Physical characterization of katG, encoding catalase HPI of Escherichia coli

Triggs-Raine, Barbara; Loewen, P.C.

doi:10.1016/0378-1119(87)90038-2

Cited by 59 publications

(24 citation statements)

References 37 publications

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“…Strains and Plasmids-The plasmids pAH1 (23), pBpKatG (24), and pBT22 (25) were used as the source of catalase-peroxidases from M. tuberculosis, Burkholderia pseudomallei, and Escherichia coli, respectively. All of the plasmids were transformed into the catalasedeficient E. coli strain UM262 (pro leu rpsL hsdM hsdR endI lacY katE1 katG17::Tn10 recA) (26) and grown in Luria broth containing 10 g/liter tryptone, 5 g/liter yeast extract, and 5 g/liter NaCl for expression of the catalase-peroxidases.…”

Section: Methodsmentioning

confidence: 99%

Catalase-peroxidases (KatG) Exhibit NADH Oxidase Activity

Singh

Wiseman

Deemagarn

et al. 2004

Journal of Biological Chemistry

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Catalase-peroxidases (KatG) produced by Burkholderia pseudomallei, Escherichia coli, and Mycobacterium tuberculosis catalyze the oxidation of NADH to form NAD ؉ and either H 2 O 2 or superoxide radical depending on pH. The NADH oxidase reaction requires molecular oxygen, does not require hydrogen peroxide, is not inhibited by superoxide dismutase or catalase, and has a pH optimum of 8.75, clearly differentiating it from the peroxidase and catalase reactions with pH optima of 5.5 and 6.5, respectively, and from the NADH peroxidase-oxidase reaction of horseradish peroxidase. B. pseudomallei KatG has a relatively high affinity for NADH (K m ‫؍‬ 12 M), but the oxidase reaction is slow (k cat ‫؍‬ 0.54 min ؊1 ) compared with the peroxidase and catalase reactions. The catalase-peroxidases also catalyze the hydrazinolysis of isonicotinic acid hydrazide (INH) in an oxygen-and H 2 O 2 -independent reaction, and KatG-dependent radical generation from a mixture of NADH and INH is two to three times faster than the combined rates of separate reactions with NADH and INH alone. The major products from the coupled reaction, identified by high pressure liquid chromatography fractionation and mass spectrometry, are NAD ؉ and isonicotinoyl-NAD, the activated form of isoniazid that inhibits mycolic acid synthesis in M. tuberculosis. Isonicotinoyl-NAD synthesis from a mixture of NAD ؉ and INH is KatG-dependent and is activated by manganese ion. M. tuberculosis KatG catalyzes isonicotinoyl-NAD formation from NAD ؉ and INH more efficiently than B. pseudomallei KatG.

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Section: Methodsmentioning

confidence: 99%

Catalase-peroxidases (KatG) Exhibit NADH Oxidase Activity

Singh

Wiseman

Deemagarn

et al. 2004

Journal of Biological Chemistry

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“…katA expression has been detected during the exponential growth phase of free-living bacteria only, and katA is the unique catalase gene inducible by exogenous H 2 O 2 . In Escherichia coli, inducibility by H 2 O 2 and expression in exponential phase have been observed for the katG catalase gene encoding the catalase-hydroperoxidase HPI (38). The H 2 O 2 induction of katG requires the positive activator OxyR (26), which directly senses oxidative stress (18,41).…”

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The katA Catalase Gene Is Regulated by OxyR in both Free-Living and Symbiotic Sinorhizobium meliloti

et al. 2005

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“…The katG gene has been cloned (22,42) and sequenced (41) and predicts an amino acid sequence that bears no resemblance to any of the known catalase sequences. Instead, a striking resemblance to a peroxidase from Bacillus stearothermophilus has been observed (23), suggesting that the bifunctional HPI is more closely related to the family of peroxidases than catalases.…”

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Nucleotide sequence of Escherichia coli katE, which encodes catalase HPII

et al. 1991

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A 3,466-bp nucleotide sequence containing the katE gene of Escherichia coli has been determined. An open reading frame of 2,259 bp was found and was preceded by a potential ribosome-binding site. The predicted N-terminal sequence agreed with the sequence determined by direct amino acid sequencing, and the predicted direction of transcription was confirmed by expression of the gene cloned in both directions behind a T7 promoter. The start site of transcription was determined to be 127 bp upstream from the start of the open reading frame, and a potential RNA polymerase-binding site similar to a sequence preceding the xthA gene, which is also controlled by the KatF protein, was identified. The predicted sequence of the 753-amino-acid protein was comnpared with known sequences of other catalases, revealing significant similarity to the shorter catalases, including the residues in the putative active site and residues involved in heme binding.Escherichia coli produces two catalases, a bifunctional catalase-peroxidase (hydroperoxidase I [HPI]) and a monofunctional catalase (HPII). Both enzymes have been purified and characterized to be quite different from each other and from the typical catalase, which is active as a tetramer of 65,000-Da subunits and four protoheme IX groups. HPI was found to be a tetramer of 78,000-Da subunits with just two protoheme IX groups per tetramer and with an associated broad-spectrum peroxidase activity (6). HPII was purified (7) and characterized as a hexamer of 93,000-Da subunits (18) and six heme d-like groups (5) per hexamer. The genes encoding HPI and HPII, katG and katE, respectively, are unlinked, mapping at 89.2 (21) and 37.2 (17) min, respectively. A third gene, katF, mapping at 59.0 min (20), was found to be required for expression of katE but not katG. The katF gene has been cloned (28) and sequenced (27), revealing a striking similarity between the KatF protein and known sigma transcription factors, suggesting its mechanism as a positive effector of katE (27), of xthA (32), and possibly of other genes involved in resistance to near-UV radiation (33).The katG gene has been cloned (22, 42) and sequenced (41) and predicts an amino acid sequence that bears no resemblance to any of the known catalase sequences. Instead, a striking resemblance to a peroxidase from Bacillus stearothermophilus has been observed (23), suggesting that the bifunctional HPI is more closely related to the family of peroxidases than catalases. On the basis of its very different physical structure, it seemed unlikely that HPII would have any sequence or structural similarity to the common catalases. This paper describes the sequence analysis of katE, the identification of the site of transcription initiation, and a comparison of the predicted amino acid sequence of HPII with the sequences of other catalases. MATERIALS AND METHODSBacterial strains and plasmids. The strains JM101 (45) and NM522 (25) were used for plasmid preparations and for the preparation of single-stranded DNA for sequencing. Cul-* Correspo...

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Physical characterization of katG, encoding catalase HPI of Escherichia coli

Cited by 59 publications

References 37 publications

Catalase-peroxidases (KatG) Exhibit NADH Oxidase Activity

Catalase-peroxidases (KatG) Exhibit NADH Oxidase Activity

The katA Catalase Gene Is Regulated by OxyR in both Free-Living and Symbiotic Sinorhizobium meliloti

Nucleotide sequence of Escherichia coli katE, which encodes catalase HPII

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