Heme content of recombinant catalase from Psychrobacter sp. T-3 altered by host Escherichia coli cell growth conditions

Kimoto, Hideyuki; Matsuyama, Hidetoshi; Yumoto, Isao; Yoshimune, Kazuaki

doi:10.1016/j.pep.2008.03.016

Cited by 16 publications

(5 citation statements)

References 19 publications

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“…Attempts to obtain an apo form of Orf13 for in vitro heme titration was also unsuccessful since the enzyme failed to express when 1,10-phenanthroline, a metal chelating agent, was added to the growth medium. In an effort to increase and control the in vivo incorporation of heme b in Orf13, δ-aminolevulinic acid (δ-ALA, a heme biosynthetic precursor (37)), exogenous non-heme iron with or without δ-ALA, or exogenous hemin were added to the medium for expression in BL21(DE3) E. coli . This strategy was ineffective as similar results in heme b occupancy (30-100%) were observed compared to Orf13 expression without any additives.…”

Section: Resultsmentioning

confidence: 99%

A Heme Peroxidase with a Functional Role as an l-Tyrosine Hydroxylase in the Biosynthesis of Anthramycin

2011

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We report the first characterization and classification of Orf13 (S. refuineus) as a heme dependent peroxidase catalyzing the ortho-hydroxylation of l-tyrosine to l-DOPA. The putative tyrosine hydroxylase coded by orf13 of the anthramycin biosynthesis gene cluster has been expressed and purified. Heme b has been identified as the required cofactor for catalysis and maximal l-tyrosine conversion to l-DOPA is observed in the presence of hydrogen peroxide. Pre-incubation of l-tyrosine with Orf13 prior to the addition of hydrogen peroxide is required for l-DOPA production. However, the enzyme becomes inactivated by hydrogen peroxide during catalysis. Steady state kinetic analysis of l-tyrosine hydroxylation revealed similar catalytic efficiency for both l-tyrosine and hydrogen peroxide. Spectroscopic data from a reduced-CO (g) UV-visible spectrum of Orf13 and electron paramagnetic resonance of ferric-heme Orf13 are consistent with heme peroxidases that have a histidyl-ligated heme-iron. Contrary to the classical heme peroxidase oxidation reaction with hydrogen peroxide that produces coupled aromatic products such as o,o'-dityrosine, Orf13 is novel in its ability to catalyze aromatic amino acid hydroxylation with hydrogen peroxide, in the substrate addition order and for its substrate specificity for l-tyrosine. Peroxygenase activity of Orf13 for the ortho-hydroxylation of l-tyrosine to l-DOPA by a molecular oxygen dependent pathway in the presence of dihydroxyfumaric acid is also observed. This reaction behavior is consistent with peroxygenase activity reported with horseradish peroxidase for the hydroxylation of phenol. Overall, the putative function of Orf13 as a tyrosine hydroxylase has been confirmed and establishes the first bacterial class of tyrosine hydroxylases.

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Section: Resultsmentioning

confidence: 99%

A Heme Peroxidase with a Functional Role as an l-Tyrosine Hydroxylase in the Biosynthesis of Anthramycin

2011

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“…If the high accumulation of catalase with a significantly high specific activity is essential for metabolic modifications (discussed above) in vktA-transformed host cells, it is desirable to use other kinds of catalase that accumulate in the cells and have a high specific activity. Such catalases are the Exiguobacterium oxidotolerans catalase (EKTA catalase; Hara et al, 2007) and the Psychrobacter piscatorii catalase (Kimoto et al, 2008), whose specific activity is comparable to that of VktA, indicating that these could be used instead of VktA.…”

Section: Discussionmentioning

confidence: 99%

Innovations in biotechnology

Gartland

Beccari

Bruschi

et al. 2015

Journal of Biotechnology

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“…The isolate was identified as a new species, P. piscatorii T-3-2 T and cell extracts of strain T-3-2 T exhibit much higher catalase activity (12,000 U/mg protein) than those of other stains belonging to the same genus, including Psychrobacter nivimaris (15 U/ mg protein), Psychrobacter proteolyticus (29 U/mg protein) and Psychrobacter aquamaris (1800 U/mg protein). Strain T-3 belongs to P. piscatorii as well [34,35] and exhibits higher catalase activity (19,700 U/mg), with catalase accounting for 10% of all proteins in the cell extract. Several reports have described Psychrobacter spp.…”

Section: Characteristics Of H 2 O 2 -Resistant Bacteriamentioning

confidence: 99%

Evolutionary Strategies of Highly Functional Catalases for Adaptation to High H₂O₂ Environments

Yumoto¹,

Hanaoka²,

Hara³

2021

Antioxidants - Benefits, Sources, Mechanisms of Action

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Enzymatic evolutionary strategies for adaptation to a high H2O2 environment have been evaluated using catalases with high catalytic efficiency isolated from two H2O2-tolerant bacteria, Exiguobacterium oxidotolerans and Psychrobacter piscatori. The entrance size of the narrow main channel in catalase has been estimated by determining the formation rate of the intermediate state of peracetic acid (b), which is a larger substrate than H2O2 versus that of catalase activity with H2O2 (a) (calculated as b/a). The ratio of b/a in E. oxidotolerans catalase (EKTA) is much higher than that of P. piscatori catalase (PKTA). To elucidate the structural differences between the catalases, the amino acids present in the main channel have been compared between the two catalases and other catalases in the database. The combination of amino acid residues, which contribute high catalytic efficiency in the narrow main channel of EKTA were different from those in PKTA. In this review, we discuss strategic differences in the elimination of high concentration of H2O2 owing to differences in the phylogenetic positions of catalases. In addition, we describe the relationships between the environmental distributions of genera involved in H2O2-resistant bacteria and their catalase functions based on the main channel structure of catalase.

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Heme content of recombinant catalase from Psychrobacter sp. T-3 altered by host Escherichia coli cell growth conditions

Cited by 16 publications

References 19 publications

A Heme Peroxidase with a Functional Role as an l-Tyrosine Hydroxylase in the Biosynthesis of Anthramycin

A Heme Peroxidase with a Functional Role as an l-Tyrosine Hydroxylase in the Biosynthesis of Anthramycin

Innovations in biotechnology

Evolutionary Strategies of Highly Functional Catalases for Adaptation to High H₂O₂ Environments

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Heme content of recombinant catalase from Psychrobacter sp. T-3 altered by host Escherichia coli cell growth conditions

Cited by 16 publications

References 19 publications

A Heme Peroxidase with a Functional Role as an l-Tyrosine Hydroxylase in the Biosynthesis of Anthramycin

A Heme Peroxidase with a Functional Role as an l-Tyrosine Hydroxylase in the Biosynthesis of Anthramycin

Innovations in biotechnology

Evolutionary Strategies of Highly Functional Catalases for Adaptation to High H2O2 Environments

Contact Info

Product

Resources

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Evolutionary Strategies of Highly Functional Catalases for Adaptation to High H₂O₂ Environments