Polyhydroxynaphthalene Reductase Involved in Melanin Biosynthesis in Magnaporthe Grisea. Purification, cDNA Cloning and Sequencing

Eur J Biochem. 1994 Feb 1;219(3):985-92. doi: 10.1111/j.1432-1033.1994.tb18581.x.

Abstract

During the biosynthesis of fungal melanin, tetrahydroxynaphthalene reductase catalyzes the NADPH-dependent reduction of 1,3,6,8-tetrahydroxynaphthalene (T4HN) into (+)-scytalone and 1,3,8-trihydroxynaphthalene into (-)-vermelone. The enzyme from Magnaporthe grisea, the fungus responsible for rice blast disease, has been purified to homogeneity. It is a tetramer of four identical 30-kDa subunits. A full-length cDNA clone of about 1 kb encoding T4HN reductase has been isolated from a cDNA library constructed in the lambda ZAP II vector and characterized. The clone contains a 846-bp open reading frame. Translation of the DNA sequence gave a 282-residue amino acid sequence with a calculated molecular mass of 29.9 kDa. Sequences corresponding to the amino-terminal part and three internal proteolytic peptides were present in the translated sequence. T4HN reductase exhibits characteristics of the short-chain alcohol dehydrogenase family. The reductase shares 56% identity with a putative ketoreductase involved in aflatoxin biosynthesis in Aspergillus parasiticus.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Ascomycota / enzymology*
  • Ascomycota / genetics
  • Base Sequence
  • Chromatography, High Pressure Liquid
  • Cloning, Molecular
  • Culture Media
  • DNA, Complementary / chemistry
  • DNA, Fungal / chemistry
  • DNA, Fungal / genetics
  • Electrophoresis, Polyacrylamide Gel
  • Fungal Proteins*
  • Melanins / biosynthesis*
  • Molecular Sequence Data
  • Oxidoreductases / chemistry
  • Oxidoreductases / genetics
  • Oxidoreductases / isolation & purification*
  • Oxidoreductases / metabolism
  • Oxidoreductases Acting on CH-CH Group Donors*
  • Sequence Alignment

Substances

  • Culture Media
  • DNA, Complementary
  • DNA, Fungal
  • Fungal Proteins
  • Melanins
  • Oxidoreductases
  • tetrahydroxynaphthalene reductase
  • Oxidoreductases Acting on CH-CH Group Donors

Associated data

  • GENBANK/L22309