Cloning, Sequencing, and Heterologous Expression of a Gene Coding for Arthromyces Ramosus Peroxidase

Haruyo Sawai-Hatanaka, Toshihiko Ashikari, Yoshikazu Tanaka, Hiroyuki Minakata, Naoki Kunishima, Teruo Amachi, Yasuhiko Asada, Toru Nakayama, Keiichi Fukuyama, Hideaki Yamada, Yuji Shibano

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42 Citations (Scopus)

Abstract

To understand the relationship between the structure and functions of the peroxidase of Arthromyces ramosus, a novel taxon of hyphomycete, and the evolutionary relationship of the A. ramosus peroxidase (ARP) with the other peroxidases, we isolated complementary and genomic DNA clones encoding ARP and characterized them. The sequence analyses of the ARP and cDNA coding for ARP showed that a mature ARP consists of 344 amino acids with a N-terminal pyroglutamic acid preceded by a signal peptide of 20 amino acid residues. The amino acid sequence of ARP was 99% identical to that of the peroxidase of Coprinus cinereus, a basidiomycete, and also had very high similarities (41-43% identity) to those of basidiomycetous lignin peroxidases, although we could find no lignin peroxidase activities for ARP when assayed with lignin model compounds. We could identitified His184 and His56 as proximal and distal ligands to heme, respectively, and Arg52 as an essential Arg. Comparison of the sequences of complementary and genomic DNAs found that protein-encoding DNA is interrupted by 14 intervening sequences. The ARP cDNA was expressed in the yeast Saccharomyces cerevisiae under the promoter of the glyceraldehyde 3-phosphate dehydrogenase gene, yielding 0.02 units/ml of a secreted active peroxidase.

Original languageEnglish
Pages (from-to)1221-1228
Number of pages8
JournalBioscience, Biotechnology, and Biochemistry
Volume59
Issue number7
DOIs
Publication statusPublished - 1995 Jan 1
Externally publishedYes

ASJC Scopus subject areas

  • Biotechnology
  • Analytical Chemistry
  • Biochemistry
  • Applied Microbiology and Biotechnology
  • Molecular Biology
  • Organic Chemistry

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    Sawai-Hatanaka, H., Ashikari, T., Tanaka, Y., Minakata, H., Kunishima, N., Amachi, T., Asada, Y., Nakayama, T., Fukuyama, K., Yamada, H., & Shibano, Y. (1995). Cloning, Sequencing, and Heterologous Expression of a Gene Coding for Arthromyces Ramosus Peroxidase. Bioscience, Biotechnology, and Biochemistry, 59(7), 1221-1228. https://doi.org/10.1271/bbb.59.1221