Catalase from the silkworm, Bombyx mori: Gene sequence, distribution, and overexpression

Kohji Yamamoto, Yutaka Banno, Hiroshi Fujii, Fumio Miake, Nobuhiro Kashige, Yoichi Aso

研究成果: ジャーナルへの寄稿記事

39 引用 (Scopus)

抄録

Living organisms require mechanisms regulating reactive oxygen species (ROS) such as hydrogen peroxide and superoxide anion. Catalase is one of the regulatory enzymes and facilitates the degradation of hydrogen peroxide to oxygen and water. Biochemical information on an insect catalase is, however, insufficient. Using mRNA from fat body of the silkworm, Bombyx mori, a cDNA encoding a putative catalase was amplified by reverse transcriptase-polymerase chain reaction and sequenced. The deduced amino acid sequence comprised 507 residues with more than seventy residues forming a scaffold for a heme cofactor conserved. The sequence showed 71% and 66% identities to those of the Drosophila melanogaster and Apis mellifera catalases, respectively; the catalase from B. mori was estimated to be phylogenetically close to that from A. mellifera. The transcripts of the gene and the catalase activity were distributed in diverse tissues of B. mori, suggesting its ubiquitous nature. Using the gene, a recombinant catalase (rCAT) was functionally overexpressed in a soluble form using Escherichia coli, purified to homogeneity, and characterized. The pH-optimum of rCAT was broad around pH 8.0. More than 80% of the original rCAT activity was retained after incubation in the following conditions: at pH 8-11 and 4°C for 24 h; at pH 7 and temperatures below 50°C for 30 min. The Michaelis constant for hydrogen peroxide was evaluated to be 28 mM at pH 6.5 and 30°C. rCAT was suggested to be a member of the typical catalase family.

元の言語英語
ページ(範囲)277-283
ページ数7
ジャーナルInsect Biochemistry and Molecular Biology
35
発行部数4
DOI
出版物ステータス出版済み - 1 1 2005

Fingerprint

Bombyx
Bombyx mori
silkworms
Catalase
catalase
Genes
nucleotide sequences
Hydrogen Peroxide
hydrogen peroxide
Apis mellifera
Fat Body
Bees
Polymerase chain reaction
RNA-Directed DNA Polymerase
heme
fat body
Drosophila melanogaster
Reverse Transcriptase Polymerase Chain Reaction
Heme
Scaffolds

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Insect Science

これを引用

Catalase from the silkworm, Bombyx mori : Gene sequence, distribution, and overexpression. / Yamamoto, Kohji; Banno, Yutaka; Fujii, Hiroshi; Miake, Fumio; Kashige, Nobuhiro; Aso, Yoichi.

:: Insect Biochemistry and Molecular Biology, 巻 35, 番号 4, 01.01.2005, p. 277-283.

研究成果: ジャーナルへの寄稿記事

Yamamoto, Kohji ; Banno, Yutaka ; Fujii, Hiroshi ; Miake, Fumio ; Kashige, Nobuhiro ; Aso, Yoichi. / Catalase from the silkworm, Bombyx mori : Gene sequence, distribution, and overexpression. :: Insect Biochemistry and Molecular Biology. 2005 ; 巻 35, 番号 4. pp. 277-283.
@article{204b8041c64b49b2aca03889e49ec8d9,
title = "Catalase from the silkworm, Bombyx mori: Gene sequence, distribution, and overexpression",
abstract = "Living organisms require mechanisms regulating reactive oxygen species (ROS) such as hydrogen peroxide and superoxide anion. Catalase is one of the regulatory enzymes and facilitates the degradation of hydrogen peroxide to oxygen and water. Biochemical information on an insect catalase is, however, insufficient. Using mRNA from fat body of the silkworm, Bombyx mori, a cDNA encoding a putative catalase was amplified by reverse transcriptase-polymerase chain reaction and sequenced. The deduced amino acid sequence comprised 507 residues with more than seventy residues forming a scaffold for a heme cofactor conserved. The sequence showed 71{\%} and 66{\%} identities to those of the Drosophila melanogaster and Apis mellifera catalases, respectively; the catalase from B. mori was estimated to be phylogenetically close to that from A. mellifera. The transcripts of the gene and the catalase activity were distributed in diverse tissues of B. mori, suggesting its ubiquitous nature. Using the gene, a recombinant catalase (rCAT) was functionally overexpressed in a soluble form using Escherichia coli, purified to homogeneity, and characterized. The pH-optimum of rCAT was broad around pH 8.0. More than 80{\%} of the original rCAT activity was retained after incubation in the following conditions: at pH 8-11 and 4°C for 24 h; at pH 7 and temperatures below 50°C for 30 min. The Michaelis constant for hydrogen peroxide was evaluated to be 28 mM at pH 6.5 and 30°C. rCAT was suggested to be a member of the typical catalase family.",
author = "Kohji Yamamoto and Yutaka Banno and Hiroshi Fujii and Fumio Miake and Nobuhiro Kashige and Yoichi Aso",
year = "2005",
month = "1",
day = "1",
doi = "10.1016/j.ibmb.2005.01.001",
language = "English",
volume = "35",
pages = "277--283",
journal = "Insect Biochemistry and Molecular Biology",
issn = "0965-1748",
publisher = "Elsevier Limited",
number = "4",

