M. Gumerov ◽  
Nikolai V. Ravin ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Alcohol Dehydrogenase ◽  
Substrate Specificity ◽  
Organic Solvents ◽  
Short Chain ◽  
Chain Alcohol ◽  
Hyperthermophilic Archaeon ◽  
Broad Substrate Specificity ◽  
Short Chain Alcohol

ABSTRACT Short-chain alcohol dehydrogenase, encoded by the gene Tsib_0319 from the hyperthermophilic archaeon Thermococcus sibiricus, was expressed in Escherichia coli, purified and characterized as an NADPH-dependent enantioselective oxidoreductase with broad substrate specificity. The enzyme exhibits extremely high thermophilicity, thermostability, and tolerance to organic solvents and salts.

scholarly journals Biochemical Characterization of the POM-1 Metallo-β-Lactamase from Pseudomonas otitidis

2014 ◽  
Vol 59 (3) ◽  
pp. 1755-1758 ◽  
Author(s):  
Luisa Borgianni ◽  
Filomena De Luca ◽  
Maria Cristina Thaller ◽  
Yunsop Chong ◽  
Gian Maria Rossolini ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Functional Analysis ◽  
Substrate Specificity ◽  
Biochemical Characterization ◽  
Content Type ◽  
Broad Substrate Specificity ◽  
Catalytic Activities

ABSTRACTThe POM-1 metallo-β-lactamase is a subclass B3 resident enzyme produced byPseudomonas otitidis, a pathogen causing otic infections. The enzyme was overproduced inEscherichia coliBL21(DE3), purified by chromatography, and subjected to structural and functional analysis. The purified POM-1 is a tetrameric enzyme of broad substrate specificity with higher catalytic activities with penicillins and carbapenems than with cephalosporins.

scholarly journals Functional Reconstitution of SdcS, a Na+-Coupled Dicarboxylate Carrier Protein from Staphylococcus aureus

2006 ◽  
Vol 189 (3) ◽  
pp. 880-885 ◽  
Author(s):  
Jason A. Hall ◽  
Ana M. Pajor
Keyword(s):  
Escherichia Coli ◽  
Staphylococcus Aureus ◽  
Substrate Specificity ◽  
Transport Properties ◽  
Family Member ◽  
Partial Purification ◽  
Carrier Protein ◽  
Electrochemical Gradient ◽  
Structural Aspects

ABSTRACT In Staphylococcus aureus, the transport of dicarboxylates is mediated in part by the Na+-linked carrier protein SdcS. This transporter is a member of the divalent-anion/Na+ symporter (DASS) family, a group that includes the mammalian Na+/dicarboxylate cotransporters NaDC1 and NaDC3. In earlier work, we cloned and expressed SdcS in Escherichia coli and found it to have transport properties similar to those of its eukaryotic counterparts (J. A. Hall and A. M. Pajor, J. Bacteriol. 187:5189-5194, 2005). Here, we report the partial purification and subsequent reconstitution of functional SdcS into liposomes. These proteoliposomes exhibited succinate counterflow activity, as well as Na+ electrochemical-gradient-driven transport. Examination of substrate specificity indicated that the minimal requirement necessary for transport was a four-carbon terminal dicarboxylate backbone and that productive substrate-transporter interaction was sensitive to substitutions at the substrate C-2 and C-3 positions. Further analysis established that SdcS facilitates an electroneutral symport reaction having a 2:1 cation/dicarboxylate ratio. This study represents the first characterization of a reconstituted Na+-coupled DASS family member, thus providing an effective method to evaluate functional, as well as structural, aspects of DASS transporters in a system free of the complexities and constraints associated with native membrane environments.

scholarly journals Molecular Cloning and Characterization of Fengycin Synthetase Gene fenB from Bacillus subtilis

1998 ◽  
Vol 180 (5) ◽  
pp. 1338-1341 ◽  
Author(s):  
Guang-Huey Lin ◽  
Chyi-Liang Chen ◽  
Johannes Scheng-Ming Tschen ◽  
San-San Tsay ◽  
Yu-Sun Chang ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Bacillus Subtilis ◽  
Amino Acid ◽  
Substrate Specificity ◽  
Affinity Chromatography ◽  
Molecular Mass ◽  
Optimum Temperature ◽  
Turnover Number ◽  
Optimum Ph

ABSTRACT A fengycin synthetase gene, fenB, has been cloned and sequenced. The protein (FenB) encoded by this gene has a predicted molecular mass of 143.6 kDa. This protein was overexpressed inEscherichia coli and was purified to near homogeneity by affinity chromatography. Experimental results indicated that the recombinant FenB has a substrate specificity toward isoleucine with an optimum temperature of 25°C, an optimum pH of 4.5, aKm value of 922 μM, and a turnover number of 236 s−1. FenB also consists of a thioesterase domain, suggesting that this protein may be involved in the activation of the last amino acid of fengycin.

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