Purification and characterization of two isoforms of glucose 6-phosphate dehydrogenase (G6PDH) from Chlorella vulgaris C-27

Ken-ichi Honjoh, Ayano Mimura, Eiko Kuroiwa, Takahiro Hagisako, Koushirou Suga, Hideyuki Shimizu, Rama Shanker Dubey, Takahisa Miyamoto, Shoji Hatano, Masayoshi Iio

Research output: Contribution to journalArticle

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Abstract

Two kinds of isoforms of glucose 6-phosphate dehydrogenase (G6PDH) were purified from cells of a freezing-tolerant strain, Chlorella vulgaris C-27, by sequential steps of chromatography on five kinds of columns, including a HiTrap Blue column which showed excellent separation of the isoforms from each other. The two isoforms (G6PDH1 and G6PDH2) were purified up to 109-fold and 197-fold with specific activity of 14.4 and 26.0 U/mg-protein, respectively. G6PDH1 showed an apparent Mr of 200,000 with a subunit Mr of about 58,000, whereas G6PDH2 showed an apparent Mr of 450,000 with a subunit Mr of about 52,000. The kinetic parameters were measured and several enzymatic features of the isoforms, such as effects of metal ions on the enzyme activity, were clarified, which showed that the two isoforms were different from each other in many respects. Among the effective ions, Cd2+ showed marked stimulating effects on both isoforms. G6PDH1 and G6PDH2 seem to be a cytosolic and a chloroplastic type, respectively, as judged by their sensitivity to DTT, and also from the results of sequence similarity searches using their N-terminal and internal amino acid sequences.

Original languageEnglish
Pages (from-to)1888-1896
Number of pages9
JournalBioscience, Biotechnology and Biochemistry
Volume67
Issue number9
DOIs
Publication statusPublished - Sep 1 2003

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Chlorella vulgaris
Glucosephosphate Dehydrogenase
Purification
Protein Isoforms
Ions
Enzyme activity
Chromatography
Kinetic parameters
Freezing
Metal ions
C 27
Amino Acid Sequence
Metals
Amino Acids
Enzymes

All Science Journal Classification (ASJC) codes

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

Cite this

Purification and characterization of two isoforms of glucose 6-phosphate dehydrogenase (G6PDH) from Chlorella vulgaris C-27. / Honjoh, Ken-ichi; Mimura, Ayano; Kuroiwa, Eiko; Hagisako, Takahiro; Suga, Koushirou; Shimizu, Hideyuki; Dubey, Rama Shanker; Miyamoto, Takahisa; Hatano, Shoji; Iio, Masayoshi.

In: Bioscience, Biotechnology and Biochemistry, Vol. 67, No. 9, 01.09.2003, p. 1888-1896.

Research output: Contribution to journalArticle

Honjoh, Ken-ichi ; Mimura, Ayano ; Kuroiwa, Eiko ; Hagisako, Takahiro ; Suga, Koushirou ; Shimizu, Hideyuki ; Dubey, Rama Shanker ; Miyamoto, Takahisa ; Hatano, Shoji ; Iio, Masayoshi. / Purification and characterization of two isoforms of glucose 6-phosphate dehydrogenase (G6PDH) from Chlorella vulgaris C-27. In: Bioscience, Biotechnology and Biochemistry. 2003 ; Vol. 67, No. 9. pp. 1888-1896.
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