}

TY - JOUR

T1 - Catalase from the silkworm, Bombyx mori

T2 - Gene sequence, distribution, and overexpression

AU - Yamamoto, Kohji

AU - Banno, Yutaka

AU - Fujii, Hiroshi

AU - Miake, Fumio

AU - Kashige, Nobuhiro

AU - Aso, Yoichi

PY - 2005/1/1

Y1 - 2005/1/1

N2 - Living organisms require mechanisms regulating reactive oxygen species (ROS) such as hydrogen peroxide and superoxide anion. Catalase is one of the regulatory enzymes and facilitates the degradation of hydrogen peroxide to oxygen and water. Biochemical information on an insect catalase is, however, insufficient. Using mRNA from fat body of the silkworm, Bombyx mori, a cDNA encoding a putative catalase was amplified by reverse transcriptase-polymerase chain reaction and sequenced. The deduced amino acid sequence comprised 507 residues with more than seventy residues forming a scaffold for a heme cofactor conserved. The sequence showed 71% and 66% identities to those of the Drosophila melanogaster and Apis mellifera catalases, respectively; the catalase from B. mori was estimated to be phylogenetically close to that from A. mellifera. The transcripts of the gene and the catalase activity were distributed in diverse tissues of B. mori, suggesting its ubiquitous nature. Using the gene, a recombinant catalase (rCAT) was functionally overexpressed in a soluble form using Escherichia coli, purified to homogeneity, and characterized. The pH-optimum of rCAT was broad around pH 8.0. More than 80% of the original rCAT activity was retained after incubation in the following conditions: at pH 8-11 and 4°C for 24 h; at pH 7 and temperatures below 50°C for 30 min. The Michaelis constant for hydrogen peroxide was evaluated to be 28 mM at pH 6.5 and 30°C. rCAT was suggested to be a member of the typical catalase family.

AB - Living organisms require mechanisms regulating reactive oxygen species (ROS) such as hydrogen peroxide and superoxide anion. Catalase is one of the regulatory enzymes and facilitates the degradation of hydrogen peroxide to oxygen and water. Biochemical information on an insect catalase is, however, insufficient. Using mRNA from fat body of the silkworm, Bombyx mori, a cDNA encoding a putative catalase was amplified by reverse transcriptase-polymerase chain reaction and sequenced. The deduced amino acid sequence comprised 507 residues with more than seventy residues forming a scaffold for a heme cofactor conserved. The sequence showed 71% and 66% identities to those of the Drosophila melanogaster and Apis mellifera catalases, respectively; the catalase from B. mori was estimated to be phylogenetically close to that from A. mellifera. The transcripts of the gene and the catalase activity were distributed in diverse tissues of B. mori, suggesting its ubiquitous nature. Using the gene, a recombinant catalase (rCAT) was functionally overexpressed in a soluble form using Escherichia coli, purified to homogeneity, and characterized. The pH-optimum of rCAT was broad around pH 8.0. More than 80% of the original rCAT activity was retained after incubation in the following conditions: at pH 8-11 and 4°C for 24 h; at pH 7 and temperatures below 50°C for 30 min. The Michaelis constant for hydrogen peroxide was evaluated to be 28 mM at pH 6.5 and 30°C. rCAT was suggested to be a member of the typical catalase family.

UR - http://www.scopus.com/inward/record.url?scp=14844311938&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=14844311938&partnerID=8YFLogxK

U2 - 10.1016/j.ibmb.2005.01.001

DO - 10.1016/j.ibmb.2005.01.001

M3 - Article

C2 - 15763464

AN - SCOPUS:14844311938

VL - 35

SP - 277

EP - 283

JO - Insect Biochemistry and Molecular Biology

JF - Insect Biochemistry and Molecular Biology

SN - 0965-1748

IS - 4

ER -